An Antithrombogenic Citric Acid-Crosslinked Gelatin with Endothelialization Activity

A novel citric acid-crosslinked gelatin matrix with endothelialization activity and anti-adhesive properties for platelets is prepared. The matrix is characterized using an endothelial cell culture and an antithrombogenic activity test. The number of endothelial cells cultured on the surface of the trisuccinimidyl citrate (TSC)-crosslinked gelatin increases as the concentration of TSC increases to 20 m M, and then decreases with further increases in TSC concentration. Compared with glutaraldehyde-crosslinked gelatin, platelet number and fibrin network formation on the TSC-crosslinked gelatin are minimal at high TSC concentration. The biocompatibility of the matrix is evaluated by bioluminescence imaging. This indicates that the inflammation reaction of the TSC-crosslinked gelatin is lower than that of glutaraldehyde-crosslinked gelatin. Physicochemical analysis of TSC-crosslinked gelatin with different TSC concentrations shows that the high concentration of the cell adhesion sequence, arginine-glycine-aspartic acid, contributes to the promotion of endothelial cell adhesion and subsequent endothelial cell growth. Analysis of the carboxyl groups in the TSC-crosslinked gelatin showed that the antithrombogenic activity is due to the increased negative charge derived from the hydrolyzed active ester groups of TSC. These findings show that TSC-crosslinked gelatin has the potential for use in biomedical devices in contact with blood, such as stents, artificial blood vessels, and artificial heart valves.