Monica Montesi1,Giada Bassi1,2,Silvia Panseri1,Arianna Rossi1,3,Denis Cochonneau4,5,Elisabetta Campodoni1,Monica Sandri1,Dominique Heymann4,5
Institute of Science, Technology and Sustainability for Ceramics (ISSMC)_ National Research Council1,University of Studies “G. D’Annunzio"2,University of Studies of Messina3,Institut de Cancérologie de l'Ouest (ICO), Tumor Heterogeneity and Precision Medicine Lab4,CNRS, Nantes Université5
Monica Montesi1,Giada Bassi1,2,Silvia Panseri1,Arianna Rossi1,3,Denis Cochonneau4,5,Elisabetta Campodoni1,Monica Sandri1,Dominique Heymann4,5
Institute of Science, Technology and Sustainability for Ceramics (ISSMC)_ National Research Council1,University of Studies “G. D’Annunzio"2,University of Studies of Messina3,Institut de Cancérologie de l'Ouest (ICO), Tumor Heterogeneity and Precision Medicine Lab4,CNRS, Nantes Université5
INTRODUCTION<br/>Capturing and reproducing the biological complexity of cancer <i>in vitro</i> is an exciting challenge for scientists. Osteosarcoma (OS) constituted the main entity among primary bone sarcomas with a poor prognosis due to chemoresistance and early metastasis. Following the tumor engineering approach, we developed a more predictive 3D scaffold-based <i>in vitro</i> model reproducing the OS microenvironment heterogeneity. Promising results were obtained by our group in 2020, confirming the higher predictivity of in vitro 3D bone-like scaffold-based models of enriched-CSCs compared to scaffold-free conditions<sup>1</sup>. Using a biomimetic hybrid scaffold, that recapitulates the bone extracellular matrix (ECM)<sup>2</sup>, 3D cell cultures of OS enriched-Cancer Stem Cells (CSCs)<sup>3</sup> and co-culture of RGB human MNNG-HOS OS (HOS) cells with mesenchymal stem cells (MSCs) or HUVEC cells, will be developed and characterized.<br/>EXPERIMENTAL METHODS<br/>The MgHA/Coll scaffolds derive from a bio-inspired biomineralization process<sup>2</sup>. The enriched-CSCs were obtained by sarcosphere-forming culture<sup>3 </sup>and RGB human MNNG-HOS OS (HOS) were kindly provided by Nantes University. The cell seeding conditions have been defined and the cell/biomaterial interaction was studied by looking at the scaffold colonization by fluorescence analysis, the cell viability and morphology (live&dead assay, histology), the expression of cell-specific markers (immunohistochemistry, Western blot analysis and qPCR), the Sarcosphere-Forming Efficiency, the Stem Cell Frequency, the migration and invasion ability (transwell inserts 8 µm pores), and looking in detail at the mechanism of cell-cell and cell-ECM crosstalk.<br/>RESULTS AND DISCUSSION<br/>The results demonstrate that cells can grow and colonize the surface of the scaffold; CSCs, seeded by injection onto the core of the scaffold, expressed a higher level of stemness marker compared to the 2D in vitro culture. The mimetic structure of the scaffold allows to creation a bioactive environment stimulating HOS-MSCs and HOS-HUVEC crosstalk.<br/>CONCLUSION<br/>Overall, the results demonstrate that the proposed 3D scaffold-based in vitro model creates a permissive and inductive microenvironment able to mimic more in detail the OS complexity.<br/> <br/>REFERENCES<br/>1. Bassi G. et al., Sci Rep 10, 22294 (2020)<br/>2. Krishnakumar GS, et al. Int J Biol Macromol. (2018) <br/>3. Brown, H.K. et al., Cancer Lett. 386,189-195 (2017)