注射式软骨缺损微创修复技术的大动物实验研究

Experimental Research of Injected Minimally Invasive Repair of Cartilage Defect on Large Animals

目的:在现有关节镜下microfracture技术的基础上,应用现代干细胞技术修复猪关节软骨缺损,探索注射式软骨缺损微创修复技术用于软骨再生治疗的可行性。方法:抽取6只猪的骨髓体外扩增培养至三代,动物随机分2组,每组6膝,每膝制备1个软骨缺损,左膝行缺损部位microfracture治疗后将3×107/mL浓度的骨髓间充质干细胞注射于关节腔内,右膝为单纯microfracture或空白对照。术后8、16周各3只动物,行大体观察、组织学检测,评价其对关节软骨缺损的再生修复效果。结果:术后8周观察见软骨缺损的修复髿表面平整,色泽渐趋正常,与周围组织整合良好;术后16周,修复组织具有透明软骨样结构,并产生大量GAGs和Ⅱ型胶原,单纯microfracture治疗组为纤维软骨修复,而空白组为少量纤维组织覆盖缺损底面,观察期内未见毛细血管长入及免疫排斥反应发生。结论:注射式软骨缺损微创修复技术创伤小,操作简便,能显著促进关节软骨缺损的再生修复的特点,具有较高的科学价值及临床应用前景。

Objective：Based on microfracture technology underthe present arthroscope, modern stem cell technology is applied to repair arthrodial cartilage defect of pigs and to investigate the feasibility of injected minimally invasive repair of cartilage defect in cartilage regeneration therapy.Methods：Bone marrow of six pigs was taken to culture to three generations.Animals were randomly divided into two groups with six knees in each group.There was one cartilage defect in each knee.Cartilage defect of the left knee was performed microfracture technology and then injected with bone marrow stem cells with a concentration of 3×107mL into the articular cavity; while right knee was made forsingle microfracture technology orblank group.In the eighth and sixteenth weeks aftersurgery, three ani-mals were respectively taken for general observation and histological detection to evaluate the effect of regeneration repairing on arthrosis cartilage defect.Results：Eight weeks after the surgery, the cartilage defect part was found of smooth surface and normal color, with integration to the surrounding tissue.Sixteen weeks aftersurgery, there was transparent cartilage in repairpart, with GAGs and Ⅱpattern collagen.Single microfracture therapy group were with repair of fibercartilage; while blank group were covered with fibertissues on the defect undersurface.During the whole phase, there was no blood capillary growing or any immunological rejection happened.Conclusion：Injected minimally invasive repair of cartilage defect, with little trauma and easy operation, can significantly promote the regeneration repair of arthrosis cartilage defect, so it has a high scientific value and clinical application.