Adipose-derived mesenchymal stem cells (ASCs) therapy is emerging as a novel therapeutic option for the treatment of a variety of diseases including diabetes and diabetic wound healing. Multiple studies indicate that ASCs could promote wound healing and reverse diabetes. However, whether ASCs from diabetic donors retain their therapeutic functions and the mechanisms of how ASCs contribute to wound healing remain largely unknown. In this study, we explored the cutaneous wound healing ability of ASCs collected from C57BL/6 mice that had been rendered diabetic with streptozotocin (STZ). ASCs were harvested from adipose tissues of type 1 diabetic (T1D) or normal C57BL/6 mice. Cell phenotypes were evaluated by flow cytometry analysis, and cell differentiation into adipocytes, chondrocytes, and osteocytes was compared. Secretions of transforming growth factor β (TGF-β1), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) by ASCs were assessed by ELISA. Migration and proliferation of fibroblasts co-cultured with T1D ASCs or control ASCs were also compared. The therapeutic effects of T1D and control ASCs in promoting wound closure were measured in vivo in a T1D wound mouse model. Granulation tissues were collected and stained with H&E at 14th day. CD34 and collagen I were detected by immunohistochemistry. Expressions of IL-6, α-SMA, CD31, collagen I, and collagen III were quantified by real-time PCR. GFP-expressing ASCs were used to trace in vivo cell differentiation. T1D ASCs and control ASCs showed similar expression of cell surface markers (CD29, CD34, CD105) and proliferation pattern. They can both differentiate into different cell types. T1D ASCs secreted similar amounts of VEGF and bFGF, but less TGF-β compared with control ASCs. Like control ASCs, T1D ASCs promoted the proliferation and migration of skin fibroblast cells. When injected in cutaneous wound of T1D mice, T1D ASCs increased wound closure and hair follicle regeneration at a comparable extent as ASCs. Mice receiving T1D ASCs or ASCs exhibited significantly higher expressions of collagen I, collagen III, and CD31 and reduced expression of IL-6 in wound tissues. Immunohistochemistry staining showed increased angiogenesis in mice receiving ASCs as was evident by increased CD34+ cells and collagen I staining. GFP+ ASCs injection showed that ASCs differentiated into fibroblasts and endothelial cells in vivo. Our results suggest that T1D ASCs could accelerate cutaneous wound healing. Mechanisms may include increasing fibroblast growth and migration, skin angiogenesis, and differentiation into fibroblasts and endothelial cells. This study provides evidence that diabetic ASCs may be used as a therapeutic option in cutaneous wound healing in diabetic recipients.

中文翻译：

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从糖尿病小鼠分离的脂肪干细胞可改善链脲佐菌素诱导的糖尿病小鼠的皮肤伤口愈合。

脂肪来源的间充质干细胞（ASCs）治疗正在成为治疗包括糖尿病和糖尿病伤口愈合在内的多种疾病的新型治疗选择。多项研究表明，ASCs可以促进伤口愈合和逆转糖尿病。然而，来自糖尿病供体的ASCs是否保留其治疗功能以及ASCs如何促进伤口愈合的机制仍是未知的。在这项研究中，我们探索了从C57BL / 6小鼠收集的ASC的皮肤伤口愈合能力，这些小鼠已被链脲佐菌素（STZ）赋予糖尿病。从1型糖尿病（T1D）或正常C57BL / 6小鼠的脂肪组织中收获ASC。通过流式细胞仪分析评估细胞表型，并比较细胞分化为脂肪细胞，软骨细胞和骨细胞。通过ELISA评估转化生长因子β（TGF-β1），碱性成纤维细胞生长因子（bFGF）和血管内皮生长因子（VEGF）的分泌。还比较了与T1D ASC或对照ASC共培养的成纤维细胞的迁移和增殖。在T1D伤口小鼠模型中体内测量了T1D和对照ASC在促进伤口闭合中的治疗作用。收集肉芽组织并在第14天用H＆E染色。通过免疫组织化学检测CD34和胶原蛋白I。通过实时PCR定量IL-6，α-SMA，CD31，胶原蛋白I和胶原蛋白III的表达。表达GFP的ASC用于追踪体内细胞分化。T1D ASC和对照ASC显示相似的细胞表面标志物表达（CD29，CD34，CD105）和增殖模式。它们都可以分化成不同的细胞类型。T1D ASC分泌相似量的VEGF和bFGF，但与对照ASC相比分泌较少的TGF-β。像对照ASC一样，T1D ASC促进皮肤成纤维细胞的增殖和迁移。当将T1D ASC注射到T1D小鼠的皮肤伤口中时，其伤口闭合和毛囊再生的程度与ASC相当。接受T1D ASC或ASC的小鼠在伤口组织中的胶原蛋白I，胶原蛋白III和CD31的表达明显较高，而IL-6的表达降低。免疫组织化学染色显示，接受ASC的小鼠血管生成增加，CD34 +细胞和胶原蛋白I染色增加证明了这一点。GFP + ASCs注射显示，ASCs在体内可分化为成纤维细胞和内皮细胞。我们的结果表明，T1D ASCs可以促进皮肤伤口愈合。机制可能包括增加成纤维细胞的生长和迁移，皮肤血管生成以及分化为成纤维细胞和内皮细胞。这项研究提供的证据表明，糖尿病ASC可以用作糖尿病患者皮肤伤口愈合的治疗选择。