Studies on a Novel Gelatin Sponge: Preparation and Characterization of Cross-Linked Gelatin Scaffolds Using 2-Chloro-1-Methylpyridinium Iodide as a Zero-Length Cross-Linker

Abstract

We prepared a novel porous gelatin (GEL) sponge which was cross-linked (CL) with a zero-length crosslinker of 2-chloro-1-methylpyridinium iodide (CMPI), and compared CPMI with 1-ethyl-3,3-dimethylaminoproplycarbodiimide (EDC). The ninhydrin assay indicated that the CMPI-CL-GEL sponge had a higher degree of cross-linking than the EDC-CL-GEL sponge at cross-linking saturation. In contrast, the EDC-CL-GEL sponge demonstrated poor water uptake and a much slower enzymatic degradation rate than the CMPI-CL-GEL sponge. Scanning electron microscopy (SEM) images of the gelatin sponge fabricated using a gradient frozen-lyophilization method showed uniformly distributed and interconnected pores. Human 3T3 fibroblasts were successfully seeded onto the scaffolds, and cell proliferation was sustained on all CL-GEL sponges. CMPI-CL-GEL sponges demonstrated significantly increased cell numbers after day 1, and cell numbers steadily rose from day 1 to 12. Meanwhile, the CMPI-CL-GEL sponge had a higher cell number than the EDC-CL-GEL sponge (P < 0.05) by day 4. In vitro studies with 3T3 fibroblasts demonstrated an increased cell viability for those cells grown on sponges cross-linked with CMPI compared to those cross-linked with EDC. SEM images revealed attachment and spreading of cells, the CMPI-CL-GEL sponges had more cells that had elongated, migrated, and formed interconnected networks with neighboring cells.