组织工程化兔耳廓软骨的实验研究

Tissue-engineered auricled cartilage: an experimental study

目的 探讨采用表面修饰过的壳聚糖 (chitosan) 聚乳酸 (polylacticacid ,PLA) 聚乙内酯(polycrylactone ,PCL)及牛跟腱胶原海绵作为支架培养组织工程化耳廓软骨的可行性 ,比较动态与静态两种不同培养方法对软骨生长的影响。方法 将 4周龄新西兰大耳白兔耳廓软骨细胞接种在 2 0个壳聚糖 PLA PCL及牛跟腱Ⅰ型胶原海绵支架上 ,将细胞支架复合物分为 2组 ,动态组 (样本量为 10 )采用旋转培养 ,静态组 (样本量为 10 )采用静置培养。所有标本体外培养 1周 ,裸鼠或兔皮下培养 8周。分别于体外 1周、体内 4周及 8周时取样行扫描电镜检查、大体及组织学观察以及免疫组化分析 ,测定软骨细胞数及细胞外基质中的Ⅱ型胶原含量。结果 表面修饰过的壳聚糖支架上软骨细胞贴壁率高 ,细胞分化与增殖好 ;每个时段动态组生成的软骨细胞量及Ⅱ型胶原均较静态组多 ,两组间差异有显著性 (P <0 0 5 ) ;2种生物支架上均长出了组织工程软骨 ;裸鼠皮下较兔皮下形成的软骨更接近天然软骨。结论 壳聚糖 PLA PCL及牛跟腱胶原海绵是较理想的组织工程耳廓软骨的支架材料 ,动态培养更有利于新生软骨组织的生成 ;

Objective To study the feasibility of engineering auricled neocartilage with chitosan/polylacticacid-polycrylactone (PLA-PCL) network scaffolds and to search the difference between dynamic and silent chondrocytal culture techniques. Methods Chondrocytes from auricled cartilage of 4 weeks old New Zealand White rabbit were seeded onto chitosan/PLA-PCL network. Ten cell-polymer scaffolds were divided into two groups: dynamic group (n1=5) with rotating bioreactors and silent group (n2=5) with ordinary dishes to culture. Using scan electroscope, grossly histological and immunohistological techniques, the morphological evaluation was done individually at 1st week in vitro, 4th and 8th week in vivo. Results Chondrocytes adhered and grew up well on the network, but more quantities of chondrocytes, Glycoaminoglygan (GAG) and type Ⅱ chollagen were found in dynamic group. There was an obvious difference between dynamic and silent group (P<0.05). Conclusion Chitosan/PLA-PCL network scaffold is good for adhesions and growth of chondrocytes. Furthermore, dynamic cell culture method is better than silent method for formation of neocartilage.