人脐带间充质干细胞联合玻璃酸钠注射液治疗膝骨关节炎

Treatment of knee osteoarthritis with human umbilical cord-derived mesenchymal stem cells combined with sodium hyaluronate injection

背景:膝骨关节炎严重影响患者的正常生活,目前仍无最佳治疗方案。目的:评价人脐带间充质干细胞联合玻璃酸钠注射液治疗膝骨关节炎的有效性和安全性。方法:①细胞学实验观察人脐带间充质干细胞对淋巴细胞增殖和调节性T细胞比例的影响;②油红O染色、阿尔新蓝染色观察预混玻璃酸钠对人脐带间充质干细胞成脂和成软骨的影响;③对新西兰白兔行右膝前交叉韧带切断术和内侧半月板切除术造模,术后第6,9和12周关节内分别注射人脐带间充质干细胞、玻璃酸钠注射液、人脐带间充质干细胞联合玻璃酸钠注射液,术后第16周处死全部新西兰白兔,取心、肝、脾、肺、肾、脑、关节腔周围肌肉和膝关节,用qPCR方法检测人特异基因FOXP2表达;取股骨髁用于形态学、组织学和免疫组化分析。结果与结论:①人脐带间充质干细胞抑制T淋巴细胞增殖和上调调节性T细胞比例;②预混玻璃酸钠的人脐带间充质干细胞依然保持成脂和成软骨的能力;③术后第16周,新西兰白兔心、肝、脾、肺、肾、脑、关节腔周围肌肉和膝关节中人特异基因FOXP2全部为阴性;④人脐带间充质干细胞联合玻璃酸钠组大体观察关节面较平整光滑,ICRS评分较玻璃酸钠组显著升高;苏木精-伊红染色显示软骨厚度增加,Pineda评分明显低于玻璃酸钠组和人脐带间充质干细胞组;软骨全层含较多蛋白聚糖,OARSI评分明显低于玻璃酸钠组和人脐带间充质干细胞组;免疫组化显示关节软骨Ⅱ型胶原表达增加,基质金属蛋白酶13表达减少;⑤结果显示,人脐带间充质干细胞联合玻璃酸钠注射液在关节软骨修复方面优于单纯玻璃酸钠注射液或单纯人脐带间充质干细胞,且无明显不良反应。

BACKGROUND:Knee osteoarthritis seriously affects the living quality of patients and there is still no optimal treatment.OBJECTIVE:To evaluate the efficacy and safety of human umbilical cord-derived mesenchymal stem cells combined with hyaluronic acid on the treatment of knee osteoarthritis.METHODS:(1)Cytological experiments were carried out to observe the effect of human umbilical cord-derived mesenchymal stem cells on lymphocyte proliferation and proportion of regulatory T cells.(2)The effects of premixed sodium hyaluronate on adipogenesis and chondrogenesis of human umbilical cord-derived mesenchymal stem cells were observed by oil red O staining and alcian blue staining.(3)Rabbit osteoarthritis was induced surgically by anterior cruciate ligament transection and medial meniscectomy of right knee joints.The intra-articular injection was given at 6,9 and 12 weeks after surgery of human umbilical cord-derived mesenchymal stem cells,sodium hyaluronate injection,umbilical cord-derived mesenchymal stem cells combined with sodium hyaluronate injection.All rabbits were sacrificed at 16 weeks after surgery.The specific FOXP2 gene was measured in the heart,liver,spleen,lung,kidney,brain,muscles around the joint cavity,and knee joint with qPCR method.The morphological,histological and immunohistochemical analyses were carried out to detect the cartilage repair of femoral condyles.RESULTS AND CONCLUSION:(1)Human umbilical cord-derived mesenchymal stem cells suppressed T lymphocyte proliferation and upregulated proportion of regulatory T cells.(2)Human umbilical cord-derived mesenchymal stem cells premixed with sodium hyaluronate still retained the property of adipogenesis and chondrogenesis.(3)At 16 weeks after treatment,the human specific gene FOXP2 was all negative in the heart,liver,spleen,lung,kidney,brain and muscles around the joint cavity of rabbits.(4)The articular surface of human umbilical cord-derived mesenchymal stem cells+sodium hyaluronate group was flat and smooth,and the ICRS score was significantly higher than that of the sodium hyaluronate group.Hematoxylin-eosin staining results showed that cartilage thickness was increased and the Pineda score of human umbilical cord-derived mesenchymal stem cells+sodium hyaluronate group was significantly lower than that of the sodium hyaluronate group and the human umbilical cord-derived mesenchymal stem cells group.The whole cartilage layer of human umbilical cord-derived mesenchymal stem cells+sodium hyaluronate group contained more proteoglycans.The OARSI score of human umbilical cord-derived mesenchymal stem cells+sodium hyaluronate group was significantly lower than that of the sodium hyaluronate group and the human umbilical cordderived mesenchymal stem cells group.Immunohistochemical analysis showed that the expression of type II collagen in articular cartilage was increased and the expression of matrix metalloproteinase-13 was decreased.(5)The results indicate that human umbilical cord-derived mesenchymal stem cells combined with sodium hyaluronate treatment were superior to either sodium hyaluronate treatment alone or human umbilical cord-derived mesenchymal stem cells treatment alone in articular cartilage repair and there was no significant adverse reaction.