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突变型缺氧诱导因子-1α修饰的骨髓间充质干细胞分泌的外泌体可减轻兔早期类固醇诱导的股骨头缺血性坏死。

Exosomes secreted from mutant-HIF-1α-modified bone-marrow-derived mesenchymal stem cells attenuate early steroid-induced avascular necrosis of femoral head in rabbit.

作者信息

Li Haile, Liu Danping, Li Chen, Zhou Shanjian, Tian Dachuan, Xiao Dawei, Zhang Huan, Gao Feng, Huang Jianhua

机构信息

Department of Orthopedics, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121001, P.R. China.

Biobank, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121001, P.R. China.

出版信息

Cell Biol Int. 2017 Dec;41(12):1379-1390. doi: 10.1002/cbin.10869. Epub 2017 Sep 25.

Abstract

Mesenchymal stem cells (MSCs)-derived exosomes exhibit protective effects on damaged or diseased tissues. Hypoxia-inducible factor 1α (HIF-1α) plays a critical role in bone development. However, HIF-1α is easily biodegradable under normoxic conditions. The bone-marrow-derived mesenchymal stem cells (BMSCs) were transfected with adenovirus carrying triple point-mutations (amino acids 402, 564, and 803) in the HIF-1α coding sequence (CDS). The mutant HIF-1α can efficiently express functional proteins under normoxic conditions. To date, no study has reported the role of exosomes secreted by mutant HIF-1α modified BMSCs in the recovery of the early steroid-induced avascular necrosis of femoral head (SANFH). In this study, we firstly analyzed exosomes derived from BMSCs modified by mutant (BMSC-Exos ) or wild-type HIF-1α (BMSC-Exos ). In vitro, we investigated the osteogenic differentiation capacity of BMSCs modified by BMSC-Exos or BMSC-Exos , and the angiogenesis effects of BMSC-Exos and BMSC-Exos on human umbilical vein endothelial cells (HUVECs). Besides, the healing of the femoral head was also assessed in vivo. We found that the potential of osteogenic differentiation of BMSCs treated with BMSC-Exos was higher than the wild-type group in vitro. In addition, BMSC-Exos stimulated the proliferation, migration, and tube formation of HUVECs in a dose-dependent manner. Compared with the BMSC-Exos or PBS control group, the injection of BMSC-Exos into the necrosis region markedly accelerated the bone regeneration and angiogenesis, which were indicated by the increased trabecular reconstruction and microvascular density. Taken together, our data suggest that BMSC-Exos facilitates the repair of SANFH by enhancing osteogenesis and angiogenesis.

摘要

间充质干细胞(MSCs)衍生的外泌体对受损或患病组织具有保护作用。缺氧诱导因子1α(HIF-1α)在骨骼发育中起关键作用。然而,HIF-1α在常氧条件下易于被生物降解。用携带HIF-1α编码序列(CDS)中三点突变(氨基酸402、564和803)的腺病毒转染骨髓间充质干细胞(BMSCs)。突变型HIF-1α能够在常氧条件下高效表达功能性蛋白质。迄今为止,尚无研究报道突变型HIF-1α修饰的BMSCs分泌的外泌体在早期类固醇诱导的股骨头缺血性坏死(SANFH)恢复中的作用。在本研究中,我们首先分析了由突变型(BMSC-Exos )或野生型HIF-1α(BMSC-Exos )修饰的BMSCs衍生的外泌体。在体外,我们研究了BMSC-Exos或BMSC-Exos 修饰的BMSCs的成骨分化能力,以及BMSC-Exos和BMSC-Exos 对人脐静脉内皮细胞(HUVECs)的血管生成作用。此外,还在体内评估了股骨头的愈合情况。我们发现,体外经BMSC-Exos 处理的BMSCs的成骨分化潜能高于野生型组。此外,BMSC-Exos 以剂量依赖性方式刺激HUVECs的增殖、迁移和管腔形成。与BMSC-Exos 或PBS对照组相比,将BMSC-Exos 注射到坏死区域显著加速了骨再生和血管生成,这通过小梁重建增加和微血管密度得以体现。综上所述,我们的数据表明BMSC-Exos 通过增强成骨作用和血管生成促进SANFH的修复。

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