Surface Topography And Composition Properties Effect On Adhesion And Growth Behavior Of MC3T3-E1Cells

Many studies have demonstrated the topography and bioactive of biomaterial surface to regulate cellular behavior. To investigate the effect of bioactive properties and surface topography of biomaterial surface on the adhesion and growth behavior of MC3T3-E1cells,4different surfaces were prepared. Poly lactic-co-glycolic acid (PLGA) electrospinning fibers (ES) were produced for porous roughness surface with a pore sizes.3,4-Dihydroxyphenylalanine (DOPA) is identified as one of the key chemical signature of adhesions. And Type Ⅰ collagen (COL Ⅰ) is the most abundant protein in the extracellular matrix. DOPA and COL I were immersed into cover slips as chemical surface and bioactive molecules surface respectively. And COL I immobilized onto porous electrospinning fibers surfaces (E-COL) was used to investigate the combine effect of bioactive molecules and topography. The cover slips without any treatments were set as control. Early adhesion and growth behavior of MC3T3-E1cells cultured on the different surfaces were studied. Cell adhesion and morphological changes were observed with FESEM and CLSM. CCK-8assay and Flow Cytometry were observed for the proliferation of cell. Immunofluorescence staining, western Blot and RT-PCR were performed to determine the expression levels of integrin β1at6,12and24h. RT-PCR determined the expression levels of adhesion markers, such as integrin, α1, α2, α5, α10and all. MC3T3-E1was observed to adhere to and fully extend on ES, E-COL and COL by FESEM. Under serum-free condition, bioactive molecules had a major influence on cell adhesion. But under serum condition, the surface topography played an important role in the early adhesion time but the effect of chemical and bioactive molecules were more important in the later time. Combine effect best supported the adhesion of cells.