新型壳聚糖基仿生网络复合膜诱导骨髓间充质干细胞定向成骨分化

Objective To study the osteogenic differentiation capacity of mesenchymal stem cells(MSCs)on chitosan based biomimetic network composites films.Methods The chitosan-based biomimetic network composites films were prepared using chitosan,gelatin and pectin in certain ratio by biomimetic approach.The experiment was designed as follow:experimental group 1 (composites films+routine medium),control group 1 (routine medium),experimental group 2(composites films+osteogenic medium),control group 2(osteogenic medium).The growth and proliferation of MSCs were observed by inverted phase contrast microscope and scanning electronmicroscopy(SEM) and detected through methyl thiazolyl tetrazolium(MTT) assay.Alkaline phosphatase(ALP) activity was used to investigate the osteogenic differentiation capacity.The formation of mineral was determined by special staining and the energy dispersive X-ray apparatus (EDX).Results MSCs could well adhere,grow and proliferate on the network composites films.The MTT assay demonstrated that there was no significant difference in cell viability between the experimental and control groups (P＞0.05).SEM observation showed that MSCs grew assembly on the surface of the films secreted a large number of extracelluar matrix,and formed scattered nodules.The cells on the experimental group 1 showed statistically higher ALP activity level than those cells on the other groups (P＜0.01).Moreover,the calcified nodules can be seen in the experimental groups by Alizarin red and Von Kossa staining.ALP staining showed blue-staining granules within the cytoplasm,and the deposition of Ca and P was detected on the films by SEM-EDX.Conclusion The novel chitosan-based biomimetic network composites films not only have good biocompatibility,but can also improve MSCs differentiation toward the osteogenic lineage.

Osteogenic differentiation behaviors of mesenchymal stem cells on chitosan based network composites films