Fibrin Glue Cross-Linked With EDC As Injectable Scaffold For Cartilage Engineering

OBJECTIVESCartilage is an important support connective organization of human body.But cartilage deficiency has limited self-recovery capacity.The tissue engineering provides a new development direction for the cartilage repair.The fibrin glue is a main material of cartilage tissue engineering for its good plasticity, adhesively and biocompatibility.As a scaffold for cartilage,fibrin glue has the shortcomings of intensity difference and fast degradation.This experiment try to enhance fibrin glue's strength and anti-biodegradation by using N-(3-dimethylaminopropyl)-N'-ethyl carbodiirnide(EDC) to increase its cross-link degree.In order to build the injectable cartilage better.MATERIALS and METHODSPart 1,Explore the reaction conditions of EDC with fibrinogen 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.Part 2,Study the effect of EDC cross-linked fibrin gel as scaffold material on cell survival activity in vitro 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.Culture the samples in constant temp incubator for 10 days. In the 1^th,4^th,7^th,10^th days of the experiment,using the LIVE/DEAD Viability/Cytotoxicity Kit to detect the survival situation of the cells in the saffcolds.RESULTSPart 1,EDC and thrombin with fibrinogen cannot result to a sol-gel reaction.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. Part 2,Cartilage cells grow well in the control group(fibrinogen 200mg/ml+ thrombin 100U/ml) and the experimental group(fibrinogen 200mg/ml+ EDC 100mg/ml).But most of the cells in the experimental groups with higher concentration of EDC were dead within one week.CONCLUSIONSFibrin 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.