Exosomes secreted by endothelial progenitor cells accelerate bone regeneration during distraction osteogenesis by stimulating angiogenesis
Exosomes secreted by endothelial progenitor cells accelerate bone regeneration during distraction osteogenesis by stimulating angiogenesis
复制标题
内皮祖细胞分泌的外泌体通过刺激血管生成加速牵引成骨过程中的骨再生
DOI：
10.1186/s13287-018-1115-7
复制
发表时间：
2019-01-11
影响因子：
7.5
通讯作者：
Chai, Yimin
中科院分区：
文献类型：
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作者：
Jia, Yachao;Zhu, Yu;Chai, Yimin
背景牵引成骨（DO）是一种有效但漫长的手术，可以完全诱导大面积骨缺损的骨再生。越来越多的证据支持内皮祖细胞分泌的外泌体（EPC-Exos）在刺激血管生成中的作用，这与骨生成密切相关。本研究旨在探讨EPC-Exos是否促进骨再生过程中DO在rats.MethodsExosomes大鼠骨髓EPCs通过ultracentrugation和其特征在于，通过透射电子显微镜，可调电阻脉冲传感分析，和western印迹分析的上清液分离。采用68只SD大鼠，以0.5mm/d的牵张速率，连续牵张10 d，建立单侧胫骨DO模型。将EPC-Exos局部注射到牵张间隙后，通过X线、显微CT、生物力学和组织学分析评估EPC-Exos对骨再生和血管生成的治疗作用。促血管生成的作用和潜在的机制EPC-Exos对人脐静脉内皮细胞的影响，随后通过体外试验，包括细胞计数试剂盒-8，伤口愈合，管形成，和western blot assays.ResultsEPC-Exos是球形或杯状囊泡，直径范围从50至150 nm，并表示标记物，包括CD 9，阿利克斯，和TSG 101进行了评估。X射线、显微CT和组织学分析显示，用EPC-Exos处理的大鼠的骨再生显著加速。Exos组的牵引胫骨也显示出增强的机械性能。此外，Exos组的血管密度高于对照组。此外，体外分析显示EPC-Exos以miR-126依赖性方式增强内皮细胞的增殖、迁移和血管生成能力。此外，EPC-Exos下调SPRED 1和激活Raf/ERK signaling.ConclusionsThe目前的结果表明，EPC-Exos加速骨再生DO通过刺激血管生成，这表明它们作为一种新的方法，以缩短治疗持续时间的DO。
BackgroundDistraction osteogenesis (DO) is an effective but lengthy procedure to fully induce bone regeneration in large bone defects. Accumulating evidence supports the role of exosomes secreted by endothelial progenitor cells (EPC-Exos) in stimulating angiogenesis, which is closely coupled with osteogenesis. This study aimed to investigate whether EPC-Exos promote bone regeneration during DO in rats.MethodsExosomes were isolated from the supernatants of rat bone marrow EPCs via ultracentrifugation and characterized via transmission electron microscopy, tunable resistive pulse sensing analysis, and western blot analysis. Unilateral tibial DO models were generated using 68 Sprague-Dawley rats with a distraction rate of 0.5mm per day for 10days. After local injection of EPC-Exos into the distraction gaps after distraction, the therapeutic effects of EPC-Exos on bone regeneration and angiogenesis were assessed via X-ray, micro-computed tomography(micro-CT), and biomechanical and histological analyses. Pro-angiogenic effects and the potential mechanism underlying the effects of EPC-Exos on human umbilical vein endothelial cells were subsequently evaluated via in vitro assays including Cell Counting Kit-8, wound healing, tube formation, and western blot assays.ResultsEPC-Exos were spherical or cup-shaped vesicles ranging from 50 to 150nm in diameter and expressed markers including CD9, Alix, and TSG101. X-ray, micro-CT, and histological analyses revealed that bone regeneration was markedly accelerated in rats treated with EPC-Exos. The distracted tibias from the Exos group also displayed enhanced mechanical properties. Moreover, vessel density was higher in the Exos group than in the control group. In addition, in vitro analyses revealed that EPC-Exos enhanced the proliferation, migration, and angiogenic capacity of endothelial cells in an miR-126-dependent manner. Further, EPC-Exos downregulated SPRED1 and activated Raf/ERK signaling.ConclusionsThe present results show that EPC-Exos accelerate bone regeneration during DO by stimulating angiogenesis, suggesting their use as a novel method to shorten the treatment duration of DO.