Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Feb 26, 2019; 11(2): 84-99
Published online Feb 26, 2019. doi: 10.4252/wjsc.v11.i2.84
Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model
Laura E Sidney, Andrew Hopkinson, Owen D McIntosh, Nagi M Marsit, Mariana Lizeth Orozco Morales
Mariana Lizeth Orozco Morales, Nagi M Marsit, Owen D McIntosh, Andrew Hopkinson, Laura E Sidney, Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom
Author contributions: Orozco Morales ML performed the majority of the data acquisition and analysis; Marsit NM processed all amniotic membrane used and helped with acquisition of data; McIntosh OD contributed to conception of the study and acquisition of data; Hopkinson A contributed to conception of the study; Sidney LE contributed to conception of the study, design of experiments, data acquisition, data analysis, and interpretation of data; all authors contributed to drafting the article, making revisions, and had final approval of the manuscript.
Institutional review board statement: Human corneoscleral rims and human AM were used with approval by the Nottingham Research Ethics Committee (Ethics approval numbers: 07/H0403/140 and OY110101, respectively).
Informed consent statement: All corneal tissue was supplied anonymously through Manchester Eye Bank, and all informed consent forms are held at this institution. For amniotic membrane, all donors provided written informed consent prior to tissue collection. During research studies all tissue was anonymised to the researchers.
Conflict-of-interest statement: There are no potential conflicts of interest relevant to this study reported by the authors.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Laura E Sidney, MSc, PhD, Senior Research Fellow, Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Queen’s Medical Centre Campus, Nottingham NG7 2UH, United Kingdom.
Telephone: +44-115-8230459
Received: July 24, 2018
Peer-review started: July 24, 2018
First decision: October 5, 2018
Revised: November 1, 2018
Accepted: January 1, 2019
Article in press: January 1, 2019
Published online: February 26, 2019

An in vitro injury model mimicking a corneal surface injury was optimised using human corneal epithelial cells (hCEC).


To investigate whether corneal-stroma derived stem cells (CSSC) seeded on an amniotic membrane (AM) construct manifests an anti-inflammatory, healing response.


Treatment of hCEC with ethanol and pro-inflammatory cytokines were compared in terms of viability loss, cytotoxicity, and pro-inflammatory cytokine release, in order to generate the in vitro injury. This resulted in an optimal injury of 20% (v/v) ethanol for 30 s with 1 ng/mL interleukin-1 (IL-1) beta. Co-culture experiments were performed with CSSC alone and with CSSC-AM constructs. The effect of injury and co-culture on viability, cytotoxicity, IL-6 and IL-8 production, and IL1B, TNF, IL6, and CXCL8 mRNA expression were assessed.


Co-culture with CSSC inhibited loss of hCEC viability caused by injury. Enzyme linked immunosorbent assay and polymerase chain reaction showed a significant reduction in the production of IL-6 and IL-8 pro-inflammatory cytokines, and reduction in pro-inflammatory cytokine mRNA expression during co-culture with CSSC alone and with the AM construct. These results confirmed the therapeutic potential of the CSSC and the possible use of AM as a cell carrier for application to the ocular surface.


CSSC were shown to have a potentially therapeutic anti-inflammatory effect when treating injured hCEC, demonstrating an important role in corneal regeneration and wound healing, leading to an improved knowledge of their potential use for research and therapeutic purposes.

Keywords: Cornea, Corneal injuries, Injury model, Corneal epithelium, Corneal stroma-derived stem cells, Amnion, Anti-inflammatory, Cell therapy

Core tip: We designed a novel in vitro inflammation model of the human corneal surface using human corneal epithelial cells treated with 20% (v/v) ethanol, followed by stimulation with 1 ng/mL interleukin-1β. We then used this model to demonstrate the anti-inflammatory and regenerative healing properties of human cornea stroma-derived stem cells seeded on an amniotic membrane substrate in a co-culture model. This study is the first step in building a topical regenerative therapy for the treatment of inflammatory disorders of the front of the eye.