The effect of human osteoblasts on proliferation and neo-vessel formation of human umbilical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds
The effect of human osteoblasts on proliferation and neo-vessel formation of human umbilical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds
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DOI：
10.1016/j.biomaterials.2008.07.024
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发表时间：
2008-11-01
期刊：
影响因子：
14
通讯作者：
Rommens, Pol Maria
中科院分区：
文献类型：
--
作者：
Hofmann, Alexander;Ritz, Ulrike;Rommens, Pol Maria
血管生成是伤口早期愈合的关键因素，被认为对组织再生和将炎性细胞引导到伤口部位非常重要。通过应用内皮细胞或血管生成生长因子来改善血管形成可能是解决大尺寸工程化骨构建的关键。在这项研究中，我们描述了一个长期的培养环境，以支持人脐静脉内皮细胞(HUVEC)的存活、增殖和体外血管生成。这一条件可以在HUVEC和原代人类成骨细胞(HOB)的共培养模型中实现，该模型使用聚氨酯支架和富含血小板的血浆在静态微环境中进行。我们清楚地表明，HOB支持内皮细胞标记CD31和vWF阳性的HUVEC增殖和自发形成多个管状结构。相反，在单一培养中，大多数人脐静脉内皮细胞既没有增殖，也没有形成任何明显的血管样结构。免疫组织化学和定量聚合酶链式反应分析显示，HOB和HUVEC在长期共培养中细胞分化稳定。这种三维的、无FCS的共培养体系为开发具有高再生和血管生成能力的复合组织工程构建物提供了新的基础。(C)2008爱思唯尔有限公司。保留所有权利。
Angiogenesis is a key element in early wound healing and is considered important for tissue regeneration and for directing inflammatory cells to the wound site. The improvement of vascularization by implementation of endothelial cells or angiogenic growth factors may represent a key solution for engineering bone constructs of large size. In this study, we describe a long-term culture environment that supports the survival, proliferation, and in vitro vasculogenesis of human umbilical vein endothelial cells (HUVEC). This condition can be achieved in a co-culture model of HUVEC and primary human osteoblasts (hOB) employing polyurethane scaffolds and platelet-rich plasma in a static microenvironment. We clearly show that hOB support cell proliferation and spontaneous formation of multiple tubelike structures by HUVEC that were positive for the endothelial cell markers CD31 and vWF. In contrast, in a monoculture, most HUVEC neither proliferated nor formed any apparent vessel-like structures. Immunohistochemistry and quantitative PCR analyses of gene expression revealed that cell differentiation of hOB and HUVEC was stable in long-term co-culture. The three-dimensional, FCS-free co-culture system could provide a new basis for the development of complex tissue engineered constructs with a high regeneration and vascularization capacity. (C) 2008 Elsevier Ltd. All rights reserved.