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血小板源性生长因子-BB与脂肪来源干细胞联合通过调节PTEN/AKT通路加速伤口愈合。

Combination of PDGF-BB and adipose-derived stem cells accelerated wound healing through modulating PTEN/AKT pathway.

作者信息

Lai Fangyuan, Dai Shijie, Zhao Ye, Sun Yi

机构信息

Center for Plastic & Reconstructive Surgery, Department of Plastic & Reconstructive Surgery, Zhejiang Provincial People's Hospital (Affiliated People's Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, China.

College of Pharmacy, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.

出版信息

Injury. 2023 Feb 13. doi: 10.1016/j.injury.2023.02.027.

Abstract

Adipose-derived stem cells (ADSCs) have been widely proven to facilitate wound healing. Our study aimed to estimate the influence of combined ADSCs and platelet-derived growth factor-BB (PDGF-BB) on wound healing. We utilized 4 healthy SD rats to isolate ADSCs. Platelet-rich plasma (PRP) was acquired utilizing a two-step centrifugation technology. The role of PRP, PDGF-BB, and PDGF-BB combined with a PI3k inhibitor LY294002 on the viability, migration, and PTEN/AKT pathway in ADSCs were examined utilizing CCK-8, Transwell, and western blot assays. Then, we constructed an open trauma model in SD rats. Effects of ADSCs treated with PDGF-BB on pathological changes, CD31, and PTEN/AKT pathway of wound closure were assessed by hematoxylin & eosin (H&E) staining, Masson staining, immunohistochemical, and western blot assays, respectively. PRP and PDGF-BB intensified the viability and migration of ADSCs by modulating the PTEN/AKT pathway. Interestingly, LY294002 reversed the role of PDGF-BB on ADSCs. In vivo experiments, combined intervention with ADSCs plus PDGF-BB/PRP facilitated wound closure and ameliorated histological injury. Moreover, combined intervention with ADSCs and PDGF-BB attenuated the PTEN level and elevated the CD31 level as well as the ratio of p-AKT/AKT in the skin tissues. A combination of ADSCs and PDGF-BB facilitated wound healing might associate with the regulation of the PTEN/AKT pathway.

摘要

脂肪来源干细胞(ADSCs)已被广泛证明可促进伤口愈合。我们的研究旨在评估ADSCs与血小板衍生生长因子-BB(PDGF-BB)联合应用对伤口愈合的影响。我们使用4只健康的SD大鼠分离ADSCs。利用两步离心技术获取富血小板血浆(PRP)。采用CCK-8、Transwell和蛋白质免疫印迹分析检测PRP、PDGF-BB以及PDGF-BB与磷脂酰肌醇-3激酶(PI3k)抑制剂LY294002联合应用对ADSCs活力、迁移及PTEN/AKT信号通路的作用。然后,我们在SD大鼠中构建开放性创伤模型。分别通过苏木精-伊红(H&E)染色、Masson染色、免疫组织化学和蛋白质免疫印迹分析评估经PDGF-BB处理的ADSCs对伤口愈合的病理变化、CD31以及PTEN/AKT信号通路的影响。PRP和PDGF-BB通过调节PTEN/AKT信号通路增强了ADSCs的活力和迁移能力。有趣的是,LY294002逆转了PDGF-BB对ADSCs的作用。在体内实验中,ADSCs与PDGF-BB/PRP联合干预促进了伤口闭合并改善了组织学损伤。此外,ADSCs与PDGF-BB联合干预降低了皮肤组织中PTEN水平,提高了CD31水平以及p-AKT/AKT比值。ADSCs与PDGF-BB联合应用促进伤口愈合可能与PTEN/AKT信号通路的调节有关。

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