Functional assessment of cross-linked porous gelatin hydrogels for bioengineered cell sheet carriers.

An efficient carrier for corneal endothelial cell therapy should deliver and retain the cell sheet transplants at the site of injury without causing adverse effects. Here we introduced a simple stirring process combined with freeze-drying (SFD1) method for the development of gelatin hydrogels with enlarged pore structure that can improve the aqueous humor circulation. Samples fabricated by air-drying (AD) or freeze-drying method were used for comparison. After cross-linking with 1 mM 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC), the discs were investigated to assess their functionality. The simultaneous presence of ice crystals and gas bubbles resulted in large pore size (461 +/- 85 mum) and high porosity (48.0 +/- 1.9%) of SFD1 carriers. Among all of the samples studied, the SFD1 hydrogels showed the most appropriate swelling characteristics without squeezing effect on the anterior segment tissues of the eye. The enlarged pore structure also allowed carriers to contain the highest fraction of mobile water and exhibit the lowest resistance to the glucose permeation. In comparison with AD samples, the SFD1 materials had better cytocompatibility and biocompatibility and more effectively prevented a drastic change of intraocular pressure. Rheological measurements showed that the SFD1 hydrogels behaved like an elastic solid and had a tough (rigid and deformable) texture. As a temporary supporter, the biodegradable gelatin hydrogel could facilitate cell sheet transfer and avoid long-term residence of foreign carriers in the body. Our findings suggest that the gelatin discs with enlarged pore structure have potential as cell sheet carriers for intraocular delivery and corneal tissue engineering.