可注射性壳聚糖/β-甘油磷酸二钠凝胶复合同种异体软骨细胞修复兔膝关节软骨缺损及威灵仙的干预效应

背景:组织工程技术已经成为关节软骨缺损修复的研究热点,生长因子是其中的重要部分,但是,生长因子疗效和安全性还不明确.研究发现,威灵仙能够维持和促进软骨细胞合成蛋白多糖与Ⅱ型胶原,并且能促进软骨细胞增殖及转化生长因子β1mRNA的表达.目的:探讨可注射性壳聚糖/β-甘油磷酸二钠凝胶复合软骨细胞凝胶修复关节软骨缺损的可行性及威灵仙的干预效应.方法:于新西兰白兔膝关节的股骨滑车面用牙科钻制成深度为0.4 mm的软骨缺损,随机分为3组,威灵仙组、普通培养基组造模后注入壳聚糖/β-甘油磷酸二钠复合软骨细胞混悬液1 mL,在凝胶注入后第2天,分别给予关节腔注射威灵仙培养基或普通培养基1 mL,1次/d,连续给药7 d.单纯造模组不作特殊处理.术后6,12周后分别行大体、组织学(苏木精-伊红染色、TB染色)、Ⅱ型胶原免疫组织化学观察,并进行Wakitani评分.结果与结论:威灵仙组、普通培养基组可见缺损关节面被较好地填充,并形成透明软骨样结构,威灵仙组表面平整度及与周围组织整合程度较普通培养基组好,组织学切片上可见类软骨形成并分泌软骨基质和特异性Ⅱ型胶原.单纯造模组未见修复,可见组织增生退变.威灵仙组修复组织与周围组织整合程度、组织学及Ⅱ型胶原分泌量均好于普通培养基组.威灵仙组、普通培养基组Wakitani评分显著低于单纯造模组(P<0.01),且威灵仙组分值明显低于普通培养基组(P<0.05).结果提示可注射性壳聚糖/β-甘油磷酸二钠凝胶复合同种异体软骨细胞能够修复关节软骨缺损,且威灵仙能够促进其对软骨缺损的修复,威灵仙在组织工程技术修复关节软骨缺损中可能起到类生长因子作用.

Repair of rabbit knee articular cartilage defect by the injectable chitosan/beta-glycerophosphate gel encapsulating allograft chondrocytes and the intervention of Weilingxian

BACKGROUND: At present, studies on repair of cartilage defect have been focused on tissue engineering technique. Growth factors are one of the most important parts. However, the effect and security of growth factors have not been confirmed. Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan, collagen Ⅱof chondrocyte, and it also can promote proliferation of chondrocyte and expression of transforming growth factor (TGF)-β1 mRNA.OBJEGTIVE: To investigate the feasibility of injectable chitosan/β-glycerol phosphate (C/β-GP) encapsulating allograft chondrocytes on the repair of articular cartilage defects and the intervention effect and possible mechanisms of Weilingxian.METHODS: A 0.4-mm defect was established on knee articular cartilage. Expeirmental New Zealand rabbits were randomly divided into three groups: Weilingxian, common culture media, and model groups. In the common culture media group, the samples were treated with C/β-GP and chondrocyte suspension (1 mL); at 2 days after gel injection, Weilingxian or common culture media (1 mL) were respectively given into joint cavity, once a day, for 7 successive days. The samples in the model group were not treated. Gross, histological (HE staining, TB staining), type Ⅱ collagen immunohistochemical, and Wakitani score examinations were performed on 6 and 12 weeks after surgery.RESULTS AND CONCLUSION: Defects of articular surface were well filled in Weilingxian and common culture media groups, and hyaline cartilage-like structure was formed. The surface flatness and degree of integration with surrounding tissue of Weilingxian group was better than common culture media group. Formation of cartilage-like and secretion of cartilage matrix and specificity of collagen type Ⅱ were found in histological slices. Defects in the model group were not repaired, while tissue proliferative degeneration was observed. Integration of.repair tissue with surrounding tissue, histology and amount of type Ⅱ collagen secretion in Weilingxian group were better than common culture media group. Wakitani scores of Weilingxian group and common culture media group were significantly lower than model group (P < 0.01), andscores of Weilingxian group was significantly lower than common culture media group (P<0.05). Injectable chitosan/β-glycerolphosphate gel encapsulating allograft chondrocytes could repair articular cartilage defects, and Weilingxian was able to promote the process of it, this manifested the role like growth factor in tissue engineering technique repairing articular cartilage defects.