title: Amniotic membrane immobilized poly(vinyl alcohol) hybrid polymer as an artificial cornea scaffold that supports a stratified and differentiated corneal epithelium.
abstract: Poly(vinyl alcohol) (PVA) is a biocompatible, transparent hydrogel with physical strength that makes it promising as a material for an artificial cornea. In our previous study, type I collagen was immobilized onto PVA (PVA-COL) as a possible artificial cornea scaffold that can sustain a functional corneal epithelium. The cellular adhesiveness of PVA in vitro was improved by collagen immobilization; however, stable epithelialization was not achieved in vivo. To improve epithelialization in vivo, we created an amniotic membrane (AM)-immobilized polyvinyl alcohol hydrogel (PVA-AM) for use as an artificial cornea material. AM was attached to PVA-COL using a tissue adhesive consisting of collagen and citric acid derivative (CAD) as a crosslinker. Rabbit corneal epithelial cells were air-lift cultured with 3T3 feeder fibroblasts to form a stratified epithelial layer on PVA-AM. The rabbit corneal epithelial cells formed 3-5 layers of keratin-3-positive epithelium on PVA-AM. Occludin-positive cells were observed lining the superficial epithelium, the gap-junctional protein connexin43-positive cells was localized to the cell membrane of the basal epithelium, while both collagen IV were observed in the basement membrane. Epithelialization over implanted PVA-AM was complete within 2 weeks, with little inflammation or opacification of the hydrogel. Corneal epithelialization on PVA-AM in rabbit corneas improved over PVA-COL, suggesting the possibility of using PVA-AM as a biocompatible hybrid material for keratoprosthesis.
authors.affiliation: Department of Ophthalmology, Tokyo Dental College, Chiba, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Uchino
authors.forename: Shigeto
authors.initials: S
authors.lastname: Shimmura
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Miyashita
authors.forename: Tetsushi
authors.initials: T
authors.lastname: Taguchi
authors.forename: Hisatoshi
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Jun
authors.initials: J
authors.lastname: Shimazaki
authors.forename: Junzo
authors.initials: J
authors.lastname: Tanaka
authors.forename: Kazuo
authors.initials: K
authors.lastname: Tsubota
mesh_terms: D016475:3T3 Cells; D000650:Amnion / Q000737:chemistry*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D024141:Collagen Type IV / Q000032:analysis; D018031:Connexin 43 / Q000032:analysis; D003315:Cornea / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D004848:Epithelium / Q000737:chemistry / Q000187:drug effects; D053558:Keratin-3 / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D062793:Occludin; D011108:Polymers / Q000737:chemistry; D011142:Polyvinyl Alcohol / Q000737:chemistry / Q000494:pharmacology*; D019736:Prostheses and Implants; D011817:Rabbits
chemical_list: D001672:Biocompatible Materials; D024141:Collagen Type IV; D018031:Connexin 43; D053558:Keratin-3; D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse; D011108:Polymers; D011142:Polyvinyl Alcohol; D003094:Collagen