Key Laboratory for Advanced Technologies of Materials, Ministry of Education, Southwest Jiaotong University, 610031 Chengdu, PR China.
Cytokine. 2011 Nov;56(2):208-17. doi: 10.1016/j.cyto.2011.06.020. Epub 2011 Jul 26.
Immobilization of biomolecules with a variety of biological functions has been a promising method to improve the biocompatibility of biomaterials. However, little is known about their inflammatory property and cytotoxicity, which are both key aspects to most biomaterials designed for tissue engineering applications and in vivo implantation. In this in vitro study, heparin/fibronectin complex (Hep/Fn) was coimmobilized onto titanium surface (HF-Ti), which had been proven to have the properties of both anticoagulation and endothelialization in our previous study. Fourier transform infrared (FTIR) spectroscopy and water contact angle measurement were utilized to determine the surface chemical compositions and physical properties. Toluidine Blue O (TBO) and immunochemistry methods were performed to quantify the surface-immobilized heparin and fibronectin. The early inflammatory responses elicited by pristine Ti and HF-Ti were investigated by proinflammatory cytokine secretion of tumor necrosis factor-alpha (TNF-α) released by attached peritoneal macrophages, monocyte chemoattractant protein-1 (MCP-1) and interleukin-1β (IL-1β) released by attached human umbilical vein endothelial cells (ECs), respectively. Scanning electronic microscopy (SEM) and immunofluorescence were employed to investigate the changes in macrophages and ECs morphologies. The incubation period for both cells was 24h and the results showed that HF-Ti revealed a weaker inflammatory response than pristine Ti, which provoked a stronger inflammatory response and higher activation of macrophages. Our data suggest that Hep/Fn coimmobilized biomaterials surface may develop to be a new generation of biomaterials with both biocompatibility and anti-inflammatory properties, especially for used as cardiovascular implants and in tissue engineering applications.
将具有各种生物功能的生物分子固定化已成为提高生物材料生物相容性的一种很有前途的方法。然而，对于大多数用于组织工程应用和体内植入的生物材料，其炎症特性和细胞毒性知之甚少，而这两者都是关键方面。在这项体外研究中，肝素/纤维连接蛋白复合物（Hep/Fn）被共固定在钛表面（HF-Ti）上，在我们之前的研究中已经证明该表面具有抗凝血和内皮化的特性。傅里叶变换红外（FTIR）光谱和水接触角测量用于确定表面化学成分和物理性质。甲苯胺蓝 O（TBO）和免疫化学方法用于定量表面固定化的肝素和纤维连接蛋白。通过附着的腹腔巨噬细胞释放的肿瘤坏死因子-α（TNF-α）、附着的人脐静脉内皮细胞（ECs）释放的单核细胞趋化蛋白-1（MCP-1）和白细胞介素-1β（IL-1β）来研究原始 Ti 和 HF-Ti 引起的早期炎症反应。扫描电子显微镜（SEM）和免疫荧光用于研究巨噬细胞和 ECs 形态的变化。两种细胞的孵育期均为 24 小时，结果表明 HF-Ti 比原始 Ti 表现出较弱的炎症反应，后者引起更强的炎症反应和巨噬细胞的更高激活。我们的数据表明，共固定化 Hep/Fn 的生物材料表面可能会开发成为具有生物相容性和抗炎特性的新一代生物材料，特别是用于心血管植入物和组织工程应用。
