December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
A Collagen-based Scaffold for a Tissue Engineered Human Cornea: Physical and Physiological Properties
Author Affiliations & Notes
  • MA Watsky
    Physiology University of Tennessee Health Science Center Memphis TN
  • CJ Doillon
    Oncology and Molecular Endocrinology Research Centre Laval University Quebec City PQ Canada
  • M Hakim
    University of Ottawa Eye Institute University of Ottawa Ottawa ON Canada
  • J Wang
    Memphis TN
  • R Munger
    Ottawa ON Canada
  • N Laycock
    Ottawa ON Canada
  • M Griffith
    Ottawa ON Canada
  • Footnotes
    Commercial Relationships    M.A. Watsky, Biowhittaker P; C.J. Doillon, Biowhittaker P; M. Hakim, None; J. Wang, None; R. Munger, None; N. Laycock, None; M. Griffith, Biowhittaker P. Grant Identification: MRC-PMAC PA13983, P&G IPAA, NSERC studentship
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 2996. doi:
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      MA Watsky, CJ Doillon, M Hakim, J Wang, R Munger, N Laycock, M Griffith; A Collagen-based Scaffold for a Tissue Engineered Human Cornea: Physical and Physiological Properties . Invest. Ophthalmol. Vis. Sci. 2002;43(13):2996.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract: : Purpose: To develop and characterize a stabilized collagen-glycosaminoglycans (GAG) based matrix that serves as a scaffold for a tissue engineered human corneal equivalent. Methods: Immortalized human corneal cells similar to low passage primary cells were used. Hydrated matrices were constructed by blending neutralized type I collagen (bovine dermis or rat tail tendon) with one of the following: chondroitin sulphates A, B, C or collagen type III, and then stabilizing by glutaraldehyde crosslinking. Glycine was added to deactivate excess glutaraldehyde, and stromal cells were mixed into the matrix. For some experiments, epithelial and endothelial cells were added. Constructs were supplemented with growth media, with or without ascorbic acid. Some were treated with chemicals to cause different degrees of wounding to evaluate their ability to recover. Results: Native, non-crosslinked gels were soft and exhibited limited longevity. Glutaraldehyde cross-linking strengthened the matrix while still permitting cell growth. FTIR spectra indicated that bovine collagen was more easily cross-linked by glutaraldehyde than rat tail collagen. Chondroitin sulphate addition increased transparency. Keratocytes grown within the matrix had a higher frequency of K+ channel expression than identical keratocytes grown on a culture dish. Ascorbic acid addition increased degradation rate of the uncrosslinked matrix in the presence of keratocytes, but enhanced keratocyte proliferation and endogenous collagen synthesis. Wounded constructs showed recovery from chemical exposure. Conclusion: Presence of keratocytes and ascorbic acid influences the scaffold properties. A very low concentration of glutaraldehyde carries out most of the biophysical and biological characteristics required for a reliable stroma substitute.

Keywords: 374 cornea: stroma and keratocytes • 370 cornea: basic science • 403 extracellular matrix 

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