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Decellularized Human Cornea for Reconstructing the Corneal Epithelium and Anterior Stroma

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Title: Decellularized Human Cornea for Reconstructing the Corneal Epithelium and Anterior Stroma
Author(s): A. Shafiq, Maryam; A. Gemeinhart, Richard; Y.J.T. Yue, Beatrice; R. Djalilian, Ali
Abstract: In this project, we strived to develop a decellularized human cornea to use as a scaffold for reconstructing the corneal epithelium and anterior stroma. Human cadaver corneas were decellularized by five different methods, including detergent- and nondetergent-based approaches. The success of each method on the removal of cells from the cornea was investigated. The structural integrity of decellularized corneas was compared with the native cornea by electron microscopy. The integrity of the basement membrane of the epithelium was analyzed by histology and by the expression of collagen type IV, laminin, and fibronectin. Finally, the ability of the decellularized corneas to support the growth of human corneal epithelial cells and fibroblasts was assessed in vitro. Corneas processed using Triton X-100, liquid nitrogen, and poly(ethylene glycol) resulted in incomplete removal of cellular material. Corneas processed with the use of sodium dodecyl sulfate (SDS) or with sodium chloride (NaCl) plus nucleases successfully removed all cellular material; however, only the NaCl plus nuclease treatment kept the epithelial basement membrane completely intact. Corneas processed with NaCl plus nuclease supported both fibroblast and epithelial cell growth in vitro, while corneas treated with SDS supported the growth of only fibroblasts and not epithelial cells. Decellularized human corneas provide a scaffold that can support the growth of corneal epithelial cells and stromal fibroblasts. This approach may be useful for reconstructing the anterior cornea and limbus using autologous cells.
Issue Date: 2011
Publisher: Mary Ann Liebert
Citation Info: Shafiq, M. A., Gemeinhart, R. A., Yue, B. Y., & Djalilian, A. R. 2011. Decellularized Human Cornea for Reconstructing the Corneal Epithelium and Anterior Stroma. Tissue Engineering Part C Methods Volume 18, Number 5, 340-348. DOI: 10.1089/ten.TEC.2011.0072
Type: Article
Description: This is a copy of an article published in the Tissue Engineering Part C Methods © 2011 Mary Ann Liebert DOI: 10.1089/ten.TEC.2011.0072
ISSN: 1937-3384
Sponsor: This research was supported by a Fight for Sight Grantin- Aid, Illinois Society for the Prevention of Blindness, unrestricted grant from Research to Prevent Blindness, a career development grant K08EY017561-A1 to ARD, and a core grant EY01792 both fromthe National Institutes of Health, Bethesda, Maryland. ARD is also the recipient of a Career Development Award from Research to Prevent Blindness. This is also supported by a grant fromthe Eye Bank Association of America to MAS. The authors thank Ruth Zelkha, MS, for technical assistance in preparation of the images and the Illinois Eye Bank for generously providing human corneal tissue.
Date Available in INDIGO: 2012-08-15

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