Understanding Ocular Surface Inflammation in Tears Before and After Autologous Cultivated Limbal Epithelial Stem Cell Transplantation

Gustavo S Figueiredo; Jeffry Hogg; Arthur Okonkwo; Oliver J Baylis; Monica Berry; Simi Ali; Majlinda Lako; Francisco C Figueiredo

doi:10.1007/s40123-023-00656-6

Understanding Ocular Surface Inflammation in Tears Before and After Autologous Cultivated Limbal Epithelial Stem Cell Transplantation

Ophthalmol Ther. 2023 Apr;12(2):1097-1107. doi: 10.1007/s40123-023-00656-6. Epub 2023 Jan 28.

Authors

Gustavo S Figueiredo^{1

2}, Jeffry Hogg^{1

2}, Arthur Okonkwo^{1

2}, Oliver J Baylis^{1

2}, Monica Berry³, Simi Ali⁴, Majlinda Lako², Francisco C Figueiredo^{5

6}

Affiliations

¹ Department of Ophthalmology, Royal Victoria Infirmary, Newcastle upon Tyne Hospitals NHS Foundation Trust, Queen Victoria Road, Newcastle upon Tyne, NE1 4LP, UK.
² Faculty of Medical Sciences, Biosciences Institute, Newcastle University, Newcastle upon Tyne, UK.
³ HH Wills Physics Laboratory, University of Bristol, Bristol, UK.
⁴ Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, UK.
⁵ Department of Ophthalmology, Royal Victoria Infirmary, Newcastle upon Tyne Hospitals NHS Foundation Trust, Queen Victoria Road, Newcastle upon Tyne, NE1 4LP, UK. Francisco.figueiredo@newcastle.ac.uk.
⁶ Faculty of Medical Sciences, Biosciences Institute, Newcastle University, Newcastle upon Tyne, UK. Francisco.figueiredo@newcastle.ac.uk.

Abstract

Introduction: We aimed to determine the expression of inflammatory cytokines in the tears of patients with unilateral total limbal stem cell deficiency (TLSCD) caused by chemical burns before and after autologous cultivated limbal epithelial stem cell transplantation (CLET).

Methods: Tear samples were collected from both eyes of 23 patients with unilateral TLSCD and 11 healthy controls, at fixed timepoints before and after CLET. Dissolved molecules were extracted from Schirmer's strips using a standardised method and analysed on an array plate of ten inflammatory cytokines (V-Plex Proinflammatory Panel 1 Human Kit, MSD).

Results: IL1β expression was significantly elevated in the TLSCD eye compared with the unaffected eye at baseline (p < 0.0001) but decreased to normal 3 months post-CLET (p = 0.22). IL6 and IL8 were unaffected at baseline but significantly elevated in the TLSCD eyes at 1 month post-CLET (p = 0.001 and p < 0.0001, respectively). IL6 returned to normal at 3 months and IL8 at 6 months post-CLET. There was a significant renewed increase in IL1β, IL6 and IL8 expression at 12 months post-CLET (p < 0.0001, p = 0.0001 and p = 0.0003, respectively). IFNγ, IL10 and IL12p70 expression were significantly reduced in both eyes of patients with unilateral TLSCD at all timepoints.

Conclusion: IL1β is a specific marker of inflammation in TLSCD eyes that could be therapeutically targeted pre-CLET to improve stem cell engraftment. At 12 months post-CLET the spike in levels of IL1β, IL6 and IL8 coincides with cessation of topical steroids, suggesting ongoing subclinical inflammation. We therefore recommend not discontinuing topical steroid treatment in cases where penetrating keratoplasty is indicated; however, further investigation is needed to ascertain this.

Trial registration: European Union Drug Regulating Authorities Clinical Trials Database (EuDRACT 2011-000608-16); ISRCTN (International Standard Randomised Controlled Trial Number (isrctn51772481).

Keywords: Autologous limbal stem cell transplant; Cytokines; Limbal stem cell deficiency; Ocular surface inflammation; Tears.

Abstract

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