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The suppression of local cytokine production in experimental models of injured cornea after stem cell treatment
Author(s) -
Kossl J.,
Hermankova B.,
Javorkova E.,
Bohacova P.,
Zajicova A.,
Holan V.
Publication year - 2017
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2017.04431
Subject(s) - cornea , stem cell , mesenchymal stem cell , transplantation , proinflammatory cytokine , cytokine , bone marrow , immunology , endothelial stem cell , tumor necrosis factor alpha , inflammation , biology , medicine , pathology , microbiology and biotechnology , in vitro , surgery , ophthalmology , biochemistry
Purpose Limbal stem cell deficiency ( LSCD ) is associated with the break of immune privilege of the cornea, a harmful local inflammatory reaction, production of cytokines and a failure of corneal regeneration. The corneal transplantation is not sufficient treatment. In the case of bilateral LSCD the only therapeutic option is an allogenic transplantation of limbal tissue or cultured limbal stem cells ( LSC s) with a systemic administration of immunosuppressive medication. As an alternative, mesenchymal stem cells ( MSC s) turned out to be a suitable source of autologous stem cells ( SC s). Methods In this study MSC s were obtained from murine bone marrow. Flow cytometry was used to characterize the phenotypic markers of such MSC s. LSC s were isolated from limbal tissue by trypsin digestion. For in vitro experiments SC s were cultured in 48‐well plates for 24 hours. Excised murine corneas were added into cultures and stimulated by proinflammatory cytokines, interleukin‐1 β ( IL ‐1 β ), interferon gamma and by lipopolysaccharide or were pretreated with 0.25M Na OH for 20 seconds. After 48 hours of cocultivation of corneas with SC s, the expression of genes for inflammation cytokines and growth factors in the cornea was detected using real‐time PCR . In in vivo experiments, SC s were seeded onto nanofiber scaffolds (allowed to adhere for 24 hours) and transplanted on the damaged (0.25M Na OH , 20 seconds) cornea. Results We showed that LSC s and MSC s have the ability to suppress the local expression of genes for IL ‐1 β , tumour necrosis factor‐ α , vascular endothelial growth factor and inducible NO synthase in the damaged cornea in both in vitro and in vivo models and that transplantation of SC s supports corneal regeneration. Conclusions We suggest that MSC s can be a suitable substitute for ocular surface regeneration in cases when autologous LSC s are absent or difficult to obtain.