title: Topical application of culture supernatant from human amniotic epithelial cells suppresses inflammatory reactions in cornea.
abstract: Human amniotic epithelial cells (HAEC) may be a source of soluble anti-inflammatory factors. The purpose of this study is to determine the effect of topically applied HAEC culture supernatant on corneal inflammatory reactions. HAEC were obtained from a placenta and cultured for 48 hr, and the supernatant was collected. The conditioned medium from HAEC contained small amounts of human interleukin-1 receptor antagonist (IL-1ra). Intrastromal sutures were placed in the cornea of BALB/c mice to induce corneal neovascularisation. Superficial cauterisation was applied to induce recruitment or activation of antigen presenting cells (APCs) in the cornea without neovascularisation. HAEC conditioned medium, placebo, or recombinant human IL-1ra was topically applied three times daily for 2 weeks. Suture-induced corneal neovascularisation was evaluated microscopically for 8 weeks. The cauterised corneas were harvested at 2 weeks, and the MHC class II(+) APCs were quantified by immunofluorescent staining and confocal microscopy. Inflammatory cytokine gene expression in the cauterised corneas was analyzed by a multiprobe ribonuclease protection assay. Conditioned medium from HAEC led to a profound suppression of corneal neovascularisation and fewer MHC class II(+) APCs in the epithelium. In contrast, human IL-1ra was only slightly effective in suppressing corneal inflammatory reactions. mRNA expression of murine IL-1ra and IL-1beta in the cauterised corneas was markedly suppressed after application of the conditioned medium. These results suggest that HAEC are a source of soluble anti-inflammatory factors and that conditioned medium from HAEC contains factors other than IL-1ra that suppress corneal inflammation.
authors.affiliation: Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan.
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Kamiya
authors.forename: Mingcong
authors.initials: M
authors.lastname: Wang
authors.forename: Saiko
authors.initials: S
authors.lastname: Uchida
authors.forename: Shiro
authors.initials: S
authors.lastname: Amano
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Oshika
authors.forename: Norio
authors.initials: N
authors.lastname: Sakuragawa
authors.forename: Junko
authors.initials: J
authors.lastname: Hori
mesh_terms: D000287:Administration, Topical; D000650:Amnion / Q000276:immunology*; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D002478:Cells, Cultured; D003315:Cornea / Q000276:immunology*; D016510:Corneal Neovascularization / Q000276:immunology*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004847:Epithelial Cells / Q000276:immunology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D006801:Humans; D007375:Interleukin-1 / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D023421:Models, Animal; D011247:Pregnancy; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors / Q000276:immunology
chemical_list: D017077:Culture Media, Conditioned; D000949:Histocompatibility Antigens Class II; D007375:Interleukin-1; D017472:Receptors, Interleukin-1