Department Tissue Engineering and Regenerative Medicine (TERM), University Hospital Wuerzburg, Wuerzburg, Germany.
Fraunhofer Translational Center Regenerative Therapies TLC-RT, Fraunhofer Institute for Silicate Research ISC, Wuerzburg, Germany.
Clin Hemorheol Microcirc. 2019;73(2):317-328. doi: 10.3233/CH-190559.
Prolyl hydroxylase inhibitors (PHIs) are promising compounds to promote angiogenesis by stabilizing hypoxia-inducible factor-1α (HIF-1α), a master regulator of angiogenesis. Increased HIF-1α presence induces expression of proangiogenic genes such as vascular endothelial growth factor (VEGF).
We investigated the pharmacological induction of hypoxia via the PHI ciclopirox olamine (CPX) as angiogenesis strategy on human dermal microvascular endothelial cell (hd-mvEC) spheroids directly and indirectly via activating human mesenchymal stem cells (hMSCs).
HMSCs were isolated from bone marrow and hd-mvECs from foreskin biopsies. MSC-conditioned medium after CPX stimulation (MSC-CM CPX) was analyzed by VEGF ELISA and Proteome Profiler™ Human Angiogenesis Array. Direct stimulation with CPX and indirect stimulation via MSC-CM CPX were compared in sprouting assays of hd-mvEC spheroids.
Direct stimulation with CPX significantly increased sprouting of hd-mvEC spheroids. MSC-CM CPX also induced sprouting from hd-mvEC spheroids, which was mediated by angiogenic VEGF and other proangiogenic factors that had been produced by stimulated hMSCs.
The stimulation with CPX increased the proangiogenic response of hd-mvECs and hMSCs. The direct stimulation of hd-mvECs with CPX has the potential to replace external VEGF supplementation. Thus, CPX can induce angiogenesis in ECs even in the absence of auxiliary cells demonstrating a promising proangiogenic approach.
脯氨酰羟化酶抑制剂（PHIs）通过稳定缺氧诱导因子-1α（HIF-1α），是一种促进血管生成的有前途的化合物，HIF-1α 是血管生成的主要调节因子。增加 HIF-1α 的存在会诱导血管生成基因如血管内皮生长因子（VEGF）的表达。
我们通过 PHI 环吡酮胺（CPX）诱导缺氧来研究血管生成策略，CPX 作为一种血管生成策略，直接作用于人真皮微血管内皮细胞（hd-mvEC）球体，间接作用于激活人间充质干细胞（hMSC）。
从骨髓中分离 hMSC，从包皮活检中分离 hd-mvEC。用 VEGF ELISA 和 Proteome Profiler™ Human Angiogenesis Array 分析 CPX 刺激后的 MSC 条件培养基（MSC-CM CPX）。在 hd-mvEC 球体的发芽实验中比较 CPX 的直接刺激和 MSC-CM CPX 的间接刺激。
CPX 的直接刺激显著增加了 hd-mvEC 球体的发芽。MSC-CM CPX 也诱导了 hd-mvEC 球体的发芽，这是由被刺激的 hMSC 产生的血管生成 VEGF 和其他促血管生成因子介导的。
CPX 刺激增加了 hd-mvEC 和 hMSC 的促血管生成反应。CPX 直接刺激 hd-mvEC 具有替代外部 VEGF 补充的潜力。因此，CPX 可以诱导 EC 中的血管生成，即使在没有辅助细胞的情况下，这表明 CPX 是一种有前途的促血管生成方法。
