Department of Plastic and Hand Surgery, University Hospital of Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erlangen, Germany.
Department of Transfusion Medicine, University Hospital of Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erlangen, Germany.
J Cell Mol Med. 2020 Jun;24(12):7034-7043. doi: 10.1111/jcmm.15374. Epub 2020 May 12.
The microvascular endothelial network is essential for bone formation and regeneration. In this context, endothelial cells not only support vascularization but also influence bone physiology via cell contact-dependent mechanisms. In order to improve vascularization and osteogenesis in tissue engineering applications, several strategies have been developed. One promising approach is the coapplication of endothelial and adipose derived stem cells (ADSCs). In this study, we aimed at investigating the best ratio of human umbilical vein endothelial cells (HUVECs) and osteogenic differentiated ADSCs with regard to proliferation, apoptosis, osteogenesis and angiogenesis. For this purpose, cocultures of ADSCs and HUVECs with ratios of 25%:75%, 50%:50% and 75%:25% were performed. We were able to prove that cocultivation supports proliferation whereas apoptosis was unidirectional decreased in cocultured HUVECs mediated by a p-BAD-dependent mechanism. Moreover, coculturing ADSCs and HUVECs stimulated matrix mineralization and the activity of alkaline phosphatase (ALP). Increased gene expression of the proangiogenic markers eNOS, Flt, Ang2 and MMP3 as well as sprouting phenomena in matrigel assays proved the angiogenic potential of the coculture. In summary, coculturing ADSCs and HUVECs stimulates proliferation, cell survival, osteogenesis and angiogenesis particularly in the 50%:50% coculture.
微血管内皮网络对于骨形成和再生至关重要。在这种情况下，内皮细胞不仅支持血管生成，还通过细胞接触依赖性机制影响骨生理学。为了在组织工程应用中提高血管生成和成骨作用，已经开发了几种策略。一种很有前途的方法是内皮细胞和脂肪来源的干细胞（ADSCs）的共同应用。在这项研究中，我们旨在研究人类脐静脉内皮细胞（HUVEC）和成骨分化的 ADSCs 的最佳比例，以研究其增殖、凋亡、成骨和血管生成。为此，我们进行了 ADSCs 和 HUVEC 的共培养，其比例为 25%:75%、50%:50%和 75%:25%。我们能够证明共培养支持增殖，而通过 p-BAD 依赖性机制，共培养的 HUVEC 中的凋亡呈单向降低。此外，ADSCs 和 HUVEC 的共培养刺激基质矿化和碱性磷酸酶（ALP）的活性。血管生成标志物 eNOS、Flt、Ang2 和 MMP3 的基因表达增加以及在 Matrigel 测定中的发芽现象证明了共培养的血管生成潜力。总之，ADSCs 和 HUVEC 的共培养特别在 50%:50%的共培养中刺激增殖、细胞存活、成骨和血管生成。
