Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model

Morales, Mariana Lizeth Orozco; Marsit, Nagi M.; McIntosh, Owen D.; Hopkinson, Andrew; Sidney, Laura E.

Abstract

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

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

METHODS
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.

RESULTS
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.

CONCLUSION
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.

Citation

Morales, M. L. O., Marsit, N. M., McIntosh, O. D., Hopkinson, A., & Sidney, L. E. (2019). Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model. World Journal of Stem Cells, 11(2), 84-99. https://doi.org/10.4252/wjsc.v11.i2.84

• Journal Article Type: Article
• Acceptance Date: Jan 1, 2019
• Online Publication Date: Feb 26, 2019
• Publication Date: Feb 26, 2019
• Deposit Date: Mar 12, 2019
• Publicly Available Date: Mar 13, 2019
• Journal: World Journal of Stem Cells
• Electronic ISSN: 1948-0210
• Publisher: Baishideng Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 2
• Pages: 84-99
• DOI: https://doi.org/10.4252/wjsc.v11.i2.84
• Keywords: Cornea; Corneal injuries; Injury model; Corneal epithelium; Corneal stromaderived stem cells; Amnion; Anti-inflammatory; Cell therapy
• Public URL: https://nottingham-repository.worktribe.com/output/1633259
• Contract Date: Mar 12, 2019