Published online Feb 26, 2019. doi: 10.4252/wjsc.v11.i2.84
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.
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.