【摘要】 目的用胶原蛋白和人血纤维蛋白混合物为载体支架在体外进行软骨细胞三维立体培养,构建人工软骨组织,利用此人工软骨修复关节软骨缺损。方法取2周龄兔关节软骨,经消化、分离的软骨细胞体外培养。培养第3周时,进行免疫组织化学分析;利用培养3周的人工软骨对异体成兔的关节软骨损伤进行修复。结果培养物内软骨细胞均得到较好存活,形成软骨陷窝,出现同源性细胞簇,分泌软骨基质;DNA和糖胺多糖(GAG)在培养第24天时达到高峰,分别为(5.18±0.19)、(214.3±2.8)μg/块;对异体关节软骨损伤的移植修复结果良好,在12周时,移植物与周边正常组织结合紧密、齐高,移植的软骨细胞趋于柱状排列,为透明软骨组织。结论用胶原蛋白和人血纤维蛋白为载体支架体外培养软骨细胞,可构建较大的组织工程软骨,能较好地修复同种异体关节软骨缺损。更多还原

【Abstract】 Objective To construct articular cartilage by three-dimensional culturing chondrocytes using collagen and human fibrin as supporter. Methods Chondrocytes were isolated from articular cartilage of 2-week-old rabbit, digested by collagenase II, and mixed with bovine type I collagen, human blood freeze-dry fibrinogen and thrombosin according to a certain proportion to culture in vitro. Stained with HE, toluibin, and detected with the methods of immunohistochemistry at in vitro culture times of 3 weeks, and analyzed the content of DNA and GAG at different times. Using in vitro cultured tissue engineering cartilage repaired articular cartilage defect in knee of rabbit. Results After 3 weeks, all cells were viable, formed cartilage lacuna and homologous cell cluster, and secreted cartilage matrix stained by toluidine blue in culture and secreted type Ⅱcollagen detected by immunohistochemistry method. The content of DNA and GAG peak appeared at in vitro culture 24 days. The articular defect repair showed good results. The defect filled with hyaline-like cartilage, characterized by its smooth surface and well-fusion with its surrounding tissues, and the high degree similarity to surrounding normal cartilage, the chondrocytes in repaired tissue arranged in column after grafting for 12 weeks. Conclusions Larger artificial cartilage tissue can be constructed by three-dimensional culturing hondrocytes using collagen and human fibrin as supporter and this tissue engineered cartilage have good repair effect in large full-thickness repair models.更多还原