摘要
目的制备负载TGF-β1壳聚糖缓释微球的三维多孔壳聚糖支架,检测TGF-β1缓释载体对软骨细胞功能的影响。方法乳化交联法制备TGF-β1壳聚糖缓释微球并将其与壳聚糖支架复合,检测其体外降解并通过ELISA法检测微球的载药、释药性能。将软骨细胞置于复合载体中立体培养,通过苏木素伊红染色、Ⅱ型胶原免疫组化染色、扫描电镜观察复合载体对细胞的增殖、功能的影响。结果缓释微球的TGF-β1包封率达90.1%,并具有良好的药物缓释性能,软骨细胞在复合载体中增殖良好,并能够保持其表型及Ⅱ型胶原分泌功能。结论负载TGF-β1壳聚糖缓释微球的壳聚糖支架作为软骨细胞的载体在组织工程软骨的构建及软骨损伤的修复中有良好的应用前景。
Objective To assess the feasibility of using biodegradable chitosan microspheres as carriers for controlled TGF-β1 delivery and the effect of released TGF-β1 on the chondrogenic potential of chondrocytes. Methods Chitosan scaffolds and chitosan microspheres loaded with TGF-β1 were prepared by the phase-separation and the emulsion-crosslinking method respectively. In vitro drug release kinetics,as measured by enzyme-linked immunosorbent assay （ELISA） , was monitored for 7 days. Lysozyme degradation was performed for 4 weeks to detect in vitro degradability of the scaffolds and the microspheres. Rabbit ehondrocytes were seeded on the scaffolds and the effects of released TGF-β1 on cell adhesivity,proliferation,morphological changes,and synthesis of the extracellular matrix were observed by light and scanning electron microscopy. Results TGF-β1 was encapsulated into chitosan microspheres and the encapsulation efficiency of TGF-β1 was high（90. 1% ）. During 4 weeks of incubation in lysozyme solution for in vitro degradation,the mass of both the scaffolds and the microspheres decreased continuously and significant morphological changes was noticed. From the release experiments,it was found that TGF-β1 could be released from the microspheres continuously. Both proliferation rate and production of collagen type Ⅱ were significantly higher in the TGF-β1 microsphere incorporated scaffolds than in the control group,indicating that the activity of TGF-β1 was retained during microsphere fabrication and after release. Conclusion Chitosan microspheres can serve as delivery vehicles for controlled release of TGF-β1. Chitosan scaffolds incorporated with chitosan microspheres loaded with TGF-β1 possess a promising potential to be applied for controlled cytokine delivery and cartilage tissue engineering.
出处
《中华显微外科杂志》
CSCD
北大核心
2005年第4期324-327,共4页
Chinese Journal of Microsurgery
基金
国家自然科学基金(30270393)
广东省科技计划项目(2003A302102)
