title: Conjunctiva reconstruction by induced differentiation of human amniotic epithelial cells.
abstract: In this study, we aimed to investigate the feasibility of directed differentiation of human amniotic epithelial cells into conjunctival epithelium under specific conditions as well as of constructing tissue-engineered conjunctiva for ocular surface reconstruction. Human amniotic epithelial cells were cultured with induced denuded conjunctival matrix and conjunctival homogenate. Immunohistochemistry of cytokeratin-4, cytokeratin-13, and muc5ac as well as PAS staining were performed. The concentration of muc5ac at different times was measured using ELISA. The differentiated cells with quantum dots were transferred onto a denuded amniotic membrane to establish tissue-engineered conjunctiva and transplanted into a rabbit model with a conjunctival defect. After induction of human amniotic epithelial cells, differentiated cells showed conjunctival epithelium phenotype, while trace amounts of mu5ac in the culture medium measured by ELISA increased gradually within 1 to 7 days. Successfully tissue-engineered conjunctiva had similar structure as normal conjunctiva and was transplanted into a rabbit model with conjunctiva defect. After 2 weeks post-surgery, conjunctiva grafts survived and were integrated. Immunohistochemistry showed conjunctival epithelium phenotype, positive cells were found in PAS staining. Thus, human amniotic epithelial cells could differentiate into conjunctival epithelium-like cells and goblet cells with partially physiological function, and we successfully restored ocular surface integrity in the rabbit model using tissue-engineered conjunctiva.
authors.affiliation: Department of Ophthalmology, People's Hospital of Taizhou City, Taizhou, China.
authors.forename: S P
authors.initials: SP
authors.lastname: Yang
authors.affiliation: Department of Ophthalmology, People's Hospital of Taizhou City, Taizhou, China.
authors.forename: X Z
authors.initials: XZ
authors.lastname: Yang
authors.affiliation: Division of Liver Disease, People' s Hospital of Taizhou, Jiangsu, China.
authors.forename: G P
authors.initials: GP
authors.lastname: Cao
mesh_terms: D000650:Amnion / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D003228:Conjunctiva; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D023421:Models, Animal; D011817:Rabbits; D012038:Regeneration; D023822:Tissue Engineering
chemical_list: D015415:Biomarkers