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Bone Marrow Stromal Cell-Derived Exosomes Promote Muscle Healing Following Contusion Through Macrophage Polarization
Stem Cells and Development ( IF 4 ) Pub Date : 2021-01-27 , DOI: 10.1089/scd.2020.0167
Zhiwen Luo 1 , Jinrong Lin 1 , Yaying Sun 1 , Chenghui Wang 1 , Jiwu Chen 1
Affiliation  

Skeletal muscle contusion is among the most common injuries in traumatology and clinics of sports medicine. The injured muscle is vulnerable to re-injury owing to fibrosis formation. Given that the bone marrow stromal cell-derived exosomes (BMSC-Exos) displayed promising therapeutic effect for various tissues, we used BMSC-Exos to treat skeletal muscle contusion and investigated its effects on muscle healing. In this study, the in vivo model of skeletal muscle contusion was established by subjecting the tibialis anterior of young male mice to hit injury, and the in vitro inflammation model was established by lipopolysaccharide treatment on macrophages. Macrophage depletion model was built by intraperitoneal injection with clodronate-containing liposomes. Exosomes were isolated and purified from the supernatant of BMSCs using gradient centrifugation. Nanoparticle tracking analysis, transmission electron microscope, and western blot were used to identify the exosomes. HE stain, Masson stain, immunofluorescence, and biomechanical testing were carried out on the muscle tissue. In addition, enzyme-linked immunosorbent assay (ELISA) assays, real-time qPCR, flow cytometry, and PKH67 fluorescence trace were conducted in vitro. Intramuscular injection of BMSC-Exos to mice after muscle contusion alleviated inflammation level, reduced fibrosis size, promoted muscle regeneration, and improved biomechanical property. After macrophages depletion, the effects of BMSC-Exos were inhibited. In vitro, PKH-67 fluorescence was internalized into macrophages. BMSC-Exos promoted M2 macrophages polarization both in vivo and in vitro. At the same time, BMSC-Exos reduced the production of inflammatory cytokines under the inflammatory microenvironment and upregulated anti-inflammatory factors expression. In conclusion, BMSC-Exos attenuated muscle contusion injury and promoted muscle healing in mice by modifying the polarization status of macrophages and suppressing the inflammatory reaction.

中文翻译:

骨髓基质细胞衍生的外泌体通过巨噬细胞极化促进挫伤后的肌肉愈合

骨骼肌挫伤是创伤学和运动医学临床中最常见的损伤之一。由于纤维化的形成,受伤的肌肉很容易再次受伤。鉴于骨髓基质细胞衍生的外泌体(BMSC-Exos)对各种组织显示出良好的治疗效果,我们使用 BMSC-Exos 治疗骨骼肌挫伤并研究其对肌肉愈合的影响。本研究通过对年轻雄性小鼠胫骨前肌进行撞击损伤建立骨骼肌挫伤的体内模型,并通过脂多糖处理巨噬细胞建立体外炎症模型。通过腹腔注射含氯膦酸盐的脂质体建立巨噬细胞耗竭模型。使用梯度离心从 BMSCs 的上清液中分离和纯化外泌体。纳米粒子追踪分析、透射电子显微镜和蛋白质印迹用于鉴定外泌体。对肌肉组织进行HE染色、Masson染色、免疫荧光和生物力学测试。此外,还进行了酶联免疫吸附测定 (ELISA) 测定、实时 qPCR、流式细胞术和 PKH67 荧光跟踪。肌肉挫伤后向小鼠肌肉注射 BMSC-Exos 可减轻炎症水平,减少纤维化大小,促进肌肉再生,并改善生物力学性能。巨噬细胞耗尽后,BMSC-Exos 的作用被抑制。在体外,PKH-67 荧光被内化到巨噬细胞中。BMSC-Exos 在体内和体外均促进 M2 巨噬细胞极化。同时,BMSC-Exos减少炎症微环境下炎症细胞因子的产生,上调抗炎因子的表达。总之,BMSC-Exos 通过改变巨噬细胞的极化状态和抑制炎症反应来减轻小鼠的肌肉挫伤损伤并促进肌肉愈合。
更新日期:2021-01-28
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