GLYCOSAMINOGLYCAN SYNTHESIS OF CHONDROCYTES IN FIBRIN GLUE WITH GHC6S PARTICLES

Abstract

Articular cartilage provides functions of lubrication to shear stress and protection from compressive force, but it has poor ability to repair itself after suffering damage. The advanced method of tissue engineering is developed and used to maintain cell functions for tissue regeneration. In order to improve the ECM synthesis for the regeneration, many materials have been examined on chondrocytes or other cell sources. In this study, fibrinogen was concentrated from plasma cryoprecipitation and then polymerized by thrombin into fibrin. Gelatin/hyaluronic acid/chondroitin-6-sulfate (GHC6S) was prepared by the cross-linking reaction with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and ground in liquid nitrogen to particles. The GHC6S particles were mixed with fibrin glue as the tissue engineering scaffold. Porcine articular cartilage chondrocytes were expanded and seeded into the scaffolds. The engineered constructs were cultured and harvested after cultured for 1 and 2 weeks. Morphology of GHC6S particle was examined by scanning electron microscopy (SEM). Total glycosaminoglycans (GAGs) and sulfated GAGs were quantified by p-dimethylaminobenzaldehyde reaction and 1,9-dimethymethylene blue (DMMB) assay, respectively. The results demonstrated that the total GAGs, especially the content of nonsulfated GAGs, hyaluronic acid, were increased with time in chondrocytes growing in fibrin glue with GHC6S particles. It suggested that the GHC6S in fibrin glue chondrocyte kept the GAGs synthesis, which could help resist the compressive force. Therefore, the GHC6S particles mixed within fibrin glue can be used as a promising scaffold for articular tissue engineering.