Qi Xin, Zhang Jieyuan, Yuan Hong, Xu Zhengliang, Li Qing, Niu Xin, Hu Bin, Wang Yang, Li Xiaolin
1. Department of Orthopaedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
1. Department of Orthopaedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.; 2. Institute of Microsurgery on Extremities, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Int J Biol Sci. 2016 May 25;12(7):836-49. doi: 10.7150/ijbs.14809. eCollection 2016.
Bone defects caused by trauma, severe infection, tumor resection and skeletal abnormalities are common osteoporotic conditions and major challenges in orthopedic surgery, and there is still no effective solution to this problem. Consequently, new treatments are needed to develop regeneration procedures without side effects. Exosomes secreted by mesenchymal stem cells (MSCs) derived from human induced pluripotent stem cells (hiPSCs, hiPSC-MSC-Exos) incorporate the advantages of both MSCs and iPSCs with no immunogenicity. However, there are no reports on the application of hiPSC-MSC-Exos to enhance angiogenesis and osteogenesis under osteoporotic conditions. HiPSC-MSC-Exos were isolated and identified before use. The effect of hiPSC-MSC-Exos on the proliferation and osteogenic differentiation of bone marrow MSCs derived from ovariectomized (OVX) rats (rBMSCs-OVX) in vitro were investigated. In vivo, hiPSC-MSC-Exos were implanted into critical size bone defects in ovariectomized rats, and bone regeneration and angiogenesis were examined by microcomputed tomography (micro-CT), sequential fluorescent labeling analysis, microfil perfusion and histological and immunohistochemical analysis. The results in vitro showed that hiPSC-MSC-Exos enhanced cell proliferation and alkaline phosphatase (ALP) activity, and up-regulated mRNA and protein expression of osteoblast-related genes in rBMSCs-OVX. In vivo experiments revealed that hiPSC-MSC-Exos dramatically stimulated bone regeneration and angiogenesis in critical-sized calvarial defects in ovariectomized rats. The effect of hiPSC-MSC-Exos increased with increasing concentration. In this study, we showed that hiPSC-MSC-Exos effectively stimulate the proliferation and osteogenic differentiation of rBMSCs-OVX, with the effect increasing with increasing exosome concentration. Further analysis demonstrated that the application of hiPSC-MSC-Exos+β-TCP scaffolds promoted bone regeneration in critical-sized calvarial defects by enhancing angiogenesis and osteogenesis in an ovariectomized rat model.
由创伤、严重感染、肿瘤切除和骨骼异常引起的骨缺损是常见的骨质疏松病症，也是骨科手术中的主要挑战，目前对此问题仍没有有效的解决方案。因此，需要开发新的治疗方法来实现无副作用的再生程序。人诱导多能干细胞（hiPSC）来源的间充质干细胞（MSC）分泌的外泌体（hiPSC-MSC-Exos）兼具MSC和iPSC的优点且无免疫原性。然而，尚无关于hiPSC-MSC-Exos在骨质疏松条件下促进血管生成和成骨作用的应用报道。在使用前对hiPSC-MSC-Exos进行了分离和鉴定。研究了hiPSC-MSC-Exos对去卵巢（OVX）大鼠来源的骨髓间充质干细胞（rBMSCs-OVX）体外增殖和成骨分化的影响。在体内，将hiPSC-MSC-Exos植入去卵巢大鼠的临界尺寸骨缺损处，并通过微型计算机断层扫描（micro-CT）、连续荧光标记分析、微丝灌注以及组织学和免疫组织化学分析来检测骨再生和血管生成情况。体外实验结果表明，hiPSC-MSC-Exos可增强rBMSCs-OVX的细胞增殖和碱性磷酸酶（ALP）活性，并上调成骨细胞相关基因的mRNA和蛋白表达。体内实验显示，hiPSC-MSC-Exos可显著促进去卵巢大鼠临界尺寸颅骨缺损处的骨再生和血管生成。hiPSC-MSC-Exos的作用随浓度增加而增强。在本研究中，我们表明hiPSC-MSC-Exos可有效刺激rBMSCs-OVX的增殖和成骨分化，且作用随外泌体浓度增加而增强。进一步分析表明，在去卵巢大鼠模型中，应用hiPSC-MSC-Exos+β-TCP支架通过增强血管生成和成骨作用促进了临界尺寸颅骨缺损处的骨再生。
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