Regeneration of Corneal Epithelium With Dental Pulp Stem Cells Using a Contact Lens Delivery System

• Published in: Investigative ophthalmology & visual science Vol. 57; no. 13; p. 5192
• Main Authors: Kushnerev, Evgeny, Shawcross, Susan G, Sothirachagan, Shankari, Carley, Fiona, Brahma, Arun, Yates, Julian M, Hillarby, M Chantal
• Format: Journal Article
• Language: English
• Published: United States 01.10.2016
• Subjects: Adult; Aged; Aged, 80 and over; Burns, Chemical - diagnosis; Burns, Chemical - surgery; Cells, Cultured; Contact Lenses; Dental Pulp - cytology; Dental Pulp - transplantation; Epithelium, Corneal - physiology; Eye Burns - diagnosis; Eye Burns - surgery; Female; Humans; Male; Microscopy, Confocal; Middle Aged; Regeneration - physiology; Stem Cell Transplantation - methods; Stem Cells - cytology; Wound Healing
• ISSN: 1552-5783; 1552-5783
• DOI: 10.1167/iovs.15-17953

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 In the case of injury or disease, LSCs can be lost or damaged to a point at which the corneal epithelial layer is no longer maintained.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. 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. 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. In this study we show that DPSCs, delivered using CLs, can be used to enhance repair and regeneration of the human corneal epithelium.