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Abstract
A bi-functional epoxy-based cross-linker, 1,4-Butanediol diglycidyl ether (BDDGE), was investigated in the fabrication of collagen based corneal substitutes. Two synthetic strategies were explored in the preparation of the cross-linked collagen scaffolds. The lysine residues of Type 1 porcine collagen were directly cross-linked using l,4-Butanediol diglycidyl ether (BDDGE) under basic conditions at pH 11. Alternatively, under conventional methodology, using both BDDGE and 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) as cross-linkers, hydrogels were fabricated under acidic conditions. In this latter strategy, Cu(BF4)2·XH2O was used to catalyze the formation of secondary amine bonds. To date, we have demonstrated that both methods of chemical cross-linking improved the elasticity and tensile strength of the collagen implants. Differential scanning calorimetry and biocompatibility studies indicate comparable, and in some cases, enhanced properties compared to that of the EDC/NHS controls. In vitro studies showed that human corneal epithelial cells and neuronal progenitor cell lines proliferated on these hydrogels. In addition, improvement of cell proliferation on the surfaces of the materials was observed when neurite promoting laminin epitope, IKVAV, and adhesion peptide, YIGSR, were incorporated. However, the elasticity decreased with peptide incorporation and will require further optimization. Nevertheless, we have shown that epoxy cross-linkers should be further explored in the fabrication of collagen-based hydrogels, as alternatives to or in conjunction with carbodiimide cross-linkers.
Highlights
Organization priority disease, in the developing world [1]
In addition to improving the mechanical properties, we explored methods for enhancing cell and nerve in-growth into our Butanediol diglycidyl ether (BDDGE) cross-linked collagen-based corneal implants. To enhance both cell and nerve in-growth, we examined the incorporation of two different laminin peptides, YIGSR and IKVAV, into the collagen hydrogel via Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/NHS and BDDGE
We conducted a series of experiments to determine the feasibility of directly cross-linking collagen using BDDGE
Summary
Organization priority disease, in the developing world [1]. While many causes of corneal blindness are treatable by donor human corneal transplantation, there is a severe shortage of high quality donor tissues despite innovations such as split grafts [2] and the use of gamma sterilization techniques to allow for processing of otherwise suboptimal tissue implants. In addition to improving the mechanical properties, we explored methods for enhancing cell and nerve in-growth into our BDDGE cross-linked collagen-based corneal implants. To enhance both cell and nerve in-growth, we examined the incorporation of two different laminin peptides, YIGSR and IKVAV, into the collagen hydrogel via EDC/NHS and BDDGE cross-linking. YIGSR promoted rapid epithelial cell overgrowth and both stromal cell and neurite extension into the implants through an acrylamide backbone [20] In this project, we incorporated YIGSR and IKVAV laminin peptides using EDC/NHS coupling, directly into cell-free collagen hydrogels cross-linked with. BDDGE and examined their effect on corneal epithelial cell attachment and neurite outgrowth
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