关节液组分分析在膝骨关节炎康复中的应用研究

Study on the Application of Component Analysis of Joint Fluid in the Rehabilitation of Knee Osteoarthritis

目的:研究膝骨关节炎患者康复程度与关节液成分变化之间的关系,寻找特异性及敏感性高的生化标志物,为准确诊断早期膝骨关节炎提供新的方法,探寻康复程度与关节液微环境变化的关系。方法:选择膝骨关节炎患者120例,对所有患者进行膝关节镜检查,按照Outerbridge分级标准将患者分为4组,Ⅰ度到Ⅳ度,每组30例。再将每组分成3组,分别进行自体富血小板血浆(PRP)+ 干细胞,自体PRP,以及臭氧灌注 + 透明质酸钠(HA)治疗。治疗前和治疗后8个月收集膝骨关节炎患者关节液标本各120份。标本均采用ELISA双抗夹心法对关节液中的印度刺猬蛋白、尿激酶纤维蛋白溶酶原激活剂(uPA)及基质金属蛋白酶-13(MMP-13)含量进行测定。比较治疗前后印度刺猬蛋白、uPA、MMP-13在关节液中的表达差异,以Spearman等级相关分析Outerbridge分级与关节液组分的关系。采用Shahariarre评分标准评价疗效。结果:印度刺猬蛋白、uPA、MMP-13表达水平与Outerbridge分级之间均呈正相关(r=0.790、r=0.476、r=0.684,P < 0.01);预后良好组与预后不良组印度刺猬蛋白、uPA、MMP-13表达水平比较,差异有统计学意义(P < 0.01);根据Outerbridge分级标准,Ⅰ度患者自体PRP + 干细胞治疗优良率与自体PRP组比较,差异无统计学意义(P > 0.05);Ⅱ~Ⅳ度患者自体PRP +干细胞组治疗优良率明显高于自体PRP组,差异有统计学意义(P < 0.05);Ⅰ~Ⅳ度患者自体PRP + 干细胞组治疗优良率均高于臭氧灌注 + HA组,差异有统计学意义(P < 0.05);Ⅱ度患者自体PRP组治疗优良率与臭氧灌注 + HA组比较,差异无统计学意义(P > 0.05),在Ⅰ度以及Ⅲ~Ⅳ度的患者中2组之间比较,差异有统计学意义(P < 0.05)。结论:自体PRP联合间充质干细胞治疗膝骨关节炎较常规HA + 臭氧灌注治疗效果显著,且膝骨关节炎患者的康复程度与关节液中印度刺猬蛋白、uPA、MMP-13的表达水平相关,说明膝骨关节炎患者的软骨损伤作用机制与印度刺猬蛋白、uPA、MMP-13的表达水平相关。

Objective:To study the relationship between the degree of rehabilitation of knee osteoarthritis and changes in the composition of joint fluid,to find biochemical markers with high specificity and sensitivity,to provide a new method for accurate diagnosis of early knee osteoarthritis,and to explore the relationship between the degree of rehabilitation and changes in the microenvironment of joint fluid.Methods:One hundred and twenty patients with knee osteoarthritis were selected.All patients were examined by knee arthroscopy.According to the Outerbridge grading standard,the patients were divided into four groups from grade Ⅰ to Ⅳ,with 30 patients in each group.Then,each group was divided into three groups,which were treated with PRP plus stem cells,autologous PRP,and ozone perfusion plus HA.One hundred and twenty samples of knee osteoarthritis were collected before treatment and 8 months after treatment.The contents of Indian hedgehog protein,uPA and MMP-13 in joint fluid were determined by ELISA double antibody sandwich method.The expressions of Indian hedgehog protein,uPA and MMP-13 in joint fluid were compared before and after operation,and the relationship between Outerbridge grade and joint fluid components was analyzed by Spearman rank correlation.The therapeutic effect was evaluated by the Shahariarre scoring standard.Results:The expression levels of Indian hedgehog protein,uPA and MMP-13 were positively correlated with Outerbridge grading(r=0.790,r=0.476,r=0.684,P < 0.01).The expression levels of Indian hedgehog protein,uPA and MMP-13in the good prognosis group were significantly different from those in the poor prognosis group(P < 0.01).According to the Outerbridge grading standard,the therapeutic excellent rate of autologous PRP plus stem cells in patients with grade I was not significantly different from that in the autologous PRP group(P > 0.05).The therapeutic excellent rate of autologous PRP plus stem cell group was significantly higher than that of autologous PRP group in patients with grade Ⅱ~Ⅳ(P < 0.05).The therapeutic excellent rate of theⅠ~Ⅳ degree autologous PRP plus stem cell group was higher than that of the ozone perfusion plus HA group,and the difference was statistically significant(P < 0.05).The therapeutic excellent rate of autologous PRP treatment group in patients with grade Ⅱ was not statistically significant compared with that of the ozone perfusion plus HA group(P > 0.05),but there was statistically significant difference between the two groups in patients with grade Ⅰ andⅢ~Ⅳ(P < 0.05).Conclusion:The effect of autologous PRP combined with mesenchymal stem cells in the treatment of knee osteoarthritis is more significant than that of conventional HA plus ozone perfusion,and the degree of rehabilitation of knee osteoarthritis patients is related to the expression levels of Indian hedgehog protein,uPA and MMP-13 in joint fluid.The difference is significant,which indicates that the mechanism of cartilage damage in knee osteoarthritis patients is related to the expression levels of Indian hedgehog protein,uPA and MMP-13.