iPSC-MSCs与hUC-MSCs治疗大鼠膝骨关节炎的实验研究

Experimental study of iPSC-MSCs and hUC-MSCs in the treatment of knee osteoarthritis in rats

目的基于动物实验,研究诱导多能干细胞来源间充质干细胞(iPSC-MSCs)与人脐带来源间充质干细胞(hUC-MSCs)对大鼠膝骨关节炎的治疗作用及可能机制。方法按照随机数字法将50只SD膝骨关节炎大鼠模型(Hulth法建立)分为模型组、iPSC-MSCs组、hUC-MSCs组、玻璃酸钠组、假手术组,每组10只。模型组与假手术组通过关节腔注射50μL的生理盐水,iPSC-MSCs组与hUC-MSCs组分别通过关节腔注射50μL的iPSC-MSCs与hUC-MSCs(均为4×108个细胞/mL),玻璃酸钠组通过关节腔注射5次玻璃酸钠注射液(1次/周,50μL/次)。比较五组大鼠的膝关节直径差值、膝关节腔Kellgren-Lawrence分级、膝关节Pelletier评分、膝关节病理组织学评分、基质金属蛋白酶(MMP)-13、Ⅱ型胶原(CollagenⅡ)及解聚蛋白样金属蛋白酶(ADAMTS)-5的水平。结果iPSC-MSCs组、hUC-MSCs组与玻璃酸钠组的膝关节直径差值均明显低于模型组(P<0.05);iPSC-MSCs组、hUC-MSCs组与玻璃酸钠组的膝关节腔Kellgren-Lawrence分级均明显优于模型组(P<0.05),iPSC-MSCs组明显优于hUC-MSCs组与玻璃酸钠组(P<0.05);iPSC-MSCs组、hUC-MSCs组与玻璃酸钠组的膝关节Pelletier评分均明显低于模型组(P<0.05),iPSC-MSCs组明显低于hUC-MSCs组与玻璃酸钠组(P<0.05);iPSC-MSCs组、hUC-MSCs组的膝关节病理组织学评分均明显低于模型组(P<0.05),iPSC-MSCs组明显低于hUC-MSCs组与玻璃酸钠组(P<0.05);iPSC-MSCs组及玻璃酸钠组的CollagenⅡ水平明显高于模型组,MMP-13与ADAMTS-5水平均明显低于模型组(P<0.05),iPSC-MSCs组的CollagenⅡ水平明显高于hUC-MSCs组及玻璃酸钠组(P<0.05),MMP-13与ADAMTS-5水平均明显低于hUC-MSCs组及玻璃酸钠组(P<0.05);hUC-MSCs组的CollagenⅡ水平明显高于玻璃酸钠组(P<0.05)。结论iPSC-MSCs与hUC-MSCs对大鼠膝骨关节炎具有明显治疗作用,iPSC-MSCs优于hUC-MSCs,其作用机制可能与上调CollagenⅡ水平及下调MMP-13与ADAMTS-5水平有关。

Objective To study the therapeutic effects and possible mechanism of induced pluripotent stem cell derived mesenchymal stem cells(iPSC-MSCs)and human umbilical cord derived mesenchymal stem cells(hUC-MSCs)on knee osteoarthritis based on the animal experiments.Methods According to the random number method,50 SD rat models of knee osteoarthritis(established by Hulth method)were divided into model group,iPSC-MSCs group,hUC-MSCs group,sodium hyaluronate group and sham operation group,10 rats in each group.In the model group and the sham group,50μL of normal saline was injected into the articular cavity once.In the iPSC-MSCs group and hUC-MSCs group,50μL of iPSC-MSCs and hUC-MSCs(both 4×108 cells/mL)were injected into the articular cavity once,respectively.In the sodium hyaluronate group,50μL of sodium hyaluronate was injected into the articular cavity 5 times(1 time/week).The difference value of knee joint diameter,knee joint cavity Kellgren-Lawrenc classification,knee joint Pelletier score,knee joint histopathological score and the level of matrix metalloproteinases(MMP)-13,ⅡCollagen type(CollagenⅡ),depolymerization protein sample metalloproteinases(ADAMTS)-5 were compared among five groups.Results The difference value of knee joint diameter of iPSC-MSCs group,hUC-MSCs group and sodium hyaluronate group was significantly lower than that of the model group(P<0.05).The knee joint cavity Kellgren-Lawrenc classification of iPSC-MSCs group,hUC-MSCs group and sodium hyaluronate group was significantly better than that of the model group(P<0.05),the iPSC-MSCs group was significantly better than that of hUC-MSCs group and sodium hyaluronate group(P<0.05).The knee joint Pelletier score of iPSC-MSCs group,hUC-MSCs group and sodium hyaluronate group was significantly lower than that of the model group(P<0.05),the iPSC-MSCs group was significantly lower than that of hUC-MSCs group and sodium hyaluronate group(P<0.05).The knee joint histopathological score of iPSC-MSCs group,hUC-MSCs group and sodium hyaluronate group was significantly lower than that of the model group(P<0.05),the iPSC-MSCs group was significantly lower than that of hUC-MSCs group and sodium hyaluronate group(P<0.05).The level of CollagenⅡof iPSC-MSCs group,hUC-MSCs group and odium hyaluronate group was significantly higher than that of the model group(P<0.05),the level of MMP-13and ADAMTS-5 of iPSC-MSCs group,hUC-MSCs group and odium hyaluronate group was significantly lower than that of the model group(P<0.05),the level of CollagenⅡof iPSC-MSCs group was significantly higher than that of hUC-MSCs group and sodium hyaluronate group(P<0.05),the level of MMP-13and ADAMTS-5 of iPSC-MSCs group was significantly lower than that of hUC-MSCs group and sodium hyaluronate group(P<0.05),the level of CollagenⅡof hUC-MSCs group was significantly higher than that of sodium hyaluronate group(P<0.05).Conclusion iPSC-MSCs and hUC-MSCs had obvious therapeutic effects on knee osteoarthritis in rats,iiPSC-MSCs was better than that of hUC-MSCs.its mechanism may be related to the up regulation of CollagenⅡand down regulation of MMP-13,ADAMTS-5.