CAJ | 학술논문

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.
배경:조직공정기술이경성위관절연골결손수복적연구열점,생장인자시기중적중요부분,단시,생장인자료효화안전성환불명학.연구발현,위령선능구유지화촉진연골세포합성단백다당여Ⅱ형효원,병차능촉진연골세포증식급전화생장인자β1mRNA적표체.목적:탐토가주사성각취당/β-감유린산이납응효복합연골세포응효수복관절연골결손적가행성급위령선적간예효응.방법:우신서란백토슬관절적고골활차면용아과찬제성심도위0.4 mm적연골결손,수궤분위3조,위령선조、보통배양기조조모후주입각취당/β-감유린산이납복합연골세포혼현액1 mL,재응효주입후제2천,분별급여관절강주사위령선배양기혹보통배양기1 mL,1차/d,련속급약7 d.단순조모조불작특수처리.술후6,12주후분별행대체、조직학(소목정-이홍염색、TB염색)、Ⅱ형효원면역조직화학관찰,병진행Wakitani평분.결과여결론:위령선조、보통배양기조가견결손관절면피교호지전충,병형성투명연골양결구,위령선조표면평정도급여주위조직정합정도교보통배양기조호,조직학절편상가견류연골형성병분비연골기질화특이성Ⅱ형효원.단순조모조미견수복,가견조직증생퇴변.위령선조수복조직여주위조직정합정도、조직학급Ⅱ형효원분비량균호우보통배양기조.위령선조、보통배양기조Wakitani평분현저저우단순조모조(P<0.01),차위령선조분치명현저우보통배양기조(P<0.05).결과제시가주사성각취당/β-감유린산이납응효복합동충이체연골세포능구수복관절연골결손,차위령선능구촉진기대연골결손적수복,위령선재조직공정기술수복관절연골결손중가능기도류생장인자작용.
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.