PRP及RGD联合修饰表面改性后仿生基质对ADSCs生物学行为的调控

目的 评估富血小板血浆(platelet rich plasma,PRP)及精氨酸-甘氨酸-天冬氨酸(Arg-Gly-Asp,RGD)联合修饰表面改性后支架的细胞生物学特征,验证优化界面整合的方法.方法 兔脂肪基质干细胞成骨诱导化,nβ-TCP/Cs/PCL仿生基质Nd:YAG激光处理,表面改性及细胞接种:A组(PRP凝胶加RGD修饰表面改性基质加ADSCs)、B组(RGD修饰表面改性基质加ADSCs)、C组(表面改性基质加ADSCs)、D组(未表面改性基质加ADSCs);第1、4、8、12、16、20、24、28天,显微镜和电镜、共聚焦技术观察和检测细胞的存活率、增殖活力、Westen blot测定碱性磷酸酶及Ⅰ型胶原蛋白表达活性,分析Runx2与OPG表达.结果 A组细胞生长旺盛、细胞外基质丰富,存活率高于其它组(88.16±1.29,P＜0.05),增殖活力、碱性磷酸酶及Ⅰ型胶原水平均高于其它组(0.92±0.13,87.27±3.08,93.27±3.91,P＜0.05),Runx2和OPG表达显著.结论 PRP及RGD修饰联合支架表面改性能促进细胞增殖,为理想的骨整合方法.

The biological response of loading ADSCs to surface topography of biomimetic matrix combined PRP with RGD modification

Objective To evaluate the biological potential of surface topography of biomimetic matrix combined PRP gels with RGD modification. Methods Surface topography of nβ-TCP/Cs/PCL matrix was made by Nd: YAG laser, tissue-engineered bone was constructed in the following ways: ADSCs were loaded to nβ-TCP/Cs/PCL matrix with PRP gels plus RGD modification (group A), ADSCs were cultured to nβ-TCP/Cs/PCL matrix with RGD modification (group B), ADSCs implantation with topography-surfaced treatment of matrix (group C), ADSCs cultivation with smooth-surfaced treatment of matrix (group D). SEM and CLM were used to observe cellular pattern, survival rate, cell activity, ALP and collagen type Ⅰ level were detected at 1, 4, 8, 12, 16, 20, 24,28 days. Runx2 and OPG expressions were assayed at different interval. Results Under observation of SEM and CLM, new tissue showed more remarkable cell proliferation with abundant ECM in group A. Compared with other groups, the survival rate in group A was significantly higher (88.16 ± 1.29, P ＜ 0.05),and the level of cell activity, ALP, collagen type Ⅰ were significantly higher (0.92± 0.13, 87.27 ± 3.08, 93.27 ± 3.91, P＜ 0.05), and remarkable expression of Runx2 and OPG was also seen. Conclusion Topography-surfaced treatment of matrix combined PRP gels with RGD modification enhances the cell proliferation and acts as a feasible osteopromotive method.