BACKGROUND In athlete horses, suspensory ligament (SL) injuries are the most common cause of lameness. Healing of SL injury is still problematic, and even proper rehabilitation and pharmacological therapy do not guarantee returning to the initial performance level. In our previous studies, we have shown that a combination of 5-azacytidine (AZA) and resveratrol (RES) exerts beneficial, rejuvenating effects on metabolic syndrome derived adipose-derived stem cells (ASCs). Thus, in the presented research, we investigate whether not only rejuvenated ASC but also microvesicles (MVsAZA/RES) secreted by them possess enhanced regenerative properties in SL injury. METHODS In the presented study, a 6-year-old Dutch Warmblood gelding, working in jumping, was diagnosed with SL injury using ultrasonography, Doppler, real-time elastography and thermography. As a therapeutic strategy, the affected animal was treated with extracellular microvesicles derived from ASC treated with the combination of 5-azacytydine (AZA) and resveratrol (RES) (MVsAZA/RES). RESULTS: First, anti-apoptotic effects of MVsAZA/RES were tested in co-culture with metabolic syndrome derived ASC. The proliferation of cells and expression of pro-apoptotic genes were investigated. Then, MVsAZA/RES were injected directly into the injured SL of the Dutch Warmblood gelding. In vitro assays revealed that MVsAZA/RES enhance the proliferation of ASC and exert an anti-apoptotic effect. In the affected horse, the application of MVsAZA/RES resulted in increased lesion filling and improvement of angiogenesis and elasticity in injured tissue. CONCLUSIONS As MVsAZA/RES mimic several of the biological actions exerted by ASC, they have become an alternative for stem cell-based therapies and can be effectively applied for the treatment of SL injury in horses.

中文翻译：

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从 5-氮杂胞苷和白藜芦醇处理的间充质干细胞中分离出的微泡用于治疗马悬韧带损伤 - 病例报告。

背景技术对于运动员马来说，悬韧带（SL）损伤是跛行的最常见原因。SL损伤的治愈仍然存在问题，即使是适当的康复和药物治疗也不能保证恢复到最初的表现水平。在我们之前的研究中，我们已经证明 5-氮杂胞苷 (AZA) 和白藜芦醇 (RES) 的组合对代谢综合征衍生的脂肪干细胞 (ASC) 发挥有益的恢复活力作用。因此，在本研究中，我们不仅研究了再生的 ASC，而且还研究了它们分泌的微泡 (MVsAZA/RES) 在 SL 损伤中是否具有增强的再生特性。方法 在本研究中，一匹从事跳跃工作的 6 岁荷兰温血骟马通过超声检查、多普勒、实时弹性成像和热成像被诊断为 SL 损伤。作为一种治疗策略，受影响的动物接受了源自 ASC 的细胞外微泡治疗，该微泡经过 5-氮杂胞苷 (AZA) 和白藜芦醇 (RES) (MVsAZA/RES) 的组合处理。结果：首先，在与代谢综合征衍生的 ASC 共培养中测试了 MVsAZA/RES 的抗凋亡作用。研究了细胞增殖和促凋亡基因的表达。然后，将 MVsAZA/RES 直接注射到荷兰温血骟马受伤的 SL 中。体外实验表明，MVsAZA/RES 可增强 ASC 的增殖并发挥抗凋亡作用。在受影响的马中，MVsAZA/RES 的应用导致损伤组织的病变充盈增加，血管生成和弹性得到改善。结论 由于 MVsAZA/RES 模仿 ASC 发挥的多种生物学作用，因此它们已成为基于干细胞的疗法的替代方案，并且可以有效地应用于马 SL 损伤的治疗。