4.6 Article

Bio-fabrication of stem-cell-incorporated corneal epithelial and stromal equivalents from silk fibroin and gelatin-based biomaterial for canine corneal regeneration

Journal

PLOS ONE
Volume 17, Issue 2, Pages -

Publisher

PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0263141

Keywords

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Funding

  1. 100th Anniversary Chulalongkorn University Fund for Doctoral Scholarship
  2. 90th Anniversary Chulalongkorn University Fund
  3. Veterinary Stem Cell and Bioengineering Research Unit, Ratchadaphiseksomphot Endowment Fund of Chulalongkorn University
  4. Chulalongkorn University [CU_GR_62_98_21_15]
  5. Faculty of Veterinary Science
  6. Chulalongkorn Academic Advancement into Its 2nd Century Project
  7. Center of Excellence for Regenerative Dentistry(CERD), Faculty of Dentistry, Chulalongkorn University
  8. Veterinary Stem Cell and Bioengineering Research Unit, Ratchadaphiseksomphot Endowment Fund, Chulalongkorn University
  9. Government Research Fund

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This study aimed to generate tissue-engineered canine cornea using canine limbal epithelial stem cells (cLESCs) and canine corneal stromal stem cells (cCSSCs). The results showed that using silk fibroin/gelatin as a substrate promoted cell adhesion, viability, and proliferation, and differentiated keratocytes expressed proteins associated with native cornea.
Corneal grafts are the imperative clinical treatment for canine corneal blindness. To serve the growing demand, this study aimed to generate tissue-engineered canine cornea in part of the corneal epithelium and underlying stroma based on canine limbal epithelial stem cells (cLESCs) seeded silk fibroin/gelatin (SF/G) film and canine corneal stromal stem cells (cCSSCs) seeded SF/G scaffold, respectively. Both cell types were successfully isolated by collagenase I. SF/G corneal films and stromal scaffolds served as the prospective substrates for cLESCs and cCSSCs by promoting cell adhesion, cell viability, and cell proliferation. The results revealed the upregulation of tumor protein P63 (P63) and ATP-binding cassette super-family G member 2 (Abcg2) of cLESCs as well as Keratocan (Kera), Lumican (Lum), aldehyde dehydrogenase 3 family member A1 (Aldh3a1) and Aquaporin 1 (Aqp1) of differentiated keratocytes. Moreover, immunohistochemistry illustrated the positive staining of tumor protein P63 (P63), aldehyde dehydrogenase 3 family member A1 (Aldh3a1), lumican (Lum) and collagen I (Col-I), which are considerable for native cornea. This study manifested a feasible platform to construct tissue-engineered canine cornea for functional grafts and positively contributed to the body of knowledge related to canine corneal stem cells.

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