诱导多能干细胞来源间充质干细胞对大鼠骨关节炎的治疗作用

Therapeutic roles of induced pluripotent stem cell-derived mesenchymal stem cells on experimental model of osteoarthritis

目的研究诱导多能干细胞来源间充质干细胞(iPSC-MSCs)对大鼠骨关节炎(OA)的治疗作用。方法采用Hulth法建立大鼠OA模型,编号后运用Excel表格随机分组功能将其分为模型组、iPSC-MSCs组、人脐带来源间充质干细胞(hUC-MSCs)组、玻璃酸钠(HA)阳性对照组,另设假手术组作为对照,每组10只。除HA组动物多次给药(每周1次,共给药5次)外,其它各组均单次给药,每次每只动物在术侧关节腔注射50μL药物iPSC-MSCs和hUC-MSCs,假手术组和模型组同法给予生理盐水。给药5周后处死动物,观察iPSC-MSCs对关节直径差值、关节腔结构及评分、关节面评分、病理学评分等指标的影响;检测膝关节基质金属蛋白酶(MMP)-13、Ⅱ型胶原(CollagenⅡ)及解聚蛋白样金属蛋白酶(ADAMTS)-5表达情况。结果 iPSCMSCs和hUC-MSCs给药均可显著降低膝关节面Pelletier评分、关节直径和病理Mankin评分,改善关节腔结构,降低X线评分和关节肿胀,且iPSC-MSCs较hUC-MSCs给药在降低Pelletier评分[(1.22±0.67)比(2.00±0.71),P=0.021]方面差异有统计学意义;相比于模型组,iPSC-MSCs关节腔注射给药可显著增加软骨层CollagenⅡ表达[(35.87±8.52)比(69.90±7.65),P<0.01],降低MMP-13[(59.25±5.56)比(33.75±5.85),P<0.01]和ADAMTS-5[(52.25±10.47)比(22.25±6.13),P<0.01]表达水平,且与hUC-MSCs给药差异有统计学意义,iPSC-MSCs组与hUC-MSCs组CollagenⅡ、MMP-13、ADAMTS-5值分别为[(69.90±7.65)比(61.00±5.52),P=0.010]、[(33.75±5.85)比(43.25±6.02),P=0.028]、[(22.25±6.13)比(36.50±4.65),P=0.011]。结论关节腔注射iPSC-MSCs可显著改善OA大鼠关节病理,具有明显的软骨保护作用。

Objective To elucidate the protective effects of induced pluripotent stem cell-derived mesenchymal stem cells(i PSCMSCs) on osteoarthritis(OA) in rats.Methods The OA model was established by Hulth method and 50 rats were randomized into sham group(control group),model group,i PSC-MSCs group,human umbilical cord-derived mesenchymal stem cells(h UC-MSCs)group,and hyaluronic acid(HA) positive control group,and each group with 10 rats.Rats in the HA group were given multiple dosing(once a week,a total of 5 times),rats in sham and model groups were dealt with 50 μL saline,all the other rats were injected with 50μL drugs in the joint of the surgery side.At the end of 5 thweek,rats were sacrificed by cervical dislocation.The joint diameter difference,joint cavity structure and score,articular surface score,pathological score and other indicators were observed.The expression of matrix metalloproteinase(MMP)-13,type Ⅱ collagen(Collagen Ⅱ) and depolymerized protein-like metalloproteinase(ADAMTS)-5 were detected.Results Administration of i PSC-MSCs and h UC-MSCs markedly reduced the knee joint Pelletier score,joint diameter and pathological Mankin score,improved joint cavity structure,and decreased X-ray score and joint swelling.The Pelletier score in h UCMSCs group and i PSC-MSCs group has statistical difference [(2.00±0.71) vs.(1.22±0.67),P=0.021].Compared with model group,administration with i PSC-MSCs significantly increased the expression of Collagen Ⅱ in the cartilage layer [(35.87±8.52) vs.(69.90±7.65),P<0.01],inhibited the expression of MMP-13 [(59.25±5.56) vs.(33.75±5.85),P<0.01] and ADAMTS-5[(52.25±10.47) vs.(22.25±6.13),P<0.01].Compared with h UC-MSCs group,administration with i PSC-MSCs was more effective in increasing Collagen Ⅱ level [(69.90±7.65) vs.(61.00±5.52),P=0.010 ],and reducing the expression of MMP-13[(33.75±5.85) vs.(43.25±6.02),P=0.028] and ADAMTS-5 [(22.25±6.13) vs.(36.50±4.65),P=0.011].Conclusion i PSC-MSCs could improve the joint pathology of OA rats and may exert the obvious protective effects on cartilage.