万古霉素藻酸盐微球对共培养的间充质干细胞成骨分化的影响

Osteogenic differentiation effect of vancomycin alginate beads on con-cultured MSCs

目的检测纤维蛋白凝胶包裹的万古霉素藻酸盐微球对骨髓间充质干细胞(MSCs)成骨分化的影响,探讨其在骨组织工程中可能的应用。方法应用滴注法制备万古霉素藻酸盐微球,然后用纤维蛋白凝胶包裹,通过检测碱性磷酸酶的活性以及成骨基因的表达来评价不同浓度的微球对共培养的MSCs诱导成骨的影响。结果MSCs可在不同浓度微球溶液中生长,浓度为0.02、0.2和2.0g/L的藻酸盐微球溶液中的MSCs的碱性磷酸酶的活性依次增强,在成骨诱导的18d内,0、0.02、0.2和2.0g/L相应的最高值分别为(0.809 4±0.028 5、0.803 2±0.023 4、1.236 7±0.028 0、1.562 1±0.059 7)pg.min-1.cell-1。成骨分化的调控基因转录因子核心结合因子1以及Ⅰ型胶原基因在各实验组均有表达。结论纤维蛋白凝胶包裹的万古霉素藻酸盐微球可促进共培养的MSCs的碱性磷酸酶活性增加,转录因子核心结合因子1和Ⅰ型胶原基因的表达并未被影响。

Objective To detect the osteogenic differentiation effect of vancomycin alginate beads coated with fibrin gel （Vanco- AB-FG） on mesenchymal stem cells（MSCs）and to investigate their possible use in bone tissue engineering. Methods Alginate beads were produced by dropping and coated with fibrin gel. The activity of alkaline phosphatase （ALP） and expression of osteogenic genes were analyzed for evaluating the osteogenic differentiation effect of different concentration alginate beads on MSCs. Results MSCs grew in solution of alginate beads. The activity of ALP in MSCs cultured in medium containing 0.02g/L, 0.2g/L and 2. 0g/L Vanco-AB-FG was increased in order. During 18d osteogenic induction, the highest values of ALP activity were respectively （0. 809 4±0. 028 5）pg · min^-1· cell^-1 in 0g/L,（0. 803 2v0. 023 4）pg · min^-1· cell^-1 in 0.02g/L, （1. 236 7±0. 028 0）pg · min^-1· cell^-1 in 0.2g/L and （1. 562 ±0. 059 7）pg · min^-1· cell^-1 in 2. 0g/L. The expression of core-binding factor alpha subunit l（cbfa-1）gene and type I collagen gene appeared in all experimental groups. Conclusion The ALP activity in MSCs was increased by Vanco-AB-FG. The expression of cbfa-1 gene and type I collagen gene was not affected by Vanco-AB-FG.