背景:关节软骨的自我修复能力有限,和目前临床治疗软骨缺损的选择是不够的。然而,鹿茸软骨具有独特的再生特性,具有快速修复自身的能力。这种快速的自我修复过程与鹿茸干细胞释放的旁分泌因子密切相关。这些发现为在临床环境中开发软骨缺损的无细胞疗法提供了潜力。本研究的目的是研究一种修复软骨的新方法。
方法:通过手术建立大鼠关节软骨缺损模型。将负载有源自鹿茸干细胞(ASC-Exos)的外泌体(Exos)的水凝胶植入大鼠软骨缺损中。使用组织学方法评估软骨损伤修复的程度。ASC-Exos对软骨细胞和大鼠骨髓间充质干细胞(BMSCs)的影响,增殖试验,和划痕试验。此外,使用实时荧光定量PCR(qPCR)和蛋白质印迹分析评估ASC-Exos维持软骨细胞表型的情况.使用数据无关采集(DIA)质谱法鉴定Exos所含的蛋白质组分。
结果:ASC-Exos显著促进软骨组织损伤的修复。实验组(ASC-Exos)的软骨修复水平高于阳性对照(人脂肪源性干细胞,hADSC-Exos)和阴性对照(dulbecco改良的鹰培养基)组(p<0.05)。体外实验表明,ASC-Exos能显著增强软骨细胞的增殖能力和骨髓间充质干细胞的增殖能力和迁移能力(p<0.05)。ASC-Exos上调Aggrecan的表达水平,胶原蛋白II(COLII),软骨细胞中的Sox9mRNA和蛋白质。对ASC-Exos蛋白成分的分析揭示了活性成分的存在,例如血清转铁蛋白(TF),S100A4和胰岛素样生长因子结合蛋白1(IGF1)。
结论:ASC-Exos对软骨损伤修复有显著作用,这可能归因于它们促进软骨细胞和BMSCs的增殖和迁移,以及软骨细胞表型的维持。这种效应可能是由TF的存在介导的,S100A4和IGF1。
BACKGROUND: Articular cartilage has limited self-repair capacity, and current clinical treatment options for cartilage defects are inadequate. However, deer antler cartilage possesses unique regenerative properties, with the ability to rapidly repair itself. This rapid self-repair process is closely linked to the paracrine factors released by deer antler stem cells. These findings present potential for the development of cell-free therapies for cartilage defects in clinical settings. The aim of this study was to investigate a novel method for repairing cartilage.
METHODS: A rat model with articular cartilage defects was established through surgery. Hydrogels loaded with exosomes (Exos) derived from antler stem cells (ASC-Exos) were implanted into the rat cartilage defects. The extent of cartilage damage repair was assessed using histological methods. The effects of ASC-Exos on chondrocytes and rat bone marrow mesenchymal stem cells (BMSCs) were evaluated using cell viability assays, proliferation assays, and scratch assays. Additionally, the maintenance of the chondrocyte phenotype by ASC-Exos was assessed using real-time fluorescence quantitative PCR (qPCR) and western blot analysis. The protein components contained of the Exos were identified using data-independent acquisition (DIA) mass spectrometry.
RESULTS: ASC-Exos significantly promoted the repair of cartilage tissue damage. The level of cartilage repair in the experimental group (ASC-Exos) was higher than that in the positive control (human adipose-derived stem cells, hADSC-Exos) and negative control (dulbecco\'s modified eagle medium) groups (p < 0.05). In vitro experiments demonstrated that ASC-Exos significantly enhanced the proliferation abilities of chondrocytes and the proliferation abilities and the migration abilities of BMSCs (p < 0.05). ASC-Exos up-regulated the expression levels of Aggrecan, Collagen II (COLII), and Sox9 mRNA and proteins in chondrocytes. Analysis of ASC-Exos protein components revealed the presence of active components such as Serotransferrin (TF), S100A4, and Insulin-like growth factor-binding protein 1 (IGF1).
CONCLUSIONS: ASC-Exos have a significant effect on cartilage damage repair, which may be attributed to their promotion of chondrocyte and BMSCs proliferation and migration, as well as the maintenance of chondrocyte phenotype. This effect may be mediated by the presence of TF, S100A4, and IGF1.