Fabrication of Gelatin Scraffold for Soft Tissue Regeneration

Abstract

Gelatin has a great potential in tissue engineering due to its excellent biocompatibility and biodegradation. However, the main issue which hindrances the use of pure gelatin as scaffold is its poor thermal stability at body temperature. In order to improve its mechanical weakness, chemical modifications of scaffold were reported. The aim of this study is to investigate the suitable methodology for cross-linking gelatin 50% w/v solution extracted from fish skin (GEL) by using 1-ethyl-3-(3-dimethyl aminopropyl)-1-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) solution (EN) as cross-linker. The results showed that, the frozen methodology (shaping frozen GEL first and immersed in EN solution) was the best method. Concentration of cross-linker affected morphology, mechanical properties, and biocompatibility of scaffold. Microscopic observation of the inner morphology of GEL scaffolds indicated that increasing of EN concentration caused the increasing of their pore size and roughness level. The chemical bond between GEL molecules to form a scaffold with cross-linking agent was confirmed by FT-IR result. The increasing of scaffold tensile strength and degradation duration due to the increasing of EN solution concentration, as confirmed by tensile strength test and biodegradation analysis. Invivo studies generally indicated that all GEL scaffolds provided a structural and chemical environment that enabled cell attachment and proliferation, support for the biological functions. H&E staining results showed the cell proliferation and cell density inside the defect replay by scaffold, focused on the edge of the area associated with the scaffold and the area biodegraded. Among the scaffolds, GEN4 was the best material for blood vessels and cells regeneration. These results indicated that a scaffold with high mechanical strength and good biocompatibility was successfully fabricated with GEL50% w/v and 130/52 mM of EN solution. Key words: gelatin, scaffold, biocompatibility, cell proliferation, degradation.