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| Effect of Ckip-1 siRNA promotes osteogenic differentiation of bone marrow mesenchymal stem cells under extreme hypoxia |
| LIU Xiangzhong1,2 NING Yu3 HU Jing4 YANG Aofei5 CAI Hantao6 LI Zhanghua2 |
1.Department of Orthopaedics, Zhongnan Hospital of Wuhan University, Hubei Province, Wuhan 430000, China;
2.Department of Orthopaedics, Tongren Hospital of Wuhan Universyity, Hubei Province, Wuhan 430000, China;
3.Department of Orthopaedics, Xiangyang Hospital of Traditional Chinese Medicine, Hubei Province, Xiangyang 441000, China;
4.Graduate School, Wuhan Sports University, Hubei Province, Wuhan 430000, China;
5.Department of Orthopaedics, Hubei Provincial Hospital of Traditional Chinese Medicine, Hubei Province, Wuhan 430000, China;
6.the First Clinical School, Hubei University of Traditional Chinese Medicine, Hubei Province, Wuhan 430000, China |
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Abstract Objective To investigate the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by Ckip-1 siRNA under extreme hypoxia. Methods BMSCs were isolated and cultured in normal oxygen (20%) and extreme hypoxia (1%) for 72 h. The expression of osteogenic gene Runt-related transcription factor 2 (Runx2), Smad5 and bone morphogenetic protein 2 (BMP2) were detected by real-time quantitative PCR (qPCR) and Western blot. Alkaline phosphatase (ALP) activity in the supernatant was detected after 3, 5, 7 days of culture. BMSCs were cultured in 1% oxygen for 72 h to observe the effect of Ckip-1 siRNA on the expression of Runx2, Smad5, BMP2 and the activity of ALP. Results The osteogenic gene expression and ALP activity of BMSCs in 1% oxygen concentration were significantly lower than those in 20% oxygen concentration group, and the differences were statistically significant (P ＜ 0.05); the osteogenic gene expression and ALP activity of BMSCs after transfection with Ckip-1 siRNA were significantly higher than those without transfection, and the differences were statistically significant (P ＜ 0.05). Conclusion Hypoxia can inhibit the osteogenic differentiation of BMSCs, and Ckip-1 siRNA can promote the differentiation of BMSCs into osteoblasts.
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