In vitro evaluation of crosslinked electrospun fish gelatin scaffolds

• Published in: Materials Science & Engineering C 2013-04, Vol.33 (3), p.1219-1227
• Main Authors: Gomes, S.R., Rodrigues, G., Martins, G.G., Henriques, C.M.R., Silva, J.C.
• Format: Article
• Language: English
• Subjects: 3T3 Cells; Animals; Cell Shape - drug effects; Cell Survival - drug effects; Cellular viability; Cold water fish skin gelatin; Cross-Linking Reagents - pharmacology; Crosslinking; Electric Conductivity; Electrospinning; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - ultrastructure; Fishes; Gelatin - pharmacology; Mice; Microscopy, Confocal; Solutions; Spectroscopy, Fourier Transform Infrared; Surface Tension - drug effects; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Viscosity - drug effects
• ISSN: 0928-4931; 1873-0191
• DOI: 10.1016/j.msec.2012.12.014

Gelatin from cold water fish skin was electrospun, crosslinked and investigated as a substrate for the adhesion and proliferation of cells. Gelatin was first dissolved in either water or concentrated acetic acid and both solutions were successfully electrospun. Cross-linking was achieved via three different routes: glutaraldehyde vapor, genipin and dehydrothermal treatment. Solution's properties (surface tension, electrical conductivity and viscosity) and scaffold's properties (chemical bonds, weight loss and fiber diameters) were measured. Cellular viability was analyzed culturing 3T3 fibroblasts plated on the scaffolds and grown up to 7days. The cells were fixed and observed with SEM or stained for DNA and F-actin and observed with confocal microscopy. In all scaffolds, the cells attached and spread with varying degrees. The evaluation of cell viability showed proliferation of cells until confluence in scaffolds crosslinked by glutaraldehyde and genipin; however the rate of growth in genipin crosslinked scaffolds was slow, recovering only by day five. The results using the dehydrothermal treatment were the less satisfactory. Our results show that glutaraldehyde treated fish gelatin is the most suitable substrate, of the three studied, for fibroblast adhesion and proliferation. ► Electrospinning of fish gelatin dissolved in both water or concentrated acetic acid ► Glutaraldehyde, genipin and dehydrothermal treatment effectively crosslink the fish gelatin fibers ► Fibroblasts effectively adhere to and propagate on all scaffolds ► Cell population is highest for glutaraldehyde crosslinked scaffolds ► Cells exhibit more filopodia and stress fibers on glutaraldehyde crosslinked scaffolds