Experimental study of IGF1 gene lentivirus transfected bone marrow mesenchyml stem cells in the treatment of osteoarthritis
XIANG Wenyuan1 DENG Yingjie2 LIAO Jun2 FANG Rui2
1.The Fourth Clinical Medical College, Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi 830000, China;
2.Department of Orthopedics, Xinjiang Medical University Affiliated Hospital of Traditional Chinese Medicine, Xinjiang Uygur Autonomous Region, Urumqi 830000, China
Abstract：Objective To investigate the effect of IGF1 gene lentivirus transfected bone marrow mesenchyml stem cells (BMSCs) in the treatment of osteoarthritis (OA) based on animal experiments. Methods A total of 40 SPF male Wistar rats (10-12 week old, weighing 280-320 g) were divided into BMSCs+IGF1 group, BMSCs group and model group according to the random number table method, with ten rats in each group. The OA model was established by anterior cruciate ligament resection of knee joint, and the other ten rats were used for BMSCs isolation. The 0.1 ml IGF1 gene lentivirus transfected BMSCs (1×107 cells) were injected into knee joint cavity in rats of BMSCs+IGF1group. The 0.1 ml BMSCs (1×107 cells) were injected into knee joint cavity in rats of BMSCs group. The 0.1 ml normal saline were injected into knee joint cavity in rats of model group. The serum levels of C-terminal peptide of type Ⅰ collagen (CTX-Ⅰ), CTX-Ⅱ, interleukin (IL)-1α, and IL-6 in rats were determined by enzyme-linked immunosorbent assay. The mRNA expression levels of osteocalcin (OC), glycogen synthase kinase-3β (GSK-3β), alkaline phosphatase (ALP), and Runt-associated transcription factor 2 (Runx2) in the knee cartilage tissues of rats were detected by real-time quantitative polymerase chain reaction. The protein expression levels of β-catenin,Wnt1, and GSK-3β in the knee cartilage tissues of rats were detected by Western blot. Results BMSCs surface antigens CD29,CD34,CD45, and CD105 were highly expressed, and the transfection efficiency of lentivirus was (90.36±2.18)%. The knee cartilage of rats was intact without obvious degeneration and the articular surface was flat in BMSCs+IGF1 group. The other two groups had varying degrees of cartilage degeneration and uneven articular surface. Compared with the model group, the serum levels of CTX-Ⅰ, CTX-Ⅱ, IL-1α, and IL-6, the mRNA expression levels of OC, GSK-3β, ALP, and Runx2, the protein expression levels of β-catenin, Wnt1, and GSK-3β were decreased in the BMSCs group, the differences were statistically significant (P ＜ 0.05). Compared with the BMSCs group, the above indexes in the BMSCs+IGF1 group were decreased, the differences were statistically significant (P ＜ 0.05). Conclusion IGF1 gene lentivirus transfected BMSCs can effectively improve the pathological injury of OA rats, and significantly promote the repair of knee joint injury.
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