Diabetic patients are characterized by long wound healing time, and adipose stem cells (ADSCs) can secrete growth factors to promote angiogenesis and improve diabetic wound healing. In this research, we attempted to interrogate the impact of platelet-rich fibrin (PRF) on ADSCs in diabetic wound healing. ADSCs were harvested from human adipose tissues and identified through flow cytometry. After pretreatment with cultured medium supplemented with different concentrations of PRF (2.5%, 5%, and 7.5%), proliferation and differentiation capacity of ADSCs were assessed by CCK-8 assay, qRT-PCR and immunofluorescence (IF), respectively. Tube formation assay measured angiogenesis. Western blot analysis analyzed expression of endothelial markers and the extracellular signal-regulated kinase (ERK) and serine/threonine kinase (Akt) pathways in PRF-induced ADSCs. The CCK-8 experiment indicated that PRF enhanced proliferation of ADSCs in dose-dependent manner, relative to normal control group. The expression of endothelial markers and the capacity of tube formation were significantly promoted by 7.5% PRF. The release of growth factors containing vascular endothelial grow factor (VEGF) and insulin-like growth factor-1 (IGF-1) from PRF was increased with the extension of detection time. When the receptors of VEGF or/and IGF-1 were neutralized, ADSCs differentiation into endothelial cells were obviously inhibited. Additionally, PRF stimulated ERK and Akt pathways, and the inhibitors of ERK and Akt attenuated PRF-induced differentiation of ADSCs into endothelial cells. In conclusion, PRF promoted endothelial cell differentiation and angiogenesis induced by ADSCs in diabetic wound healing, which appears to give guidance for treating patients.

糖尿病患者的特点是伤口愈合时间长，脂肪干细胞（ADSC）可以分泌生长因子促进血管生成，改善糖尿病伤口愈合。在这项研究中，我们试图探讨富血小板纤维蛋白 (PRF) 对糖尿病伤口愈合中 ADSC 的影响。ADSC 从人类脂肪组织中收获并通过流式细胞术进行鉴定。用添加不同浓度PRF（2.5%、5%和7.5%）的培养基预处理后，分别通过CCK-8测定、qRT-PCR和免疫荧光（IF）评估ADSC的增殖和分化能力。管形成测定测量血管生成。蛋白质印迹分析分析了 PRF 诱导的 ADSC 中内皮标记物以及细胞外信号调节激酶 (ERK) 和丝氨酸/苏氨酸激酶 (Akt) 通路的表达。CCK-8实验表明，相对于正常对照组，PRF以剂量依赖性方式增强ADSC的增殖。7.5% PRF显着促进内皮标志物的表达和管形成能力。随着检测时间的延长，PRF释放的生长因子包括血管内皮生长因子（VEGF）和胰岛素样生长因子-1（IGF-1）。当VEGF或/和IGF-1受体被中和时，ADSCs向内皮细胞的分化受到明显抑制。此外，PRF 刺激 ERK 和 Akt 通路，ERK 和 Akt 抑制剂减弱了 PRF 诱导的 ADSC 向内皮细胞的分化。总之，PRF 在糖尿病伤口愈合中促进 ADSC 诱导的内皮细胞分化和血管生成，这似乎可以为治疗患者提供指导。