Sun Yaying, Luo Zhiwen, Chen Yisheng, Lin Jinrong, Zhang Yuhan, Qi Beijie, Chen Jiwu
Department of Sports Medicine, Huashan Hospital, Fudan University, Shanghai, China.
Department of Sports Medicine, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, China.
Biomater Res. 2022 Aug 19;26(1):39. doi: 10.1186/s40824-022-00286-2.
Adhesive capsulitis is a common shoulder disorder inducing joint capsule fibrosis and pain. When combined with rotator cuff tear (RCT), treatments can be more complex. Currently, targeted therapy is lacking. Since adhesive capsulitis is reported to be related to circulating materials, we analyzed the contents and biology of circulating exosomes from RCT patients with and without adhesive capsulitis, in an attempt to developing a targeting treatment.
Samples from a consecutive cohort of patients with RCT for surgery were collected. Circulating exosomal miRNAs sequencing were used to detect differentially expressed miRNAs in patients with and without adhesive capsulitis. For experiments in vitro, Brdu staining, CCK-8 assay, wound healing test, collagen contraction test, real-time quantitative polymerase chain reaction, and western blot were conducted. Histological and immunofluorescent staining, and biomechanical analysis were applied in a mouse model of shoulder stiffness. The characteristics of liposomes loaded with siRNA were measured via dynamic light scattering or electron microscopy.
Circulating exosomal miRNAs sequencing showed that, compared to exosomes from patients without adhesive capsulitis, miR-142 was significantly up-regulated in exosomes from adhesive capsulitis (Exo-S). Both Exo-S and miR-142 could inhibit fibrogenesis, and the anti-fibrotic effect of Exo-S relied on miR-142. The target of miR-142 was proven to be transforming growth factor β receptor 1 (Tgfbr1). Then, liposomes were developed and loaded with si-Tgfbr1. The si-Tgfbr1-loading liposomes exhibited promising therapeutic effect against shoulder stiffness in mouse model with no evidence toxicity.
This study showed that, in RCT patients with adhesive capsulitis, circulating exosomes are protective and have anti-fibrotic potential. This effect is related to the contained miR-142, which targets Tgfbr1. By mimicking this biological function, liposomes loaded with si-Tgfbr1 can mitigate shoulder stiffness pre-clinically.
粘连性关节囊炎是一种常见的肩部疾病，可导致关节囊纤维化和疼痛。当与肩袖撕裂（RCT）合并时，治疗可能会更加复杂。目前，缺乏靶向治疗方法。由于据报道粘连性关节囊炎与循环物质有关，我们分析了患有和未患有粘连性关节囊炎的RCT患者循环外泌体的成分和生物学特性，试图开发一种靶向治疗方法。
收集连续队列中接受手术的RCT患者的样本。采用循环外泌体miRNA测序检测患有和未患有粘连性关节囊炎患者中差异表达的miRNA。在体外实验中，进行了Brdu染色、CCK-8检测、伤口愈合试验、胶原收缩试验、实时定量聚合酶链反应和蛋白质印迹分析。在肩部僵硬的小鼠模型中进行了组织学和免疫荧光染色以及生物力学分析。通过动态光散射或电子显微镜测量负载siRNA的脂质体的特性。
循环外泌体miRNA测序显示，与未患有粘连性关节囊炎患者的外泌体相比，粘连性关节囊炎患者（Exo-S）的外泌体中miR-142显著上调。Exo-S和miR-142均能抑制纤维化，Exo-S的抗纤维化作用依赖于miR-142。已证实miR-142的靶标是转化生长因子β受体1（Tgfbr1）。然后，开发了负载si-Tgfbr1的脂质体。负载si-Tgfbr1的脂质体在小鼠模型中对肩部僵硬表现出有前景的治疗效果，且无毒性证据。
本研究表明，在患有粘连性关节囊炎的RCT患者中，循环外泌体具有保护作用且具有抗纤维化潜力。这种作用与所含的靶向Tgfbr1的miR-142有关。通过模拟这种生物学功能，负载si-Tgfbr1的脂质体可在临床前减轻肩部僵硬。
