Proliferation and collagen production of bovine fetal fibroblasts inside 3d scaffold based on macroporous poly(n-isopropylacrylamide) hydrogel.

LIAUDAT CECILIA; BARBERO, CESAR ALFREDO; REBECA RIVERO; CAPELLA, VIRGINIA; BOSCH, PABLO; RODRIGUEZ, NANCY; CLAUDIA R. RIVAROLA

CANCUN

Congreso; XXVII International Materials Research Congress, Cancún, México. 2018.; 2019

PROLIFERATION AND COLLAGEN PRODUCTION OF BOVINE FETAL FIBROBLASTS INSIDE 3D SCAFFOLD BASED ON MACROPOROUS Poly(N-ISOPROPYLACRYLAMIDE) HYDROGELRebeca Rivero1, Virginia Capella2, A. Cecilia Liaudat3, Pablo Bosch3, Nancy Rodriguez3, Cesar Alfredo Barbero1, Claudia R.Rivarola11Instituto de Investigaciones en Tecnologias Energeticas y Materiales Avanzados (UNRC-CONICET), Chemistry, Argentina.2Instituto de Investigaciones en Tecnologias Energeticas y Materiales Avanzados (UNRC-CONICET), Molecular Biology,Argentina. 3Universidad Nacional de Rio Cuarto, Molecular biology, Argentina.Macroporous hydrogel based on poly(N-isopropylacrylamide) (PNIPAM) is studied as 3D scaffold for bovine fetal fibroblasts growing. Interconnected macropores are obtained by free radical cryopolymerization at -18 º C. Morphology of pores is followed by optical and scanning electron microscopy. Methods of cell seeding to induce the 3D proliferation inside matrix and sample treatment to histology assays are described. Cell growth, adhesion and biocompatibility are analyzed progressively inside scaffold during several culture days. Hoechst, MTT and Neutral Red uptake assays were applied to analyze the cell proliferation and then were observed by scanning confocal microscopy. Cell adhesion over hydrogel macropores walls was observed by bright-field optical microscopy throughPicrosirius Red stain. Swelling and mechanical properties of the scaffold in culture medium show that the water taken by matrix remains constant and the elastic module decreases during the days of cell grown without losing the 3D initial structure. The matrix degradation was verified by inverted brightfield microscopy. The collagen production was analyzed by collagen birefringence using Picrosirius Red and polarized light microscopy. It was demonstrated that BFF cells are able to adapt the 3D PNIPAM surface in order to proliferate and migrate inside the macroporous matrix. BFF cell growth was followed for more than 70 culture days showing that macroporous PNIPAM is highly biocompatibleand it could be applied as a 3D scaffold material for tissue generation and regenerative medicine.