Coadministration of adipose-derived stem cells and control-released basic fibroblast growth factor facilitates angiogenesis in a murine ischemic hind limb model
Coadministration of adipose-derived stem cells and control-released basic fibroblast growth factor facilitates angiogenesis in a murine ischemic hind limb model
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DOI：
10.1016/j.jvs.2015.09.054
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发表时间：
2016-12-01
影响因子：
4.3
通讯作者：
Mizuno, Hiroshi
中科院分区：
文献类型：
--
作者：
Horikoshi-Ishihara, Hisako;Tobita, Morikuni;Mizuno, Hiroshi
目的：脂肪源性干细胞（ASC）具有血管生成潜力，因为它们分化为内皮细胞并释放血管生成生长因子以引起旁分泌作用。此外，明胶水凝胶持续控制释放的碱性成纤维细胞生长因子（bFGF）也支持有效的血管生成。我们试图确定将 ASC 和对照释放的 bFGF 共同施用到小鼠缺血肢体中是否促进血管生成。方法：通过酶联免疫吸附测定来测量在有或没有对照释放的 bFGF 培养的 ASC 的条件培养基中生长因子的水平。生成小鼠缺血后肢模型并肌内注射以下物质：明胶水凝胶（第1组）、大量ASC（第2组）、对照释放的bFGF（第3组）、少量ASC和对照释放的bFGF（第4组）以及大量ASC和对照释放的bFGF（第5组）。直到第7天，分别通过激光多普勒灌注成像和组织学分析评估宏观和微观血管变化。结果：对照释放的bFGF增强了肝细胞生长因子、血管内皮生长因子和转化生长因子-β1的分泌。根据激光多普勒灌注成像，第 4 组和第 5 组的血管得到改善。苏木精和伊红染色以及 CD31 免疫组织化学染色显示第 4 组和第 5 组的血管密度、血管直径和血管壁厚度增加。在第 4 组和第 5 组的血管壁周围观察到 CD146、α-平滑肌肌动蛋白和转化生长因子-β 1 染色呈阳性的细胞。 在小鼠缺血后肢模型中，ASC 和控制释放的 bFGF 在血管成熟方面促进血管生成。
Objective: Adipose-derived stem cells (ASCs) have angiogenic potential owing to their differentiation into endothelial cells and their release of angiogenic growth factors to elicit paracrine effects. In addition, control-released basic fibroblast growth factor (bFGF) sustained with a gelatin hydrogel also supports effective angiogenesis. We sought to determine if coadministration of ASCs and control-released bFGF into murine ischemic limbs facilitates angiogenesis.Methods: Levels of growth factors in the conditioned media of ASCs cultured with or without control-released bFGF were measured by enzyme-linked immunosorbent assays. A murine ischemic hind limb model was generated and intramuscularly injected with the following: gelatin hydrogel (group 1), a high number of ASCs (group 2), control-released bFGF (group 3), a small number of ASCs and control-released bFGF (group 4), and a high number of ASCs and control-released bFGF (group 5). Macroscopic and microscopic vascular changes were evaluated until day 7 by laser Doppler perfusion imaging and histologic analyses, respectively.Results: Secretion of hepatocyte growth factor, vascular endothelial growth factor, and transforming growth factor-beta 1 was enhanced by control-released bFGF. Vascular improvement was achieved in groups 4 and 5 according to laser Doppler perfusion imaging. Hematoxylin and eosin staining and CD31 immunohistochemical staining demonstrated an increase in the vascular density, vessel diameter, and thickness of vessel walls in groups 4 and 5. Cells positively stained for CD146, alpha-smooth muscle actin, and transforming growth factor-beta 1 were observed around vessel walls in groups 4 and 5.Conclusions: These findings suggest that coadministration of ASCs and control-released bFGF facilitates angiogenesis in terms of vessel maturation in a murine ischemic hind limb model.