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
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 sign...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2016-01-01
|
| Series: | Journal of Ophthalmology |
| Online Access: | http://dx.doi.org/10.1155/2016/4805986 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832567136088752128 |
|---|---|
| author | Fawzia Bardag-Gorce Richard H. Hoft Andrew Wood Joan Oliva Hope Niihara Andrew Makalinao Jacquelyn Thropay Derek Pan Imara Meepe Kumar Tiger Julio Garcia Amanda Laporte Samuel W. French Yutaka Niihara |
| author_facet | Fawzia Bardag-Gorce Richard H. Hoft Andrew Wood Joan Oliva Hope Niihara Andrew Makalinao Jacquelyn Thropay Derek Pan Imara Meepe Kumar Tiger Julio Garcia Amanda Laporte Samuel W. French Yutaka Niihara |
| author_sort | Fawzia Bardag-Gorce |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-db8be70059d54ec2b486545a2d7d1537 |
| institution | Kabale University |
| issn | 2090-004X 2090-0058 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Ophthalmology |
| spelling | doaj-art-db8be70059d54ec2b486545a2d7d15372025-02-03T01:02:19ZengWileyJournal of Ophthalmology2090-004X2090-00582016-01-01201610.1155/2016/48059864805986The 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 DeficiencyFawzia Bardag-Gorce0Richard H. Hoft1Andrew Wood2Joan Oliva3Hope Niihara4Andrew Makalinao5Jacquelyn Thropay6Derek Pan7Imara Meepe8Kumar Tiger9Julio Garcia10Amanda Laporte11Samuel W. French12Yutaka Niihara13Los Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USALos Angeles Biomedical Research Institute (LA BioMed), Harbor UCLA Medical Center, Torrance, CA 90502, USAThe 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.http://dx.doi.org/10.1155/2016/4805986 |
| spellingShingle | Fawzia Bardag-Gorce Richard H. Hoft Andrew Wood Joan Oliva Hope Niihara Andrew Makalinao Jacquelyn Thropay Derek Pan Imara Meepe Kumar Tiger Julio Garcia Amanda Laporte Samuel W. French Yutaka Niihara 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 Journal of Ophthalmology |
| 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| url | http://dx.doi.org/10.1155/2016/4805986 |
| work_keys_str_mv | AT fawziabardaggorce theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT richardhhoft theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT andrewwood theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT joanoliva theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT hopeniihara theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT andrewmakalinao theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT jacquelynthropay theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT derekpan theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT imarameepe theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT kumartiger theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT juliogarcia theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT amandalaporte theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT samuelwfrench theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT yutakaniihara theroleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT fawziabardaggorce roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT richardhhoft roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT andrewwood roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT joanoliva roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT hopeniihara roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT andrewmakalinao roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT jacquelynthropay roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT derekpan roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT imarameepe roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT kumartiger roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT juliogarcia roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT amandalaporte roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT samuelwfrench roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency AT yutakaniihara roleofecadherininmaintainingthebarrierfunctionofcornealepitheliumaftertreatmentwithculturedautologousoralmucosaepithelialcellsheetgraftsforlimbalstemdeficiency |