氧化石墨烯纳米支架培养脐源性MSC调节关节微环境对软骨的修复作用

Experimental Study on the Regulation of Microenvironment of Joint by Umbilical MSC Cultured on 3D Graphene Oxide Nano Scaffold for Cartilage Repair Cells

目的:研究氧化石墨烯(GO)载脐带间充质干细胞(UCMSC)对两种膝骨性关节炎(KOA)动物模型的软骨修复作用。方法:选择12周雄性新西兰兔64只,随机分为通过改良Hulth+软骨缺损造模组和通过改良木瓜蛋白酶控释剂注射方法造模组,每组造模后分为四组:空白对照组、GO治疗组(GO组)、UCMSC治疗组(UCMSC组)、GO载UCMSC治疗组(GO+UCMSC组),通过ELISA法检测血清一氧化氮(NO)、白细胞介素-6(IL-6)、肿瘤坏死因子(TNF)-α、糖胺聚糖(GAG)和Ⅱ型胶原蛋白(COL-Ⅱ)的含量,并对其股骨髁进行切片,HE染色,显微镜下观察。结果:改良Hulth+软骨缺损组和改良木瓜蛋白酶控释剂关节注射组中GO+UCMSC组血清NO、COL-Ⅱ、GAG、IL-6、TNF-α与空白组相比差异有统计学意义(P<0.01);两组间NO与空白组差异有统计学意义(P<0.05);两组间的COL-Ⅱ比较,GO+UCMSC组、UCMSC组差异有统计学意义(P<0.01);两组间的GAG比较,空白组、GO+UCMSC组差异有统计学意义(P<0.01);两组间的IL-6比较,GO+UCMSC组、空白组差异均具有统计学意义(P<0.05),基于股骨髁HE染色病理学观察发现两组之间存在一定的差异,UCMSC+GO、UCMSC、GO、空白组的软骨修复效果呈梯度降低。结论:氧化石墨烯载UCMSC能促进两种KOA动物模型软骨细胞分泌,降低关节内炎性水平,起到软骨修复作用。

Objective To study the effects of umbilical cord mesenchymal stem cells(MSCS)loaded with GO on cartilage repair in two KOA animal models.Methods 64 male New Zealand rabbits of 12 weeks were selected and randomly divided into modified Hulth and cartilage defect model,which were modified papain controlled-release injection model.After the building,theywere divided into four groups:blank control group,GO treatment group(GO),UCMSC therapy group(UCMSC)and GO particles carrying UCMSC treatment group(GO+UCMSC).They were detected the serum IL-6,TNF-α,NO,GAG and COL-II by ELISA method.The femoral condyles were sliced,HE stained and they were observed under a microscope.Results The serum levels of NO,COL-II,GAG,IL-6 and TNF-αin the GO+UCMSC group were significantly different from those in the blank group(P<0.01).NO comparison between the two groups was statistically significant(P<0.05).Comparison of COL-II between the two groups showed that GO+UCMSC group and UCMSC group have significant differences(P<0.01).The comparison of GAG between the two groups showed that the blank group and the GO+UCMSC group have statistically significant differences(P<0.01).The comparison of IL-6 between the two groups showed that GO+UCMSCwas significant difference(P<0.01),while the difference between the blank group was significant(P<0.05).Based on the pathological observation of the femoral condyle HE staining,there were some differences between group A and B,but at the same time,they all had gradient of the repair effects in UCMSC+GO,UCMSC,GO and the blank group.Conclusion GO loaded UCMSC can promote the secretion of chondrocytes in two KOA animal models,which can reduce inflammatory levels in joints and play a role in cartilage repair.