摘要
目的研究RGD多肽修饰CS/HA复合多孔支架对于MSCs在支架上黏附的数量及质量。方法通过原位杂化法制备CS/HA多孔支架并应用物理吸附的方法将RGD多肽负载到支架表面制备出CS/HA-RGD复合支架并对其表征。分别将从Wistar大鼠提取并培养的MSCs种植到实验组CS/HA-RGD及对照组CS/HA两种支架各10个进行复合培养。4 h后检测其黏附率,3天后应用扫描电镜及激光共聚焦显微镜观察细胞黏附状态。结果 X射线光电子能谱(XPS)及吸光度法测定RGD的负载率为67%。应用电镜测得支架的大孔道的直径约为400μm,而小孔径为3～5μm。应用酶标仪测定两种支架4 h的骨髓基质干细胞的黏附率,负载RGD组为(80.7±1.8)%而无RGD组为(54.7±2.6)%。继续培养3天后应用激光共聚焦显微镜及扫描电镜观察,负载RGD组明显提高了MSCs的黏附数量和质量。结论 RGD多肽具有促进MSCs在短时间内迅速黏附CS/HA复合多孔支架作用。
Objective To study the regulation of RGD for chitosan/hydroxyapatite（CS/HA） scaffold with channel/spherical pore in order to detect cell adhesion rate and quality on scaffold.Methods The CS/HA scaffold was prepared via in situ hybridization.Physical adsorption method was applied to load RGD peptide on the stent surface,CS/HA-RGD scaffolds was prepared and marked.MSCs extracted and cultured from Wistar rats were planted to the experimental group CS/HA-RGD（n=10） and control group CS/HA（n=10） and given combined cultivation.After 4 h,the adhesion rate was detected,after 3 days scanning electron microscopy and laser confocal microscopy were applied to observe cell adhesion status.Results The load rate of RGD on the scaffold was 67% detected by XPS and absorption spectrophotometry.The size of channel pore and sphere pore of CS/HS scaffold were about 400 μm and 3～5 μm,respectively.Cell adhesion rate of CS/HA scaffold with RGD were（80.7±1.8）% after 4 h cultivation,which were higher than that of CS/HA group（54.7±2.6）%.The quantity and quality of adhesion of MSCs on RGD-load scaffold were significantly increased.Conclusion RGD peptides can promote adhesion of MSCs on CS/HA combined scaffold in a short time.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2011年第3期199-202,共4页
Journal of Harbin Medical University
基金
国家自然科学基金资助项目(50702017
302672126)
