Evaluation of poly(vinyl alcohol) hydrogels as a component of hybrid artificial tissues

Hydrogels are three-dimensional polymeric networks very similar to biological tissues. Many synthetic polymers can be used in preparing hydrogels. Among them poly(vinyl alcohol) (PVA), physically crosslinked by repeated freeze-thawing cycles of polymer aqueous solutions, is widely employed to make hydrogels for biomedical applications. To increase the similarity between hydrogels and natural tissues and to obtain polymeric hybrid tissues, we attempted to incorporate 3T3 cells, from a mouse fibroblast cell line, into PVA hydrogels obtained by one freeze-thawing cycle using as a solvent complete culture medium. Hydrogels were also made using eight freeze-thawing cycles from PVA solutions prepared using as a solvent either complete culture medium or water. Cell adhesion experiments were performed by seeding 3T3 and human umbilical vein endothelial cells (HUVEC) on to the hydrogel surface. The effect of the solvent and of the different number of freeze-thawing cycles on the mechanical characteristics of the PVA hydrogels were investigated by dynamic-mechanical techniques. A scanning force microscope analysis of the hydrogel surface viscoelastic properties was also carried out. Our results show that PVA is not cytotoxic. Although PVA hydrogel surface characteristics do not seem to favour the adhesion of substrate-dependent cells, encouraging results were obtained with the 3T3 cells incorporation. DMA analysis indicates that the networks prepared by eight freeze-thawing cycles possess a mechanical consistency comparable, even slightly better, than the ones prepared by only one freeze-thawing cycle and used for the cell incorporation studies.