自体富血小板血浆联合骨髓基质干细胞复合改建脱细胞真皮基质修复兔关节软骨的可行性研究

The feasibility study of acellular dermal matrix reconstructed with autologous platelet-rich plasma combined with bone marrow stem cells in the repair of cartilage of rabbit

目的:研究自体富血小板血浆(PRP)联合骨髓基质干细胞(BMSCs)复合改建脱细胞真皮基质(ADM)修复兔关节软骨的修复效果及临床价值。方法:选择60只股骨髁关节损伤的青紫蓝兔作为研究对象,随机分为研究组、对照组和空白组,各20只。首先制备PRP,再分离BMSCs,并进行体外培养,接种于ADM支架上。研究组培养基添加10%PRP,其他条件同对照组,空白组兔不做任何处理。将BMSCs-ADM复合物植入兔关节缺损处,在1、2、3个月末对构建的软骨组织进行免疫力学、组织学、生物力学检测,比较PRP-BMSCs-ADM联合修复关节缺损的效果。结果:随着培养时间延长,BMSCs的细胞数目显著增多,呈现S形生长曲线,第3天后进入对数期,第9天后进入平台期;从第3天开始,细胞数目OD值研究组均明显高于对照组(P<0.01);随着时间的延长,各组标本软骨样细胞数目增多,软骨陷窝逐渐成熟,软骨表面逐渐光滑,IRCS宏观评分和组织学评分均呈现增长趋势(P<0.01),标本压力弹性模量有上升趋势(P<0.05);同时期研究组标本评分与压力弹性模量均显著高于对照组和空白组(P<0.01);3个月后,研究组软骨缺损修复状况明显优于对照组和空白组。结论:自体PRP联合BMSCs复合改建ADM修复兔关节软骨的修复效果良好,具有可行性。

Objective： To investigate the effects and clinical value of acellular dermal matrix（ ADM） reconstructed with autologous platelet-rich plasma（ PRP） combined with bone marrow stem cells（ BMSCs） in the repair of cartilage of rabbit. Methods： Sixty rabbits with femoral condylar articular injury were randomly divided into the study group,control group and blank group（ 20 rabbits each group）. The PRP and BMSCs were isolated,cultured,and inoculated with the ADM scaffolds in vitro. The 10% of PRP was added to the culture medium in study group,and the other treatment was the same as the control group. The blank group did not give any process. The BMSCs-ADM composite was implanted into the articular defect of rabbits. The reconstructing cartilage tissue was detected using the immune mechanics,histology and biomechanics at the end of 1,2 and 3 months of implantion. The restoration effects of joints defect with PRP-BMSCs-ADM were analyzed. Results： With the prolongation of the culture time,the number of BMSCs cells increased significantly,the growth curve of cells appeared S-shaped,which arrived at logarithmic phase and plateau phase after 3th day and 9th day,respectively. From the 3th day,the OD value of cell counts in study group was significantly higher than that in control group（ P〈0. 01）. With the extension of time,the number of cartilage-like cells in all groups increased,the cartilage space matured gradually,the surface of cartilage smoothed gradually,the macro scoring of IRCS and histological scoring increased significantly（ P〈0. 01）,the elastic modulus of the specimens pressure increased（ P〈0. 05）. During the same period,the specimen scoring and pressure elastic modulus in study group were significantly higher than those in control group and blank group（ P〈0. 01）. After 3 months,the repair of cartilage defect in study group was significantly better than that in control group and blank group（ P〈0. 01）. Conclusions： The effects of ADM reconstructed with PRP combined with BMSCs in repairing the cartilage of rabbit are good,which is feasible.