摘要
目的:探讨利用碳化二亚胺(EDC)增加纤维蛋白凝胶的交联度、生物强度和抗降解能力的可能性,更好地构建可注射性软骨。方法:(1)利用不同配比的纤维蛋白原和EDC进行反应制备理想的凝胶样本,并通过检测稳定性、抗酶解能力和扫描电镜检查等分析材料的性能,将其与传统的纤维蛋白凝胶进行比较;(2)EDC和纤维蛋白原进行反应制备的支架材料与软骨细胞混合构建组织工程软骨样本,对照组为传统的纤维蛋白凝胶软骨细胞样本。利用活细胞荧光检测试剂盒检测样本中的细胞在实验的第1,第4,第7,第10天的存活情况。结果:用EDC取代凝血酶形成的纤蛋白凝胶较传统的纤维蛋白凝胶有更好的生物强度和抗酶解能力,且随着EDC浓度的增加而提高;软骨细胞在对照组(纤维蛋白原200mg.mL-1+凝血酶100U.mL-1)和实验组(纤维蛋白原200mg.mL-1+EDC100mg.mL-1)的支架材料中生长良好。结论:适当浓度的EDC对纤维蛋白原交联形成的纤维蛋白凝胶在生物强度和抗酶解能力方面较传统的纤维蛋白凝胶有明显提高,所形成的三维多孔隙网状结构能为细胞的生长提供足够的空间,软骨细胞在该支架材料中存活良好。
Objective：To enhance fibrin glue＇s strength and anti-biodegradation by using N-（3-dimethylaminopropyl）-N＇-ethyl carbodiimide（EDC） to increase its cross-link degree.In order to build the injectable cartilage better.Methods：（1）Use different ratio of fibrinogen and EDC to prepare ideal gel sample.Then analyse the materials＇ performance by stability testing、the ability of anti-enzyme detection、scanning electron microscopy,with the comparison of the traditional fibrin glue.（2）EDC reacts with fibrinogen to prepare different scaffolds,mixed with the chondrocytes to construct cartilage tissue engineering samples.Construct the control group using traditional fibrin glue.using the LIVE/DEAD Viability /Cytotoxicity Kit to detect the survival situation of the cells in the saffcolds.In the 1th,4th,7th,10th days of the experiment.Results： The fibrin glue cross-linked by EDC used to replace thrombin has a better biological strength and the ability of anti-enzyme,with more closely and orderly three-dimensional porous structure than the traditional fibrin glue,and enhanced as the EDC concentration increases.Cartilage cells grow well in the control group（fibrinogen 200mg/ml＋thrombin 100U/ml） and the experimental group（fibrinogen 200mg/ml＋EDC 100mg/ml）.Conclusions：Fibrin glue cross-linked by EDC of appropriated concentration behaves stronger biological strength and anti-biodegrability than conventional fibrin glue,with more closely and orderly spatial structure.The three-dimensional porous network structure of the modified fibrin glue can also provide adequate space for the cells＇ growth.The cartilage cells show good viability in the scaffold during the experimental observation period.But high concentrations of EDC are not suitable for cells to survive.
出处
《中国临床医学》
2010年第6期895-898,共4页
Chinese Journal of Clinical Medicine
基金
复旦大学青年科学基金(编号:No:236)
