Influence of solvent composition on the performance of carbodiimide cross-linked gelatin carriers for retinal sheet delivery
Gelatin is a protein molecule that displays bioaffinity and provides a template to guide retinal pigment epithelial (RPE) cell organization and growth. We have recently demonstrated that the carbodiimide cross-linked gelatin membranes can be used as retinal sheet carriers. The purpose of this work was to further determine the role of solvent composition in the tissue delivery performance of chemically modified biopolymer matrices. The gelatin molecules were treated with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) in the presence of binary ethanol/water mixtures with varying ethanol concentrations (70–95 vol%) to obtain the carriers with different cross-linking efficiencies and mechanical properties. Results of melting point measurements and in vitro degradation tests showed that when the cross-linking index reached a high level of around 45 %, the EDC cross-linked gelatin materials have sufficient thermal stability and resistance to enzymatic degradation, indicating their suitability for the development of carriers for retinal sheet delivery. Irrespective of the solvent composition, the chemically modified gelatin samples are compatible toward human RPE cells without causing toxicity and inflammation. In particular, the membrane carriers prepared by the cross-linking in the presence of solvent mixtures containing 80–90 vol% of ethanol have no impact on the proliferative capacity of ARPE-19 cultures and possess good efficiency in transferring and encapsulating the retinal tissues. It is concluded that, except for cell viability and pro-inflammatory cytokine expression, the retinal sheet delivery performance strongly depends on the solvent composition for EDC cross-linking of gelatin molecules.
KeywordsGelatin Solvent Composition Human Retinal Pigment Epithelial Cell Gelatin Molecule Gelatin Sample
This work was supported by Grant NSC100-2628-E-182-004-MY3 from the National Science Council of Republic of China and grant CMRPD2A0111 from Chang Gung Memorial Hospital. The author is grateful to Mr. I-Hao Tu (Institute of Biochemical and Biomedical Engineering, Chang Gung University) for technical assistance.
- 1.Lu PL, Lai JY, Tabata Y, Hsiue GH. A methodology based on the “anterior chamber of rabbit eyes” model for noninvasively determining the biocompatibility of biomaterials in an immune privileged site. J Biomed Mater Res A. 2008;86:108–16.Google Scholar