In Vitro Slow-release Scaffold Of Chitosan-gelatin-growth Factor Composite Effect On Proliferation Of Bone Marrow Mesenchymal Stem Cells

Objective:The aim of this project is to generate a 3D chitosan-gelatin porous scaffold that constantly releases basic fibroblast growth factor (bFGF) and/or hepatocyte growth factor(HGF), and to further discuss in vitro combined effect of both growth factors in Bone marrow stromal cells (BMSCs) proliferationMethod:Chitosan, gelatin, bFGF and HGF are mixed in varied proportions and dry freezed to form porous 3D scaffolds. BMSCs are isolated from SD rat femur bone marrow, expanded in vitro. Take the third generation of favourable BMSCs and seed in 96-well plate, then mixed with scaffolds. Cell proliferation was then analysed by MTT proliferation analysis 5 days later. All animal experiments meet ethical and animal welfare requirementsResult: BMSC growth is significantly increased (P<0.05) by the constant release growth factors from scaffold with 1μg/mL bFGF along or 1μg/mL bFGF combined with 1μg/mL HGF.. But there is no different between two groups. BMSC growth is little increased (P>0.05) by the constant release growth factors from scaffold with 0.1μg/mL bFGF,0.01μg/mL bFGF or 1μg/mL HGF along.Conclusion:Highly concentrated and viable BMSCs can be isolated by density gradient centrifugation combined with attachment culture method. Chitosan-glutin can form a 3D scaffold that can slowly but constant release growth factor; while bFGF promotes BMSCs proliferation in vitro which is not enhanced by HGF. The more bFGF involved, the more BMSCs prolife.