Different types of polymers and processes are utilized to prepare pharmaceutical formulations to achieve the desired drug delivery attributes from solid oral dosage forms. In this study, selected pharmaceutical grade polymers and additives were utilized to prepare films and coated formulations using different techniques. Experiments were performed to characterize the bulk and surface properties of the materials to gain insights into the functionality of the polymers. Examples of the methods that were utilized to characterize the materials included Thermogravimetric Analyzer (TGA) combined with an Infrared Spectrometer (TG-IR), X-ray diffraction, dynamic vapor sorption (DVS), Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS), and Confocal Raman Spectroscopy. Special emphasis was given to study the effect of moisture and plasticizers on the physical properties of the films and formulations. Selected formulations were developed to prepare solid oral dosage forms containing different active pharmaceutical ingredients. The presentation will provide a summary of the results obtained from characterizing the films, coated formulations and drug delivery dosage forms.

Biomaterials have been characterized by an ever increasing collection of highly advanced experimental techniques, while consistent and integrative evaluation of all related test results remains a significant, largely unmet challenge. As a remedy, we here present several recent activities.
We start with a new evaluation method (Czenek et al, J Mat Res 29, 2757ff., 2014), which uses the unique linear relationship between gray values and X-ray attenuation coefficients, together with the energy-dependence of the latter, to identify (i) the average x-ray energy used in the CT device, (ii) the X-ray attenuation coefficients, and (iii), via the x-ray attenuation average rule, the intravoxel composition, i.e., the microporosity, which, amongst others, governs the voxel-specific mechanical properties, such as stiffness and strength. The method is realized for six 3D tricalcium phosphate scaffolds, seeded with pre-osteoblastic cells and differentiated for 3, 6, and 8 weeks, respectively. The corresponding voxel-specific microporosities turn out to increase during the culturing period (resulting in reduced elastic properties, as determined from micromechanical considerations), while the overall macroporosity remains constant.
As a second example, consistent combination of nanoindentation, ultrasound, and micromechanics theory is shown by example of porous baghdadite (Ca₃ZrSi₂O₉) scaffolds (Kariem et al, Mat Sci Eng 46C, 553ff., 2015). The resulting porosity-stiffness relations further confirm a formerly detected, micromechanically explained, general relationship for a great variety of different polycrystals (Fritsch et al, J Appl Mech 80, 020905-1, 2013),which also allows for estimating the zero-porosity case, i.e. Young modulus and Poisson ratio of pure (dense) baghdadite. These estimateswere impressively confirmed by a physically and statistically independent nanoindentation campaign comprising some 1750 indents. Consequently, we can present a remarkably complete picture of porous baghdadite elasticity across a wide range of porosities, and, thanks to the micromechanical understanding, reaching out beyond classical elasticity, towards poroelastic properties, quantifying the effect of pore pressure on the material system behavior.
The new methods are expected to further foster the development of a rationally based and computer-aided design of biomaterials and tissue engineering scaffolds.

Hydrogels are three-dimensional networks of polymer that have great ability to imbibe exceptionally large volumes of water. The water uptake and hence swelling of hydrogels can be controlled and manipulated depending on hydrogel design. As such, their swelling can be exploited in sensing and controlled release platforms, with applications varying from providing a simple inert protective sensor coating, to use as an intelligent drug delivery system capable of sensing physiological changes and auto-titrating a drug. This presentation will describe the work we are doing on responsive hydrogel materials, their design, and applicability to both sensing and controlled release. The context of the work will be discussed within the application area of epidermal sensing, e.g., monitoring skin barrier function.
In relation to the hydrogel-based sensing platform, electrochemical impedance spectroscopy is used to sensitively track the swelling responsive of pH-sensitive hydrogels. Indeed, the demonstration of this system for glucose detection at the sub-micromolar levels opens up the possibility of glucose detection in sweat. Electrically-stimulated drug release from these hydrogels that are composited with reduced graphene oxide will also be discussed. Finally, our recent work on the fabrication of super-macroporous structures of these hydrogels and the dramatic influence of this induced structure on the material&’s swelling behaviour and hence its sensing characteristics will be presented.

Nanoimprint lithography (NIL) is a rapid, low cost mechanical technique to create nanoscale to microscale patterns across large surface areas. Although originally introduced as a potential next generation lithography for the semiconductor industry, the technique has found itself applied in a variety of applications including biomaterials and devices. Its advantage over other techniques is to be able to provide fully deterministic surface patterning, including pattern gradients and multiscale features, in a wide variety of materials. In this work we report on a new development to improve nanoimprint processing to provide both topographical and chemical function to current and potential future tissue scaffolding polymers.
In small amplitude oscillatory shear forming (SAOSF) [1], a small amplitude cyclic shear displacement (up to 10&’s of nm) is applied to a rigid die in contact with a soft solid film leading to a plastic-ratchet forming action (Fig. 1). As with conventional nanoimprint, nano-feature fidelity and registry are achieved, however the technique also has advantages of constant low temperature processing and hence a wide choice of materials. For solid films, the underlying mechanism of SAOSF is due to a combination of uniformly plasticizing contacted areas of the film while simultaneously activating a novel pumping action involving die-geometry-induced broken circulation of elastoplastic flow. Crucially, the process is scale independent for thin film planar imprint, since only a critical shear strain is required. A minimal applied normal load largely eliminates elastic distortions thought to be responsible for residual layer non-uniformity and other replication defects common to imprint. Finite element simulations of the process with simple elastic-elastoplastic materials agree well with experimental observations.
In this work we demonstrate the successful application of SAOSF to large area surfaces and thin films of the biodegradable polymer PLGA. Imprint dies with nanoscale patterns were imprinted with high fidelity across cm scale areas at temperatures below above and below the glass transition, greatly reducing biopolymer-die adhesion and thus increasing die lifetime, as well as reducing overall imprint time due to a short thermal cycle. We demonstrate the efficacy of this technique in influencing stem cell response to surface patterns. While currently implemented in conventional hot embossing and nanoimprint setups, steps toward adapting the approach high throughput to roll-to-roll processing will be described, including a lamination strategy allowing fully 3D porous tissue scaffolds to be fabricated with complete determinism of internally patterned surfaces. The advantages that SAOSF provides may become particular great in the continuous processing environment.
[1] G. L. W. Cross, B. S. O'Connell, H. O. Ozer and J. B. Pethica, Nano Lett. 7, 357 (2007).

Low permeability is one of the key challenges that exists in the area of drug delivery system. Due to the nature of polymers, micro drug carriers made of polymers are usually porous which leads to the failure of retaining small molecules and narrows the applications of the polymeric micro containers. Thus, our group has been trying to overcome this shortage.
In this work, low permeable Poly(methyl methacrylate) shells which are not permeable by small molecules were fabricated via surface initiated Atom Transfer Radical Polymerization (ATRP).
The kinetics of PMMA brush growth was first studied on flat substrates on which the pre-synthesized macroinitiator was deposited using Layer-by-Layer technique. The thicknesses of the PMMA films were measured with ellipsometer. The surface hydrophobicities of different substrates with and without PMMA layer were then compared by measuring the water contact angles. Increased surface hydrophobicity was witnessed from the significantly increased water contact angle on the surface with PMMA monolayer.
Further to this, low permeable PMMA shells were synthesized on the surface of CaCO₃ templates from the pre-deposited macroinitiators via ATRP polymerization. Fourier Transform Infrared Spectroscopy (FTIR) and Thermogravimetric Analysis (TGA) spectrums showed that PMMA was successfully coated on the inorganic cores. Transmission Electron Microscope (TEM) was applied to observe the thicknesses of PMMA shells and they were found to be nearly in accordance with the kinetics on planar surfaces. The results of the Energy-dispersive X-ray Spectroscopy (EDS) together with those obtained from Scanning Electron Microscope (SEM) and TEM demonstrated that there were no differences in the element compositions as well as surface and inner structures of PMMA coated particles before and after ethylenediaminetetraacetic acid (EDTA) treatment. This indicates that PMMA shells are non-permeable to the dissolution agent, which has low molecular weight.
After having proven that hydrophobic polymer shells synthesized by “brush growth” technique are able to form low permeable micro containers, the same strategy was applied to fabricate the low permeable and biodegradable Poly(lactic acid) microcapsules, the properties of which are being investigated. We believe that our work could contribute to the biomedical applications.

Piezoelectric inkjet printing is a materials deposition technique that relies on pressure waves from a piezoelectric actuator to precisely pattern picoliter volumes of liquids onto a surface. We have recently utilized this approach for modifying the surfaces of microneedles, which are microscale projections that may be used for transdermal delivery of many types of pharmacologic agents. Precise patterning of several pharmacologic agents was demonstrated. The mechanical and chemical properties of the inkjet-modified devices with compared against those of the unmodified devices. In addition, the functionality of the inkjet-printed drugs was validated using in vitro assays. The results suggest that piezoelectric inkjet printing is an appropriate approach for incorporating pharmacologic agents within medical devices since it is associated with rapid processing times, low cost, and facile scalability.

Dinoflagellates are protists, mostly found as marine plankton. Some species produce a calcareous shell around their exterior as a cyst for protection during the dormant, zygotic stage of their life. Using this core-shell architecture as an inspiration, we have developed a methodology for encapsulating “soft” liquid droplets within a “hard” mineral shell. The polymer-induced liquid-precursor (PILP) mineralization process was used to deposit a calcium carbonate (CaCO₃) mineral shell on the surface of surfactant-stabilized fluidic particles comprised of either oil-in-water emulsion droplets or liposomes. Size and morphological analyses were performed by various microscopic techniques, where it was observed that the PILP droplets preferentially adsorb to anionic surfactants/lipids that stabilize the suspension, and due to their liquid-like character, coalesce to form a smooth and continuous mineral coating. The processing conditions for forming these CaCO₃-coated microcapsules are benign, so they can encapsulate virtually any active agent of interest. For example, hydrophobic compounds can be dissolved in oil-in-water emulsions, while hydrophilic and/or hydrophobic compounds can be dissolved within liposomes, which can then be coated with the CaCO₃ mineral shell. Confocal microscopy was used to demonstrate an oil-soluble fluorescent dye is encapsulated in the interior of the emulsion, or both hydrophilic and hydrophobic compounds can be incorporated with the aqueous and hydrocarbon regions of liposomes, respectively. The metastable morphology of the CaCO₃ shell enables a pH dependent degradation of the particles, allowing for release of the active agent of interest. These CaCO₃-coated microcapsules can be dried down to a powder, while retaining the active agent within the fluidic core, thereby saving in storage and transportation expenses because the large solution phase of the suspension is removed. In conclusion, while dinoflagellates utilize this encapsulation strategy as a protective shell, these biomimetic core-shell microcapsules have a myriad of potential applications because of their inherent environmentally-benign composition and biocompatibility, such as in release of pesticide or fertilizer, in chemical reactions such as release of catalyst of specific reagents, in hair/skin care products for release of conditioner or other compounds, in health care for release of pharmaceutics, or in self-healing composites for release of a sealant during capsule fracture.

We report herein a facile approach to synthesize processable bimetallic nanoparticles (Pd-Au/Au-Pd/Ag-Au/Au-Ag) decorated Prussian blue nanocomposite (PB-AgNP). The presence of cyclohexanone/formaldehyde facilitates the formation of functional bimetallic nanoparticles from 3-aminopropyltrimethoxysilane (3-APTMS) capped respective noble metal ions under desired ratio of hetero noble metal ions. The use of aforementioned reducing agents (3-APTMS and cyclohexanone) also enables the synthesis of polycrystalline Prussian blue nanoparticles (PBNPs). As synthesized PBNPs, Pd-Au/Au-Pd/Ag-Au/Au-Ag enable the formation of nano-structured composites displaying better catalytic activity than that recorded with natural enzyme. The nanomaterials have been characterized by UV-vis, FT-IR and Transmission Electron Microscopy (TEM) with following major findings: (1) 3-APTMS capped noble metal ions in the presence of suitable organic reducing agents [3-Glycidoxypropyltrimethoxysilane (GPTMS), Cyclohexanone and Formaldehyde] are converted into respective nanoparticles under ambient conditions, (2) the time course of synthesis and dispersibility of the nanoparticles are found as a function of organic reducing agents, (3) the use of formaldehyde and cyclohexanone in place of GPTMS with 3-APTMS outclasses the other two in imparting better stability to amphiphilic nanoparticles with reduced silanol content, (4) an increase in 3-APTMS concentrations causes decrease in nanogeometry of nanoparticles (5) simultaneous synthesis of bimetallic nanoparticles under desired ratio of palladium/gold and silver/ gold cations are recorded, (6) cyclohexanone mediated synthesis nanoparticles enable the formation of homogeneous nanocomposite with PBNP as peroxidase mimetic representing potential substitute of peroxidase enzyme. The peroxidase mimetic ability has been found to vary as a function of 3-APTMS concentration revealing the potential role of functional metal nanoparticles in bioanalytical applications.

Introduction
Most active implantable medical devices (AIMD) cause contraindication for implanted patients with magnetic resonance imaging (MRI) due to safety reasons (force, heating, imaging artefacts). Artefact reduction is especially relevant to enable imaging in the direct vicinity of implanted devices e.g. for postoperative placement verification and continuous checkup examinations [1].
Many AIMDs rely on thin film technology, where, depending on the deposition process, the edge of a structure shows various slopes. While the far-field of the magnetic distortion will just be dominated by the dipole that is proportional to the volume, the artefacts of thin films may also highly depend on edge effects. These will depend on the film thickness and at least to higher order also on the slope. While one can easily simulate the magnetic field itself, it is non-trivial how this translates into MR artefacts. Hence, we took an experimental approach to investigate this systematically at the example of Pt/Ir electrodes. For this purpose we developed a process to laser-structure trapezoidal cross sections and compared the artefact size of electrodes featuring various edge slopes.
Methods
For sample preparation, we laminated 25 µm thick Pt/Ir foil (Goodfellow) on silicone substrate. Thereafter, we used a Rapid10 picosecond-laser (Lumera) to structure 750 µm in diameter, disc shaped, electrode-like samples with trapezoidal cross section of various angles. Then, the samples underwent MRI examination in a 7 T Bruker Biospec 70/20. For comparison, we fabricated samples with the same slope angles but with constant volume (larger diameter). Finally, samples with rectangular cross section but varying thickness were investigated.
Results
Laser ablation proved to be a reproducible method for structuring Pt/Ir discs with trapezoidal cross section and various slope angles. The MRI examination of these samples (non-constant volume) showed distinct differences in MRI artefact size. However, samples with the same slopes but constant volume showed hardly any difference in MRI artefact size. The samples with rectangular cross section and varying thickness suggested a linear dependency of volume to MRI artefact size.
Conclusion
We show that implant electrode structures with trapezoidal cross section can be fabricated with a laser ablation process. Furthermore, these preliminary investigations suggest that sloped edges have no significant influence on MRI artefact size, whereas the sample volume (thickness) seems crucial, as expected. We conclude, for MRI artefact reduction purposes more effort should be dedicated to reducing metal volume in implants, whereas the structure of the edges and hence the choice of manufacturing process is less critical.
Acknowledgements: This work was funded within the Cluster of Excellence “BrainLinks-BrainTools” by the German Research Foundation (DFG ExC1086).
References
[1]Zrinzo et al.:WorldNeurosurgery;76(1):164-172(2011)

Polymer microfluidic devices are an emerging technology, enabling low-cost and quick clinical diagnostics. The polymer materials used in the devices are generally hydrophobic, and surface modification is required for stable flow in the micro-size channels. There are several techniques for obtaining a hydrophilic surface. Plasma surface modification, especially oxygen plasma treatment, is widely used for this application; however hydrophobic recovery quickly starts after the treatment. Therefore, it is not easy to keep the surface hydrophilic over long time periods. UV/ozone treatment is also a well-known technique for the surface modification. The disadvantage of UV/ozone treatment is that the process is more time-consuming compared to oxygen plasma treatment. Therefore, rapid and effective surface modification processes by UV/ozone treatment need to be developed.
In this presentation, we will describe parameters for rapid and effective surface modification by UV/ozone treatment and then will compare the subsequent hydrophobic recovery with oxygen plasma treatment. The polymer materials chosen in this study were: (1) polymethyl methacrylate (PMMA), (2) cyclic olefin copolymer (COC), (3) cyclic olefin polymer (COP) and (4) polyether ether ketone (PEEK). A UV/ozone cleaner (Model UV-2, SAMCO Inc.) was used for the surface modification. The system is equipped with an ozone generator for higher ozone concentration (30-160 g/m³) and a sample stage heater capable of controlling from ambient to a maximum temperature of 300°C. For evaluation of the wettability of the surface, the static contact angle was measured by the sessile drop method. Then, subsequent hydrophobic recovery was investigated by comparing UV/ozone treated polymer samples with identical samples treated using oxygen plasma. Function groups on the surface before and after UV/ozone treatment were characterized using X-ray photoelectron spectroscopy (XPS).
It was found that employment of ex-situ generated ozone and stage temperature control both contributed to the rapid and effective surface modification of each material. Also, the UV/ozone treatment was optimized for each material and the UV/ozone treated surfaces showed a long-term stable hydrophilic surface for up to 6 months, which was in contrast to the hydrophobic recovery that was seen following oxygen plasma treatment. For the XPS characterization of COP, the sample processed with UV/ozone using ex-situ generated ozone and stage temperature control showed a larger amount of ester functional groups (-COOR), compared to the sample processed with UV/ozone but without ex-situ generated ozone and stage temperature control. We believe that the UV/ozone cleaner equipped with an ex-situ, high-concentration ozone generator and controlled stage heating is superior to traditional UV/ozone cleaners for obtaining a long-term stable hydrophilic surface of polymer materials.

Research in microfluidic biosensors has led to dramatic improvements in sensitivities. Very few examples of these devices have been commercially successful, keeping this methodology out of the hands of potential users. In this study, we developed a method to fabricate a flexible microfluidic device containing electrowetting valves and electrochemical transduction. The device was designed to be amenable to a roll-to-roll manufacturing system, allowing a low manufacturing cost. Microchannels with high fidelity were structured on a PET film using UV-NanoImprint Lithography (UV-NIL). The electrodes were inkjet-printed and photonically sintered on second flexible PET film. The film containing electrodes was bonded directly to the channel-containing layer to form sealed fluidic device. Actuation of the multivalve system with food dye in PBS buffer was performed to demonstrate automated fluid delivery. The device was then used to detect Salmonella in a liquid sample.

Anti-inflammatory agents have been used widely for drug delivery to neural tissue. A considerable amount of research has been dedicated to improve the biocompatibility and efficacy of the neural microelectrodes. However, there are some challenges such as (1) the burst effect of drug in the brain and (2) the huge amount of neuronal killing zone around the implanted electrode in the brain tissue. To overcome these challenges, we have proposed a method for fabrication and sustained release of drug using anti-inflammatory agent dexamethasone (DEX)-loaded within hybrid biodegradable nanofibers and conducting polymers.
The objective of this study was to develop a novel method for encapsulation of DEX within aligned conducting polymer nanotubes using electrojetting of aligned poly (lactic-co-glycolic acid) (PLGA) nanofibers and electrochemical polymerization of conducting polymer poly(pyrrole) (PPy). Briefly, DEX-PLGA solution was electrospun on Au substrates followed by electrochemical polymerization of PPy. After fabrication of aligned DEX-loaded PLGA nanofibers, PPy was electrodeposited on Au substrates and around nanofibers from a solution containing 0.2 M pyrrole and 0.2M sodium-p-styrenesulfonate (PSS) using and applied current density 0.5mA.cm^-2 in 6 different deposition time from 1min to 6min. Surface morphology of aligned PLGA nanofibers and PPy nanotubes were characterized using optical microscopy and field emission scanning electron microscopy. Cyclic Voltammetry (CV) has been employed to actuate the DEX-loaded PPy nanotubes in order to increase the amount of DEX. We hypothesize that the parameters of CV such as voltage (from 0.2V to 1V), scan rate (from 50mv/s to 400mv/s, and number of cycles (from 1 to 50 cycles) have direct influence on the rate of DEX release. DEX-loaded PPy nanotubes were compared for both stimulated and unstimulated conditions. For quantification Dex release, we used UV spectrophotometry and detected DEX at wavelength 242nm. For the future study we will control the growth of PPy around the aligned PLGA nanofibers. It is anticipated that by increasing the thickness of PPy coating around the PLGA nanofibers, the amount of DEX release will be modulated. Our developed technique can be potentially applied in neural microelectrode for neural recording and/or stimulation.

In the field of clinical diagnostics, highly sensitive sensing system is strongly required to quickly obtain diagnostics results during medical treatments or to continuously monitor a very small amount of specimens of subcutaneous tissue fluid. In the present study, highly sensitive optical waveguide biosensors were designed by using the combination of dye and polymer-enzyme complex. Optical light waveguide can detect the optical change in the vicinity of the guide surface with high sensitivity due to the evanescent wave scattering. The optical change depends on the quantity of color development of dye in the sensing membranes formed on the optical waveguide. The sensing membranes, composed of dye, enzymes, and biocompatible polymers, were prepared by solution-coating on the optical waveguide. Herein, we used 3, 3&’, 5, 5&’-tetoramethylbenzidine (TMBZ) as a dye, glucose oxidase (GOD) and peroxidase (POD) as enzymes, and carboxymethyl cellulose (CMC) as a binder, and phosphatide polymer for protection of biological activity of enzymes. Then we investigated effects of the composition and structure of sensing membranes on the enhancement of sensitivity and response speed. The developed glucose sensors are up to 20 times more sensitive than the conventional light waveguide glucose sensors and can detect 0.1g/L glucose in 60 seconds. For the further improvement in sensitivity, microporous sensing membranes were fabricated by using electrospraying techniques. The electroprayed sensing membranes showed 40 % higher sensitivity than nonporous sensing membranes. These results show that both the composition (e.g., polymer-enzyme complex) and structure (e.g., active surface area) of sensing membrane are crucial factors for highly sensitive and high-throughput optical waveguide biosensors.

A failure of Titanium (Ti) implant is often caused by a lack of strong interaction and between implant surfaces and surrounding bones. Therefore, there have been attempts to improve Ti surface properties including modification of surface roughness and hydrophilicity. Several surface modification techniques, such as sandblasting and acid etching (SLA), anodizing and laser, have been used to improve cell adhesion and osteointegration of the implants. These techniques, however, are expensive, time-consuming, and some procedures require chemical process.
Here we demonstrated a new approach to modify Ti surface using thermal annealing in air. The Ti plates were annealed at 300,400, 500 and 600 #870; C and investigated their surface structures and chemical compositions as a function of temperatures. Annealed Ti surface were observed alterations of both morphologies and chemical compositions as a TiO₂ layer formed on surface. This leaded to an improvement of wettability of annealed Ti surface compared to untreated Ti surface, evidenced by a reducing of a contact angle from 89±5 #870; of untreated Ti specimens to 22 -30 #870; of the annealed Ti surfaces. The phase compositions and the oxidation depth of TiO₂ layers were found to be strongly depended on oxidation temperature. At 300 #870; C, Ti metal was a dominant phase of the Ti surface, and an increasing temperature increased the rutile phase composition.
In addition, mouse osteoblasts (MC3T3) exhibited a normal growth on these modified Ti surface compared to the polylysine coated tissue culture plates. The influences of these surface structures and chemistry on the cell proliferation and adhesion are being investigated in order to determine the optimum surface treatment condition. The thermal annealing process could be a potential technique to prepare Ti implant due to its low cost, simplicity and capability to modify surface structure, chemical compositions and hydrophilicity.

Nowadays, smart contact lens have attracted significant interest for diagnostic and drug delivery applications to ocular diseases as a minimally invasive platform. Here, we fabricated a smart contact lens consist of biosensor, drug delivery system, communication circuitry for the treatment of ocular diseases or diabetes. The biosensing, drug delivery systems, communication circuitry were integrated on a wireless powered contact lens. For the diagnosis of ocular diseases, tear glucose content was measured as a non-invasive alternative for the blood glucose content monitoring. The biosensor belong to electrochemical amperometric sensor contained Pt electrode immobilized with glucose oxidase. When glucose binds to glucose oxidase, hydrogen peroxide is produced and oxidized on the electrode generating electrons. From the current, we measured the amount of glucose in the tear. For the drug delivery system, we fabricated thin film Au membrane on flexible SU-8 drug reservoirs and molded to contact lens. Applying pulse of electrical current can remove each Au membrane on reservoir by exploit the electrochemistry of Au to AuCl₄^- in NaCl solution. Finally, the WiTricity power transfer system fabricated on smart contact lens to operate the sensors, drug delivery systems, and communication circuitry. This smart contact lens can be exploited for various ubiquitous healthcare applications.

Multiphoton ablation lithography is used for material surface modification and treatment in order to control biofouling by cells. We fabricated isometric and gradient ablated crater patterns with a variety of geometries and spatial arrangements on tissue culture polystyrene (TCPS) using direct laser writing technique. Varying the shape (i.e. width and depth) and the spacings (i.e. the spatial density) of ablated features could alter the focal adhesion (FA) dimensions and distributions, hence effectively guiding the cell migration and patterning. On isometric patterns, we were able to identify the surface ligand density threshold for repellent migration . Below this threshold, FAs of fibroblast do not exhibit specific interaction with ablated patterns. In the low nanocrater density region, cells could establish stable FAs. In contrast, in the high nanocrater density region, they formed nascent FAs. On the gradient pattern, this difference enhanced migration from lower spacing (i.e. denser) pattern toward more favorable region with larger spacing (i.e. more sparse) patterns providing increased planar surface area for stable focal adhesion formation. This persistent migration created different cell patterns, such as linear stripes. To further investigate the interaction between TCPS substrate and cells, we showed the effect of extracellular protein absorption on cell patterning using quartz crystal microbalance with dissipation (QCM-D).. Different patterned arrays of RGD peptide ligands induced cell migration in a density dependent manner. This laser-treated technique can be applied to various materials and a wide range of applications such as breast implants, electrode leads on pacemakers and defibrillators and orthopedic implants representing a significant health care cost and risk. Especially noteworthy is that the surface modification using femtosecond laser on TCPS does not induce chemical alteration. Cell patterning on TCPS is in essence a simple method, that is applicable to diverse experiments as an effective tool for mechanotrasduction studies in the context of regenerative medicine.

The production of heterogeneous nanoparticle systems in a single process step is of profound interest for various applications. Multifunctional nanomaterials are attractive for biomedics, where multiple diagnostic modalities or the possibility of combined therapy and diagnosis (theranostics) via a single agent are sought out. Although the production of various heterogeneous systems has been reported previously their large scale synthesis still represents a key challenge. This gains even more importance if the materials which are to be combined heterogeneously also form homogeneous alloyed or doped states. Here, the one-step synthesis of a heterogeneous Zn_0.4Fe_2.6O₄ / Gd₂O₃ multi-component systems via dry and scalable multi-spray aerosol technology is reported. A precise process control is shown, which allows the fine tuning of the resulting material composition. The produced materials are compared to their homogeneously mixed counterpart. Special emphasis is put on improved magnetism and increased efficiency as magnetic resonance imaging contrast agents.

More than 387 million people suffer from diabetes making it one of the most widespread chronic diseases worldwide.¹ For 2035, it is projected that their number will increase to 592 million.¹ Breath acetone is a promising marker that could indicate diabetes already in an early-stage and help monitoring its progression. In fact, exhaled acetone levels are elevated significantly from 300 - 900 ppb² for healthy to above 1800 ppb for uncontrolled diabetes.³ Chemo-resistive gas sensors offer great advantages for sensing such breath markers such as simple operation, high miniaturization potential, low power consumption and production cost.⁴ Quite promising for acetone is ZnO for its high analyte sensitivity.⁵ However, drawbacks that hinder its application are the lack of selectivity towards other breath compounds⁵ and the high cross-sensitivity to relative humidity (RH),⁶ i.e. for breath ~90% RH.⁷
Here, ZnO was doped during its synthesis to overcome these drawbacks. So pure and doped ZnO nanoparticles were produced by flame spray pyrolysis (FSP) and directly deposited onto sensor substrates forming a highly porous sensing film. Applied dopants improved the acetone selectivity against breath-relevant ethanol, NH₃, CO and NO. Additionally, the strong interference of humidity was reduced. This optimized sensor was capable to selectively detect even ultra-low acetone levels of 5 ppb, more than sufficient for breath analysis. Furthermore, response times were lower than 1 min enabling real-time monitoring. In conclusion, doped ZnO sensors showed high potential for implementation in an easy, inexpensive, portable and non-invasive breath acetone analyzer.
(1) IDF Diabetes Atlas: Update 2014, International Diabetes Federation, Brusseles, 2014.
(2) Diskin, A.M.; Spanel, P.; Smith, D. Physiol Meas.2003, 24, 107-119.
(3) Deng, C.; Zhang, J.; Yu, X.; Zhang, W.; Zhang, X. J Chromatogr B2004, 810, 269-275.
(4) Righettoni, M.; Amann, A.; Pratsinis, S.E. Mater. Today2015, 18, 163-171.
(5) Zeng, Y.; Zhang, T.; Yuan, M.; Kang, M.; Lu, G.; Wang, R.; Fan, H.; He, Y.; Yang, H. Sens. Actuators B2009, 143, 93-98.
(6) Zhang, Y.; Yu, K.; Jiang, D.; Zhu, Z.; Geng, H.; Luo, L. Appl. Surf. Sci.2005, 242, 212-217.
(7) Zehentbauer, G.; Krick, T.; Reineccius, G.A. J. Agric. Food. Chem.2000, 48, 5389-5395.

As the pathogenic viruses are always major threat to the human society, various kinds of antiviral agents are widely studied by many researchers. Of the various kinds of antiviral agent available, silver nanoparticles have received significant attention. However, inhibition mechanism of silver nanoparticles against viruses is not thoroughly understood yet. Understanding the inhibition mechanism of silver nanoparticle is essential for their medical uses. With conventional studies, silver nanoparticles used in antiviral studies are usually used. Colloidal silver nanoparticles are usually coated with surfactants to control their size and stability in solution system, which make difficult to discuss their surface effect against viruses. Also, effect of free silver nanoparticles on virus-infected cells sometimes make difficult to understand the mechanism.
This paper presents a new technique for synthesizing silver nanoparticles immobilized on textile fabrics using a radiochemical process. As no surfactants or polymers are applied in the synthesis process, the silver nanoparticles on textile fabrics have bared surface, which would interact with viruses directly. The silver nanoparticles are firmly immobilized on support textile fabrics and their effects on virus-infected cells are negligible. The antiviral activities of silver nanoparticles with bared surfaces were investigated.
Samples of the textile fabrics were then immersed in AgNO₃ solution. Following immersion, any excess solution was removed by centrifugal dewatering. The resulting AgNO₃-soaked textile fabrics were irradiated with a high-energy electron beam (4.8 MeV, 40 kGy). Radiochemical species generated by the irradiation reduce Ag ions to form metallic Ag nanoparticles. The resulting immobilized silver nanoparticles were characterized by TEM, ICP-AES and XAS, and their antiviral properties are discussed. These nanoparticles are firmly immobilized on the surface of a support textile fabric without the need for any binder or surfactant. Ag on textile fabrics exist as metallic state, which was confirmed by XAS analysis. TEM observation revealed that small Ag particles of about 2-4 nm were observed together with relatively large particles of more than 10 nm.
The antivirus test was performed basically following the ISO18184. Influenza virus and feline calicivirus were used. Concentration of EMEM (Eagle's Minimal Essential Medium) buffer was controlled as experimental parameter. With low medium concentration, Ag nanoparticles showed antiviral activity depending on the amount of Ag on textile fabrics. However, antiviral activities of Ag nanoparticles were not observed with high medium concentrations. Antiviral activity of the Ag nanoparticles are discussed on the basis of Ag nanoparticle amount and their chemical states.

Integrated microfluidic devices with nanosized array electrodes and microfiltration capabilities can greatly increase sensitivity and enhance automation in immunoassay devices. In this contribution, we utilize the edge-patterning method of thin aluminum (Al) films in order to form nano- to micron-sized gaps. Evaporation of high work-function metals (i.e., Au, Ag, etc.) on these gaps, followed by Al lift-off, enables the formation of electrical uniform nanowires from low-cost, plastic-based, photomasks. By replacing Al with chromium (Cr), the formation of high resolution, custom-made photomasks that are ideal for low-cost fabrication of a plurality of array devices were realized. To demonstrate the feasibility of such Cr photomasks, SU-8 micro-pillar masters were formed and replicated into PDMS to produce micron-sized filters with 3-4 mu;m gaps and an aspect ratio of 3. These microfilters were capable of retaining 6 mu;m beads within a localized site, while allowing solvent flow. The combination of nanowire arrays and micro-pillar filtration opens new perspectives for rapid R&D screening of various microfluidic-based immunoassay geometries, where analyte pre-concentration and highly sensitive, electrochemical detection can be readily co-localized.

Understanding biological pathways is essential for developing strategies for treating diseases and developing new drugs. The mainstay for probing these pathways is the microwell plate, a standardized array of containers in which a number of biological tests can be conducted and analyzed in parallel, thereby increasing throughput and accelerating scientists&’ ability to collect and analyze data. As technology has advanced, and the cost of reagents has increased, the size of the wells has decreased, such that 1536 wells, molded into a single plastic carrier, are commercially available. Microarrays are gaining in popularity as they are able to further increase the density of test sites and reduce the quantities of reagents required. However as microplates and microarrays advance, costs are rapidly increasing. Cost drivers include the increased precision required to dispense the small volumes of solutions consumed, the tolerances of both the plates and dispensing systems used, and the microfluidic systems needed for microarrays.
In this paper, we describe a new microplate technology which we call the nanoliter-droplet based Virtual-Well Microplate (nVWMP). Using hydrophilic glass pedestals, the surface works in conjunction with a simple dispensing system to precisely deposit nanoliter volumes of biological solutions in precise locations on the surface. Because wetting is controlled by a combination of surface chemistry and surface geometry, the nVWMP design relieves the tight tolerances required in conventional dispensing systems, lowering costs and increasing precision. Droplets that measure 28 nL can be dispensed with a precision of 2% using a simple robot costing less than $1,000.
The glass surface of the nVWMP can be chemically modified to selectively absorb peptides and proteins that are subsequently detected either by fluorescence microscopy or MALDI-TOF. We describe the modification of the surface with a combination of polyethylene glycol (to reduce non-specific protein absorption) and biotin moieties to selectivity bind NeutrAvidin from a solution containing BSA at concentrations as low as 0.1 micro-grams/mL (total of 8.33 x 10^.7 nanomoles of NeutrAvidin). MALDI-TOF was used with this geometry to detect NeutrAvidin with a sensitivity of 4 attomoles of protein. In another example we used a glass surface coated with a Ni+-chelate resin to selectively bind the KcsA ion channel protein. The nVWMP so prepared was shown to selective bind the TX7335 peptide from snake venom.

The design of sensitive diagnostic biosensor is of crucial significance for early detection of cardiac diseases. This is achieved by measuring the concentration of the biomarkers of interest. The binding of target antigens with the antibodies immobilized on the electrodes and the subsequent changes in the impedance are the driving factors for the detection of the biomarkers. The interactions at the electrode-electrolyte interface result in the modulation of the height of the electrical double layer (EDL). This theory of tuning the EDL is enhanced with the use of conformal electrodes. Hence, we use the porous structures as the substrates for electrode material deposition. The material properties of the electrode has a significant effect on the sensor performance. Thus, it is imperative to characterize the material properties of the electrode material and the sensor substrate.
Our initial experimental approach was focused on characterization of the surface of the sensor substrate after the electrode material deposition. We chose Molybdenum as the electrode material to investigate its properties for biosensing applications as there have been prior research works supporting the use of Molybdenum in biosensing applications. We analyzed the changes in the impedance at every step of the immunoassay which was built on the electrode substrates. The performance of the sensor was tested for multiple concentrations of the target antigen in ranges required for early disease detection. The ranges of the doses of the proteins were tested from ng/ml to µg/ml. The choice of buffer was varied and its effect on the sensor performance would be analyzed. The performance of the conformal electrode sensor was compared with that of planar electrodes. The electrode dimensions were varied and its effect were analyzed. The effect of variation of electrode dimensions and its corresponding correlation to electric field distribution were analyzed through finite element modelling (FEM) methods. The feedback obtained from the FEM results was incorporated in the design of experiments to validate the FEM analysis.

Plasmon enhancement of luminescence close to noble metal nanoparticles films is a powerful tool largely employed in many different areas. Here we investigate the possibility of using the plasmonic properties of silver nanoparticles (AgNP) films to get plasmon enhanced optically stimulated luminescence (OSL), which is a technique largely employed for radiation detection and dosimetry in many medical procedures. Silver nanoparticles/chitosan films were produced using the layer-by-layer technique and characterized by UV-Vis spectroscopy, Atomic Force Microscopy, and Fourier Transform Infrared Spectroscopy. We collected the Optically Stimulated Luminescence signal from irradiated NaCl deposited over nanoparticles films with 3, 5 and 10 AgNP/Chitosan layers. There is a strong enhancement of the optically stimulated luminescence intensity for NaCl deposited over the AgNP films compared to NaCl deposited over glass, and the maximum enhancement is obtained for the film with 10 AgNP/Chitosan layers. The enhancement can also be controlled by varying the distance between the luminescent crystals and the silver nanoparticle film, which leads to a maximum OSL intensity for a 15 nm distance. Results will be further discussed in terms of the different film properties. These findings may lead to a new class of highly sensitive and miniaturized radiation detectors.

Elastically deformable membranes have found different applications in biomedical devices such as micro-valves or Organs-on-a-chip. The elastic modulus of these membranes is an important parameter to determine their functionality. Typically, the elastic modulus is characterized by bulging tests, interferometry, or indentation techniques [1, 2]. However, all these techniques require specialized tools (e.g. microscopes, interferometers, or atomic force microscopes, respectively) to carry out the characterization. In this work, we present a simple, microfluidic platform to rapidly and accurately measure the elastic modulus of polymeric membranes.
The device is composed of two compartments. One of the compartments is connected to a microfluidic channel. The microfluidic channel is designed such that any movement of a dye solution is visible with the naked eye. The second compartment is connected to a syringe and a pressure transducer. The membrane under consideration can be placed and sealed between the two compartments. The first compartment is partially filled with a dye solution. The membrane is deformed by applying pressure through the second compartment using the syringe. The applied pressure is recorded by the pressure transducer. As the membrane is deformed due to the applied pressure, it displaces the fluid/dye inside the microfluidic channel. The extent of deformation (i.e. bulging) of the membrane is indicated by the displacement of the dye meniscus inside the microfluidic channel. The width of the microfluidic channel was set to 550 mu;m for the ease of tracking the meniscus displacement. Measurement of elastic modulus was demonstrated using a 35 mu;m Polydimethylsiloxane (PDMS) membranes and 10 micron Polyurethane (PU) membranes. The displacement of the meniscus changed linearly with the applied pressure. Young&’s elastic modulus was calculated for multiple independently fabricated membranes. The sensitivity of this platform can be tailored to measure the elastic modulus of membranes with very small deflection. For example, by designing the device with a membrane diameter of 8 mm, and microfluidic channel height of 250 mu;m, it is possible to measure membrane deflection as small as 10 mu;m with the naked eye.
We have demonstrated the ability to characterize the elastic modulus of elastic membranes using a simple microfluidic platform. In spite of its simplicity, our platform should be able to perform sensitive measurements with a wide range of polymeric membranes applicable for many biomedical devices.
Ref: [1] Ping Du, Hongbing Lu and Xin Zhang (2009). Measuring the Young&’s Relaxation Modulus of PDMS Using Stress Relaxation Nanoindentation. MRS Proceedings, 1222, 1222-DD02-03 doi:10.1557/PROC-1222-DD02-03. [2] Y. Xiang, X. Chen and J.J. Vlassak (2005). Plane-strain Bulge Test for Thin Films. Journal of Materials Research, 20, pp 2360-2370. doi:10.1557/jmr.2005.0313.

Many studies have shown that graphene has ability to enhance the stem cell proliferation and mediate differentiation linages. Graphene-polymer composite materials have been popularized in tissue engineering due to the outstanding thermal, electrical and mechanical properties of graphene. Currently, however, most of the studies focus on 2-D structured films which hardly represent the real conditions of scaffolds in vivo environment. In addition, dispersion of graphene in polymer matrix has always been challenging since the graphene tends to aggregate. In our study, we have successfully introduced graphene nanoplatelets (GNPs) into poly(4-vinphylpridine) (P4VP) matrix and fabricated nano- and micro-scale size fibers by using a cost-effective technique, electrospinning. SEM and TEM reveal uniform defect-free fiber structures and good dispersion of graphene inside the fibers. DSC and AFM indicate the enhancement of physical properties. We have also examined the biocompatibility of electrospun 3-D scaffolds of GNPs-P4VP nano- and micro-fibers with dental pulp stem cells (DPSCs). Our results show that the cells can accelerate proliferation to respond to the existence of GNPs even though cells have no direct contact with GNPs. EDAX together with SEM reveals a deposition of mineralized calcium matrix on the fiber scaffolds after 28-day incubation, which has possibly been caused by cell differentiation induced by fibrous scaffolds. In this study, we demonstrated an efficient approach to manufacture the graphene loaded polymer scaffolds which showed great dispersion of graphene, good physical properties and promising potential to promote DPSCs proliferation and differentiation.
The research was supported in part by the NSF-INSPIRE program. Part of research was achieved in ThINC facility at Advanced Energy Center.

Endovascular drug delivery system based on drug eluting balloon (DEB) is an innovative approach to reduce stenosis or neointimal formation as well as to avoid the possibility of in-stent-restenosis (ISR). Even though DEB technology has demonstrated safety and efficacy in preclinical and clinical studies, they are still limited in terms of the efficiency of drug delivery. DEBs are likely to lose drug due to blood flow during the delivery of the DEBs to target region in blood vessels. This loss of drug to the blood stream might induce the unwanted side effect of anti-proliferative drug such as paclitaxel at other tissue locations. Furthermore, in case of ISR, reliable contact is extremely difficult to make between the drug-coated surface of DEBs and diseased tissue under stent struts. Therefore, local and concentrated drug delivery to the tunica media of blood vessel with minimal drug loss and reliable contact to the vascular tissue region with ISR are highly desired. Microneedles (MNs) have been extensively investigated in the field of drug delivery especially for transdermal or ocular administration. Recently, we have demonstrated the multi-fold enhancement of drug delivery efficiency of perivascular drug delivery by developing perivascular MN cuffs that contained an array of MNs in our rabbit abdominal aorta models. The results showed two orders of magnitude enhancement in drug delivery and homogeneous distribution of drugs within tunica media layers. In this work, we developed MN drug eluting balloon (MNDEB) that can effectively deliver drug to the endothelium from the interior of blood vessels without any surgical procedure. An array of 40 mu;m tall MNs was fabricated by KOH chemical wet etching. Then, the MN shape was replicated to PDMS and finally SU-8 MNs shaped on the surface of MNDEB. This dual-step replication molding on the curved surface of DEB was performed by wrapping the PDMS mold on a pressurized balloon and curing the SU-8. The MNDEB was uniformly coated with a drug/carrier formulation containing a fluorescent model drug, rhodamine B. Afterwards, MNDEB angioplasty was performed with rabbit artery in vivo. No deformation or damage of the structure of MN array was observed after the animal study. No visible inflammation was observed when the MNDEB was applied. The animal study was performed with DEB only, DEB with drug coating, MNDEB with drug coating cases. Drug distribution within the vascular tissue samples was analyzed with the cryo-sectioned tissue samples after 2 day follow-up. Higher and more concentrated fluorescent signal was observed for the samples treated with MNDEBs. This confirms the enhanced endovascular drug delivery of MNDEB.

Calcium phosphate such as dicalcium phosphate anhydrate (DCPA), dicalcium phosphate dihydrate (DCPD) or hydroxyapatite (HAp) shows good biocompatibility and is a candidate of drug delivery carriers. A reverse micro-emulsion method has been investigated to control their crystal morphology in the nanometer region and to increase specific surface area. This study investigated the effect of mixing ratio of two surfactants on crystalline phases and specific surface area of calcium phosphate synthesized by a reverse micro-emulsion method. The nonionic surfactant with hydroxyl groups, tween80 (T) and the cationic surfactant, aliquate336 (A) were mixed into kerosene as an oil phase at the different T/A ratios, in which the amount of aliquate336 was fixed. A di-ammonium hydrogen phosphate solution including phosphoric acid to control pH at 4.0 and a calcium nitrate solution were adjusted, which were separately added into the kerosene. Both the emulsions were then mixed at the same volume and the Ca/P ratio of 1.0 and stirred for 24 hours. The products were centrifuged and washed with ethanol and distilled water. The crystalline phase was characterized with XRD and the specific surface area was measured by a BET method. The crystalline phases were depended on the T amounts; pure DCPD with the specific surface area of 6.7 to 12 m²/g was obtained at the T/A ratio of 4, the mixture of DCPD and DCPA with that of 48 to 162 m²/g was at the ratio of 5 to 8, and a low crystalline HAp with 163 m²/g was at the ratio of 9. These specific surface areas of calcium phosphate were apparently higher than those prepared with a wet method. The change of crystalline phases would be explained as follows; the increase of T amount decreased the micro-emulsion sizes to reduce bulk water to be DCPA, and increased the pH to precipitate HAp nanocrystals.

Biomolecular self-assembly at water/solid interface is being paid wide attention due to the applications in biosensors, nano-devices and medical treatments. Among many kinds of biomolecules, peptide is a unique candidate to modify properties of solid surfaces because of its short sequence and specific binding affinity to solid surfaces. Some specific peptides have been demonstrated to have an ability of self-assembly into long-range ordered structures on solid surfaces, which results into control of solid surface characteristics. The self-assembly is carried out by just placing a droplet of peptide aqueous solution on a solid surface at room temperature. In order to have a good understanding about peptide self-assembly process, the physical parameters relevant to the self-assembly behavior including concentration, incubation time, solvents have been studied so far. Temperature is also an important factor, but not fully understood yet.
In this work, we employed graphite binding peptide and its mutants ^[1-2] to gain a knowledge of the temperature effect on their self-assembly on graphite surface. We also investigated on MoS₂, another layered material which is more suitable for biosensors due to its semiconducting property. We found that temperature sensitively affects the adsorption and diffusion rate of peptides on the surfaces, which can be seen in the ratio of ordered phase and disordered phase of self-assembled peptides. The coverage of peptides on solid surfaces was adjusted as well. Furthermore, we found that the temperature can modify the conformation of peptides observed by the various heights of peptides and multiple kinds of ordered structures with different molecular recognitions.
Reference
Controlling Self-Assembly of Engineered Peptides on Graphite by Rational Mutation. Christopher R. So, Yuhei Hayamizu, Mehmet Sarikaya. ACS Nano, 6, 1648 (2012).
Controlling the surface chemistry of graphite by engineered self-assembled peptides. Dmitriy Khatayevich, Yuhei Hayamizu Mehmet sarikaya, Langmuir, 28, 8589 (2012)

Nanotechnology applied to the Medicine is providing new tools to the current therapeutic and diagnostic approaches because it offers the opportunity of winning some of the open challenges in the field. Currently, the demand is for dual or multiple-mode uptaking, protecting, stabilizing, enhancing and/or transporting functional payloads, such as two or more therapeutics, or therapeutics and contrast agents. Liposomes and in general vesicles, have shown to be one of the most promising supramolecular assemblies for delivering bioactives and some liposome-based-formulations are already present on the market. However, their overspread has been mainly limited by the lack of manufacturing strategies reliable at industrial level.^1,2 Recently, a compressed fluid-based technology, DELOS-SUSP process (Depressurization of an Expanded Liquid Organic Solution-Suspension), has been established for generating “tailor-made” lipid-based nanovesicles. DELOS-SUSP, which comprises the depressurization of a CO₂-expanded solution of lipids into an aqueous phase, is a simple, one-pot, versatile and clean manufacturing process, viable at industrial level and able to grant high control over the product properties. A new family of unilamellar nanovesicles, named Quatsomes, has been synthetized by exploiting DELOS-SUSP. More in details, Quatsomes are generated by the spontaneous self-assembly of a quaternary ammonium surfactant and a Cholesterol-like lipid. In general, Quatsomes show a very narrow particle size distribution (PDI 0.15-0.2) and an outstanding homogeneity in terms of morphology. The small size of these nanovesicles (70-100 nm) coupled with the high value of zeta;-potential (+70-80 mV) grants an excellent long-term colloidal stability.^3,4 Recently, it has been observed, that those nanovesicles have “built-in” anti-microbial and anti-biofilm activity.⁵ Furthermore, the inherent structure of such nanovesicles shows the advantage of offering multiple “strategies” for loading bio-actives. Dyes and biomolecules have been incorporated in Quatsomes by exploiting their surface, the hydrophobic environment offered by their membrane, their aqueous inner core and the functional groups available on their surface. Virtually infinite and controlled structures can be generated tuning the experimental parameters and using the generated nanovesicles as “building blocks” of multifunctional bio-active nanodevices. The favorable properties-by-process of Quatsomes together with the inherent advantages of DELOS-SUSP encourage us to further develop and evaluate this versatile “nanomaterial platform” as potential drug delivery carriers.
1. J. Shi et al., Nano Lett., 2010, 10 (9), 3223-3230.
2. H.I C.hang and M.K. Yeh, Int. J. Nanomedicine 2012, 7, 49-60.
3. L. Ferrer et al., Langmuir, 2013, 29 (22), 6519-6528.
4. I. Cabrera et al., Nano Lett., 2013, 13 (8), 3766-3774.
5. N. Thomas et al., J. Mater. Chem. B, 2015, 3, 2770-2777.

Nanostructured materials are of interest for a variety of applications. This talk describes the design and functionalization of nanostructured materials for biological and biomedical applications. Specifically, we have synthesized metallic, metal oxide, semiconducting and organic nanoparticles and nanocomposites for bioimaging, bioseparation, biosensing, theranostic, drug delivery and tissue engineering applications. The synthesis, unique properties and biocompatibility of these nanostructured biomaterials will be presented in this lecture.
We have also fabricated nanofluidic systems for drug screening, in vitro toxicology, clinical sample preparation and diagnostic applications. The principles behind the design and creation of nanoscale patterns, building blocks and surface functionality to tackle various challenges in biomedical applications will be discussed.

The synthesis of gold nanoparticles (AuNPs) having better dispersibility and catalytic ability in variable buffering systems containing desired salt concentrations is reported herein. The fact that aldehydes and ketones results in the formation of catalytic hybrid material with amino functionalized silanes directed the use of carbonyl functional group (aldehydes and ketones) specifically formaldehyde, acetaldehyde, acetone and t-butyl methyl ketone alongwith 3-aminopropyltrimethoxysilane (3-APTMS) to synthesis such nanomaterial. Accordingly, a comparative study on the synthesis of 3-APTMS and variable organic reducing agents mediated synthesis of AuNPs are reported herein. The findings reveal that 3-APTMS capped gold ions are converted into AuNPs with precise control of pH- and salt- sensitivity. The major findings reveal the following: (1) 3-APTMS being amphiphilic, dispersibility of as prepared AuNPs largely depends on the organic reducing agents. (2) An increase in the hydrocarbon content of the reducing agent facilitate the dispersibility of AuNPs in organic solvent whereas decrease of the same increases the dispersibility in water, (3) AuNPs made through aldehydic reducing agents (formaldehyde and acetaldehyde) have relatively better salt and pH tolerance as compared to ketonic reducing agents (acetone, t-butyl methyl ketone), and (4) an increase in 3-APTMS concentrations imparts better salt- and pH- resistant property to AuNPs irrespective of organic reducing agents. A typical example on the role of AuNPs in homogeneous catalysis and as peroxidase mimetic will be discussed.

Modifying the nanostructure/chemistry of materials allows to optimize their properties [1]. Our strategy rests on creating nanopatterns that act as surface cues [2,3], affecting cell behavior. Chemical oxidation creates unique topographies [4], becoming a general strategy to improve biocompatibility. Our treatment selectively inhibits fibroblast growth while promoting osteogenic cell activity [5] in vitro. Enhancement of mechano-biocompatibility may occur by coating with spider silk [6,7]. Improvement of antibacterial properties using plasma strategies will also be discussed [8,9].
[1]F Rosei, J Phys Cond Matt 16, 1373 (2004)
[2]F Variola et al, Small 5, 996 (2009)
[3]F Variola et al, Biomaterials 29, 1285 (2008)
[4]F Vetrone et al, Nanolett 9, 659 (2009)
[5]L Richert et al, Adv Mater 20, 1488 (2008)
[6]C Brown et al, Nanoscale 3, 3805 (2011)
[7]C Brown et al, ACS Nano 6, 1961 (2012)
[8]O Seddiki et al, Appl Surf Sci 308, 275 (2014)
[9] M Cloutier et al, Diam. Rel. Mater. 48, 65 (2014)

Nano-carbons have greatly been interested in various fields of research including biomedical area. We believe that the large scale synthesis of nano-carbon should be free from using excess energies for firing, sintering, melting,expensive equipments and/or multple steps. We, propose here Soft processing(=Eco-Processing) of functionalized Graphenes at ambient conditions. The Soft processing provides number of advantages which includes (a) simple reaction set up,(b) at ambient temperature and presure conditions, (c) simple ad short-cut procedures and (d)less operating costs.
In the present study, we have utilized “Submerged Liquid Plasma [SLP]” and “Electrocemical Exfoliation[ECE] methods. SLP methods resulted the direct synthesis of Nitrogen functionalized Graphene Nano-sheets from Graphene suspension and/or Graphite electrode in acetonitrile liquids. Products contains few layers (< 5) Graphene nanosheets. Unsaturated or high energy functional group (e.g. C#65309;C, C#65309;N and Cequiv;N) have formed in the products. We could confirm those functionalized Graphenes are electrochemically active. Using pencil rods instead of Graphite rods we have also succeeded to prepare the Nano-clay/Graphene hybrids by this SLP methods. Reduction and functionalization of Graphene oxides and Synthesis of Graphene/Au Hybrids also realized by SLP.
In the ECE, graphite anode is exfoliated electrochemically by H₂O₂-NaOH or Glycine-H₂SO₄ aqueous solutions under ambient temperature and pressure,for 5-30 min with +1-+5 volt, into 3-6 layers Graphene Nanosheets[GNs]. Those conditions are much milder than those reported before using other chemicals like ionic liquids and/or H₂SO₄-KMnO₄,etc.,because O₂^2- ions or ionic complex like Glycine-HSO₄^- would assist the exfoliation of graphite layers. Our products:GNs suspended in solutions can be transformed in the 2nd step in the same container using BrCH₂CN/dioxane into N-FG, further into Au-Hybridized N-FG by the sonification with Au nanoparticles. We have confirmed the excellent catalytic performance of those hybrids. It should be noted that Soft Processing can directly produce “Graphene Ink”;Graphenes dispersed in various liquids, under mild conditions..
Key words: Graphene, Solution,Liquid Plasma, Exfoliation,Hybrid
References J. Mater Chem A,(2014) 2, 3332: Sci. Rep., 4(2014),04395: Carbon,78 (2014),446: Sci. Rep. 4 ,4237-4242 (2014): Nanoscale(2014) 6,12758: Adv. Funct. Mater. 25,298-305(2015): J. Mater Chem A,3,3035(2015)

The Néel relaxation of magnetic nanoparticles (MNP), subject to alternating magnetic fields, depends exponentially on their core diameter whilst the Brownian relaxation depends critically on their hydrodynamic volume[1]. Recent developments[2] in the synthesis of highly monodisperse and phase-pure magnetite nanoparticles allows for reproducible control of the former, even in biological environments, enabling novel imaging[3]^,[4] and spectroscopy[5], under ac excitations such as magnetic particle imaging/spectroscopy (MPI/MPS). With appropriate functionalization of the monodisperse core, and utilizing the latter relaxation, it also allows for the development of sensitive, relaxation based assays.
Following a brief review of the physics of magnetic relaxation, we describe the development of a high-throughput, sensitive and rapid method for detection of proteases, crucial in diagnostics and therapeutic applications, using a facile magnetic assay based on magnetic particle spectroscopy (MPS). We used a peptide labeled with biotin at N- and C- terminals, with a middle part containing a selective site to be recognized and cleaved by a specific protease. When this peptide is added to the neutravidin functionalized nanoparticles the MNPs aggregate, resulting in well-defined changes of the MPS signal (both peak intensity and position - in principle, eliminating false positives). However, in the presence of protease, this peptide is cleaved and as a result, nanoparticles get redispersed in the solution again, and the MPS signal(s) returns to its pre-aggregation state (before addition of the peptide). These variations help to detect a specific protease by MPS, only relying on the magnetic relaxation characteristics of the nanoparticles. The utility of this assay is demonstrated by the detection of two proteases -- Trypsin and Matrix Metalloproteinase.
Further, surface functionalization of the nanoparticles with PEG of appropriate length and density results in favorable biodistribution, avoiding the kidney (of critical concern in patients with chronic kidney disease) and promoting enhanced circulation time in animal models. The latter has demonstrated the first blood volume images in rodents using magnetic particle imaging[6]. Finally, conjugation of dyes turns these MNPs into multimodal contrast agents[7] (MRI, MPI and NIRF) and which we have used to further enhance studies of the MNP biodistribution, with significant anatomical detail, in rodent models[8].
[1] Kannan M. Krishnan, IEEE Trans. Mag.46, 2523-2558 (2010
[2] R. Hufschmid et al, Nanoscale (accepted, in press), DOI: 10.1039/C5NR01651G
[3] B. Gleich & J. Weizenecker, Nature435, 1214 (2005).
[4] R.M. Ferguson et al, IEEE Trans. Med. Imag. 34, 1077 (2015)
[5] S. A. Shah et al, Phys. Rev. B (submitted)
[6] P. Goodwill et al, Proc. 5^th IWMPI (2015)
[7] Hamed Arami et al, Biomaterials52, 251 (2015)
[8] This work was supported by NIH/NIBIB grants 1RO1EB013689-01, 1R41EB013520-01 & 2R42EB013520-02A1

RF plasma nanotexturing technique is utilized to create dense, vertically aligned metallic nanowire arrays on metallic substrate structures such as coronary stents and neural electrodes. Antibiofouling neural electrodes were produced from a 316L stainless steel substrate by nanotexturing the surface with repetitive RF-plasmas processing and selectively coating the tips of the resultant nanopillars through oblique incidence sputter deposition of hydrophobic PTFE. Such a nanowire array structure induces a significant reduction of surface impedance, thus allowing a room to add antibiofouling nano-island arrays without sacrificing the desired electrical conductivity much. Human aortic endothelial cells were cultured on the antibiofouling electrode surface and cell viability was examined by immunofluorescent imaging of cytoskeletal actin, FDA assay, MTT assay, and area-based evaluations of cell spreading. Data indicates notable antibiofouling properties for the nanotextured surface which is enhanced by the selective addition of a hydrophobic coating. The textured and coated samples exhibited an 8-fold decrease in cell adhesion/coverage, thus enhancing the antibiofouling tendency. The proposed nano processing can be beneficial to chronic electrode implants for the purpose of retaining electrical conductivity while decreasing cellular biofouling.

The ability to pattern biomaterials in planar and three-dimensional forms is of critical importance for several applications, including drug safety screening, tissue engineering and repair. 3D printing enables one to rapidly design and fabricate soft materials in arbitrary patterns without the need for expensive tooling, dies, or lithographic masks. In this talk, our efforts to creating vascularized living tissues via 3D bioprinting will be described. I will present recent advances in the design of cell-laden inks, extracellular matrices and fugitive (vascular) inks for 3D bioprinting of vascularized, heterogeneous cell-laden tissue constructs with as well as ongoing efforts to characterize these 3D living tissues.

Fluorescently labeled nanoparticles (NPs) are emerging as a versatile tool in biomedical applications and nanotoxicological studies. At present, the simultaneous colocalization of accumulated fluorescent NPs within cellular components is still a challenge due to spectral overlay of NP-fluorescence and multiple fluorescent stained cellular components. To tackle this problem we have developed different synthesis routes to create multi-fluorescent silica NPs with controllable sizes, low polydispersity index, high labeling efficiency and enhanced fluorescence photostability.
By using a water in oil microemulsion method or amino acid catalyzed seeds regrowth technique (ACSRT), core/shell NPs with multiple fluorescence characteristics have been obtaining by direct incorporation of organic fluorescent dyes. Such particles can be easily colocalized and clearly distinguished from multiple stained cellular components using conventional fluorescent microscopes, and offer several advantages for in vitro studies at single-cell-level.
Besides particle size, surface charge plays an important role in cellular accumulation and possible cytotoxicity of NPs. Therefore, we synthesized five different single- or multi-functionalized fluorescent silica NPs (FFSNPs) by introducing various ratios of amino and sulfonate groups into their surface and to tailor particles zeta potential from positive to negative values. We show that particle uptake does not only depend on the surface charge, but also by the presence of serum proteins. In the absence of serum, positively charged NPs are stronger accumulated by cells than negatively charged NPs. In contrast, in serum-containing medium anionic FFSNPs are internalized by cells more strongly. This shows that at physiological condition, when the presence of proteins is inevitable, sulfonate-functionalized silica NPs are feasible choice to achieve a desired high rate of NP internalization.
In conclusion, we show that by creating multiple-doped fluorescent NPs and by tuning surface chemistries, it is possible to design particles with unique properties with high potential as probes for fundamental biological studies as well as for molecular imaging.

While there have been significant advances in stem cell therapy and tissue engineering, the search for an ideal substrate for stem cell culture and differentiation continues. Human bone marrow derived mesenchymal stem cells (hMSCs) have been widely recognized to have significant biological and clinical importance due to their ability to differentiate into multiple cell types. While stem cell differentiation depends largely on the various soluble factors and biomolecules, lineage commitment of stem cells can also be governed by their interaction with nanoscale features of the extracellular matrix (ECM). Nanotechnology has emerged as a promising tool to design and create materials with nano-sized features and architectures which are capable of biomimicking the native ECM environment for stem cell culture and differentiation. In particular, graphene anf graphene oxide have generated great interest because of its unique chemical, mechanical and electrical properties. The synergy of the properties of graphene and graphene oxide and differentiation potential of stem cells could provide exciting opportunities for new and novel therapeutic applications. However, little is known about the effect of these graphene coatings on stem cell fate. Here, we report the interaction between stem cells and graphene and graphene oxide coating in terms of stem cell growth and differentiation. Our results show that these coatings promoted differentiation of MSCs and their unique differentiation profiles are dependent on the π- π interactions between the graphene/graphene oxide coatings and the soluble biomolecules involved during the differentiation process.

Regenerative medicine aims to regenerate tissues and organs by utilizing the highly regenerative potentials of various stem cells, such as embryonic stem cells, multipotent adult stem cells, tissue specific stem cells, and induced pluripotent stem cells. There is a growing recognition of 3D matrix microenvironments on the fate and function of stem cells. A key challenge facing regenerative medicine is to rationally design and create 3D microenvironments that can recapitulate those in a developmental or healing program to maintain stemness, to accelerate proliferation, or to direct stem cells to differentiate along a specific therapeutic lineage. Our lab takes a biomimetic approach to design biomaterial-based 3D microenvironments on the nano- and micro-scales (matrix, signals, and supporting cells etc.) for stem cells to regenerate tissues. Several examples of regenerating dental/craniofacial, orthopaedic, and cardiofascular tissues will be presented to demenstrate the effectiveness of biomimetic approaches on the nano- and micro-scales.

Tooth caries are among the most prevalent of human chronic diseases. Incipient caries, e.g., are the earliest detectable signs of tooth decay clinically and radiographically. At this stage, the lesion has not penetrated the dentin, limited to enamel tissue with an intact DEJ (dentin-enamel junction). If left untreated, caries often result in pulp damage requiring pulp extirpation, complex restorative procedures or tooth extraction. A long-standing practical challenge associated with caries and other dental diseases, therefore, is associated with incorporating a functional mineral layer fully integrated into the molecular structure of the tooth for use in repairing damaged tooth surfaces. Both in teeth and bone, hydroxyapatite (HAp) is the primary mineral component which possess microstructures resulting in distinct mechanical properties relevant for the functional performance of the specific mineralized tissue (enamel, dentin, cementum and bone). Formation and morphogenesis of HAp in dental tissues and bones are controlled by proteins specific to each tissue. Herein, using directed evolution approaches, we identified peptide domains that bind to HAp and showed that they can control formation and morphogenesis of the apatite crystals. Using knowledge-based bioinformatics, and based on a critical discovery that HABPs have sequence similarity to amelogenin (the key protein in enamel formation), we identified amelogenin derived peptides (ADPs). Using ADPS in various solutions and gel formulations, we demonstrated that they can direct calcium phosphate min formation with controlled structural characteristics deposited as a thin mineral layer on dentin, enamel and cementum on extracted human teeth in vitro and rat in vivo. Integrating the recent advances in peptide design for directing biomineralization we address one of the key problems in dental tissue restoration through achieving remineralization on the surface of demineralized teeth. Establishing the scientific foundations for remineralization on teeth and developing novel biomimetic therapies have enormous potential to empower clinicians and practicing dentists to address a wide variety of dental problems and, thereby, disruptively change the current “drill-and-fill,” dental practices

Endotracheal tubes (ETTs) are a widely used medical device for patients in need of assisted breathing. While in use, biofilm formation occurs as a result of mucus accumulation on the surface of the device which can lead to obstruction of airflow and respiratory infection. In addition, the recent emerging of antibiotic-resistant bacterium further elevates the danger. Therefore, there is a need to modify existing medical devices with alternative therapeutic agents and surface chemistries in order to prevent biofilm formation from bacterium.
In this study, endotracheal tubes were modified by applying a multi-layered, polymer coating to yield anti-microbial and anti-fouling properties. The coating system was composed of three distinctive layers: (1) a reservoir layer composed of an anti-microbial peptide (AMP) for drug release with efficacy for 72 hr to kill bacteria above the minimum inhibitory concentration (MIC). 2) A capping layer, which controls release rate of AMP during the time scale and provide anchorage for 3) a non-fouling hydrogel layer, which accommodates AMP on the surface and provide sputum resistance.
Two different polymers were designed and synthesized based on their functions. A variety of analytical and in vitro methods were carried out to assess the dual functionality of the modified ETTs. AMP release was quantified using high-performance liquid chromatography (HPLC) and resulted in a controlled release pattern after 72 hr. The anti-fouling coating was able to reduce mucus attachment by 25-30% determined by radio-labeled porcrine mucin. In vitro cultures of S. aureus (MRSA) and P. aeruginosa were used to assess the efficacy of the AMP. The AMP was able to maintain a 3.5-3.7 log reduction against MRSA for 72 hr. A decrease in efficacy was observed in P. aeruginosa cultures. These results show great promise for manufacturing medical devices capable of greatly reducing respiratory complications.

We propose that the next generation of bone substitutes could be load-bearing if they are engineered as a bioactive material which not only emulates the complex hierarchical organization of the interpenetrating protein and mineral phases, but is also capable of maintaining the mechanical properties throughout bioresorption. The polymer-induced liquid-precursor (PILP) process has been previously shown to result in the intrafibrillar mineralization of different types of type I collagen substrates, which has reproduced the fundamental nanostructural organization of native mineralized tissues. In order to achieve bone-like mechanical properties, we are now targeting the lamellar microstructure of bone by preparing laminated films of dense, collagen scaffolds that are assembled with cholesteric order. These liquid-crystalline (LC) collagen scaffolds are made by concentrating acidic collagen solutions up to physiological levels, and then stabilizing the assembly through fibrillogenesis brought on by neutralization. We are currently mineralizing these dense, LC collagen constructs via the PILP process to create a bioresorbable material with a high degree of mineral loading, mimicking bone&’s hierarchical organization from the nanoscale to the microscale. It is hypothesized that this biomimetic material will enable us to emulate the high strength and toughness of bone, while also providing a bone-like matrix that can stimulate cellular activity to regenerate natural bone tissue through the concerted action of the bone remodeling unit.

Often glass, which is made traditionally by quenching a melt, is considered a homogeneous solid for many applications. This has been true certainly in the biomedical field, where, for example, the most widely used 45S5 sodium phosphosilicate glass is assumed to be a single phase bioactive material. However, now we are learning that it is not only phase separated at nanoscale but in fact cell response is significantly affected by the nature of its nanostructure. We have also discovered that more than ten thousand times larger cells respond differently on nano-macro porous calcium silicate bioactive scaffolds that differ only in the size of nanopores, for example, 4 nm vs. 18 nm. These observations demonstrate the importance of nanostructure for the performance of glass products in dental and bone regeneration applications. Then it becomes crucial to develop glass fabrication processes for a given chemical composition, which will yield desired nanostructure of the ultimate product. In this presentation we will review and discuss these specific processes, which are broadly based on the conventional melt-quench and sol-gel methods of glass making. We will show how the variables like melt temperature, cooling rate, sample size of the former method affect the nanostructure (for example, interconnected vs. droplet-type distribution of phases), or gelation time, type/amount of catalyst, drying conditions, sample configuration, etc. affect the nano/macro porosity that results from the latter process. Examples of the correlation between processing conditions with the nanostructure and the performance of the final product (such as attachment and proliferation of MC3T3-E1 pre-osteoblast cells) will be presented.

Bulk metallic glasses (BMGs) have been advanced as a means to achieve durable multiscale, nanotextured surfaces with desirable properties dictated by topography for a multitude of applications, including cellular response-manipulating patterns. One barrier to this is the lack of a bridging technique between macroscale thermoplastic forming processes and nanoimprint lithography. This arises from the difficulty and cost of generating controlled nanostructures on complex geometries by conventional top-down approaches, compounded by the necessary destruction of any resulting reentrant structures (such as any feature patterned on a vertical wall) during rigid demolding. We have developed a generalized method to overcome this limitation by sacrificial template imprinting using zinc oxide nanostructures. It is established that such structures can be grown cheaply and quickly with tunable morphologies on a wide variety of substrates out of solution, which we exploit to generate the nanoscale portion of the multiscale pattern through this bottom-up approach. In this way, we achieve metallic structures simultaneously demonstrating features from the macroscale down to the nanoscale requiring only the top-down fabrication of macro/microstructured molds. Upon detachment of the formed part from the multiscale molds, the zinc oxide remains embedded in the surface and can be removed by etching in mild conditions to both regenerate the mold and render the surface of the BMG nanoporous. Further, by skipping the etching step on the BMG, we have generated surface nanocomposites of antimicrobial nanorod arrays embedded in a metallic glass. The ability to pattern metallic surfaces in a single step on length scales from cm down to nm is a critical step towards fabricating devices with complex shapes which rely on multiscale topography for their function, such as medical implants or antimicrobial components.

Electrophoretic deposition method has been developed for the deposition of biopolymers, such as chitosan, hyaluronic acid, alginic acid and chiral polymers, such as poly-l-lysine and polyl-ornithine. The electrodeposition mechanism is triggered by the pH changes at the cathode or anode surface. It was found that polymers can be used as dispersing, charging and film forming agents for electrophoretic deposition of nanostructured hydroxyapatite, titania, zirconia and other bioceramics, halloysite nanotubes and bioglass. The composite films containing bioceramics and bioglass nanoparticles in a polymer matrix were prepared on stainless steel, Ti and NiTi alloy substrates. The films provided corrosion protection of the metallic implants in simulated body fluid solutions. The results of thermogravimetric analysis showed that hydroxyapatite content in the polymer matrix was varied in the range of 35-80 wt% by variation of hydroxyapatite or polymer concentration in the solutions. The hydroxyapatite particles showed preferred crystallographic orientation in the chitosan matrix. It was shown that film thickness can be varied in the range of 0.1-200 microns. The composite films were prepared as monolayers, multilayers or materials of graded composition. Electrochemical strategies have been developed for the deposition of composite films containing heparin and albumin in a biopolymer matrix. It was found that composite films can be used for controlled release of drugs and antimicrobial agents. The co-deposition of biopolymers, enzymes and carbon nanotubes alowed for the fabrication of composite films for application in biosensors. It was discovered that bile acids can be used for efficient dispersion and electrophoretic deposition of carbon nanotubes for application in biosensors. New electrochemical strategies have been developed for the anodic electropolymerization of polypyrrole and fabrication of adherent films on stainless steel. The approach was based on the use of dopants from dopamine and salicylic acid families, which reduced electropolymerization potential, promoted charge transfer and allowed for the fabrication of adherent films of polypyrrole and composites. The films showed promising performance for aplications in biomedical implants and biosensors.

Mechanical assessment of soft biological tissues and organs has broad relevance in clinical diagnosis and treatment of disease. However, at present it relies on characterization methods that are invasive, lack micro-scale spatial resolution, and are only tailored for specific regions of the body under quasi-static conditions. Here, we develop conformal and piezoelectric nanoribbons of lead zirconate titanate (PZT) that enable in vivo measurements of soft tissue viscoelasticity in the near surface regions of the epidermis. Soft, reversible lamination onto the skin enables rapid, quantitative assessment of viscoelastic moduli, with ability for spatial mapping. These systems achieve conformal contact with the underlying complex topography and texture of the targeted skin, as well as other organ surfaces, under both quasi-static and dynamic conditions. Experimental and theoretical characterization of the responses of piezoelectric actuator-sensor pairs laminated on a variety of soft biological tissues and organ systems in animal models provides information on the operation of the devices. Studies on human subjects establish the clinical significance of these devices for rapid and non-invasive characterization of skin material properties. Applications in vitro with mock and ex vivo skin preparations under varying conditions and in vivo on human subjects, collected at various locations over all main regions of body, under both normal conditions and following administration of pharmacological and cosmetic (moisturizing) agents, demonstrate the capabilities.

Cell guides and cell traps consist of microstructured surfaces featuring selective cell affinity within calibrated designs. Among the bunch of techniques being used for their fabrication, direct laser writing emerges as a reliable, fast and flexible technique. In the present work, an IR laser has been used to selectively write anatase and graphite coatings on Si. The laser parameters were optimized to induce local damage with features of circa 10 mu;m and minimized ablation searching for the sharper edges possible. The writen features were characterized by both microscopic and spectroscopic techniques. Field emission scanning electron microscopy shows that hierarchichal structures are formed at particular processing conditions with nanofiber decorated microparticles emerging from irradiated rails. Energy disperse X-ray maps show that the underlying Si becomes the dominant element in the irradiated areas. Thus, the micropatterns can be described as hybrid chemo-topographic structures. This feature is additionaly confirmed by micro-Raman spectroscopy. The variation of the surface properties was further tested by making contact angle measurements on densely irradiated surfaces. The relatively hydrophobic behaviour of the anatase and graphite films became a strongly hydrophilic surface after irradiation. Particular micropatterns were designed for the culture of human mesenchymal stem cells (hMSCs). Adhesion studies confirmed that the irradiated rails become barriers for the hMSCs, which presented particularly eleongated shapes on confluent edges with the micropatterned features. Particular designs with non confluent hexagonal corrals show that cells can migrate around the hexagons while they become mostly trapped within the corrals. Such devices are relevant tools for the study of hMSCs differentiation under non-conventional substrate conditions.

The need for new even smarter microengineering materials for biomedical applications is driven by a need for enhancement of their performance in real biomedical applications. Therefore, we developed new advanced nanocomposite materials based on biocompatible SU8 resist (standard negative tone photoresist). In order to control and engineer the properties of the nanocomposite materials, we used different fillers like carbon nanotubes (CNTs) previously optimized for this purpose, carbon black nanoparticles or TiO₂.
At first, we have optimized the inks preparation conditions (including the appropriate solvents, surfactants and all steps of the nanoparticle-based inks preparation process). Nanoparticles dispersion was characterized in the ink and in the SU8 matrix in composites. Processing of the obtained composites was successfully optimized for the ink jet printing process, photolithography and screen printing on various substrates (like different wafers, plastic, paper, fabric). Especially, photo-patterning was optimized at each processing step in order to minimize their drawbacks. In this way, we achieved to develop new biocompatible nanocomposites having high absorption (SU8-carbon black nanocomposites), high refraction (SU8-TiO₂ composites), tuneable electrical resistivity, thermal conductivity and mechanical properties (SU8-CNTs nanocomposites). As an example, the increase in hardness of 122% and in the Young&’s modulus of 56% (for 0.8 wt% of CNTs in SU8) and thermal conductivity of 3.7 times (for 10wt% of CNTs in SU8) was achieved for smart nanocomposites containing randomly oriented CNTs.

Titanium-based alloys are common materials for orthopedic implants. The fast development in additive manufacturing has broadened their applications in biomedical area, because of its high geometrical freedom in fabricating customized implants. Direct Metal Laser Sintering is an additive manufacturing technique. Due to high localized thermal input and fast cooling rate, the as-sintered materials contain non-equilibrium phase. In Ti-6Al-4V, the as-fabricated microstructure is acicular martensite with poor ductility. The major failure mode for Ti-6Al-4V is by fretting corrosion at the metallic interface in body fluid. So, proper post-treatments are introduced to ensure better properties. In this work, a variety of solutions, stress relieving and annealing heat treatments were conducted on Ti-6Al-4V fabricated by direct metal laser sintering. The effect of heat treatment temperature on microstructure, mechanical properties and corrosion behavior were experimentally investigated. The microstructure, phase percentage and lattice parameters were measured by X-ray diffraction with Rietveld refinement. The corrosion behavior of post heat treated samples were characterized electrochemically in simulated body fluid. It was found that β solution treatment, annealing and stress relieving at elevated temperature can improve the corrosion resistance by reducing the α/β phase boundaries which are vulnerable to corrosion. Rutile phase with different degree of crystallinity was detected on the Ti-6Al-4V/electrolyte interface after various heat treatments. X-ray photoelectron spectroscopy (XPS) was utilized to examine the interface properties.

Silicon nitride is a ubiquitous nanofabrication material that offers favorable mechanical and electrical properties in a multitude of device types. We favor thin-film silicon nitride membranes with well-defined micro- and nanoscale pores to make chemical sensing devices, including single-molecule nanopore sensors. Nanopore sensors pass solution-phase molecules, usually with electrophoretic driving, through ~1-100nm diameter channels in thin, <100nm membranes. Silicon nitride has favorable physical properties for supporting nanopores, but its complex surface chemistry poses challenges for these devices in terms of chemical stability and in the strong interactions between the chemically-diverse surface and highly constrained molecules within the pores. Pore clogging is only one possible challenge we are hoping to circumvent. We use electroless gold plating to controllably tailor the nanopore dimensions, and to allow thiol monolayer self-assembly to give us tunable surface chemistry for improved chemical stability, performance, and reliability. We show control over the thin gold film nanostructure and surface decoration, and can use the extreme nature of the zeptoliter nanopore volume to probe chemical-surface interactions and improve sensing. The deposition of electroless gold films onto silicon nitride opens up a myriad of applications, and can provide a base for electrical connections. Furthermore, this process can be used in combination with spatial patterning of the gold for the fabrication of optically active surfaces, including substrates for surface enhanced Raman spectroscopy.

Self-assembling peptides (SAPs) have the ability to spontaneously assemble into hierarchical structures in solutions such as ribbons, fibrils and fibres. The reversible molecular association of SAPs has been shown to macroscopically lead to self-supporting gels, offering great promise in therapeutics e.g. bone and enamel regeneration[1]. Self-assembled gels typically display weak mechanical properties. Although these gels could have been applied as e.g. injectable scaffold in treatment of small size bone/tooth defects[2], their mechanical stability is insufficient for large load-bearing bones and there are challenges in implanting them into tissue defects. Incorporation of SAPs into polymeric matrices is a common strategy to enhance their mechanical stability[3]; however integrated methods enabling fibre assembly at both micro and molecular levels are lacking. To address the above mentioned challenge, this research investigated the encapsulation of two types of tape forming 11 amino acid residue SAP within a poly(ε-caprolactone) (PCL) fibrous web via. an electrospinning process. With regards to electrostatic charges during spinning process, P₁₁-4 (CH3COQQRFEWEFEQQNH2) with -2 net charge and P₁₁-8 (CH3COQQRFOWOFEQQNH2, where O is ornithine) with +2 net charge were selected as model SAPs. Resulting webs exhibited interesting biphasic nano- and micro-structural features resulting from simultaneous fabrication processes at molecular and microscopic level. The novel mechanism of fabricating electrospun PCL sub-micron fibres containing self assembled peptide nano-fibrils in a simple one-step process is resulting from the SAP monomeric assembly and the spinning process. FTIR and CD spectroscopy results on PCL-peptide solutions suggested that monomeric peptide self-assembly takes place during or immediately after electrospinning, while FTIR spectroscopy highlighted a β-sheet secondary conformation across the fibres. Peptide concentration and its net charge were found to be highly influential in the fibrous morphology and specifically in determining the nano-scale structure of the fabric. The direct and indirect cell culture studies on the electrospun self-assembled matrices demonstrated a spread-like morphology of L929 mouse fibroblasts. The biphasic structure combines the benefits of nano-fibres, suitable for cell attachment and submicron fibres with open pore areas suitable for cell penetration and differentiation [4].Considering the current challenges of 3D scaffolds such as lack of surface bioactivity or poor biomechanical stabilities, the investigations of these materials for mineralization of hard tissues are underway.
[1] Stephanopoulos, N., et al. Acta materialia, 2013. 61(3): p. 912-930.
[2] Firth, A., et al. Nanomedicine, 2006. 1(2): p. 189-199.
[3] Andukuri, A., et al. Acta biomaterialia, 2011. 7(1): p. 225-233.
[4] Gentsch, R., et al. Macromolecular rapid communications, 2010. 31(1): p. 59-64.

Carbon nanotubes (CNTs) have been studied and utilized in a variety of biomedical applications including drug delivery devices, cellular interfacing, biosensors and molecular detection owing to their unique chemical, mechanical, and electrical properties and hollow structure. Template-based chemical vapor deposition (CVD) is one of the most efficient ways of CNT synthesis. The CNTs manufactured by this process are the most versatile for a wide range of applications due to the possibility of integrating them into nanoscale devices without the need for nano-assembly. In this process, decomposition of carbon carrying gas results in deposition of carbon over a choice of template. The template dictates certain physical features of the CNT, including length and outer diameter, while the process itself affects features, such as tube wall thickness, carbon deposition rate and carbon morphology. These CNTs has been used previously for injecting sub-attoliters of fluid and particles into cells and electrically measure cell membrane kinetics. Also, electrochemical sensors have been fabricated by array of aligned CNTs in an integrated chip. In order to properly employ CNTs for aforementioned applications, tailoring the CNT features and properties for any specific application is highly desirable.
To understand how process parameters affect the resultant CNTs manufactured by template-based CVD and to control their geometrical and structural features, the results of a systematic parametric study will be presented. The effect of three main process parameters of deposition time, temperature, and gas flow rate, on properties of CNTs resulted from deposition of carbon over commercially available anodized aluminum oxide membranes will be discussed. The results show that temperature and deposition time can be used for fabricating CNTs with very thick walls and creation of carbon layer for interfacing with CNTs. For changes in precursor gas flow rate, carbon deposition reaches a maximum where mass transport or reaction kinetics limits the deposition below and above that threshold. Moreover, deposition time shows three regimes of nucleation, normal deposition, and carbon infiltration, over the period of 20 hours. Importantly, carbon morphology does not change over the range of the three parameters tested. These findings show that process parameters can be independently utilized to produce CNTs with similar or differing cross-sectional dimensions and other useful features, each with distinct advantages.
The results of the work provide a means of tailoring the properties of free or embedded CNTs that are useful for bio applications including implantable sensors, bio-fluid transport, electrochemical sensing and cellular biology. Preliminary work will be presented to highlight how tailoring CNT properties are useful for these applications.

Due to the low melting point of gallium (29.8 #730;C), it can be melted in warm water/organic liquids/aqueous organic solution which are not too volatile, and thus produce two immiscible liquid phases. Irradiation of such a liquid system with ultrasonic energy induces cavitation in the liquid, which results in dispersion of the molten gallium as micro/nano-spheres. We attempted to entrap organic substances in metallic Ga particles during their ultrasonic formation, rather than by homogeneous reduction of metal ions in solution. Chiral imprinting in molten Ga can be attained by ultrasonic irradiation of molten Ga overlayed by an aqueous solution of D- or L-tryptophan. The enantiomeric excess (~12%) was determined using polarimetry, Chiral HPLC and CD measurements. When the reaction was done with other low melting point metals (Sn, In, Ga and Zn) under sonication. Binary combinations of bismuth and one of these metals were melted together in hot silicone oil and irradiated with ultrasonic energy to form micro/nano alloy particles. It was found that bismuth forms metal matrix composite with tin and zinc, intermetallic compounds with indium and an alloy with gallium. The extension of this work was studied the combined effect of ultrasonic cavitation and heterogeneous reduction in binary systems that included molten gallium and ionic solutions of one of several metals. It was revealed that in all the cases of the reducible ions, the free metals (Pt, Ag, Cu, and Au) were obtained together with some intermetallic compounds with the gallium (PtGa₃, Ag₂Ga, CuGa₂ and AuGa₂). It was found that Ga-Pt embedded in graphene showed better MOR and ORR activity than a commercial Pt/C catalyst. The same process was followed for fabrication of carbon dots (C-dots) doped with Ga atom (Ga@C-dots). Ga@C-dots showing the high photosensitization with respect to that of pristine C-dots. Ga@C-dots was found very high antimicrobial activity.
References:
V. B. Kumar, G. Kimmel, Z. Porat, A. Gedanken, New J. Chem.2015, (Accepted) DOI: 10.1039/C5NJ00781J
V. B. Kumar, Z. Porat, A. Gedanken, UltrasonicsSonochemistry 2015, 26, 340-4
V. B. Kumar, I. Perelshtein, G. Kimmel, Z. Porat, A. Gedanken Journal of Alloys and Compounds, 2015, 637, 538-544
V. B. Kumar, Y. Mastai, Z. Porat, A. Gedanken, , New J. Chem.2015, 39, 2690-6
V. B. Kumar, I. Perelshtein, Z. Porat, A. Gedanken, RSC Advance, 2015, 5, 25533.
V. B. Kumar, A. Gedanken, Z. Porat, J. of Therm. Anal. Cal. 2015 119, 1587-1592.
V. B. Kumar, Y. Koltypin, A. Gedanken, Z. Porat,J. Mater. Chem. A, 2014, 2, 1309-17.
V. B. Kumar, G. Kimmel, Z. Porat, A. Gedanken, UltrasonicsSonochemistry 2014, 21(3):1166-73.

Nanostructured electrode platforms have gained significant importance in design of electrochemical biosensor devices due to sensitivity, selectivity and amplification of output signal response that they offer. Electrical/electrochemical characterization techniques involve simple methods to measure charge perturbations that occur on electrode surface due to physiochemical changes, resulting from capture probe and target analyte interaction. In this work we study the dynamic changes at electrode-buffer interface on nanostructured zinc oxide (ZnO) using Mott-Schottky (MS) analysis and have segmented the effects of the semiconductor (ZnO) and (fluidic buffer analyte) contribution towards measured signal. We have used cardiac biomarker troponin-T (cTnT) as a study model to understand this behavior.
An electrode in contact with buffer medium (phosphate buffered saline - PBS in this case) experience excess of electronic charge distribution under the influence of applied electric field. Factors such as electrode type, material properties, conductivity of buffer and amount of biomolecular binding influence charge distribution at the electrode-buffer interface in addition to applied electric potential. The conductivity of PBS is much higher than semiconducting ZnO electrodes and thus polarization results in localized charge distribution in ZnO known as space-charge.
Experiments were performed using two planar metallic gold/ZnO electrodes fabricated on glass substrates. Nanostructures of ZnO were synthesized using RF-magnetron sputtered ZnO seed at working electrode using hydrothermal synthesis. Mott-Schottky is a technique using AC voltage signal at fixed frequency and varying DC bias. A copper wire was used as third dipping electrode. Using the described setup, we observed modulation to space charge capacitance in ZnO nanostructures as a function of biomolecular binding. Control experiments were performed on bare gold electrodes. Space-charge capacitance behavior was not observed with metallic gold electrodes as charge distribution occurs only on solution side of the electrode-buffer interface. However, in presence of ZnO electrodes, the width of space charge region gets modulated with biomolecular interaction which can be used as detection modality for quantification of biomarker. Results demonstrated decrease in space-charge capacitance with increasing concentration of cTnT biomarker with limit of detection at 100 fg/mL. The non-linear behavior in Mott-Schottky plots was observed due to presence of surface states in nanostructured ZnO. Hence we have shown here, that the physically and chemically tailored ZnO surfaces enhances sensitivity of detection taking advantage of the smaller size, high surface area and electrical transport occurring at its surface.

Drug eluting coatings represent a relatively new class of medical products that incorporate controlled release technologies to improve functionality and performance. The primary example is the drug eluting stent (DES), which has demonstrated marked improvement in therapeutic outcomes compared to traditional stents. The successful implementation of these coatings depends on the complex microstructures that evolve during coating fabrication and subsequent impact on drug release. We employ a diffuse interface model, in concert with all-atom molecular dynamics simulations, to predict structure evolution in these systems. We have found that not only can molecular dynamics be used to successfully predict the relevant thermodynamic and kinetic parameters, but also that the diffuse interface predictions are consistent with experimental observations. Finally, we demonstrate that these same computational tools can be extended to inform biocompatibility assessments of device polymers that contain potentially toxic additives.

Innovative manufacturing methods with mass production capabilities for making nanofibrous structures are in great demand worldwide. Scale-up using the currently available production techniques is extremely challenging and current technologies do not fulfil the demand. For example, electrospinning solutions and suspensions, which are widely used in many investigations, can generate uniform nanofibers of tailored morphologies and functionalities from a diverse range of materials, but suffer from low productivity and are mostly limited to small-batch operations. If melt electrospinning is used, fibers generated are largely limited to the submicrometer diameter range. It is widely agreed that novel manufacturing routes should have capabilities to process multi-functional nanofibers and nanofibrous structures that can safely, consistently and cost-effectively be scaled-up, and this is offered by combining gyration with pressure, and now infusion.
The pressurized gyration process, invented in 2013, is essentially a one-pot process and has generated exceptional interest (http://onlinelibrary.wiley.com/doi/10.1002/marc.201300339/citedby) for the production of functionalized fibers. It simply involves rotation and simultaneous blowing of the feedstock, containing all the constituents of the desired composition, in a sealed cylindrical vessel with circumferential orifices. It relies on application of centrifugal force, dynamic fluid flow and evaporation to jet-out (spray) fibers.The product fiber size distribution and morphology depends on the two process control parameters which are rotating speed of the vessel and working pressure, and also on polymer solution concentration. Optimizing these parameters is significant in order to obtain continuous nanofibers with well-defined morphologies and properties. The selection of primary solvents and their mixtures for the materials from which nanofibers are to be prepared is one of main factors which can influence gyratory spinnability, and recent work has enabled the more informed selection of solvents. The process has also been modelled to predict the generated fiber diameter. It has also been recently modified to prepare microbubbles with antimicrobial and biosensing capabilities.
One drawback of the pressurised gyration process is that it does not allow control of fluid flow through the fiber generating orifices in the vessel where the infusion rate of polymer solution influences fiber size distribution and the morphology of the spun fibres. Therefore, recently, we have created another novel method, infusion gyration (http://onlinelibrary.wiley.com/doi/10.1002/marc.201500174/abstract) which does not require external pressure, but relies on controlling the infusion rate of the polymer solution into the vessel. This allows the generation of nanofibers which can be integrated with other nanoassemblies, such as nanoparticles, peptides and proteins.

The capability of an implant material to successfully operate inside the human body is based on numerous materials properties that are in general referred to as “Biocompatibility”. In scrutiny, requirements regarding a material and its surface, like composition/chemistry, structural integrity, release of ions or adhesion to proteins and cells are diverse and to a large extend dictated by site-specific conditions of the human body and the designated function of an implant. A common approach for adjusting materials surfaces to the wide range of specific requirements concerning implantation site and function is applying coatings, i.e. using additional materials or/and drugs covering the implant material&’s surface. Although successfully applied in many instances, coating tends to give unsatisfactory results in case of pseudoelastic Ni-Ti alloys, a unique material for fabricating self-deploying minimally invasive implants. The main reason is that the uniquely large reversible (pseudoelastic) deformation of the material of >6% during implantation causes fracture and delamination of virtually any artificial coating. Accordingly, modifying the native surface of NiTi may represent a more promising route for tailoring the material to specific biomedical applications.
In general, Ni-Ti alloys exhibit a strong tendency to form a thin but compact surface oxide layer that mainly consists of titanium oxides. Recently it was demonstrated that features like crystallinity of the oxide and Ni content at the surface of the oxide layer strongly depend on the annealing conditions. Accordingly, precisely controlled heat treatments represent a new tool for optimizing the material&’s surface. Limitations of this approach arise from the required mechanical integrity of the oxide that was shown to deteriorate with increasing thickness of the oxide layer. In the present work, the perspectives for tailoring the properties of native surfaces of Ni-Ti alloys will be presented and discussed concerning aspects such as the amount of Ni at the surface, long- and short-term Ni release from the material and crystallinity of the oxide. The options for generating oxide surfaces with a Ni content ranging from zero to above 20 at.% are presented, based on the recently investigated mechanisms of oxide layer growth. The limitations of the tailored oxide layers lie in the crack initiation and flaking of the oxide during deformation, depending on the thickness of the layer. The question to what degree the various surfaces of the material may promote different biological responses like hemocompatibility or enhanced adhesion of proteins like fibrinogen will be discussed.

Classical antibacterial surfaces usually involve antiadhesive and/or biocidal strategies. Glycosilated surfaces are usually used to prevent biofilm formation via antiadhesive mechanisms. We show the first example of a glycosilated surface with biocidal properties created by the covalent grafting of sophorolipids (a sophorose unit covalently grafted to an oleic acid chain) through a self-assembled monolayer (SAM) of short aminothiol on gold (111) surfaces. The biocidal effect of such surfaces on Gram+ bacteria was assessed by a wide combination of techniques including microscopy observations, fluorescent staining and bacterial growth tests. About 50% of the bacteria are killed via membrane lysis. In addition, the role of the sophorose unit and aliphatic chain configuration are highlighted by the lack of activity of substrates modified respectively with sophorose-free oleic acid and stearic acid derivatives of sophorolipids. This system demonstrates thus the direct implication of a carbohydrate in a cell envelope destabilization and disruption.

The immunoisolation membrane is the essential component that determines the performance of a macroencapsulation device. A good immunoisolation membrane needs to shield the transplanted cells from the immune attack, facilitate the nutrient transport and minimize the host response. In particular, the topographical characteristics of the membrane has been implicated in the foreign body reaction and the tissue remodeling process. The electrospun membrane can be fabricated from a facile setup with tunable topography. It is therefore desirable to investigate whether the electrospun membrane can be used for immunoisolation purposes. In this study, the thermoplastic polyurethane (TPU) membranes with nanofibrous or microfibrous structures were fabricated via an electrospun process and the biocompatibility and immunoisolation properties of the two membranes were compared.
When implanted alone in a subcutaneous rat model in vivo, the two types of membranes induced distinctively different host reactions. The microfibrous membranes were surrounded with a large number of macrophages and foreign body giant cells; in contrast, the macrophages on the surface of the nanofibrous membranes were scarce following the 4-week implantation. In addition, significantly more blood vessels and thinner and less dense fibrotic capsules were found surrounding the nanofibrous membranes compared with the microfibrous ones. No host cells were found to infiltrate into the nanofibrous membranes after a 2-month implantation. These results indicate the nanofibrous structure processes superior biocompatibility and at the same time serves as a barrier prohibiting cell penetration. The results were also consistent with our in vitro observation: 1) The macrophages (RAW 264.7) cultured on the nanofibrous membrane polarized into a more anti-inflammatory phenotype; 2) The fibroblasts penetrated the microfibrous membrane but only formed a planar cell sheet on the surface of the nanofibrous membrane.
To further investigate whether the nanofibrous membrane can be exploited as an immunoisolation membrane, a planar macroencapsulation device was fabricated from two layers of the TPU membranes supported by a polyester mesh. In the allogenic models, primary cells including mesenchymal stem cells and hepatocytes isolated from the SD rats were loaded into the device and transplanted subcutaneously into the SD rat hosts. In another syngeneic model, the islets isolated from the C57BL/6J mice were transplanted into a C57BL/6J mice. After the three-week transplantation, the histological analysis showed that the devices were resided with viable cells with specific morphology and structures.
In conclusion, the TPU nanofibrous membrane can function as a robust immunoisolation barrier showing good biocompatibility and properties for nutrient-transport and cell-isolation. The macroencapsulation devices based on the TPU nanofibrous membrane warrant further study for immunoisolation applications.

In the present study, we report a new method of expanding electrospun nanofiber mats in the third dimension by making use of a modified gas foaming technique. The expanded nanofiber scaffolds form layered structures with controllable gap widths and layer thicknesses on the order of microns. Expanded nanofiber scaffolds possess significantly higher porosity than unexpanded nanofiber mats, while simultaneously maintaining nanotopographic cues. The distributions of gap widths and layer thicknesses are directly dependent on the processing time of nanofiber mats within the gas bubble forming solution. In vitro testing demonstrates robust cellular infiltration and proliferation within expanded nanofiber scaffolds as compared to limited cellular proliferation on the surface of traditional nanofiber mats. The presented method was further applied to fabricate tubular scaffolds composed of expanded nanofibers. This novel class of scaffolds holds significant promise for applications in regenerative medicine and building 3D in vitro tissue models for drug screening and biological study.

The electrospinning technique is a most versatile tool able to produce nanofibrous membranes that provide: (a) a wide variety of materials; (b) control of fiber morphologies and dimensions; (c) extremely high surface area-to-volume ratio; (d) highly porous membranes with controllable pore size.
Multi-axial electrospinning enhances its versatility by combining different material properties in separate core-sheath layers. This provides encapsulation functionality and enables the controlled release of molecules from the core. Because encapsulated macromolecules are released through the outer layers, manipulating the properties of intermediate and sheath layers controls the release kinetics of the encapsulated material.
In recent years, heavy use of antibiotic drugs has led to an increasing risk of antimicrobial resistance to patients, due to multi-drug resistant bacterial strains against antibiotics. As an alternative, natural biomaterials such as bacteriocins have emerged in antimicrobial applications. Among bacteriocins, nisin is the most attractive peptide because it provides a broad spectrum against many gram-positive bacteria and is also used as an FDA approved food additive. In the battlefield, soldiers exposed to the bacterial infection cannot change their clothes frequently. Safe antimicrobial textiles which last up to 7 days will be highly beneficial for their health. Therefore, the formation of electrospun nisin-containing membranes is very attractive for various applications, such as protective textiles, wound dressing, food packaging, etc.
We report on the use of triaxially electrospun membranes encapsulating nisin in the core with hydrophobic PCL intermediate and hygroscopic cellulose acetate sheath layers that combine a sustained release for a long time period (up to 7 days) with a skin-friendly material. Excellent antimicrobial performance of the nisin-encapsulated triaxial membranes have been demonstrated using the antimicrobial textile test (AATCC 100) and modified soft agar overlay (AATCC 147) with Staphylococcus. aureus bacteria. From AATCC 147 test, we have qualitatively demonstrated obvious antimicrobial activities up to 7 days. Also, the quantitative analysis from AATCC 100 has indicated that our triaxially electrospun membranes provide > 99.999% bacteria kill (5 log kill) up to three days. Compared with other type of electrospun membranes, triaxial membranes provided stronger and more sustained antimicrobial activities. Single fibers with nisin showed relatively weak activities and only in the first day. Coaxial fiber membranes exhibited antimicrobial activities for longer periods, but their effect is weaker than that of triaxial fiber membranes that still generate a removal of >99% (2 log kill) of bacteria on the third day of exposure.

Natural materials, such as bone, teeth, shells and wood, exhibit outstanding properties despite being porous and made of weak constituents. Frequently, they represent a source of inspiration to design strong, tough and lightweight materials. Although many techniques have been introduced to create such structures, a long-range order of the porosity as well as a precise control of the final architecture remains difficult to achieve. These limitations severely hinder the scale-up fabrication of layered structures aimed for larger applications. Here, we report on a new bidirectional freezing-casting technique to successfully assemble ceramic particles into scaffold with large-scale aligned lamellar porous, nacre-like structure, and long-range order at the centimeter scale. This was achieved by modifying the cold finger with a Polydimethylsiloxane (PDMS) wedge to control the nucleation of ice crystals under dual temperature gradients. Our approach could provide an effective way of manufacturing novel bioinspired structural materials, in particular advanced materials such as composites, where a higher level of control over the structure is required.

For tissue regeneration, tissue engineering principles have been utilized to generate tissue mimetic models that induce new human tissue formation. Scaffold design and predictable degradation are critical to successful application of tissue engineering. These tissue engineering material models rely on degrading scaffolds and growth of new tissue working in tandem. Scaffolds are often designed from polymeric (and biopolymeric) and polymer-composite materials. The degradation mechanisms of polymers (and biopolymers) is well understood over a large time scale; however polymer nanocomposite systems present a challenging problem towards ability to accurately predict mechanical response to loading over the biological and physiochemical mechanisms of scaffold degradation and tissue formation. The multifaceted degradation characteristics of these nanocomposites arise from degradation characteristics of the polymer, nanoparticles, nanoscale interfaces and the altered regions of the polymer due to nanoparticle addition. We have designed a biomimetic nanoclay system with synthetic polymers that is induces bone growth and stimulates human mesenchymal stem cells appropriately. A unique biomimetic biomineralization route is utilized to develop porous scaffolds of polycapralactone with nanoclays. Here, we represent a mechanics-based methodology to predict degradation of nanoclay polycaprolactone nanocomposites seeded with human osteoblast cells. The experimentally obtained degradation behavior using micro CT is used to design degradation models of the nanocomposite in a multiscale framework using finite element modeling. The finite element models include material models designed from molecular behavior using steered molecular dynamics. The degradation over the time scale of tissue attachment and growth is numerically captured into design of robust models. This experiment-modeling synergistic methodology allows for a robust predictive capability for simulating response and mechanical behavior of the tissue-scaffold interface. Accurate prediction of tissue growth and regeneration is vital to potential application of bone tissue engineering for large bone defects.

In the field of tissue engineering, design and fabrication of precisely and spatially patterned, highly porous scaffolds/matrixes are required to guide overall shape of tissue growth and replacement. Although Rapid Prototyping fabrication techniques have been used to fabricate the scaffolds with desired design characteristics, controlling the interior architecture of the scaffolds has been a challenge due to CAD constrains. Moreover, thick engineered tissue scaffolds show inadequate success due to the limited diffusion of oxygen and nutrients to the interior part of the scaffolds. These limitations lead to improper tissue regeneration. In this work, in order to overcome these design and fabrication limitations, research has been expanded to generation of scaffolds which have inbuilt micro and nanoscale fluidic channels. Branching channels serve as material delivery paths to provide oxygen and nutrients for the cells. These channels are designed and controlled with Lindenmayer Systems (L-Systems) which is an influential way to create the complex branching networks by rewriting process. In this research, through the computational modeling process, to control the thickness, length, number and the position of the channels/branches, main attributes of L-Systems algorithms are characterized and effects of algorithm parameters are investigated. After the L-System based branching design is completed, 3D tissue scaffolds were fabricated by “UV-Maskless Photolithography”. In this fabrication technique, Polyethylene (glycol) Diacrylate (PEGDA), which is biodegradable and biocompatible polymer, was used as a fabrication material. Our results show that L-System parameters can be successfully controlled to design of 3D tissue engineered scaffolds. Our fabrication results also show that L-System based designed scaffolds with internal branch structures can be fabricated layer-by-layer fashion by Maskless Photolithography. This technology can be easily applied to engineering living systems.

Biological tissues/organs consist of hierarchical organization of various extracellular matrix (ECM) molecules with spatial arrangement of cells and soluble factors. Highly hydrated ECM molecules assemble to form fibrils, microfibrils that organize into regional tissue structures such as muscle fiber bundles or osteons, tightly adapted to the specific tissue functions. Tissue architecture also defines spatial organization of biochemical and mechanical cues, in turn creating highly complex and heterogeneous cellular microenvironments. There are many examples where form/architecture is instrumental for the function of the specific tissue. To generate the functional tissue equivalents in vitro, engineered scaffolds should mimic the structural, mechano-chemical, and cellular complexity by recapitulating the unique microenvironments observed during the tissue development and morphogenesis. Inspired by hierarchical natural ECM organization and enabled by various micro/nanotechnologies, we strive to engineer synthetic ECM-mimics with intricate structural features observed in vivo. For example, we have developed highly aligned fibrous hydrogels using microfluidics that show nano- to micro- to macroscale hierarchy of natural collagen. Interestingly, these hydrogels are able to sequester inorganic ions from simulated body fluids and guide biomineralization in a manner similar to collagen. In another study, we demonstrate that both nano- as well as micro-scale architecture of the scaffolds is important for their bioactivity. Specifically, nanoscale functionalization of carbon-based porous scaffolds by carbon nanotubes promotes myoblast differentiation into myocytes; however, they fall short of promoting myocyte fusion into multinucleated myotubes even after 21 days. Surprisingly, when the microstructure of carbon-based scaffolds is changed to aligned carbon fibers along with carbon nanotube functionalization, differentiated myotubes fuse forming multinucleated myotubes within 14 days. Both these examples highlight importance of form/architecture of scaffolds in making them highly bioactive. Such bioinspired approaches to build biomimetic materials can be highly promising for functional tissue engineering.

The apatite deposition on material surfaces under the body conditions has mostly been considered according to the Kokubo model. Here, the material surface should be rich in M-OH (M: metal atoms like Si, Ti, Zr, Nb, or Ta) to form a hydrated oxide layer, which induces the heterogeneous nucleation of a calcium phosphate similar in composition and structure to bone mineral apatite (hydroxycarbonate apatite). This model is derived from in vivo and in vitro experiment, and has been employed intensively in the ceramic fields. However, models involving nano-clusters with or without organic oligomers have gained extensive supports these days. Thus, the Kokubo model and the nano-cluster models will be compared each other, and the use of varied simulated body fluids is discussed.

Endoscopy has become an essential procedure in contemporary medicine. However, its effectiveness is routinely compromised due to staining of the camera lens by body fluids. Current maneuvers to clear the lens include wiping on the walls of tissue, vigorous suction/irrigation, or complete removal of the scope for cleaning before reinsertion. These tactics pose risks such as tissue abrasions due to wiping, luminal collapse of narrow airways when using suction, or dislodging previously formed clots due to irrigation leading to additional bleeding. These interruptions result in unnecessary risks to the patient and longer surgical times. We propose the use of an omniphobic coating on the lens of the scope to prevent the wetting and adhesion of body fluids thereby eliminating the need for any clearance methods currently used by physicians. The proposed coating method relies on a layer-by-layer technique to assemble a nanostructured surface that traps a lubricant within the structure through capillary forces and matching surface chemistry, forming a smooth immobilized liquid interface (Sunny, S., Vogel, N., Howell, C., Vu, T.L., Aizenberg, J. Advanced Functional Materials 2014, 24 (42), 6658-6667) capable of repelling body fluids. This simple and scalable strategy results in a transparent, conformable, mechanically robust, and biocompatible coating. The efficacy of this material was assessed using a bronchoscope and a porcine lung model. Coated scopes were able to retain nearly 100% visibility after one hundred dips in whole porcine blood and mucus while the currently used control scope failed instantly. By providing continuous visual clarity for camera-guided instruments, we anticipate that liquid-based repellent coatings can improve outcomes for disease diagnoses and treatment.

Freeze casting, the controlled, directional solidification of aqueous solutions and slurries, is ideally suited for the manufacture of biomaterials, because: (i) the ice crystal growth in solutions and slurries can be carefully controlled so that scaffolds can be created that have highly aligned, unidirectional pore structures with additional microstructural features within the pores, (ii) the geometry and surface structure of the ice crystal, and thus the ice-templated scaffold, can be controlled through materials composition, additives and freezing conditions, and (iii) the cell wall material self-assembles into structures during solidification with features and properties that cannot be achieved by any other manufacturing technique. Very attractive is also that, because freeze casting is a cold process, (iv) growth factors can easily be added before and during processing, both directly and in encapsulated form, and that. additionally, (v) the scaffold surfaces can be modified and functionalized post processing. Collating empirical and theoretical results obtained for scaffolds made from polymers, ceramics, and metals, and plotting them in structure-property-processing charts aids an objective comparison of scaffold properties and performance of the different compositions and designs, pointing out property profiles which are of particular promise for applications that range from nerve and bone regeneration to cancer therapies.

Restoring the antibacterial properties of existing antibiotics is of great concern. Herein, we present, for the first time, the formation and deposition of stable antibiotic nanoparticles (NPs) on graphene oxide (GO) sheets by a facile one-step sonochemical technique. Sonochemically synthesized graphene oxide/tetracycline (GO/TET) composite shows enhanced activity against both sensitive and resistant Staphylococcus aureus (S. aureus). The size and deposition of tetracycline (TET) nanoparticles on GO can be controlled by varying the sonication time. The synthesized NPs ranged from 21 to 180 nm. Moreover, ultrasonic irradiation does not cause any structural and chemical changes to the TET molecule as confirmed by Fourier transform infrared spectroscopy (FTIR). The virtue of π-π stacking between GO and TET additionally facilitate the coating of TET NPs upon GO. A time dependent release kinetics of TET NPs from the GO surface is also monitored providing important insights regarding the mechanism of antibacterial activity of GO/TET composites. Our results show that the GO/TET composite is bactericidal in nature, resulting in similar values of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). This composite is found to be active against TET resistant S. aureus at a concentration four times lower than the pristine TET. The sensitive S. aureus follows the same trend showing six times lower MIC values compared to pristine TET. GO shows no activity against both sensitive and resistant S. aureus even at a concentration as high as 1 mg/mL but influences the biocidal activity of the GO/TET composite. We propose that the unique structure and composition manifested by GO/TET composites may be further utilized for different formulations of antibiotics with GO. The sonochemical method used in this work can be precisely tailored for the stable deposition of a variety of antibiotics on the GO surface to reduce health risks and increase the spectrum of applications.

In the tissue engineering field, different approaches are investigated and adopted for human body tissue regeneration and scaffold-based tissue engineering has been predominantly used since “tissue engineering” emerged two decades ago. With scaffold-based tissue engineering, both porous scaffolds and materials making the scaffolds play important roles in tissue regeneration. The basic requirement for a tissue engineering scaffold is the provision of a conducive microenvironment for cells to adhere, proliferate and differentiate, leading to new tissue formation. Recent advances in the field require scaffolds to also have the ability to induce the differentiation of stem cells into targeted tissue cells. There are many techniques for fabricating tissue engineering scaffolds and additive manufacturing has attracted great attention recently. Traditional materials for tissue engineering are biodegradable biopolymers such as PLLA, PLGA and PCL, which show various limitations, and hence developments have been made in using biodegradable bioceramics and even biocompatible and biodegradable metals for scaffolds. Nanocomposites, largely bioceramic-polymer nanocomposites, are intensively investigated as new tissue engineering materials - after all, many human body tissues are natural nanocomposites. In recent years, multifunctionality for tissue engineering scaffolds has been a focus in research. For example, multifunctional scaffolds may function to assist both angiogenesis and osteogenesis when they release suitable growth factors at different stages. Growth factors can be encapsulated in tissue engineering scaffolds during scaffold manufacture and released later in vitro or in vivo in a controlled manner. Furthermore, using the hybrid approach, multicomponent scaffolds can be constructed, with each component in the scaffold acting separately and independently as a delivery vehicle for respective bioactive agent. Among the techniques for fabricating tissue engineering scaffolds, not all of them are suitable for encapsulating biomolecules in scaffolds. Moreover, for some scaffolds produced by popular techniques such as electrospinning, a critical issue is cell infiltration and migration in scaffolds. New ways of incorporating specific cells directly in scaffolds and achieving high cell viability are thus investigated. Both cells and growth factors can be incorporated in scaffolds for enhancing tissue regeneration. For combating infection at the wound site, antibiotics can be incorporated in scaffolds for their later release in the body. For cancer patients, novel theranostics can be incorporated in scaffolds for achieving tissue regeneration at the tumor site after surgery and also detection and treatment of cancer recurrence. This presentation will introduce our designs of different multifunctional tissue engineering scaffolds, their fabrication and evaluation (physical and biological). Some critical issues will also be discussed.

Interfacing biological cells and solid-state devices is crucial in many applications, ranging from well-established fields, such as electrophisiology, to the newly developed areas of optogenetics and mechanobiology. Most biological cells are anchored to substrates with elastic modulus, E, in the range of ~1 to 100 kPa, the moduli of brain-tissue and osteoid, respectively. On the other hand, bulk semiconductor substrates have ~6 orders of magnitude higher elastic modulus. This large elastic mismatch between devices and the natural microenvironment of cells is an issue for bio-device integration, as cells are highly sensitive to mechanical cues. Specifically, cells exert traction forces on their surroundings and adjust their adhesion mechanism, cytoskeleton, locomotion and overall state according to the stiffness of the substrate they are anchored to. This type of behavior makes it a significant challenge to integrate semiconductor and photonic devices with biological cells without altering the cell state.
We demonstrate a new family of culture platforms that enables the integration of biological cells and electronic/photonic devices from a mechanical perspective. These platforms are referred to as effectively compliant layered substrates (ECLS). ECLS are based on inorganic nanomembranes (NMs) partially suspended or bonded to compliant substrates. The unique characteristic of ECLS is in that the constitutive material of the NM provides the electrical and optical functionality necessary to a device operation, while the NM geometry and the nature of the supporting substrate can be tailored to match the mechanical response of biological tissues. We present fabrication and bio-interfacing of ECLS including device-grade, single-crystal Si NMs on compliant substrates with tunable elastic moduli from ~kPa to ~MPa. NMs with thicknesses in the range of 30-300 nm and ~ 1x1 cm² lateral areas are used in this study. ECLS are obtained using a two-step process, including synthesis of the compliant supporting substrate and fabrication, release and transfer of the NM onto the compliant host. Characterization of the mechanical properties of the ECLS and of the bare compliant substrate is performed by instrumented nanoindentation.
We culture 3T3 fibroblast on the fabricated ECLS, as well as on bare compliant substrates, to investigate the effect of the SiNM on cellular behavior, particularly with respect to response of the cells to mechanical cues. Results of these studies are benchmarked against data acquired for 3T3 cells cultured in identical conditions on glass and polystyrene substrates, i.e., standard culture substrates. Specifically we investigate cytotoxicity, morphology, adhesion mechanisms and migration for 3T3 fibroblasts on all culture substrates mentioned above. Bright-field and confocal fluorescence microscopy are used for this study.

Despite huge success of silicon-based Micro Electro Mechanical Systems (MEMS), substituting rigid inorganic materials by soft organic ones offers promising economical and functional benefits. Indeed, a wide panel of functional properties given by polymers allows elaboration of low-cost, tailor-made organic MEMS for targeted applications. However, in order to take advantage of downscaling offered by microsystems that enables portability, an integrated electrical transduction of mechanical motion remains a challenging issue yet to be solved.
When MEMS are used as biosensors, they generally operate either in dynamic or static mode. In dynamic mode, analytes bind to the sensitive coating increasing mass and hence decreasing the resonant frequency. In static deflection mode, analyte binding causes unbalanced surface stress resulting in a measurable mechanical deflection. For both modes of operation, it is clear that integrated electro mechanical transduction allows a direct observation of biological events. In this context, we have evaluated several transduction schemes in an all-organic approach. First, piezoresistive gauges have been elaborated, where gauge factors up to 200 have been observed for relatively large strain values (>3%), in both static and dynamic modes. However, in dynamic mode, external actuation of MEMS remains mandatory. For achieving simultaneous actuation and detection, piezoelectric polymers based on polyvinylidene fluoride (PVDF) are of particular interest. Using reversibility of piezoelectric effect, integrated organic piezoelectric resonators have been fabricated and used in liquid media. Indeed, through an optimized fabrication process, it was made possible to monitor resonant frequency of organic cantilevers in aqueous and organic solvents, a key step for real-time biosensing assays. Piezoelectric polymers have also been included recently in Organic Field Effect Transistors (OFETs) as active dielectric layers. In this case, applied strain to the piezoelectric film results in changes of the charge density (within the channel). Such advanced transduction scheme is extremely sensitive, since gauge factors higher than 300 are commonly observed for low strain values (<1%). These advances on strain sensitive piezoelectric transistors pave the way for the development of original MEMS biosensors in an all organic approach.
The aforementioned organic MEMS have been functionalized by Molecularly Imprinted Polymers (MIPs), biomimetic polymers capable of binding analytes specifically and selectively. Already demonstrated as efficient sensitive coatings when combined with MEMS resonators, work is now in progress to consider swelling effects that occur after analyte rebinding. Such configuration will result in the development of integrated MEMS sensors operating in static mode. The final objective is to evaluate the most sensitive mode of operation of such organic microsystems fabricated at low-cost in an all organic approach.

New digital technologies are needed to compliment lithography for low volume manufacturing of personalised biosensors and companion diagnostics. These new technologies will enable ultra-precise patterning of advanced functional materials that are often sensitive to the lithographic process, while also minimizing expensive biological and nanomaterial waste. This presentation focuses on the underpinning science of a hybrid approach combining two non-contact digital techniques to process these materials.
(I) Inkjet Printing: biological elements, advanced functional materials for sensing and catalysts for nanomaterial growth
(II) Ultrafast Laser Ablation: accurate structuring of sensing elements to sub-micron resolution, controlling the surface chemistry and delivering quantitative sensing
We report the application of this technique to biological sensing through two collaborations. Firstly, as well as examining the standard model systems, the work looks at specific lipid-antigen systems which play an important role in detection of pathogens. Secondly, the application of this technique to electrochemical sensing is studied by deposition and tuning of carboxyl functionalised carbon nanotube mats and patterned growth of carbon nanotube forests by controlled printing of iron oxide catalyst nanoparticles. We explore the effect of laser parameters on the functional materials through analysis with a suite of characterisation techniques, such as AFM, SEM/FIB, BET, Raman and TGA. This approach can tackle the challenges of digital patterning while also providing a route towards 3D microscale fabrication of biosensing devices.

Accurate, cheap and fast glucose sensing is of paramount importance in several technology areas such as health care, food industry and biotechnology. Non-enzymatic glucose sensors have recently attracted a lot of scientific attention and have shown reliable and fast glucose sensing, with sensitivity in the order of few mA mM^-1 cm^-2. [1,2]. The use of Ni takes advantage of the Ni²⁺/Ni³⁺ red-ox shuttle which is capable of fast and effective glucose oxidation [2]. However, the preparation of nanostructured Ni is still a challenge, involving high temperature, pressure and vacuum systems, as well as exotic substrates. As a result, the manufacturing cost of potential Ni based glucose sensors is expected to be high. Here we present a simple route for the preparation of high surface area Ni nanostructures which is compatible with low-cost fabrication of flexible (plastic substrate) glucose biosensors with high efficiency (up to 3 mA mM^-1 cm^-2).
We report on high sensitivity, non-enzymatic glucose sensors based on a novel Ni nanofoam (NF) electrode fabricated via a facile method. Ni(OH)₂ nanosheets are obtained through room temperature chemical bath deposition. FTO (Fluorine doped Tin oxide) covered glass or ITO (Indium Tin oxide) covered UPILEX plastic (50 µM thick) were used as substrates. The as prepared material is converted to a packed ensemble of small (20 nm) Ni particles (nanofoam) by forming gas annealing at 350°C. Scanning electron microscopy, X-ray Diffraction, temperature programmed reduction, and BET surface area analyses were used for chemical and structural characterization, while a VersaStat 4 potentiostat was used for electrochemical studies in a three electrodes set-up. The NF electrode (both on plastic and glass substrates) was conditioned for glucose sensing by cyclic voltammetry (CV) between -0.1 and 0.8 V (vs SCE) in 0.1 M NaOH electrolyte. Changes in current density with increasing amount of glucose concentration of the electrolyte was monitored by chronoamperometry analyses at 0.5 V. The results showed a linear response in the 10-700 mM glucose concentration range, a sensitivity of 3 mA mM^-1 cm^-2, and a fast response time of 1 second. Long-term stability study indicated that there was 5 % decrease in signal after 60 days. The electrodes were also tested against some common interfering species (as uric acid, ascorbic acid and acetaminophen), and they showed high selectivity for glucose. Finally, the sensors exhibited excellent resistance to chloride poisoning when tested in chloride electrolyte (0.2 M KCl).
The reported data show that the novel electrode based on Ni NF offers many advantages for low-cost, portable and wearable bio-sensing applications

Understanding spatial and temporal neuronal activities is crucial for finding a cure for brain related ailments and advancement of our knowledge about the brain itself. This paper discusses our recent finding of the patternable rough textured gold microwire for neurochemical sensing. We have successfully fabricated the ~1 µm wide and ~ 60 nm thick gold microwires based electrochemical sensor. We produced these microwires along the edge of lithohraphically patterned nickel thin film. Nickel thin film edge was shadowed by the photoresist overhang during electrochemical growth only to allow gold deposition along the edges. Our electrochemical growth conditions yielded very rough textured sensor. Rough textures biosensors are highly desirable for increasing surface/volume ratio for efficient electrochemical sensing. These rough-textured microwires were transformed into the functional neurochemical sensor to detect dopamine. Our voltammetry and chronoamperometry studies on rough textured microwires based sensor confirmed the successful detection of dopamine.

NiTi alloys are highly promising candidate materials for use in orthopedic implants because of their preeminent mechanical properties and compatibility with the human physiological system. To ensure long term stability, the biomedical implants need to have high wear resistance to avoid foreign body inflammatory responses generated by wear debris, and sufficient fatigue performance to withstand repetitive contact stresses. Many efforts have been aimed at improving the mechanical performance and biocompatibility of biomedical implants. In this study, an innovative surface processing technique, Ultrasonic Nanocrystal Surface Modification (UNSM), is investigated with an objective to improve the mechanical properties of a NiTi alloy. UNSM utilizes low amplitude ultrasonic frequency vibrations superimposed on a static load to induce high strain rate plastic deformation on NiTi surface, which causes surface amorphization through multiple dynamic loading, and eliminates crystalline phases and grain boundaries at the top surface of the material. Adipose tissue-derived Stem cell culture in normal growth medium demonstrated that UNSM processing did not deteriorate the biocompatibility compared to the non-processed samples. Taken together, the results demonstrated that UNSM method can significantly improve mechanical properties without compromising biocompatibility, and would contribute significantly towards the efficient and rational design of future implant materials for bone tissue engineering.

Seamless and minimally invasive three-dimensional interpenetration of electronics with artificial and/ natural structures, such as hydrogel and biological tissues, could allow for continuous monitoring and manipulation of their properties. Flexible electronics provide a means for conforming electronics to non-planar surfaces such as tissue surfaces, yet targeted delivery and integration of flexible electronics to internal regions in a minimally-invasive manner has not been possible. Here we overcome this challenge by demonstrating the syringe injection (and subsequent unfolding) of submicrometer-thick, centimeter-scale macroporous mesh electronics through needles with a diameter as small as 100 µm. Our results show that electronic components can be injected into man-made and biological cavities, as well as dense gels and tissue, with >90% device yield. We demonstrate several applications of syringe-injectable electronics as a general approach for interpenetrating flexible electronics with three-dimensional structures, including (1) monitoring internal mechanical strains in polymer cavities, (2) tight integration and low chronic immunoreactivity with cavity region of rodent brain tissue such as lateral ventricle region to promote the migration of neural progenitor cells migrate along the injected macroporous electronics and (3) implantation with low chronic immunoreactivity in hippocampal region with in vivo multiplexed neural recording. Moreover, we demonstrate syringe injection enables the delivery of flexible electronics through a rigid shell, the delivery of large-volume flexible electronics that can fill internal cavities, and co-injection of electronics with other biomaterials and primary cultured cells into host structures, opening up unique potential applications for flexible electronics.
J. Liu, et al. “Syringe-injectable electronics,” Nature Nanotechnol. DOI: 10.1038/NNANO.2015.115,

Current protocols for immune system monitoring involve collecting cells from blood or cerebrospinal fluid, or via invasive sampling and biopsies. However, since major populations of immune cells reside within tissues, these invasively-obtained body fluid samples are, at best, indirect indicators of the status of the immune system. Further, these methods are difficult to incorporate into long-term, repetitive, longitudinal immune monitoring. On the other hand, direct interrogation of the immune system, as currently employed by simple delayed-type hypersensitivity tests (e.g., Mantoux tuberculin test), does not provide information about the phenotype and functional characteristics of responding immune cells. We report here a technology that addresses these challenges simultaneously, with the synergistic goals of providing enhanced diagnostic methods for sampling and analyzing the function of the immune system, and providing a greater insight into the status of the immune system than state of the art assays. We have fabricated alginate-coated, cell-sampling microneedles capable of sampling cells and interstitial fluid and permitting the quantification of constituents like IgG. Cell recruitment was enhanced by incorporating chemoattractants in the alginate coating within 8 hours of application. Cells obtained from retrieved microneedle arrays could be subjected to phenotype analysis and stained for surface markers. We employed a subcutaneous alginate gel injection model to determine the optimal time of sampling and the chemoattractant dosing, Alginate gels injected subcutaneously into the dorsal flanks of mice showed enhanced infiltration of CD4+ T cells under the influence of inflammatory chemoattractants and showed increasing infiltration of cells with time, over 48 hours. Microneedle arrays retrieved at different points of time, under the influence of different chemoattractants and antigens, are currently being analyzed to further understand the temporal characteristics of immune cell recruitment towards sites of inflammation. This work allows ex vivo analysis of cells retrieved directly from the local tissue environment and enables the investigation of antigen-specific cell interrogation for diagnostic purposes as well as answering spatio-temporal questions related to immunology in local tissue environments.

Biomacromolecular materials with fibrous architecture are of high technological and scientific interest in order to e.g. mimic the hierarchical organization of biological tissues and accomplish material systems with unexpected properties and functionalities [1]. However, the design of defined material geometries is challenging since the molecular material architecture can be irreversibly altered due to the manufacturing route [2]. To explore this challenge, we investigated the wet-spinnability of varied biomimetic building blocks aiming at wet#8209;stable fibres and fabric-like 3D assemblies with defined protein conformation. Gelatin and hydrolyzed fish collagen were selected as high and low molecular weight polypeptide, whilst non-hydrolyzed rat tail collagen was employed as simple mimetic of the extracelular matrix of tissues. A range of fibre morphologies were observed using varied wet-spinning solvents and coagulating media. Circular dichroism (CD) demonstrated nearly complete triple helix retention in wet-spun non-hydrolyzed collagen fibres. Post-spinning diacid-based crosslinking route led to preserved fibrous morphology in phosphate buffer solution (PBS). Fibre tensile and swelling properties could be adjusted depending on the covalent network architecture at the molecular scale, i.e. based on either inter- or intramolecular crosslinks, respectively. In light of the remarkable hydrated tensile moduli, crosslinked fibres are currently being investigated for the formation of macro-porous fibrous materials for e.g. dental applications or chronic wound care.
[1] C. Meier, M.E. Welland (2011). “Wet-spinning of amyloid protein nanofibers into multifunctional high performance biofibres”. Biomacromol. (12) 3453-3459
[2] P.R. Stoessel, R.N. Grass, A. Sánchez-Ferrer, R. Fuhrer, T. Schweizer, R. Mezzenga, W.J. Stark (2014). “Spinning angora rabbit wool-like porous fibers from a non#8209;equilibrated gelatin/water/2-propanol mixture”. Adv. Funct. Mater. (24) 1831-1839

The development of methods to achieve precise spatiotemporal control over biomolecular gradients could enable significant advances in areas including biomedical device design, synthetic tissue engineering, and bio-nano-interfaces. Cells that detach from a primary tumor and enter the bloodstream are known as circulating tumor cells (CTCs), driving metastasis of cancers throughout the body. While strong evidence for CTCs as valuable prognostic biomarkers has been reported, extremely low concentrations (1-10 cells per 10 ml in the peripheral blood) of CTCs in early-stage cancer patients limit their clinic applications. Numerous methods have been developed to detect and capture CTCs with various microfluidic and nanostructured devices. However, these technologies typically rely on random motion of suspended cells. Active CTC enrichment based on directional cell migration remains a critical challenge. To mimic the dynamic microenvironment surrounding tumor cells, highly orchestrated gradients of molecular factors (e.g. epidermal growth factor (EGF) supplied by macrophages) are required to lead cellular chemotaxis.
To address this issue, we developed a method to 3D print stimuli-responsive core/shell capsules for programmable release of multiplexed molecular gradients. These capsules are comprised of an aqueous core with payload biomolecules and a biocompatible polymer shell functionalized with gold nanorods (AuNRs). The aqueous cores have been demonstrated to maintain the activity of CTC-related payloads, while the photothermal response of AuNRs permits selective laser-rupturing of capsules. Furthermore, CTC-capture agents can also form highly ordered patterns by 3D printing of avidin-modified gold nanoparticles and subsequent self-assembly of biotinylated antibodies. The advantages of this 3D printing-based method include: 1) highly monodisperse capsules with efficient encapsulation of active payloads, 2) “on the fly” programmable reconfiguration of gradients, and 3) precise spatial patterning of both capsule and capture-agent arrays.
This precise control over space, time, and selectivity allows for the ability to pattern 2D and 3D multiplexed arrays of capsules and antibodies, along with tunable and sequential laser-triggered release of EGF, cell markers or/and drugs. Here, EGF-loaded capsules simulate microphages to build controllable EGF gradients, which lead to chemotaxis of tumor cells and enrichment of CTCs at patterned antibody arrays. Furthermore, in addition to active attraction and specific capture of CTCs, effective identification of tumor cell types and corresponding targeted treatment can also be achieved via subsequent rupturing of capsules loaded with cell markers and drugs, respectively. Therefore, 3D printing of programmable release capsule arrays may represent a powerful new tool to enable spatiotemporal control over biomolecular gradients, enabling effective integration of cancer diagnostics and therapeutics on a single chip.

A key element for the successful implementation of nanoscale materials in clinical applications is multi-functionality [1]. Here, plasmonic (e.g. Ag, Au) nanoparticles are combined with a magnetic material (Fe₂O₃) and in-situ coated with a nanothin layer of amorphous SiO₂ forming, therefore, hybrid nanoparticles. In fact, the small size of the magnetic component renders it superparamagnetic, i.e. it exhibits magnetic properties only in the presence of a magnetic field, allowing therefore their magnetic manipulation for targeted drug delivery or for magnetically assisted cell sorting and separation [2]. Such hybrid nanoparticles can also be detected by magnetic resonance imaging (MRI), with which small magnetic nanoparticles can be used as contrast agents. Furthermore, the hermetic SiO₂ coating on the surface of such nanoparticles facilitates their aqueous dispersion and surface bio-functionalization [3] and further prevents their magnetic particle-particle interaction and flocculation. Such multifunctional plasmonic nanoparticles can be used as superior biomarkers by specifically binding them to live HeLa and Raji cells [2]. Furthermore, by finely tuning the plasmonic interparticle distance using the SiO₂ film thickness (or content) the plasmonic coupling can be finely controlled bringing their optical absorption to the near-IR that is most important for human tissue transmittance [4]. The SiO₂ shell also prevents the reshaping or coalescence of plasmonic particles during laser irradiation, thereby allowing their use in multiple treatments. Their effectiveness as photo-thermal agents is demonstrated by killing human breast cancer cells with a short, four minute near-IR laser irradiation (785 nm) at low flux (4.9 W cm^-2).
References
[1] G. A. Sotiriou, WIREs Nanomed. Nanobiotechnol., 5, 19-30 (2013).
[2] G. A. Sotiriou, A. M. Hirt, P. Y. Lozach, A. Teleki, F. Krumeich, S. E. Pratsinis, Chem. Mater., 23, 1985-1992 (2011).
[3] G. A. Sotiriou, T. Sannomiya, A. Teleki, F. Krumeich, J. Vörös, S. E. Pratsinis, Adv. Funct. Mater., 20, 4250-4257 (2010).
[4] G. A. Sotiriou, F. Starsich, A. Dasargyri, M. C. Wurnig, F. Krumeich, A. Boss, J.-C. Leroux, S. E. Pratsinis, Adv. Funct. Mater., 24, 2818-2827 (2014).