摘要
目的 :研究人关节软骨细胞在PLGA三维支架上的贴附和生长情况 ,利用组织工程技术培养工程化关节软骨。方法 :制作PLGA三维细胞支架。分离培养成人关节软骨细胞 ,体外扩增后种植到PLGA支架中。在体外和裸鼠体内分别培养软骨细胞 -支架复合物 ,分期终止培养 ,进行组织学染色、扫描电镜、Ⅱ型胶原免疫组织化学及cDNA探针原位杂交检测。结果 :体外培养发现 ,软骨细胞支架材料内贴附生长良好 ,长期培养仍保持软骨细胞特性 ,裸鼠体内培养 8周后可形成软骨样组织。结论 :PLGA三维支架适合软骨细胞贴附生长和分泌软骨外基质 ,可作为软骨组织工程的细胞载体。利用组织工程的方法可获得适于移植的工程化关节软骨。
Objective:To verify the ability of PLGA, a three-dimensional biodegradable polymer, to support the growth of human chondrocytes, and to reconstruct human articular cartilage by tissue engineering.Method:Free chondrocytes isolated from adult articular cartilage were seeded onto sponge PLGA scaffold after expansion by in vitro culture. The complex of cell-scaffold were then cultured ex vivo and transplanted to subcutaneous of nude mouse respectively. The cell-seeded scaffolds in different culture intervals were assessed by histological staining of H-E, Safranine-O and Alcian blue. Scanning electron microscopy was utilized to verify the interactions between the cells and the biomaterial. Immunohistochemical analyses and in situ hybridization were also performed to evaluate collagen type II synthesis and mRNA expression. Result:Articular chondrocytes were well distributed and adhered to PLGA scaffolds and maintained their characteristics of articular cartilage throughout the culture period. Both the first passaged chondrocytes and seeded cells could express and product collagen type Ⅱ.After cultured in vivo for 8 weeks the cell-seeded scaffolds grew into hyaline cartilage-like tissues. Conclusion:PLGA scaffolds can support the chondrocytes' proliferation with maintenance of their phenotype, and can be suitable carriers for tissue engineering of articular cartilage.
出处
《中国现代医学杂志》
CAS
CSCD
2003年第11期1-5,共5页
China Journal of Modern Medicine
基金
国家自然科学基金资助项目 ( 3 9970 744 )
