Corneal regeneration by induced human buccal mucosa cultivated on an amniotic membrane following alkaline injury

Rohaina Che Man, Then Kong Yong, Min Hwei Ng, Wan Haslina Wan Abdul Halim, Aida Zairani Mohd Zahidin, Roszalina Ramli, Aminuddin Bin Saim, Ruszymah Idrus

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Various clinical disorders and injuries, such as chemical, thermal, or mechanical injuries, may lead to corneal loss that results in blindness. PURPOSE: The aims of this study were to differentiate human buccal mucosa (BMuc) into corneal epithelial-like cells, to fabricate engineered corneal tissue using buccal mucosal epithelial cells, and to reconstruct a damaged corneal epithelium in a nude rat model. Methods: BMuc were subjected to 10 d of induction factors to investigate the potential of cells to differentiate into corneal lineages Results: Corneal stem cell markers β1-integrin, C/EBPδ, ABCG2, p63, and CK3 were upregulated in the gene expression analysis in induced BMuc, whereas CK3 and p63 showed significant protein expression in induced BMuc compared to the uninduced cells. BMuc were then left to reach 80% confluency after differential trypsinization. The cells were harvested and cultivated on a commercially available untreated air-dried amniotic membrane (AM) in a Transwell system in induction medium. The corneal constructs were fabricated and then implanted into damaged rat corneas for up to 8 weeks. A significant improvement was detected in the treatment group at 8 weeks post-implantation, as revealed by slit lamp biomicroscopy analysis. The structure and thickness of the corneal layer were also analyzed using histological staining and time-domain optical coherence tomography scans and were found to resemble a native corneal layer. The protein expression for CK3 and p63 were continuously detected throughout the corneal epithelial layer in the corneal construct. Conclusions: In conclusion, human BMuc can be induced to express a corneal epithelial-like phenotype. The addition of BMuc improves corneal clarity, prevents vascularization, increases corneal thickness and stromal alignment, and appears to have no adverse effect on the host after implantation

Original languageEnglish
Pages (from-to)810-822
Number of pages13
JournalMolecular Vision
Volume23
Publication statusPublished - 21 Nov 2017

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Amnion
Mouth Mucosa
Regeneration
Wounds and Injuries
Epithelial Cells
Nude Rats
Corneal Epithelium
Cheek
Optical Coherence Tomography
Blindness
Integrins
Cornea
Proteins
Stem Cells
Hot Temperature
Air
Staining and Labeling
Phenotype
Gene Expression

ASJC Scopus subject areas

  • Ophthalmology

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Corneal regeneration by induced human buccal mucosa cultivated on an amniotic membrane following alkaline injury. / Man, Rohaina Che; Yong, Then Kong; Ng, Min Hwei; Wan Abdul Halim, Wan Haslina; Mohd Zahidin, Aida Zairani; Ramli, Roszalina; Saim, Aminuddin Bin; Idrus, Ruszymah.

In: Molecular Vision, Vol. 23, 21.11.2017, p. 810-822.

Research output: Contribution to journalArticle

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