title: In vitro characterization of electrochemically compacted collagen matrices for corneal applications.
abstract: Loss of vision due to corneal disease is a significant problem worldwide. Transplantation of donor corneas is a viable treatment option but limitations such as short supply and immune-related complications call for alternative options for the treatment of corneal disease. A tissue engineering-based approach using a collagen scaffold is a promising alternative to develop a bioengineered cornea that mimics the functionality of native cornea. In this study, an electrochemical compaction method was employed to synthesize highly dense and transparent collagen matrices. We hypothesized that chemical crosslinking of electrochemically compacted collagen (ECC) matrices will maintain transparency, improve stability, and enhance the mechanical properties of the matrices to the level of native cornea. Further, we hypothesized that keratocyte cell viability and proliferation will be maintained on crosslinked ECC matrices. The results indicated that uncrosslinked and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-N-hydroxysuccinimide (EDC-NHS) crosslinked ECC matrices were highly transparent with light transmission measurements comparable to native cornea. Stability tests showed that while the uncrosslinked ECC matrices degraded within 6 h when treated with collagenase, EDC-NHS or genipin crosslinking significantly improved the stability of ECC matrices (192 h for EDC-NHS and 256 h for genipin). Results from the mechanical tests showed that both EDC-NHS and genipin crosslinking significantly improved the strength and modulus of ECC matrices. Cell culture studies showed that keratocyte cell viability and proliferation are maintained on EDC-NHS crosslinked ECC matrices. Overall, results from this study suggest that ECC matrices have the potential to be developed as a functional biomaterial for corneal repair and regeneration.
authors.affiliation: Department of Chemical Engineering, Florida Institute of Technology, Melbourne, FL 32901, USA. Department of Biomedical Engineering, Florida Institute of Technology, Melbourne, FL 32901, USA. Author to whom any correspondence should be addressed. Department of Chemical Engineering, Florida Institute of Technology, 150 W. University Blvd, Melbourne, FL 32901, USA.
authors.forename: Vipuil
authors.initials: V
authors.lastname: Kishore
authors.forename: Ranjani
authors.initials: R
authors.lastname: Iyer
authors.forename: Athela
authors.initials: A
authors.lastname: Frandsen
authors.forename: Thuy-Uyen
authors.initials: TU
authors.lastname: Nguyen
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry; D057005:Bioengineering / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000737:chemistry*; D017364:Collagenases / Q000378:metabolism; D003315:Cornea / Q000473:pathology*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004563:Electrochemistry / Q000379:methods*; D006801:Humans; D039823:Iridoids / Q000737:chemistry; D015603:Keratinocytes / Q000166:cytology; D008422:Materials Testing; D012038:Regeneration; D013314:Stress, Mechanical; D013388:Succinimides / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*
chemical_list: C000625275:1-ethyl-3-(3-dimethylaminopropyl)carbodiimide N-hydroxysuccinimide; D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D039823:Iridoids; D013388:Succinimides; D003094:Collagen; C007834:genipin; D017364:Collagenases; C001426:N-hydroxysuccinimide