title: The use of an IL-1 receptor antagonist peptide to control inflammation in the treatment of corneal limbal epithelial stem cell deficiency.
abstract: Corneal limbal stem cell deficiency (LSCD) may be treated using ex vivo limbal epithelial stem cells (LESCs) derived from cadaveric donor tissue. However, continuing challenges exist around tissue availability, inflammation, and transplant rejection. Lipopolysaccharide (LPS) or recombinant human IL-1β stimulated primary human keratocyte and LESC models were used to investigate the anti-inflammatory properties of a short chain, IL-1 receptor antagonist peptide for use in LESC sheet growth to control inflammation. The peptide was characterized using mass spectroscopy and high performance liquid chromatography. Peptide cytotoxicity, patterns of cell cytokine expression in response to LPS or IL-1β stimulation, and peptide suppression of this response were investigated by MTS/LDH assays, ELISA, and q-PCR. Cell differences in LPS stimulated toll-like receptor 4 expression were investigated using immunocytochemistry. A significant reduction in rIL-1β stimulated inflammatory cytokine production occurred following LESC and keratocyte incubation with anti-inflammatory peptide and in LPS stimulated IL-6 and IL-8 production following keratocyte incubation with peptide (1 mg/mL) (P < 0.05). LESCs produced no cytokine response to LPS stimulation and showed no TLR4 expression. The peptide supported LESC growth when adhered to a silicone hydrogel contact lens indicating potential use in improved LESC grafting through suppression of inflammation.
authors.affiliation: Biomaterials and Medical Devices Research Group, School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton BN2 4GJ, UK ; Blond McIndoe Research Foundation, Queen Victoria Hospital, East Grinstead RH19 3DZ, UK.
authors.forename: E
authors.initials: E
authors.lastname: Fok
authors.affiliation: Biomaterials and Medical Devices Research Group, School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton BN2 4GJ, UK.
authors.forename: S R
authors.identifier: 0000-0001-8367-2758
authors.initials: SR
authors.lastname: Sandeman
authors.affiliation: Brighton Studies in Tissue-Mimicry and Aided Regeneration, School of Pharmacy and Biomolecular Sciences, University of Brighton, Brighton BN2 4GJ, UK.
authors.forename: A L
authors.initials: AL
authors.lastname: Guildford
authors.affiliation: Blond McIndoe Research Foundation, Queen Victoria Hospital, East Grinstead RH19 3DZ, UK.
authors.forename: Y H
authors.identifier: 0000-0003-2881-2441
authors.initials: YH
authors.lastname: Martin
mesh_terms: D002102:Cadaver; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003315:Cornea / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003316:Corneal Diseases / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D060527:Corneal Keratocytes / Q000187:drug effects / Q000473:pathology; D019573:Epithelium, Corneal / Q000187:drug effects / Q000473:pathology; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D053583:Interleukin-1beta / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D016209:Interleukin-8 / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000633:toxicity; D010455:Peptides / Q000008:administration & dosage* / Q000378:metabolism; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051197:Toll-Like Receptor 4 / Q000187:drug effects
chemical_list: D053583:Interleukin-1beta; D016209:Interleukin-8; D008070:Lipopolysaccharides; D010455:Peptides; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4