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      Cryopreserved rabbit amniotic membrane alleviated inflammatory response and fibrosis following experimental strabismus surgery in rabbits

      1 , 2 , * , 3

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          Abstract

          We evaluate the ability of cryopreserved rabbit amniotic membrane (AM) transplantation to reduce postoperative inflammation and the extent of fibrosis following experimental strabismus surgery. Ten white rabbits underwent bilateral superior rectus (SR) muscle resection. In the left eye, the resected SR muscle was wrapped with cryopreserved rabbit AM. The right eye underwent SR resection only and served as a control. The eyes were enucleated 4 weeks after strabismus surgery. The degree of postoperative inflammatory infiltration, the extent of fibrosis, and profile of the relative expression of inflammatory mediators in the SR muscle were evaluated and compared between the two groups by histological analysis and real-time polymerase chain reaction (PCR). There were statistically meaningful differences in the degree of postoperative inflammatory infiltration and extent of fibrosis between the eyes treated with cryopreserved rabbit AM after SR resection and those underwent SR resection only. A significant decrease in the expression of inflammatory cytokines [interleukin (IL)-12a, IL-12b, IL-17f, and tumor necrosis factor- alpha (TNF-α)], and a markedly increased expression of anti-inflammatory cytokines (transforming growth factor-beta-1(TGFβ-1) and IL-10) were observed in the eyes treated with cryopreserved rabbit AM. In this study, we demonstrate that cryopreserved rabbit AM is effective in reducing postoperative inflammation and extent of fibrosis in a rabbit model of strabismus surgery. Our results imply that cryopreserved AM allograft has anti-inflammatory and anti-scarring properties that can prevent postoperative adhesions following strabismus surgery.

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          Most cited references 31

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          Amniotic membrane transplantation with or without limbal allografts for corneal surface reconstruction in patients with limbal stem cell deficiency.

          To examine whether amniotic membrane transplantation (AMT), in preparing the perilimbal stroma, enhances the success of allograft limbal transplantation (ALT). Thirty-one eyes of 26 consecutive patients had cytologically proven limbal deficiency resulting from chemical burns (14 eyes); Stevens-Johnson syndrome, toxic epidermal necrolysis, or pseudopemphigoid (5 eyes); contact lens-induced keratopathy (3 eyes); aniridia (3 eyes); multiple surgical procedures (2 eyes); atopy (2 eyes); or an unknown cause (2 eyes). Based on the severity of limbal deficiency, group A (mild), comprising 10 eyes, received AMT alone; group B (moderate), comprising 7 eyes, received AMT and ALT; and group C (severe), comprising 14 eyes, received AMT, ALT, and penetrating keratoplasty. All patients except those in group A received continuous oral cyclosporine. Except for the 2 eyes with atopy, all amniotic membrane-covered surfaces showed rapid epithelialization (in 2 to 4 weeks) and reduced inflammation, vascularization, and scarring, and the surfaces became smooth and wettable. For the mean follow-up period of 15.4 months, 25 (83%) of 30 eyes showed visual improvement, consisting of 6 or more lines (13 eyes), 4 to 5 lines (6 eyes), or 1 to 3 lines (6 eyes). Visual improvement decreased with the severity of limbal deficiency from 8 (100%) of 8 eyes in group A to 5 (71%) of 7 eyes in group B and 11 (79%) of 14 eyes in group C. In group C, corneal graft rejection occurred in 9 (64%) of 14 eyes, and reversible early limbal allograft rejection was noted in 3 (14%) of 21 eyes of groups B and C. For partial limbal deficiency with superficial involvement, AMT alone is sufficient and hence superior to ALT because there is no need to administer cyclosporine. For total limbal deficiency, additional ALT is needed, and AMT helps reconstruct the perilimbal stroma, with reduced inflammation and vascularization, which collectively may enhance the success of ALT.
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            Regulatory T cells and immune regulation of allergic diseases: roles of IL-10 and TGF-β.

            The prevalence of allergic diseases has significantly increased in industrialized countries. Allergen-specific immunotherapy (AIT) remains as the only curative treatment. The knowledge about the mechanisms underlying healthy immune responses to allergens, the development of allergic reactions and restoration of appropriate immune responses to allergens has significantly improved over the last decades. It is now well-accepted that the generation and maintenance of functional allergen-specific regulatory T (Treg) cells and regulatory B (Breg) cells are essential for healthy immune responses to environmental proteins and successful AIT. Treg cells comprise different subsets of T cells with suppressive capacity, which control the development and maintenance of allergic diseases by various ways of action. Molecular mechanisms of generation of Treg cells, the identification of novel immunological organs, where this might occur in vivo, such as tonsils, and related epigenetic mechanisms are starting to be deciphered. The key role played by the suppressor cytokines interleukin (IL)-10 and transforming growth factor (TGF)-β produced by functional Treg cells during the generation of immune tolerance to allergens is now well established. Treg and Breg cells together have a role in suppression of IgE and induction of IgG4 isotype allergen-specific antibodies particularly mediated by IL-10. Other cell types such as subsets of dendritic cells, NK-T cells and natural killer cells producing high levels of IL-10 may also contribute to the generation of healthy immune responses to allergens. In conclusion, better understanding of the immune regulatory mechanisms operating at different stages of allergic diseases will significantly help the development of better diagnostic and predictive biomarkers and therapeutic interventions.
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              Update on amniotic membrane transplantation.

              Cryopreserved amniotic membrane modulates adult wound healing by promoting epithelialization while suppressing stromal inflammation, angiogenesis and scarring. Such clinical efficacies of amniotic membrane transplantation have been reported in several hundred publications for a wide spectrum of ophthalmic indications. The success of the aforementioned therapeutic actions prompts investigators to use amniotic membrane as a surrogate niche to achieve ex vivo expansion of ocular surface epithelial progenitor cells. Further investigation into the molecular mechanism whereby amniotic membrane exerts its actions will undoubtedly reveal additional applications in the burgeoning field of regenerative medicine. This article will focus on recent advances in amniotic membrane transplantation and expand to cover its clinical uses beyond the ocular surface.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Formal analysisRole: MethodologyRole: SoftwareRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                24 October 2017
                2017
                : 12
                : 10
                Affiliations
                [1 ] Department of Ophthalmology, Kyungpook National University Hospital, Kyungpook National University School of Medicine, Daegu, Korea
                [2 ] Brain science & Engineering Institute, Kyungpook National University School of Medicine, Daegu, Korea
                [3 ] Department of Ophthalmology, Dongtan Sacred Heart Hospital, Hallym University College of Medicine, Hwaswong, Korea
                Ocular Surface Center, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Article
                PONE-D-17-09983
                10.1371/journal.pone.0187058
                5655353
                29065159
                © 2017 Chun, Rhiu

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 3, Tables: 0, Pages: 10
                Product
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004600, Kyungpook National University Hospital;
                Award ID: Grant from Biomedical Research Institute (2013)
                Award Recipient :
                This work was supported by a grant 2013 from the Biomedical Research Institute, Kyungpook National University Hospital. (BYC). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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