Corneal regeneration by conditioned medium of human uterine cervical stem cells is mediated by TIMP-1 and TIMP-2.

Abstract

The aim of the present study was to evaluate the effect and the mechanism of action of the conditioned medium from human uterine cervical stem cells (CM-hUCESC) on corneal wound healing in a rabbit dry eye model. To do this, dry eye and corneal epithelial injuries were induced in rabbits by topical administration of atropine sulfate and NaOH. Hematoxylin-Eosin (H&E) and Ki-67 immunostaining were carried out to evaluate corneal damage and cell proliferation, and real-time PCR was used to evaluate proinflammatory cytokines in the cornea. In addition, in order to investigate possible factors involved in corneal regeneration, primary cultures of rat corneal epithelial cells (rCECs) were used to evaluate cell migration, proliferation, and apoptosis before and after immunoprecipitation of specific factors from the CM-hUCESC. Results showed that CM-hUCESC treatment significantly improved epithelial regeneration in rabbits with dry eye induced by atropine and reduced corneal pro-inflammatory TNF-α, MCP-1, MIP-1α and IL-6 cytokines. In addition, metalloproteinase inhibitors TIMP-1 and TIMP-2, which are present at high levels in CM-hUCESC, mediated corneal regenerative effects by both inducing corneal epithelial cell proliferation and inhibiting apoptosis. In summary, CM-hUCESC induces faster corneal regeneration in a rabbit model of dry eye induced by atropine than conventional treatments, being TIMP-1 and TIMP-2 mediators in this process. The results indicate that an alternative CM-based treatment for some corneal conditions is achievable, although future studies would be necessary to investigate other factors involved in the multiple observed effects of CM-hUCESC.

• Authors: Juan Sendon-Lago, Samuel Seoane, Anxo Martinez-Ordoñez, Noemi Eiro, Jorge Saa, Francisco J Vizoso, Francisco Gonzalez, Roman Perez-Fernandez, Maria A Bermudez
• Journal: Experimental eye research (Exp Eye Res) Vol. 180 Pg. 110-121 (03 2019) ISSN: 1096-0007 [Electronic] England
• PMID: 30557571 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2018 Elsevier Ltd. All rights reserved.

Chemical References

• Culture Media, Conditioned
• Cytokines
• Ki-67 Antigen
• Tissue Inhibitor of Metalloproteinase-1
• Tissue Inhibitor of Metalloproteinase-2
• Sodium Hydroxide
• Atropine

Topics

• Animals
• Apoptosis
• Atropine (toxicity)
• Blotting, Western
• Cell Movement
• Cell Proliferation
• Cervix Uteri (cytology)
• Culture Media, Conditioned (pharmacology)
• Cytokines (genetics)
• Disease Models, Animal
• Dry Eye Syndromes (chemically induced, drug therapy, metabolism)
• Epithelium, Corneal (physiology)
• Female
• Ki-67 Antigen (metabolism)
• Male
• Rabbits
• Rats
• Rats, Sprague-Dawley
• Real-Time Polymerase Chain Reaction
• Regeneration (physiology)
• Sodium Hydroxide (toxicity)
• Stem Cells (cytology)
• Tandem Mass Spectrometry
• Tissue Inhibitor of Metalloproteinase-1 (metabolism)
• Tissue Inhibitor of Metalloproteinase-2 (metabolism)
• Wound Healing (drug effects)