title: Regeneration of Corneal Epithelium With Dental Pulp Stem Cells Using a Contact Lens Delivery System.
abstract: OBJECTIVE
  The corneal epithelium is sloughed off surface of the eye by the action of blinking and is continually replaced by division and maturation of the limbal stem cells (LSCs). In the case of injury or disease, LSCs can be lost or damaged to a point at which the corneal epithelial layer is no longer maintained. leading to LSC deficiencies (LSCDs). When this occurs, the opaque conjunctiva overgrows the anterior surface of the eye, leading to vision impairment or loss. Dental pulp stem cells (DPSCs) are promising candidates as autologous LSC substitutes. In this study, contact lenses (CLs) are used as a novel medical device to deliver DPSCs onto corneal surface to enhance corneal epithelium regeneration.
  
  
  METHODS
  Dental pulp stem cells labeled with green fluorescent Qtracker 525 were seeded onto the pretreated CLs, allowed to adhere, then delivered to debrided human corneas. Expression of KRT3, 12, 13, and 19 was investigated by immunostaining, then standard and confocal microscopy.
  
  
  RESULTS
  Dental pulp stem cells were successfully isolated, labeled, and delivered to the corneal surface using CLs. Following removal of CLs, confocal microscopy showed that the DPSCs had migrated onto the cornea. Coexpression of KRT12 and green fluorescent Qtracker 525 confirmed that the DPSCs had transdifferentiated into corneal epithelial progenitors. Delimitation of KRT 19 and green fluorescence provides evidence that Qtracker 525-labeled DPSCs establish a barrier to the invasion of the cornea by conjunctiva.
  
  
  CONCLUSIONS
  In this study we show that DPSCs, delivered using CLs, can be used to enhance repair and regeneration of the human corneal epithelium.
authors.affiliation: School of Dentistry, Stopford Building, University of Manchester, Manchester, United Kingdom.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Kushnerev
authors.affiliation: Blond McIndoe Laboratories, School of Biological Sciences, Faculty of Biology, Medicine, and Health, University of Manchester, Manchester, United Kingdom.
authors.forename: Susan G
authors.initials: SG
authors.lastname: Shawcross
authors.affiliation: Division of Pharmacy and Optometry, School of Health Sciences, Faculty of Biology, Medicine, and Health, University of Manchester, Manchester, United Kingdom.
authors.forename: Shankari
authors.initials: S
authors.lastname: Sothirachagan
authors.affiliation: Manchester Royal Eye Hospital, Oxford Road, Manchester, United Kingdom.
authors.forename: Fiona
authors.initials: F
authors.lastname: Carley
authors.affiliation: Manchester Royal Eye Hospital, Oxford Road, Manchester, United Kingdom.
authors.forename: Arun
authors.initials: A
authors.lastname: Brahma
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Dentistry, JR Moore Building, University of Manchester, Manchester, United Kingdom.
authors.forename: Julian M
authors.initials: JM
authors.lastname: Yates
authors.affiliation: Division of Pharmacy and Optometry, School of Health Sciences, Faculty of Biology, Medicine, and Health, University of Manchester, Manchester, United Kingdom.
authors.forename: M Chantal
authors.initials: MC
authors.lastname: Hillarby
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002057:Burns, Chemical / Q000175:diagnosis / Q000601:surgery; D002478:Cells, Cultured; D003261:Contact Lenses; D003782:Dental Pulp / Q000166:cytology* / Q000637:transplantation; D019573:Epithelium, Corneal / Q000502:physiology*; D005126:Eye Burns / Q000175:diagnosis / Q000601:surgery; D005260:Female; D006801:Humans; D008297:Male; D018613:Microscopy, Confocal; D008875:Middle Aged; D012038:Regeneration / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology*; D014945:Wound Healing