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脂肪干细胞和富含血小板的血浆在真皮再生模板中诱导血管样结构。

Adipose Stem Cells and Platelet-Rich Plasma Induce Vascular-Like Structures in a Dermal Regeneration Template.

机构信息

Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy.

Department of Medicine and Health Sciences "V. Tiberio," University of Molise, Campobasso, Italy.

出版信息

Tissue Eng Part A. 2021 May;27(9-10):631-641. doi: 10.1089/ten.TEA.2020.0175. Epub 2020 Oct 12.

Abstract

In the context of biointeractive dressings used for enhancing wound healing, the use of stromal vascular fraction (SVF) or adipose-derived stem cells (ASCs) hereof derived has not been fully exploited yet. Noncultured SVF, a heterogeneous mesenchymal population of cells, is attractive in the field of dermal regeneration because it can be instantaneously obtained, avoids genomic alterations, and is comparatively safer than cultured ASCs. Integra Dermal Regeneration Template (DRT) was sprinkled with ASCs in complete medium supplemented with 10% fetal bovine serum (FBS), or SVF, obtained from emulsified or nonemulsified fat, in medium supplemented with 2% platelet-rich plasma (PRP). The presence and differentiation of cells were evaluated by standard histochemistry and immunohistochemistry, whereas conditioned media were analyzed for vascular endothelial growth factors (VEGF) by ELISA. experiments were conducted to analyze ASC proliferation in the presence of either FBS or PRP. Deposition of ASCs in medium supplemented with FBS caused their integration into Integra DRT as early as 1 h. ASCs were found as aggregates until 6-10 days without forming organized structures. When seeded onto Integra DRT, SVF cells in medium supplemented with PRP formed aggregates at early times, which at 7 and 10 days organized into vascular-like structures, lined by CD31 and smooth muscle actin-positive cells. With nonemulsified fat, the lacunar structures did not show an organized distribution of SVF cells. PRP induced ASC proliferation although at lower level than FBS. VEGF secretion was enhanced when fat emulsification was introduced into the protocol. In conclusion, the combination of SVF cells obtained from emulsified fat, PRP, and Integra DRT exhibit synergistic effect on the formation of vessel-like structures indicating a step forward aimed at regenerative surgery for chronic wound healing.

摘要

在用于促进伤口愈合的生物交互式敷料的背景下,基质血管部分 (SVF) 或脂肪来源的干细胞 (ASC) 的使用尚未得到充分利用。未培养的 SVF 是一种异质的间充质细胞群体,因其可即时获得、避免基因组改变且比培养的 ASC 更安全,在皮肤再生领域具有吸引力。Integra 真皮再生模板 (DRT) 用完全培养基中的 ASC 进行喷洒,该培养基补充有 10%胎牛血清 (FBS),或用含有 2%富血小板血浆 (PRP) 的培养基从乳化或非乳化脂肪中获得的 SVF。通过标准组织化学和免疫组织化学评估细胞的存在和分化,而通过 ELISA 分析条件培养基中的血管内皮生长因子 (VEGF)。进行实验以分析在 FBS 或 PRP 存在下 ASC 的增殖。在补充 FBS 的培养基中沉积的 ASC 导致它们在 Integra DRT 中整合,早在 1 小时即可观察到。直到 6-10 天,ASC 仍以聚集形式存在,而没有形成有组织的结构。当接种到 Integra DRT 上时,补充 PRP 的培养基中的 SVF 细胞在早期形成聚集物,在 7 天和 10 天形成类似于血管的结构,由 CD31 和平滑肌肌动蛋白阳性细胞排列。用非乳化脂肪,腔隙结构没有显示出 SVF 细胞的有组织分布。尽管 PRP 诱导 ASC 的增殖水平低于 FBS,但仍有所增强。当引入脂肪乳化时,VEGF 分泌得到增强。总之,从乳化脂肪中获得的 SVF 细胞、PRP 和 Integra DRT 的组合对血管样结构的形成表现出协同作用,这表明在慢性伤口愈合的再生手术方面向前迈进了一步。

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