title: Seeding of corneal wounds by epithelial cell transfer from micropatterned PDMS contact lenses.
abstract: Persistent corneal wounds result from numerous eye disorders, and to date, available treatments often fail to accelerate reepithelialization, the key initial step in wound healing. To speed reepithelialization, we explored a cell-transfer transplant method utilizing polydimethylsiloxane (PDMS) contact lenses to deliver epithelial cells derived from limbal explants directly within a corneal wound. Human primary epithelial cells and an immortalized corneal epithelial cell line (HCE-SV40) grew well on PDMS contact lenses and their morphology and growth rates where similar to cells grown on tissue culture polystyrene. To initially study cell transfer from PDMS, HCE-SV40 cells were seeded onto PDMS with or without micropatterned posts. After a day in culture, HCE-SV40 cells attached to the unpatterned PDMS uniformly, whereas on micropatterned PDMS they appeared to attach primarily between posts. The cell-covered PDMS contacts were then placed cell-side down onto tissue culture plastic and, after 1, 2, or 3 days, the PDMS contact was removed and the transferred cells were trypsinized and counted. Micropatterned PDMS contact lenses with 100-microm-diameter posts and a post height of 40 microm transferred three times as many cells as unpatterned PDMS. Cell transfer to a wounded cornea was tested in a pig cornea organ culture model de-epithelialized by alkali treatment. Post micropatterned PDMS contact lenses were seeded with labeled HCE-SV40 cells at a density 50,000 cells/cm2 and applied to the wounded pig corneas. After 24, 48, or 96 h of application, PDMS contact lenses were removed, corneas fixed with formaldehyde, and sectioned. After 48 h, epithelial cells transferred from post micropatterned contact lenses to provide 35% epithelial coverage of denuded pig corneas; after 96 h coverage was 65%. We conclude that cell transfer from epithelial-coated PDMS contact lenses micropatterned with posts provides a promising approach to reepithelialize corneal surfaces.
authors.affiliation: Biomedical Engineering, Vanderbilt University, Nashville, TN 37235, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Pino
authors.forename: Frederick R
authors.initials: FR
authors.lastname: Haselton
authors.forename: Min S
authors.initials: MS
authors.lastname: Chang
mesh_terms: D002465:Cell Movement; D002478:Cells, Cultured; D003261:Contact Lenses; D003315:Cornea / Q000601:surgery; D065306:Corneal Injuries; D004129:Dimethylpolysiloxanes / Q000008:administration & dosage*; D019573:Epithelium, Corneal / Q000637:transplantation*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012828:Silicones / Q000008:administration & dosage*; D013997:Time Factors; D014945:Wound Healing
chemical_list: D004129:Dimethylpolysiloxanes; D012828:Silicones; C013830:baysilon