title: The Role of E-Cadherin in Maintaining the Barrier Function of Corneal Epithelium after Treatment with Cultured Autologous Oral Mucosa Epithelial Cell Sheet Grafts for Limbal Stem Deficiency.
abstract: The role of E-cadherin in epithelial barrier function of cultured autologous oral mucosa epithelial cell sheet (CAOMECS) grafts was examined. CAOMECS were cultured on a temperature-responsive surface and grafted onto rabbit corneas with Limbal Stem Cell Deficiency (LSCD). E-cadherin levels were significantly higher in CAOMECS compared to normal and LSCD epithelium. Beta-catenin colocalized with E-cadherin in CAOMECS cell membranes while phosphorylated beta-catenin was significantly increased. ZO-1, occludin, and Cnx43 were also strongly expressed in CAOMECS. E-cadherin and beta-catenin localization at the cell membrane was reduced in LSCD corneas, while CAOMECS-grafted corneas showed a restoration of E-cadherin and beta-catenin expression. LSCD corneas did not show continuous staining for ZO-1 or for Cnx43, while CAOMECS-grafted corneas showed a positive expression of ZO-1 and Cnx43. Cascade Blue® hydrazide did not pass through CAOMECS. Because E-cadherin interactions are calcium-dependent, EGTA was used to chelate calcium and disrupt cell adhesion. EGTA-treated CAOMECS completely detached from cell culture surface, and E-cadherin levels were significantly decreased. In conclusion, E cadherin high expression contributed to CAOMECS tight and gap junction protein recruitment at the cell membrane, thus promoting cellular adhesion and a functional barrier to protect the ocular surface.
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Fawzia
authors: 0000-0001-9433-7269
authors: F
authors: Bardag-Gorce
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Richard H
authors: RH
authors: Hoft
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Andrew
authors: A
authors: Wood
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Joan
authors: J
authors: Oliva
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Hope
authors: H
authors: Niihara
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Andrew
authors: A
authors: Makalinao
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Jacquelyn
authors: J
authors: Thropay
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Derek
authors: D
authors: Pan
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Imara
authors: 0000-0001-8291-8365
authors: I
authors: Meepe
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Kumar
authors: K
authors: Tiger
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Julio
authors: J
authors: Garcia
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Amanda
authors: A
authors: Laporte
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Samuel W
authors: SW
authors: French
authors: Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USA.
authors: Yutaka
authors: Y
authors: Niihara