Studies On Preparation And Cell Adhesion Properties Of BSA Modified DBM Scaffold

Objective:1.Surface modification of demineralized bone matrix(DBM)scaffolds with different concentrations of bovine serum albumin(BSA)to prepared DBM scaffolds with BSA coating.The surface morphology of each scaffold was observed and its performance was analyzed.2.In a sterile environment,DBM scaffolds before and after BSA modification were co-cultured with mouse fibroblasts(L929).The growth of the cells on the scaffold surface was observed,and the effect of scaffold materials on cell proliferation activity was determined.3.After co-cultured of L929 cells and scaffold materials in a sterile environment for a certain period of time,the adhesion ability of DBM scaffolds to cells before and after BSA modification was determined,and the appropriate BSA concentration was explored.Methods:1.DBM scaffolds were prepared by degreasing and decalcifying the bovine cancellous bone in vitro.In order to obtain a scaffold with BSA coating,DBM was treated with BSA solutions at concentrations of 10 mg/ml,20 mg/ml,30 mg/ml,and 40 mg/ml,respectively.The surface of the scaffold was observed under scanning electron microscope(SEM).The binding of BSA was analyzed by Fourier transform infrared spectroscopy(FTIR).The porosity,degradation rate and hydrophilicity of the scaffold were determined.2.L929 cells were resuscitated,and the cells with good growth were inoculated into the sterile DBM scaffolds and BSA-coated scaffolds.After 3 days,the growth of the cells in the scaffold material was observed under an SEM,and the effect of the scaffold on cell proliferation was examined by MTT assay.3.L929 cells were inoculated with DBM scaffolds and BSA-coated scaffolds.After 6 hours,the cells were treated with DAP I solution,and the cells adhesion were observed under confocal laser scanning microscope.After co-cultured with scaffolds for 6 h,MTT assay was used to determine the adhesion of cells to scaffolds.Results:1.The DBM scaffold had a loose porous sponge structure,and the internal pore size of the scaffold was different.The shape was mostly irregular circular or elliptical.The internal pores communicate with each other,the pore diameter was between 350?450?m,and the porosity was(69.28±2.46)%.After modification with different concentration of BSA,the pore size and porosity of the scaffolds were decreased,the degradation rate was accelerated and the hydrophilicity of the scaffolds was increased.2.Cell adhesion was observed on the surface of both DBM scaffolds and BSA-modified scaffolds,and the cell growth was good.The scaffold was co-cultured with the cells,the MTT results showed that the difference between the 20 mg/ml BSA-treated scaffold and the DBM scaffold group was statistically significant(P<0.05).3.Under the confocal laser scanning microscope,the nuclei were round blue-stained structure and distributed in three-dimensional form within the scaffolds.Compared with the DBM scaffold,when the concentration of BSA was 20 mg/ml and 30 mg/ml,the number of cells attached to BSA-coated scaffolds was increased.The results of cell adhesion showed that the absorbance value of DBM scaffolds treated with 20 mg/ml,30 mg/ml BSA in the experimental group was significantly higher than that in the other groups(P<0 05).Conclusions:1.DBM scaffolds and DBM scaffolds with BSA coating were prepared successfully.After modification,the pore size and porosity of the scaffolds were decreased,the degradation rate was accelerated,and the hydrophilicity of the scaffolds was increased.2.DBM scaffolds and scaffolds with BSA coating had good cytocompatibility,and 20 mg/ml BSA-treated scaffolds promoted cell proliferation.3.The DBM scaffolds before and after BSA modification were co-cultured with cells,and the 20 mg/ml,30 mg/ml BSA modified scaffolds facilitated the adhesion of cells to the scaffolds.