Sonic Hedgehog Promotes Angiogenesis and Osteogenesis in a Coculture System Consisting of Primary Osteoblasts and Outgrowth Endothelial Cells
Sonic Hedgehog Promotes Angiogenesis and Osteogenesis in a Coculture System Consisting of Primary Osteoblasts and Outgrowth Endothelial Cells
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DOI：
10.1089/ten.tea.2009.0493
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发表时间：
2010-04-01
影响因子：
4.1
通讯作者：
Kirkpatrick, Charles James
中科院分区：
文献类型：
--
作者：
Dohle, Eva;Fuchs, Sabine;Kirkpatrick, Charles James
许多先前的研究记录了体外和体内生长的内皮细胞的血管生成潜力，并提供了证据表明，治疗的成功可能取决于共培养或共植入策略。因此，更深入地了解这种促血管生成作用的分子机制的共培养可能提供新的翻译选择组织工程和再生医学。一个有前途的信号通路在骨修复参与新血管生成和骨形成是音刺猬（Shh）途径。在这篇文章中，我们专注于影响Shh的形成微血管样结构和成骨细胞分化的原代成骨细胞和生长的内皮细胞的共培养。在处理24小时后，与未处理的共培养物相比，Shh导致微血管样结构大量增加。血管生成结构的形成增加似乎与在mRNA和蛋白水平上研究的血管内皮生长因子或血管生成素（Ang-1和Ang-2）的上调相关。此外，用hedgehog信号传导抑制剂环巴胺处理，阻断了共培养物中微血管样结构的形成。然而，外源性Shh也导致了实时聚合酶链反应中几种成骨分化标志物的上调，以及矿化和碱性磷酸酶活性的增加。目前的数据强调了Shh途径在骨再生和血管化中的核心作用。此外，Shh在未来的临床应用中可能具有改善血管生成和成骨的潜力。
A number of previous studies documented the angiogenic potential of outgrowth endothelial cells in vitro and in vivo and provided evidence that therapeutic success could depend on coculture or coimplantation strategies. Thus, deeper insight into the molecular mechanisms underlying this pro-angiogenic effect of cocultures might provide new translational options for tissue engineering and regenerative medicine. One promising signaling pathway in bone repair involved in neoangiogenesis and bone formation is the sonic hedgehog (Shh) pathway. In this article, we focus on the effect of Shh on the formation of microvessel-like structures and osteoblastic differentiation in cocultures of primary osteoblasts and outgrowth endothelial cells. Already after 24 h of treatment, Shh leads to a massive increase in microvessel-like structures compared with untreated cocultures. Increased formation of angiogenic structures seems to correlate with the upregulation of vascular endothelial growth factor or angiopoietins (Ang-1 and Ang-2) studied at both the mRNA and protein levels. In addition, treatment with cyclopamine, an inhibitor of hedgehog signaling, blocked the formation of microvessel-like structures in the cocultures. However, exogenous Shh also resulted in the upregulation of several osteogenic differentiation markers in real-time polymerase chain reaction, as well as in an increased mineralization and alkaline phosphatase activity. The present data highlight the central role of the Shh pathway in bone regeneration and vascularization. Further, Shh might have the potential to improve both angiogenesis and osteogenesis in clinical applications in the future.