离心管技术构建的组织工程软骨细胞生物学特性的研究

Study of biological chacteristics of chondrocytes in tissue engineered articular cartilage constructed by technique of centrifuge tube

目的 观察离心管技术构建的组织工程化软骨的细胞生物学特点 ,探讨该技术构建组织工程化软骨的可行性。方法 取 3周龄兔关节软骨的表层软骨 ,经酶消化获得大量的软骨细胞 ,用离心管技术构建软骨组织。对培养 3周的组织工程软骨进行组织学、免疫组织化学、透射电镜、3 H TdR掺入量等检查。结果 组织学显示软骨细胞位于陷窝 ,基质异染 ,Ⅱ型胶原免疫组化阳性 ,细胞内富含高尔基体 ,分泌囊泡和糖原颗粒 ,3 H TdR掺入显示软骨细胞DNA较正常软骨细胞明显增强。结论 离心管内软骨细胞的增殖较正常关节软骨增强。离心管技术可用于组织工程软骨的构建。

Objective To observe the biological behavior of chondrocytes in tissue engineered articular cartilage constructed by technique of centrifuge tube and explore the feasibility of constructing tissue engineered cartilage by the technique of centrifuge tube. Methods Chondrocytes isolated by enzymic digestion from the super layer of cartilages of 3 week old rabbits and cultured in 15ml Falcon centrifuge tube after centrifugation without any scaffold meterials.The cellular morphological and functional chacteristics were observed after cultured in centrifuge tube with histological examination,typeⅡ collagen immunocytochemcal examination, 3H TdR incorporation and electronic microscopy technique. Results The histobiological observation showed the chondrocytes in tissue engineered articular cartilage are of round morphology and were surronuded by matrix. The results of biochemistry examination were strong positive, typeⅡ collagen were stained positive.Most cells appeared to contain rich golgi zones and large areas of rough endoplasmic reticulum,partly filled with dense secretory material. The chondrocytes in tissue engineered articular cartilage have more 3H TdR incorporation than that of the chondrocytes from 3 week old rabbits.Conclusion The technique of centrifuge tube is situable for constructing tissue engineered articular cartilage