碱性成纤维细胞生长因子/双层胶原基复合材料制备及其生物安全性

背景：胶原特殊的分子结构和生物活性有利于多种细胞黏附、增殖和分化，并可降解为新生组织提供足够空间。 目的：制备一种复合负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子双层胶原基复合材料，并评价其生物安全性。 方法：制备交联风干胶原膜和交联冻干胶原膜。将壳聚糖-肝素纳米粒滴于交联冻干胶原膜上，再将湿态交联风干胶原膜置于复合纳米粒子的交联冻干胶原膜上风干，即碱性成纤维细胞生长因子/双层胶原基复合材料。采用急性全身毒性试验、溶血试验、热原试验和细胞毒性试验评价其生物安全性。 结果与结论：碱性成纤维细胞生长因子/双层胶原基复合材料为双层结构，一侧表面致密，另一侧疏松多孔。在其中间负载碱性成纤维细胞生长因子的壳聚糖-肝素纳米粒子呈不规则球形分布于胶原膜内侧面；急性全身毒性试验、热原试验、溶血试验均为阴性，细胞毒性为0级。说明碱性成纤维细胞生长因子/双层胶原基复合材料具有良好的生物安全性，对机体无毒，符合ISO 10993-1评价标准。

Preparation and biological safety of basic fibroblast growth factor/double-layered collagen composite

BACKGROUND:Collagen is a common used scaffold in tissue engineering, its specific molecular structure and biological activity are conducive to a variety of cell adhesion, proliferation and differentiation, and can speed up wound healing and degradation to provide sufficient space for new tissues. OBJECTIVE: Toprepare a kind of inhomogeneous double layered collagen composite incorporated with basic fibroblast growth factor (bFGF) loaded chitosan-heparin (CS-Hep) nanoparticles, and to evaluate its biological safety. METHODS: Theair-dried dense layer collagen membranes and freeze-dried loose layer collagen membrane were prepared respectively and then cross-linked by D-ribose (termed as CAM and CFM respectively). CS-Hep nanoparticles were prepared and then dropped on the surface of CFM. The wet CAM was laid on the surface of CFM with nanoparticles, and then air dried completely at 4 ℃. The obtained material was termed as bDM. Its physical and chemical properties were investigated. In addition, its bio-safety was also examined through acute systemic toxicity, pyrogen, hemolysis and cellular toxicity tests. RESULTS AND CONCLUSION: The bFGF/double-layered collagen composite was made up of double layers with one dense layer (CAM) and another loose layer (CFM). As part of bDM, CS-Hep nanopartiles with irregular global shape were aligned in the scope of D-period structure of the inner surface of CFM; the bFGF/double-layered collagen composite had no systemic acute toxicity, no cytotoxicity and pyrogen reactions as well as no hemolytic effect. The bFGF/double-layered collagen composite is prepared and found to have good biocompatibility and safety. Moreover, this material conforms to the ISO 10993-1, and can be used as a basic scaffold material in tissue engineering.