Thimm Benjamin W, Unger Ronald E, Neumann Hans-Georg, Kirkpatrick C James
Institute of Pathology, Johannes Gutenberg-University Mainz, Langenbeckstr.1, 55101 Mainz, Germany.
Biomed Mater. 2008 Mar;3(1):015007. doi: 10.1088/1748-6041/3/1/015007. Epub 2008 Feb 6.
The bone biomaterial BONITmatrix, a nanoporous, granular scaffold composed of hydroxylapatite, calcium phosphate and SiO2, linked by a dense collagen mesh, was tested for its biocompatibility using endothelial cells (EC) in the form of macrovascular HUVEC, microvascular HDMEC and the endothelial cell line ISOHAS-1. Cells were examined for their adherence and growth on the biomaterial and this was followed by confocal laser scanning microscopy after vital staining or immunocytochemical reactions, as well as by scanning electron microscopy. Macro- and microvascular ECs predominantly spread on BONITmatrix-collagen mesh-covered surfaces and fibres and maintained their typical morphology. As ECs in vivo must build up a functional vasculature, the seeded cells were further tested for proinflammatory expression markers and cytokine expression after lipopolysaccharide stimulation. Protein-coating studies revealed that BONITmatrix-collagen scaffolds needed human blood serum coating to successfully support the growth of ECs. All cells expressed endothelium-specific surface marker proteins such as PECAM-1, VE-cadherin and vWF. The in vitro data support recent in vivo studies and indicate that this calcium phosphate/SiO2-xerogel composite could be a useful scaffold material for tissue engineering.
骨生物材料BONITmatrix是一种纳米多孔颗粒支架,由羟基磷灰石、磷酸钙和SiO₂组成,通过致密的胶原网连接。采用大血管人脐静脉内皮细胞(HUVEC)、微血管人真皮微血管内皮细胞(HDMEC)和内皮细胞系ISOHAS-1等内皮细胞对其生物相容性进行了测试。检测细胞在生物材料上的黏附与生长情况,之后通过活细胞染色或免疫细胞化学反应后的共聚焦激光扫描显微镜以及扫描电子显微镜进行观察。大血管和微血管内皮细胞主要在覆盖有BONITmatrix-胶原网的表面和纤维上铺展,并保持其典型形态。由于体内内皮细胞必须构建功能性脉管系统,因此在脂多糖刺激后,对接种的细胞进行促炎表达标志物和细胞因子表达的进一步检测。蛋白质包被研究表明,BONITmatrix-胶原支架需要人血清包被才能成功支持内皮细胞生长。所有细胞均表达内皮细胞特异性表面标志物蛋白,如血小板内皮细胞黏附分子-1(PECAM-1)、血管内皮钙黏蛋白(VE-钙黏蛋白)和血管性血友病因子(vWF)。体外数据支持了最近的体内研究,并表明这种磷酸钙/SiO₂ -干凝胶复合材料可能是一种用于组织工程的有用支架材料。