脂肪干细胞与富血小板血浆治疗大鼠膝骨关节炎疗效比较

The comparison of adipose derived stem cells and platelet rich plasma in the treatment of knee osteoarthritis in rats

目的比较富血小板血浆(PRP)和脂肪干细胞(ADSCs)对膝骨关节炎(KOA)大鼠模型关节疼痛和软骨修复的作用.方法随机选取4只雄性SD大鼠,心内采血分离制备PRP并取其腹股沟脂肪组织培养收集ADSCs.选取雄性SD大鼠32只,随机分为空白组、KOA模型组、PRP组、ADSCs组,每组各8只,除空白组外均以碘乙酸注射膝关节腔建立大鼠KOA模型.PRP组、ADSCs组分别用1×10^(6)个/ml浓度的PRP和1×10^(6)个/ml浓度的ADSCs对大鼠双侧膝关节采纳注射干预,注射1次/周,共8周,定时察看大鼠的生理行为,评估疼痛指标,8周后取膝关节进行组织病理学染色并进行OARSI评分.结果压痛实验结果表明,造模后第8周,模型组压痛阈值比空白组明显降低(P<0.01),用药干预后的PRP组和ADSCs组压痛阈值与模型组相比有所提高(P<0.01),PRP组压痛阈值与ADSCs组相比有所提高(P<0.05).热痛结果表明,造模后第8周,模型组热痛阈值比正常组明显降低(P<0.01),用药干预后的PRP组和ADSCs组压痛阈值比模型组提高(P<0.01).PRP组和ADSCs组热痛阈值差异无统计学意义(P>0.05).病理学结果表示,与空白组比较,KOA模型组大鼠膝关节软骨表面明显存在残缺,损害处软骨细胞显著缺失,软骨面明显变薄,软骨下骨出现纤维化退行性变;PRP组、ADSCs组均可改善KOA模型大鼠关节软骨损害,镜下可观察到大量的软骨细胞,软骨表面平整.其中ADSCs组软骨仍有少量软骨细胞丢失,软骨层轻度变薄,仍可见少量肥大软骨细胞,软骨下骨有少量纤维化退变;PRP组软骨基本复原,软骨面增厚.KOA模型组OARSI评分(4.30±1.63)均高于空白组(0.94±0.70)、RPR组(1.83±1.00)和ADSCs组(2.66±0.90)(P<0.01),PRP组OARSI评分高于ADSCs组(P<0.05).结论PRP和ADSCs均可改善KOA大鼠膝关节炎症疼痛,修复软骨损伤,但PRP疗效优于ADSCs.

Objective To observe and compare the efects of platelet rich plasma(PRP)and adipose derived stem cells(ADSCs)on joint pain and cartilage repair in knee osteoarthrtis(KOA).Methods The PRP was isolated from peripheral blood in 4 SD rats and the ADSCs were obtained from their fats 32 SD rats were selected and were randomly divided into blank group,KOA model group,PRP group and ADSCs group,8 cases in each group.Except the blank group,the pain models of KOA in rats were cstablished by iodoacetic acid method.The rats in PRP group,ADSCs group were administrated with intra-articular injection of 1x106 cells/ml PRP and 1x106 ell/ml ADSCs in bilateral knee joint,once a week for consecutive 8 weeks.Pressure-pain threshold,Paw withdrawal latency(PWL)and OARSI scoring of knee joint were performed regularly,and pathological results were observed after 8 injections.Results At the 8th week afer the model replication,the results of tenderness test showed that threshold of model group was signifcantly lower than that of blank group(P<0.01).The tendemess threshold of PRP group and ADSCs group was higher than that of model group(P<0.01).The tendermess threshold of PRP group was higher than that of ADSCs group(P<0.05).The results showed that the thermal pain threshold of model group was significandly lower than that of normal group(P<0.01),and that of PRP group and ADSCs group was higher than that of model group(P<0.01).There was no diference in thermal pain threshold between PRP group and ADSCs group(P>0.05).Compared with the blank group,the pathological results showed that the KOA model group had the defect of articular cartilage suface,the loss of chondrocyte,fibrosis and degeneration of subchondral bone.The PRP group and ADSCs group had the improvement efect on the articular cartilage,a large number of chondrocytes were observed and the cartilage surface was smooth.In ADSCs group,there were still loss of chondrocyte,thickness of cell bypertrophic degencration,the cartilage surface was thinner than PRP group.In PRP group,the cartilage recovered and the cartilage surface thickened.The OARSI score of KOA model group(4.30±1.63)was higher than that of blank group(0.94±0.70),RPR group(1.83±1.00)and ADSCs group(2.66±0.90)(P<0.01).The OARSI score of PRP group was higher than that of ADSCs group(P<0.05)。Conclusion Both PRP and ADSCs can ameliorate joint pain in KOA rats and repair artilage damage,but the curative effect of PRP is superior to ADSCs.