METHODS: PRP-Exo was isolated, and its impact on MSC viability was evaluated. The effects of PRP-Exo-treated MSCs (MSCPExo) on Schwann cells (SCs) from injured sciatic nerves and human umbilical vein endothelial cells (HUVECs) were assessed. Furthermore, the conditioned medium from MSCPExo (MSCPExo-CM) was analyzed using a cytokine array and validated through ELISA and Western blot.
RESULTS: PRP-Exo enhanced MSC viability. Coculturing MSCPExo with SCs ameliorated apoptosis and promoted SC proliferation following PNI. Similarly, MSCPExo-CM exhibited pro-proliferative, migratory, and angiogenic effects. Cytokine array analysis identified 440 proteins in the MSCPExo secretome, with 155 showing upregulation and 6 showing downregulation, many demonstrating potent pro-regenerative properties. ELISA confirmed the enrichment of several angiotrophic and neurotrophic factors. Additionally, Western blot analysis revealed the activation of the PI3K/Akt signaling pathway in MSCPExo.
CONCLUSIONS: Preconditioning MSCs with PRP-Exo enhanced the paracrine function, particularly augmenting neurotrophic and pro-angiogenic secretions, demonstrating an improved potential for neural repair.
方法:分离PRP-Exo,并评估其对MSC活力的影响。评估了PRP外切处理的MSC(MSCPExo)对来自受损坐骨神经和人脐静脉内皮细胞(HUVEC)的雪旺细胞(SC)的影响。此外,使用细胞因子阵列分析来自MSCPEExo的条件培养基(MSCPEExo-CM),并通过ELISA和Westernblot进行验证.
结果:PRP-Exo增强MSC活力。MSCPEExo与SCs共混改善了PNI后的细胞凋亡并促进了SC增殖。同样,MSCPEExo-CM表现出促增殖,迁徙,和血管生成效应。细胞因子阵列分析鉴定了MSCPEExo分泌组中的440种蛋白质,155个显示上调,6个显示下调,许多展示出有效的促再生特性。ELISA证实了几种血管营养和神经营养因子的富集。此外,Western印迹分析揭示了MSCPExo中PI3K/Akt信号传导途径的激活。
结论:用PRP-Exo预处理MSCs增强旁分泌功能,特别是增加神经营养和促血管生成分泌,证明了神经修复的潜力。