New craniofacial bone engineering through miR-23-27-24 cluster mediated osteogenic-angiogenic coupling

血管影响骨骼生理的方方面面，包括矿物质生长。 在新骨形成过程中。使骨骼成为我们现在拥有的血管组织 专注于将microRNA作为分子工程工具，因为它们有能力 可逆地瞄准多个监管网络。我们和其他人最近的研究 实验室已经证明，miRNAs促进骨系分化和 WNT、BMP和Notch信号通路中的新骨形成及其调控 骨吸收。在我们寻求促进偶合骨骼的新疗法方面 重塑和血管生成用于治疗我们这里的大型骨缺损 确定miR-27~23~24 microRNA簇是一个很有前途的miRNA候选 试剂。我们的初步分析揭示了多个miR-23和miR-27结合位点 Wnt拮抗剂SFRP1和NLK 3‘的非翻译区(UTR)，它在 结合血管生成抑制剂BIM非编码区上的miR-24结合位点 和Sema4a展示了miR-27-23-24 microRNA簇对 成骨和血管生成分子工程策略的结合。 在初步研究中，我们已经证明了miR-23-27-24簇成员 在炎症组织中的表达显著降低。相反，miR-27模拟 牙周病变组织合并严重垂直性骨缺损的治疗 80%的修复完全证明了意外广泛的新骨形成 大颅骨垂直骨高度与矿化组织新沉积的关系 应用miR-27 8周后出现缺陷。含有PEG-PLGA-PLL-miR-的植入物- 27纳米颗粒促进基质重塑，新血管形成，以及 在临界大小的缺损处有大量的颅骨再生。在移动电话上 水平，miR-27过表达增加碱性磷酸酶活性和骨标记物 基因表达，而抑制miR-27导致显著增加 破骨细胞生成相关基因表达与骨吸收MiR-24和 MIR-27促进血管内皮细胞增殖和血管形成。 总之，这些令人兴奋的初步数据促使我们提出了一种新的分子 将利用成骨和促血管生成特性的工程战略 MIR-23/27/24集群成员与适当的脚手架和重新平衡 牙槽骨和颅骨的动态平衡促进新的颅面骨形成。

Blood vessels affect all aspects of bone physiology, including mineral growth during new bone formation. To engineer bone as a vascularized tissue we have now focused on microRNAs as molecular engineering tools because of their ability to reversibly target multiple regulatory networks. Recent studies from our and other laboratories have demonstrated that miRNAs promote bone lineage differentiation and new bone formation through WNT, BMP, and Notch signaling pathways and also control bone resorption. In our quest for novel therapeutics that promote coupled bone remodeling and angiogenesis for the treatment of large bone defects we have here identified the miR-27~23~24 microRNA cluster as a highly promising miRNA candidate reagent. Our preliminary analysis revealed multiple miR-23 and miR-27 binding sites on the untranslated region (UTR) of the Wnt antagonist Sfrp1 and Nlk 3’, which in conjunction with miR-24 binding sites on the UTR of the angiogenesis inhibitors BIM and SEMA4A demonstrate the potential of the miR-27-23-24 microRNA cluster for coupled osteogenic and angiogenic molecular engineering strategies. In preliminary studies, we have demonstrated that miR-23-27-24 cluster member expression was greatly reduced in inflammatory tissues. Conversely, miR-27 mimic treatment of diseased periodontal tissues with severe vertical bone defects resulted in unexpectedly extensive new bone formation as evidenced by 80% complete restoration of vertical bone height and new deposition of mineralized tissue in large calvarial bone defects after 8 weeks of miR-27 application. Implants containing PEG-PLGA-PLL-miR- 27 nanoparticles promoted matrix remodeling, new blood vessel formation, and substantial amounts of calvarial bone regeneration in critical size defects. On a cellular level, miR-27 overexpression increased alkaline phosphatase activity and bone marker gene expression, while inhibition of miR-27 resulted in a dramatic increase in the expression of osteoclastogenesis related genes and bone resorption. Both miR-24 and miR-27 promoted endothelial cell proliferation and blood vessel tube formation. Together, these exciting preliminary data prompted us to propose a novel molecular engineering strategy that will harness the osteogenic and pro-angiogenic qualities of miR-23/27/24 cluster members in conjunction with suitable scaffolds and re-balance alveolar and calvarial bone homeostasis for new craniofacial bone formation.