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背景:骨髓间充质干细胞在不同诱导条件下具有向中胚层组织细胞如成骨细胞、成软骨细胞、肌细胞、脂肪细胞等分化的能力。目的:验证用组织工程方法诱导分化骨髓间充质干细胞修复兔关节软骨损伤的效果。设计、时间及地点:随机对照动物实验,于2005-05-03/2007-12-30在沈阳医学院临床中心实验室完成。材料:20只两三月龄的健康新西兰白兔,雌雄不限。方法:①诱导分化体外培养的兔骨髓间充质干细胞。实验组加入地塞米松、碱性成纤维细胞生长因子、维生素C培养1周,再将转化生长因子β替换碱性成纤维细胞生长因子培养3周;以不加诱导剂细胞做对照。②取20只兔,建立膝关节软骨缺损模型,随机分为3组。实验组10只膝关节内植入经诱导的骨髓间充质干细胞;对照组植入未经诱导的骨髓间充质干细胞;空白对照组植入生理盐水。分别于术后2,4,6,8周时处死实验组处死2只,对照组和空白对照组各处死1只进行各项指标检测。主要观察指标:①细胞形态学。②碱性磷酸酶活性的测定。③大体标本观察。④X射线观察。⑤组织切片观察。结果:①经诱导的骨髓间充质干细胞,细胞形态发生明显变化,逐渐由长梭形变为多角形,类似于软骨细胞样形态。②骨髓间充质干细胞经诱导4周后,其碱性磷酸酶活性明显增强(P<0.05)。③术后8周,实验组标本修复组织表面光滑,与周围软骨间界限模糊不清;X射线表现为关节间隙变宽,软骨下骨质囊变得到改善;组织切片观察显示与正常软骨细胞基本上一致。结论:自体间充质干细胞移植可修复关节软骨损伤。
BACKGROUND: Bone marrow mesenchymal stem cells have the ability to differentiate into mesodermal tissue cells such as osteoblasts, chondroblasts, myocytes and adipocytes under different inducing conditions. OBJECTIVE: To verify the effect of inducing the differentiation of bone marrow mesenchymal stem cells in rabbit articular cartilage by tissue engineering. DESIGN, TIME AND SETTING: The randomized controlled animal experiment was performed at the Laboratory of Clinical Center, Shenyang Medical College from May 2005 to March 2007. MATERIALS: Twenty healthy New Zealand White rabbits two to three months old, male or female. Methods: ① Induction of differentiation of rabbit bone marrow mesenchymal stem cells in vitro. Experimental group was added dexamethasone, basic fibroblast growth factor, vitamin C for 1 week, and then transforming growth factor β-basic fibroblast growth factor for 3 weeks; without the addition of inducer cells as a control. ② Take 20 rabbits to establish the knee cartilage defect model, were randomly divided into 3 groups. Ten knee joints of the experimental group were implanted with induced bone marrow mesenchymal stem cells; the control group was implanted with uninjured bone marrow mesenchymal stem cells; and the blank control group was implanted with saline. Two rabbits were sacrificed at 2, 4, 6, and 8 weeks postoperatively respectively. Two rabbits in the control group and one blank control group were killed. MAIN OUTCOME MEASURES: ① Cell morphology. ② alkaline phosphatase activity determination. ③ gross specimen observation. ④ X-ray observation. ⑤ tissue sections observed. Results: ① The induced morphological changes of bone marrow mesenchymal stem cells changed significantly from long spindle to polygon, similar to chondrocyte-like morphology. ② After 4 weeks, the alkaline phosphatase activity of bone marrow mesenchymal stem cells increased significantly (P <0.05). At 8 weeks after operation, the surface of the repaired tissue in the experimental group was smooth and the boundary between the experimental group and the surrounding cartilage was vague. The X-ray showed the widened joint space and the subchondral bone mass. The histological section showed that the repaired tissue showed no difference with the normal chondrocytes On the same. Conclusion: Autologous mesenchymal stem cell transplantation can repair articular cartilage injury.