从低氧预处理脂肪来源干细胞中分离的细胞外囊泡促进大鼠随意皮瓣缺氧诱导因子 1α 介导的血管生成。

Extracellular Vesicles Isolated From Hypoxia-Preconditioned Adipose-Derived Stem Cells Promote Hypoxia-Inducible Factor 1α-Mediated Neovascularization of Random Skin Flap in Rats.

机构信息

From the Department of Plastic Surgery, The Second Affiliated Hospital of Nanchang University, Jiangxi, People's Republic of China.

出版信息

Ann Plast Surg. 2022 Aug 1;89(2):225-229. doi: 10.1097/SAP.0000000000003266.

DOI:10.1097/SAP.0000000000003266

PMID:35943229

Abstract

BACKGROUND

Random flaps are widely used for wound repair. However, flap necrosis is a serious complication leading to the failure of operation. Our previous study demonstrated a great proangiogenic potential of hypoxia-treated adipose-derived stem cells-extracellular vesicles (HT-ASC-EVs). Thus, we aim to evaluate the effect of HT-ASC-EVs in the survival and angiogenesis of random skin flap in rats.

METHODS

Adipose-derived stem cells-extracellular vesicles were respectively isolated from adipose-derived stem cell culture medium of 3 donors via ultracentrifugation. The expression of hypoxia-inducible factor 1α (HIF-1α) and proangiogenic potential of HT-ASC-EVs and ASC-EVs were compared by co-culturing with human umbilical vein endothelial cells. Forty male Sprague-Dawley rats were randomly divided into 3 group (n = 10/group). A 9 × 3-cm random skin flap was separated from the underlying fascia with both sacral arteries sectioned on each rat. The survival and angiogenesis of flaps treated by ASC-EVs or HT-ASC-EVs were also compared. Laser Doppler flowmetry and immunohistochemistry were used to evaluate skin perfusion and angiogenesis of skin flaps on postoperative day 7.

RESULTS

Hypoxia-treated adipose-derived stem cells-extracellular vesicles further improve the proliferation, migration, tube formation with upregulated HIF-1α, and VEGF expression of human umbilical vein endothelial cells in vitro, compared with ASC-EVs. In vivo, postoperatively injecting HT-ASC-EVs suppressed necrosis rate (29.1 ± 2.8% vs 59.2 ± 2.1%) and promoted the angiogenesis of skin flap including improved skin perfusion (803.2 ± 24.3 vs 556.3 ± 26.7 perfusion unit), increased number of CD31-positive cells, and upregulated expression of HIF-1α in vascular endothelium on postoperative day 7, compared with ASC-EVs.

CONCLUSIONS

Intradermal injecting HT-ASC-EVs improve the survival of random skin flap by promoting HIF-1α-mediated angiogenesis in rat model.

摘要

背景

随意皮瓣被广泛应用于创面修复。然而，皮瓣坏死是导致手术失败的严重并发症。我们之前的研究表明，缺氧处理的脂肪来源干细胞-细胞外囊泡（HT-ASC-EVs）具有很强的促血管生成潜力。因此，我们旨在评估 HT-ASC-EVs 对大鼠随意皮瓣存活和血管生成的影响。

方法

通过超速离心法，分别从 3 位供体的脂肪来源干细胞培养物中分离出脂肪来源干细胞-细胞外囊泡。通过与人脐静脉内皮细胞共培养，比较 HT-ASC-EVs 和 ASC-EVs 的缺氧诱导因子 1α（HIF-1α）表达和促血管生成能力。40 只雄性 Sprague-Dawley 大鼠随机分为 3 组（每组 10 只）。在每只大鼠的骶动脉切断后，从筋膜下分离出 9×3cm 的随意皮瓣。比较 ASC-EVs 和 HT-ASC-EVs 处理的皮瓣的存活和血管生成情况。激光多普勒血流仪和免疫组织化学法评估术后第 7 天皮瓣的皮肤灌注和血管生成情况。

结果

与 ASC-EVs 相比，缺氧处理的脂肪来源干细胞-细胞外囊泡在体外进一步提高了人脐静脉内皮细胞的增殖、迁移、管腔形成能力，并且上调了 HIF-1α 和 VEGF 的表达。在体内，术后注射 HT-ASC-EVs 可降低皮瓣坏死率（29.1±2.8%比 59.2±2.1%），促进皮瓣血管生成，包括改善皮肤灌注（803.2±24.3 比 556.3±26.7 灌注单位）、增加 CD31 阳性细胞数量，并上调术后第 7 天血管内皮细胞中 HIF-1α 的表达，与 ASC-EVs 相比。