Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Russian Federation.
Faculty of Medicine and Health Sciences, School of Veterinary Medicine and Science, University of Nottingham, Nottingham, UK.
Cells Tissues Organs. 2024;213(1):24-39. doi: 10.1159/000526845. Epub 2022 Sep 1.
Tendons have a limited capacity to repair both naturally and following clinical interventions. Damaged tissue often presents with structural and functional differences, adversely affecting animal performance, mobility, health, and welfare. Advances in cell therapies have started to overcome some of these issues, however complications such as the formation of ectopic bone remain a complication of this technique. Regenerative medicine is therefore looking toward future therapies such as the introduction of microvesicles (MVs) derived from stem cells (SCs). The aim of the present study was to assess the characteristics of artificially derived MVs, from equine mesenchymal stem cells (MSCs), when delivered to rat tendon cells in vitro and damaged tendons in vivo. The initial stages of extracting MVs from equine MSCs and identifying and characterizing the cultured tendon stem/progenitor cells (TSCs) from rat Achilles tendons were undertaken successfully. The horse MSCs and the rat tendon cells were both capable of differentiating in 3 directions: adipogenic, osteogenic, and chondrogenic pathways. The artificially derived equine MVs successfully fused with the TSC membranes, and no cytotoxic or cytostimulating effects were observed. In addition, co-cultivation of TSCs with MVs led to stimulation of cell proliferation and migration, and cytokine VEGF and fractalkine expression levels were significantly increased. These experiments are the first to show that artificially derived MVs exhibited regeneration-stimulating effects in vitro, and that fusion of cytoplasmic membranes from diploid cell lines originating from different species was possible. The experiment in vivo demonstrated the influence of MVs on synthesis of collagen I and III types in damaged tendons of rats. Explorations in vivo showed accelerated regeneration of injured tendons after introduction of the MVs into damaged areas. The results from the studies performed indicated obvious positive modifying effects following the administration of MVs. This represents the initial successful step required prior to translating this regenerative medicine technique into clinical trials, such as for tendon repair in injured horses.
肌腱在自然和临床干预后都有有限的修复能力。受损组织通常表现出结构和功能上的差异，这会对动物的性能、运动能力、健康和福利产生不利影响。细胞疗法的进步已经开始克服其中的一些问题，但是，形成异位骨等并发症仍然是该技术的一个问题。因此，再生医学正在寻求未来的治疗方法，例如引入源自干细胞的微泡 (MVs)。本研究的目的是评估从马间充质干细胞 (MSCs) 人工衍生的 MV 在体外递送至大鼠肌腱细胞和体内损伤的肌腱中的特征。成功地进行了从马 MSC 中提取 MVs 的初始阶段，并鉴定和表征了来自大鼠跟腱的培养肌腱干细胞/祖细胞 (TSC)。马 MSC 和大鼠肌腱细胞都能够向 3 个方向分化：成脂、成骨和软骨形成途径。人工衍生的马 MVs 成功地与 TSC 膜融合，并且没有观察到细胞毒性或细胞刺激作用。此外，TSC 与 MVs 的共培养导致细胞增殖和迁移的刺激，并且细胞因子 VEGF 和 fractalkine 的表达水平显著增加。这些实验是首次表明人工衍生的 MV 在体外表现出再生刺激作用，并且来自不同物种的二倍体细胞系的细胞质膜融合是可能的。体内实验表明，MV 对大鼠损伤肌腱中 I 型和 III 型胶原合成的影响。体内探索表明，在损伤区域引入 MVs 后，受伤肌腱的再生得到加速。所进行的研究结果表明，MV 的给药具有明显的积极修饰作用。这代表了将这种再生医学技术转化为临床试验（例如受伤马的肌腱修复）之前所需的初始成功步骤。
