RESUMO
A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.
Assuntos
Quitosana/química , Condroitina/química , Gelatina/química , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Osteoblastos/citologia , Osteoblastos/fisiologia , Osteogênese/fisiologia , Animais , Substitutos Ósseos/química , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células Cultivadas , Força Compressiva , Elasticidade , Teste de Materiais , Porosidade , Ratos , Ratos Wistar , Engenharia Tecidual/métodosRESUMO
A commercial glass fiber with Al(2)O(3) (68.4%) and SiO(2) (27.6%) as major components and CaO, TiO(2), Fe(2)O(3), and CuO as minor components was used as substrate in a silica sol-gel coating process. After cleaning, fiber samples were immersed into tetraethoxysilane (TEOS) at room temperature for 1 h, and then individual fiber samples were soaked into a simulated body fluid (SBF) solution,1 and removed after 5, 10, 15, and 20 days. Zeta potential and Energy Dispersive Spectroscopy (EDS) analyses showed that the fiber surfaces were effectively coated with a silica layer, which improved the formation of an HA layer upon immersion into SBF solution for 5 days. The coating became even more continuous after 10-day immersion. Fourier Transform Infrared Spectroscopic (FTIR) analyses confirmed that the coating layer has P--O vibration bands characteristic of hydroxyapatite (HA) near 1060 and 600 cm(-1).