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Bauer D, Böhm MRR, Wu X, Wang B, Jalilvand TV, Busch M, Kasper M, Brockhaus K, Wildschütz L, Melkonyan H, Laffer B, Meyer Zu Hörste G, Heiligenhaus A, Thanos S. Crystallin β-b2 promotes retinal ganglion cell protection in experimental autoimmune uveoretinitis. Front Cell Neurosci 2024; 18:1379540. [PMID: 39318470 PMCID: PMC11419989 DOI: 10.3389/fncel.2024.1379540] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/31/2024] [Accepted: 08/22/2024] [Indexed: 09/26/2024] Open
Abstract
Crystallin βb2 (crybb2) is upregulated in regenerating retinas and in various pathological conditions of the retina, including uveoretinitis. However, the role of crybb2 in this disease is largely unknown. Therefore, we used recombinant crybb2 (rcrybb2) as intravitreal treatment of B10.RIII mice prior to immunization with human interphotoreceptor retinoid-binding protein peptide 161-180 (hIRBPp161-180) in complete Freund's adjuvant (CFA) and concomitant injection of pertussis toxin (PTX) to induce experimental autoimmune uveoretinitis (EAU). In naïve mice, more beta III-tubulin (TUBB3) + and RNA-binding protein with multiple splicing (RBPMS) + cells were found in the ganglion cell layer of the retina than in EAU eyes, suggesting a loss of retinal ganglion cells (RGC) during the development of EAU. At the same time, the number of glial fibrillary acidic protein (GFAP) + cells increased in EAU eyes. RGCs were better protected in EAU eyes treated with rcrybb2, while the number of GFAP+ cells decreased. However, in retinal flatmounts, both retinal ganglion cells and retinal endothelial cells stained positive for TUBB3, indicating that TUBB3 is present in naïve B10.RIII mouse eyes not exclusive to RGCs. A significant decline in the number of RBPMS-positive retinal ganglion cells was observed in retinal flatmounts from EAU retinas in comparison to naïve retinas or EAU retinas with intravitreal rcrybb2 treatment. Whereas no significant decrease in TUBB3 levels was detected using Western blot and RT-qPCR, GFAP level, as a marker for astrocytes, increased in EAU mice compared to naïve mice. Level of Bax and Bcl2 in the retina was altered by treatment, suggesting better cell survival and inhibition of apoptosis. Furthermore, our histologic observations of the eyes showed no change in the incidence and severity of EAU, nor was the immune response affected by intravitreal rcrybb2 treatment. Taken together, these results suggest that intravitreal injection of rcrybb2 reduces retinal RGC death during the course of EAU, independent of local or systemic autoimmune responses. In the future, treating posterior uveitis with rcrybb2 to protect RGCs may offer a promising novel therapeutic strategy.
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Affiliation(s)
- Dirk Bauer
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Michael R. R. Böhm
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
- Institute for Experimental Ophthalmology, Westfalian-Wilhelms-University of Münster, Münster, Germany
- Department of Ophthalmology, University of Duisburg-Essen, Essen, Germany
| | - Xiaoyu Wu
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Bo Wang
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Tida Viola Jalilvand
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
- Institute for Experimental Ophthalmology, Westfalian-Wilhelms-University of Münster, Münster, Germany
| | - Martin Busch
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Maren Kasper
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Katrin Brockhaus
- Institute for Experimental Ophthalmology, Westfalian-Wilhelms-University of Münster, Münster, Germany
- Institute for Physiological Biochemistry, Westfalian-Wilhelms-University of Münster, Münster, Germany
| | - Lena Wildschütz
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Harutyun Melkonyan
- Institute for Experimental Ophthalmology, Westfalian-Wilhelms-University of Münster, Münster, Germany
| | - Björn Laffer
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | | | - Arnd Heiligenhaus
- Department of Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
- Department of Ophthalmology, University of Duisburg-Essen, Essen, Germany
| | - Solon Thanos
- Institute for Experimental Ophthalmology, Westfalian-Wilhelms-University of Münster, Münster, Germany
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Gottlieb J, Hanes DA, Bustos MA, Choe J, Luu A, Seizer D, Hoon DSB, Wilson TG. Impact of Cryopreserved Placental Allografts on Biochemical Recurrence in Prostate Cancer. Cancers (Basel) 2024; 16:2973. [PMID: 39272831 PMCID: PMC11394080 DOI: 10.3390/cancers16172973] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/29/2024] [Revised: 08/16/2024] [Accepted: 08/23/2024] [Indexed: 09/15/2024] Open
Abstract
BACKGROUND Human placental allografts are widely used to promote wound healing. Placental (or amniotic membrane/umbilical cord) allografts are placed along the neurovascular bundles during radical prostatectomy to improve continence and erectile function recovery. It is unknown whether placental allografts impact biochemical recurrence (BCR). METHODS This was a single-surgeon retrospective study of 566 robotic radical prostatectomies performed from April 2015 to March 2021. The patients were divided into three groups: the negative control, Brand A, and Brand B. Brand A and Brand B were both cryopreserved amniotic membrane (CAM) allografts. A total of 324 cases were included for BCR Kaplan-Meier and risk-adjusted multivariate analyses (362 for continence analysis). In vitro analyses were performed to determine the effect of CAM allografts on prostate cancer (PCa) cell line growth. RESULTS For propensity score-matched analysis (adjusting for pre-operative PSA, tumor stage, Gleason Grade, and margin status), (1) the allograft groups did not show differences in time to BCR vs. the negative control group (p = 0.7), and (2) combined allograft treatment groups showed better continence recovery vs. the negative controls (p = 0.01). In vitro, placental allografts reduced PCa cell line growth in co-culture assays. CONCLUSIONS cryopreserved AM allografts (combined or individual brands) did not show a significant effect on BCR but improved continence recovery for PCa patients.
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Affiliation(s)
- Josh Gottlieb
- Department of Urologic Oncology, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Douglas A Hanes
- Department of Biostatistics, Providence Saint Joseph Health Center, Portland, OR 97213, USA
| | - Matias A Bustos
- Department of Translational Molecular Medicine, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Jane Choe
- Department of Urologic Oncology, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Albert Luu
- Department of Urologic Oncology, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Daniel Seizer
- Department of Urologic Oncology, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Dave S B Hoon
- Department of Translational Molecular Medicine, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
| | - Timothy G Wilson
- Department of Urologic Oncology, Providence Saint John's Cancer Institute, Santa Monica, CA 90404, USA
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Sanders FWB, Huang J, Alió Del Barrio JL, Hamada S, McAlinden C. Amniotic membrane transplantation: structural and biological properties, tissue preparation, application and clinical indications. Eye (Lond) 2024; 38:668-679. [PMID: 37875701 PMCID: PMC10920809 DOI: 10.1038/s41433-023-02777-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2023] [Revised: 07/20/2023] [Accepted: 09/25/2023] [Indexed: 10/26/2023] Open
Abstract
The amniotic membrane is a single epithelial layer of the placenta. It has anti-inflammatory, anti-scarring, anti-angiogenic and possibly bactericidal properties. The basement membrane of the amniotic membrane acts as a substrate to encourage healing and re-epithelialisation. It has been used in many ocular surface diseases including persistent epithelial defects (corneal or conjunctival), chemical or thermal burns, limbal stem cell deficiency, cicatrising conjunctivitis, ocular graft versus host disease, microbial keratitis, corneal perforation, bullous keratopathy, dry eye disease, corneal haze following refractive surgery and cross-linking, band keratopathy, ocular surface neoplasia, pterygium surgery, and ligneous conjunctivitis. This review provides an up-to-date overview of amniotic membrane transplantation including the structural and biological properties, preparation and application, clinical indications, and commercially available products.
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Affiliation(s)
- Francis W B Sanders
- Department of Ophthalmology, Singleton Hospital, Swansea Bay University Health Board, Swansea, UK
| | - Jinhai Huang
- Eye and ENT Hospital, Fudan University, Shanghai, China
| | - Jorge L Alió Del Barrio
- Division of Ophthalmology, School of Medicine, Universidad Miguel Hernández, Alicante, Spain; and Cornea, Cataract and Refractive Surgery Department, VISSUM Corporation, Alicante, Spain
| | - Samer Hamada
- Corneo Plastic Unit and Eye Bank, Queen Victoria Hospital, East Grinstead, UK
| | - Colm McAlinden
- Eye and ENT Hospital, Fudan University, Shanghai, China.
- Corneo Plastic Unit and Eye Bank, Queen Victoria Hospital, East Grinstead, UK.
- School of Optometry and Vision Sciences, Cardiff University, Maindy Road, Cardiff, UK.
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Norte-Muñoz M, Botelho MF, Schoeberlein A, Chaves J, Neto Murta J, Ponsaerts P, Agudo-Barriuso M, Costa E. Insights and future directions for the application of perinatal derivatives in eye diseases: A critical review of preclinical and clinical studies. Front Bioeng Biotechnol 2022; 10:969927. [PMID: 36425647 PMCID: PMC9679153 DOI: 10.3389/fbioe.2022.969927] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/15/2022] [Accepted: 10/17/2022] [Indexed: 09/05/2023] Open
Abstract
Perinatal derivatives (PnD) are gaining interest as a source for cell-based therapies. Since the eye is easily accessible to local administration, eye diseases may be excellent candidates to evaluate novel therapeutic approaches. With this work, we performed a systematic review of published preclinical and clinical studies addressing PnD in the treatment of ocular diseases. We have set two specific objectives: (i) to investigate the current level of standardization in applied technical procedures in preclinical studies and (ii) to assess clinical efficacy in clinical trials. Hereto, we selected studies that applied amniotic membrane (hAM) and mesenchymal stromal cells derived from amniotic membrane (hAMSC), placenta (hPMSC), umbilical cord (hUC-MSC) and Wharton's Jelly (hUC-WJ-MSC), excluding those where cells were not transplanted individually, following a systematic PubMed search for preclinical studies and consultation of clinical studies on https://clinicaltrials.gov and https://www.clinicaltrialsregister.eu/. Our bibliographic search retrieved 26 pre-clinical studies and 27 clinical trials. There was a considerable overlap regarding targeted ocular structures. Another common feature is the marked tendency towards (i) locally administered treatments and (ii) the PnD type. In the cornea/ocular surface, hAM was preferred and usually applied directly covering the ocular surface. For neuroretinal disorders, intra-ocular injection of umbilical or placental-derived cells was preferred. In general, basic research reported favourable outcomes. However, due to lack of standardization between different studies, until now there is no clear consensus regarding the fate of administered PnD or their mode of action. This might be accountable for the low index of clinical translation. Regarding clinical trials, only a minority provided results and a considerable proportion is in "unknown status". Nevertheless, from the limited clinical evidence available, hAM proved beneficial in the symptomatic relief of bullous keratopathy, treating dry eye disease and preventing glaucoma drainage device tube exposure. Regarding neuroretinal diseases, application of Wharton's Jelly MSC seems to become a promising future approach. In conclusion, PnD-based therapies seem to be beneficial in the treatment of several ocular diseases. However, much is yet to be done both in the pre-clinical and in the clinical setting before they can be included in the daily ophthalmic practice.
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Affiliation(s)
- María Norte-Muñoz
- Experimental Ophthalmology Group, IMIB-Arrixaca, University of Murcia, Murcia, Spain
| | - Maria Filomena Botelho
- Institute of Biophysics and Institute for Clinical and Biomedical Research (iCBR), Area of Environment Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, Coimbra, Portugal
- Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, Coimbra, Portugal
- Clinical and Academic Centre of Coimbra (CACC), Coimbra, Portugal
| | - Andreina Schoeberlein
- Department of Obstetrics and Feto-maternal Medicine, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland
- Department for BioMedical Research (DBMR), University of Bern, Bern, Switzerland
| | - João Chaves
- Ophthalmology Department, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal
| | - Joaquim Neto Murta
- Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, Coimbra, Portugal
- Clinical and Academic Centre of Coimbra (CACC), Coimbra, Portugal
- Ophthalmology Department, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal
- Ophtalmology Universitary Clinic and Institute for Clinical and Biomedical Research (iCBR), Faculty of Medicine, University of Coimbra, Coimbra, Portugal
| | - Peter Ponsaerts
- Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Antwerp, Belgium
| | - Marta Agudo-Barriuso
- Experimental Ophthalmology Group, IMIB-Arrixaca, University of Murcia, Murcia, Spain
| | - Esmeralda Costa
- Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, Coimbra, Portugal
- Clinical and Academic Centre of Coimbra (CACC), Coimbra, Portugal
- Ophthalmology Department, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal
- Ophtalmology Universitary Clinic and Institute for Clinical and Biomedical Research (iCBR), Faculty of Medicine, University of Coimbra, Coimbra, Portugal
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Jafari A, Rezaei-Tavirani M, Niknejad H, Zali H. Tumor Targeting by Conditioned Medium Derived From Human Amniotic Membrane: New Insight in Breast Cancer Therapy. Technol Cancer Res Treat 2021; 20:15330338211036318. [PMID: 34402329 PMCID: PMC8375331 DOI: 10.1177/15330338211036318] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023] Open
Abstract
Objectives: Traditional breast cancer treatments have challenges including inefficiency, multidrug resistance, severe side effects, and targeting non-specifically. The development of alternative treatment strategies has attracted a great deal of interest. Using the amniotic membrane has become a promising and convenient new approach for cancer therapy. This study aimed to evaluate the anti-cancer ability of conditioned medium extracted from the human amniotic membrane (hAM-CM) on breast cancer cells. Methods: Conditioned medium was collected after 48 h incubation of hAM in epithelial up manner. MTT, cell cycle, apoptosis, colony formation, and sphere assays were used to determine the impact of hAM-CM on breast cancer cell lines. The effects of hAM-CM on the migration and invasion of breast cancer cells were determined using scratch wound healing and transwell assays, respectively. Results: Based on the results, cell viability was significantly decreased by hAM-CM in a dose-dependent manner. The hAM-CM remarkably induced apoptosis and necrosis of cancer cells. Moreover, cell migration and invasion potential of cancer cells decreased after the hAM-CM treatment. Further, both the number of colonies and their morphologies were affected by the treatment. In the treated group, a significant decrease in the number of colonies along with an obvious change in their morphologies from holoclone shape to a dominant paracolone structure was observed. Conclusion: Our results indicate that the conditioned medium derived from the human amniotic membrane able to inhibit proliferation and metastasis of tumor cells and can be considered a natural and valuable candidate for breast cancer therapy.
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Affiliation(s)
- Ameneh Jafari
- Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mostafa Rezaei-Tavirani
- Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Hassan Niknejad
- Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Hakimeh Zali
- Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.,Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Jafari A, Rezaei-Tavirani M, Farhadihosseinabadi B, Zali H, Niknejad H. Human amniotic mesenchymal stem cells to promote/suppress cancer: two sides of the same coin. Stem Cell Res Ther 2021; 12:126. [PMID: 33579346 PMCID: PMC7881457 DOI: 10.1186/s13287-021-02196-x] [Citation(s) in RCA: 29] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2020] [Accepted: 01/27/2021] [Indexed: 02/08/2023] Open
Abstract
Cancer is a leading cause of death in both developed and developing countries, and because of population growth and aging, it is a growing medical burden worldwide. With robust development in medicine, the use of stem cells has opened new treatment modalities in cancer therapy. In adult stem cells, mesenchymal stem cells (MSCs) are showing rising promise in cancer treatment due to their unique properties. Among different sources of MSCs, human amniotic fluid/membrane is an attractive and suitable reservoir. There are conflicting opinions about the role of human amniotic membrane/fluid mesenchymal stem cells (hAMSCS/hAFMSCs) in cancer, as some studies demonstrating the anticancer effects of these cells and others suggesting their progressive effects on cancer. This review focuses on recent findings about the role of hAMSCs/hAFMSCs in cancer treatment and summarizes the suppressing as well as promoting effects of these cells on cancer progression and underling mechanisms.
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Affiliation(s)
- Ameneh Jafari
- Department of Basic Sciences, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
- Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
| | - Mostafa Rezaei-Tavirani
- Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | | | - Hakimeh Zali
- Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Hassan Niknejad
- Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
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Beeken LJ, Ting DS, Sidney LE. Potential of mesenchymal stem cells as topical immunomodulatory cell therapies for ocular surface inflammatory disorders. Stem Cells Transl Med 2021; 10:39-49. [PMID: 32896982 PMCID: PMC7780815 DOI: 10.1002/sctm.20-0118] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/16/2020] [Revised: 07/10/2020] [Accepted: 07/30/2020] [Indexed: 12/17/2022] Open
Abstract
Ocular surface inflammatory disorders (OSIDs) are a group of highly prevalent, heterogeneous diseases that display a variety of aetiologies and symptoms and are risk factors for serious complications, including ocular and cornea impairment. Corneal inflammation is a common factor of all OSIDs, regardless of their cause or symptoms. Current medications include over-the-counter lubricating eye drops, corticosteroids, and ciclosporin, which either do not treat the corneal inflammation or have been associated with multiple side effects leading to alternative treatments being sought. Regenerative medicine cell therapies, particularly mesenchymal stem cells (MSCs), have shown great promise for immunosuppression and disease amelioration across multiple tissues, including the cornea. However, for successful development and clinical translation of MSC therapy for OSIDs, significant problems must be addressed. This review aims to highlight considerations, including whether the source of MSC isolation impacts the efficacy and safety of the therapy, in addition to assessing the feasibility of MSC topical application to the cornea and ocular surface through analysis of potential scaffolds and cell carriers for application to the eye. The literature contains limited data assessing MSCs incorporated into scaffolds for corneal administration, thus here we highlight the necessity of further investigations to truly exploit the potential of an MSC-based cell therapy for the treatment of OSIDs.
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Affiliation(s)
- Lydia J. Beeken
- Academic Ophthalmology, Division of Clinical NeurosciencesUniversity of Nottingham, Queens Medical Centre CampusNottinghamUK
| | - Darren S.J. Ting
- Academic Ophthalmology, Division of Clinical NeurosciencesUniversity of Nottingham, Queens Medical Centre CampusNottinghamUK
| | - Laura E. Sidney
- Academic Ophthalmology, Division of Clinical NeurosciencesUniversity of Nottingham, Queens Medical Centre CampusNottinghamUK
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Wang B, Kasper M, Laffer B, Meyer zu Hörste G, Wasmuth S, Busch M, Jalilvand TV, Thanos S, Heiligenhaus A, Bauer D, Heinz C. Increased Hydrostatic Pressure Promotes Primary M1 Reaction and Secondary M2 Polarization in Macrophages. Front Immunol 2020; 11:573955. [PMID: 33154752 PMCID: PMC7591771 DOI: 10.3389/fimmu.2020.573955] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/18/2020] [Accepted: 08/26/2020] [Indexed: 12/30/2022] Open
Abstract
Patients with chronic anterior uveitis are at particularly high risk of developing secondary glaucoma when corticosteroids [e.g., dexamethasone (Dex)] are used or when inflammatory activity has regressed. Macrophage migration into the eye increases when secondary glaucoma develops and may play an important role in the development of secondary glaucoma. Our aim was to evaluate in vitro if increased hydrostatic pressure and corticosteroids could induce changes in macrophages phenotype. By using a pressure chamber cell culture system, we assessed the effect of increased hydrostatic pressure (HP), inflammation, and immunosuppression (Dex) on the M1/M2 phenotype of macrophages. Bone marrow-derived macrophages (BMDMs) were stimulated with medium, lipopolysaccharide (LPS, 100 ng/ml), Dex (200 ng/ml), or LPS + Dex and incubated with different HP (0, 20, or 60 mmHg) for 2 or 7 days. The numbers of CD86+/CD206- (M1 phenotype), CD86-/CD206+ (M2 phenotype), CD86+/CD206+ (intermediate phenotype), F4/80+/TNF-α+, and F4/80+/IL-10+ macrophages were determined by flow cytometry. TNF-α and IL-10 levels in cell culture supernatants were quantified by ELISA. TNF-α, IL-10, fibronectin, and collagen IV expression in BMDMs were detected by immunofluorescence microscopy. Higher HP polarizes macrophages primarily to an M1 phenotype (LPS, 60 vs. 0 mmHg, d2: p = 0.0034) with less extra cellular matrix (ECM) production and secondary to an M2 phenotype (medium, 60 vs. 0 mmHg, d7: p = 0.0089) (medium, 60 vs. 20 mmHg, d7: p = 0.0433) with enhanced ECM production. Dex induces an M2 phenotype (Dex, medium vs. Dex, d2: p < 0.0001; d7: p < 0.0001) with more ECM production. Higher HP further increased M2 polarization of Dex-treated macrophages (Dex, 60 vs. 0 mmHg, d2: p = 0.0417; d7: p = 0.0454). These changes in the M1/M2 phenotype by high HP or Dex treatment may play a role in the pathogenesis of secondary uveitic glaucoma- or glucocorticoid (GC)-induced glaucoma.
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Affiliation(s)
- Bo Wang
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Maren Kasper
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Björn Laffer
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Gerd Meyer zu Hörste
- Institution of Neurology and Institution for Translational Neurology, Universitätsklinikum Münster, Münster, Germany
| | - Susanne Wasmuth
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Martin Busch
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | | | - Solon Thanos
- Institution of Experimental Ophthalmology, Westfälische Wilhelms-Universität, Münster, Germany
| | - Arnd Heiligenhaus
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
- Ophthalmology, University of Duisburg-Essen, Essen, Germany
| | - Dirk Bauer
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
| | - Carsten Heinz
- Ophthalmology and Ophtha-Lab at St. Franziskus Hospital, Münster, Germany
- Ophthalmology, University of Duisburg-Essen, Essen, Germany
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Brocks D, Mead OG, Tighe S, Tseng SCG. Self-Retained Cryopreserved Amniotic Membrane for the Management of Corneal Ulcers. Clin Ophthalmol 2020; 14:1437-1443. [PMID: 32581504 PMCID: PMC7266945 DOI: 10.2147/opth.s253750] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/25/2022] Open
Abstract
Purpose To evaluate the clinical outcomes of self-retained cryopreserved amniotic membrane (cAM) for the treatment of corneal ulcers. Methods This was a single-center, retrospective review of consecutive patients with non-healing corneal ulcers that underwent treatment with self-retained cAM (PROKERA® Slim). The primary outcome measure was time to complete corneal epithelialization. Ocular discomfort, corneal staining, corneal signs, and visual acuity were assessed at 1 week, 1 month, 3 months, and 6 months. Complications, adverse events, and ulcer recurrence were also recorded. Results A total of 13 eyes (13 patients) with recalcitrant corneal ulcers were included for analysis, 9 (69%) of which progressed from neurotrophic keratitis (NK). Prior to cAM application, patients used conventional treatments such as artificial tears (n = 11), antibiotics (n = 11), ointment (n = 11), steroids (n = 6), and antivirals (n = 3). Self-retained cAMs (n = 1.5 ± 0.8) were placed for 6.8 ± 3.4 days, during which time antibiotics were continued. Four cases (31%) were subsequently treated with bandage contact lens (n = 3) and tarsorrhaphy (n = 1). All corneal ulcers healed in a median of 14 days (range: 4-43). This was accompanied by a significant improvement in ocular discomfort, corneal staining, and corneal signs at 1 week, 1 month, 3 months, and 6 months (P<.05). Recurrence was noted in one case. No adverse events were observed. Conclusion Self-retained cAM may be a valuable, in-office treatment option for healing recalcitrant corneal ulcers of various etiologies, especially those with underlying NK. Further prospective, controlled studies are warranted.
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Affiliation(s)
| | - Olivia G Mead
- Ocular Surface Center and TissueTech Inc, Miami, FL, USA
| | - Sean Tighe
- Department of Ophthalmology, Florida International University, Miami, FL, USA.,Department of Biochemistry and Molecular Biology, University of Miami, Miami, FL, USA
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Human amniotic membrane conditioned medium inhibits proliferation and modulates related microRNAs expression in hepatocarcinoma cells. Sci Rep 2019; 9:14193. [PMID: 31578445 PMCID: PMC6775050 DOI: 10.1038/s41598-019-50648-5] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2018] [Accepted: 09/17/2019] [Indexed: 12/19/2022] Open
Abstract
The placental stem cells have called the focus of attention for their therapeutic potential to treat different diseases, including cancer. There is plenty evidence about the antiproliferative, antiangiogenic and proapoptotic properties of the amniotic membrane. Liver cancer is the fifth cause of cancer in the world, with a poor prognosis and survival. Alternative treatments to radio- or chemotherapy have been searched. In this work we aimed to study the antiproliferative properties of the human amniotic membrane conditioned medium (AM-CM) in hepatocarcinoma cells. In addition, we have analyzed the regulation of pro and antiOncomiRs expression involved in hepatocarcinoma physiology. We have determined by 3H-thymidine incorporation assay that AM-CM inhibits DNA synthesis in HepG2 cells after 72 h of treatment. AM-CM pure or diluted at 50% and 25% also diminished HepG2 and HuH-7 cells viability and cell number. Furthermore, AM-CM induced cell cycle arrest in G2/M. When proliferation mechanisms were analyzed we found that AM-CM reduced the expression of both Cyclin D1 mRNA and protein. Nuclear expression of Ki-67 was also reduced. We observed that this CM was able to promote the expression of p53 and p21 mRNA and proteins, leading to cell growth arrest. Moreover, AM-CM induced an increase in nuclear p21 localization, observed by immunofluorescence. As p53 levels were increased, Mdm-2 expression was downregulated. Interestingly, HepG2 and HuH-7 cells treatment with AM-CM during 24 and 72 h produced an upregulation of antiOncomiRs 15a and 210, and a downregulation of proOncomiRs 206 and 145. We provide new evidence about the promising novel applications of human amniotic membrane in liver cancer.
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Ahmed M, Esposito M, Lovallo G. A single-center, retrospective review of robot-assisted laparoscopic prostatectomy with and without cryopreserved umbilical cord allograft in improving continence recovery. J Robot Surg 2019; 14:283-289. [PMID: 31152310 PMCID: PMC7125058 DOI: 10.1007/s11701-019-00972-9] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2019] [Accepted: 05/01/2019] [Indexed: 12/19/2022]
Abstract
The objective of this study was to evaluate the safety and effectiveness of cryopreserved umbilical cord (UC) allograft as a nerve wrap around the neurovascular bundle (NVB) in accelerating return to continence after radical prostatectomy. A single-center, retrospective study was performed on 200 patients who underwent bilateral, nerve-sparing robot-assisted radical prostatectomy (RARP) with and without placement of UC around the NVBs (n = 100/group). Patients were excluded if they had previous simple or transurethral prostatectomy or history of pelvic radiation. Post-operative continence, defined as 0 or 1 safety pad, was analyzed between groups at 1, 3, 6, and 12 months. Complications, biochemical recurrence and adverse events were assessed to determine safety. Patients who underwent RARP with UC were significantly more likely to be continent at 1 month (65% vs. 44%, p = 0.018), 3 months (83% vs. 70%, p = 0.03), and 12 months (97% vs. 87%, p = 0.009). Sample stratification revealed that UC is beneficial for obese patients and those > 60 years, both of which are high risk for post-RARP incontinence. Biochemical failure was noted in 2 (UC) and 4 (control) patients. No adverse events or complications related to UC were observed. The results suggest that UC allograft is safe and accelerates continence recovery in post-RARP patients. Prospective, randomized trials are warranted.
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Affiliation(s)
- Mutahar Ahmed
- NJ Center for Prostate Cancer and Urology, 255 W Spring Valley Ave #101, Maywood, NJ, 07607, USA.
| | - Michael Esposito
- NJ Center for Prostate Cancer and Urology, 255 W Spring Valley Ave #101, Maywood, NJ, 07607, USA
| | - Gregory Lovallo
- NJ Center for Prostate Cancer and Urology, 255 W Spring Valley Ave #101, Maywood, NJ, 07607, USA
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12
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Orozco Morales ML, Marsit NM, McIntosh OD, Hopkinson A, Sidney LE. Anti-inflammatory potential of human corneal stroma-derived stem cells determined by a novel in vitro corneal epithelial injury model. World J Stem Cells 2019; 11:84-99. [PMID: 30842807 PMCID: PMC6397805 DOI: 10.4252/wjsc.v11.i2.84] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/24/2018] [Revised: 11/01/2018] [Accepted: 01/01/2019] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND An in vitro injury model mimicking a corneal surface injury was optimised using human corneal epithelial cells (hCEC). AIM To investigate whether corneal-stroma derived stem cells (CSSC) seeded on an amniotic membrane (AM) construct manifests an anti-inflammatory, healing response. METHODS Treatment of hCEC with ethanol and pro-inflammatory cytokines were compared in terms of viability loss, cytotoxicity, and pro-inflammatory cytokine release, in order to generate the in vitro injury. This resulted in an optimal injury of 20% (v/v) ethanol for 30 s with 1 ng/mL interleukin-1 (IL-1) beta. Co-culture experiments were performed with CSSC alone and with CSSC-AM constructs. The effect of injury and co-culture on viability, cytotoxicity, IL-6 and IL-8 production, and IL1B, TNF, IL6, and CXCL8 mRNA expression were assessed. RESULTS Co-culture with CSSC inhibited loss of hCEC viability caused by injury. Enzyme linked immunosorbent assay and polymerase chain reaction showed a significant reduction in the production of IL-6 and IL-8 pro-inflammatory cytokines, and reduction in pro-inflammatory cytokine mRNA expression during co-culture with CSSC alone and with the AM construct. These results confirmed the therapeutic potential of the CSSC and the possible use of AM as a cell carrier for application to the ocular surface. CONCLUSION CSSC were shown to have a potentially therapeutic anti-inflammatory effect when treating injured hCEC, demonstrating an important role in corneal regeneration and wound healing, leading to an improved knowledge of their potential use for research and therapeutic purposes.
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Affiliation(s)
- Mariana Lizeth Orozco Morales
- Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom
| | - Nagi M Marsit
- Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom
| | - Owen D McIntosh
- Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom
| | - Andrew Hopkinson
- Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom
| | - Laura E Sidney
- Academic Ophthalmology, Division of Clinical Neuroscience, University of Nottingham, Nottingham, NG7 2UH, United Kingdom.
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Raphael A. A single-centre, retrospective study of cryopreserved umbilical cord/amniotic membrane tissue for the treatment of diabetic foot ulcers. J Wound Care 2018; 25:S10-S17. [PMID: 29027852 DOI: 10.12968/jowc.2016.25.sup7.s10] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/13/2023]
Abstract
OBJECTIVE Histopathological studies have shown a prolonged inflammatory phase in wounds of patients with diabetes, delaying formation of mature granulation tissue and reducing wound tensile strength, making these wounds difficult for physicians to heal. Cryopreserved human umbilical cord (cUC) tissues possess unique anti-inflammatory and anti-scarring properties and have been found to help improve closure of these chronic wounds. METHOD A retrospective chart review was performed to assess the efficacy of cUC as an advanced treatment modality to help promote the closure of chronic DFUs. Overall healing rate, duration to wound closure, and number of cUC applications used to achieve closure were used to assess cUC treatment efficacy. RESULTS A total of 32 wounds in 29 patients treated at a single health-care centre were included in the study population The average initial wound area for all wounds was 10.6 ± 2.15cm2. Of the 32 wounds 28 achieved complete epithelialisation for an overall healing rate of 87.5%. Average time to wound closure was 13.8 ± 1.95 weeks with a median of 9 weeks and an average of 1.68 ± 0.18 cUC applications. CONCLUSION The results suggest cUC allograft may be effective in improving the healing of DFUs ulcers as well as potentially reducing the medical costs associated with chronic DFUs due to the low number of applications needed to achieve complete wound closure. Prospective, randomised controlled trials are suggested to better understand the efficacy of cUC in chronic wound healing. DECLARATION OF INTEREST Dr Raphael is a paid speaker for Amniox Medical, Inc.
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Affiliation(s)
- A Raphael
- Village Podiatry Centers, Smyrna, GA, US
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14
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McDonald MB, Sheha H, Tighe S, Janik SB, Bowden FW, Chokshi AR, Singer MA, Nanda S, Qazi MA, Dierker D, Shupe AT, McMurren BJ. Treatment outcomes in the DRy Eye Amniotic Membrane (DREAM) study. Clin Ophthalmol 2018; 12:677-681. [PMID: 29670328 PMCID: PMC5898584 DOI: 10.2147/opth.s162203] [Citation(s) in RCA: 36] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/03/2022] Open
Abstract
Purpose To evaluate the efficacy of cryopreserved amniotic membrane (CAM) in reducing signs and symptoms of dry eye disease (DED) in a large patient population. Methods A retrospective chart review at 10 clinical sites was done of patients with refractory DED who received CAM and completed at least 3 months of follow-up. Data collected were demographics; medical history including previous and current ocular treatment, diagnosis, clinical presentations, comorbidity, duration and frequency of treatment with CAM; and concomitant medications. The primary outcome was the change in dry eye workshop (DEWS) score after treatment. Results A total of 97 eyes of 84 patients exhibited severe dry eye despite maximal medical treatments including topical artificial tears, cyclosporine-A, serum, antibiotics, and steroids. Patients manifested with superficial punctate keratitis (86%), filamentary keratitis (13%), exposure keratitis (19%), neurotrophic keratitis (2%), and corneal epithelial defect (7%). After CAM treatment for 5.4±2.8 days, 74 (88%) patients demonstrated an improved ocular surface along with a notable reduction of the severity as the overall DEWS score was significantly reduced from 3.25±0.5 at baseline to 1.44±0.6 at 1 week, 1.45±0.6 at 1 month, and 1.47±0.6 at 3 months (p<0.001). Ten eyes (10%) required repeated treatment to complete healing. Apart from discomfort during CAM placement, there were no adverse events. Conclusion Placement of CAM is promising to enhance the recovery of ocular surface health and reduce signs and symptoms in patients with moderate-to-severe DED.
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Affiliation(s)
| | - Hosam Sheha
- Ocular Surface Center and TissueTech, Inc., Miami, FL, USA.,Florida International University Herbert Wertheim College of Medicine, Miami, FL, USA.,Hofstra University School of Medicine, Hempstead, NY, USA.,Research Institute of Ophthalmology, Cairo, Egypt
| | - Sean Tighe
- Ocular Surface Center and TissueTech, Inc., Miami, FL, USA.,Florida International University Herbert Wertheim College of Medicine, Miami, FL, USA
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15
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Amniotic membrane extract differentially regulates human peripheral blood T cell subsets, monocyte subpopulations and myeloid dendritic cells. Cell Tissue Res 2018; 373:459-476. [DOI: 10.1007/s00441-018-2822-1] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/30/2017] [Accepted: 03/06/2018] [Indexed: 01/05/2023]
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16
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Cheng AMS, Tseng SCG. Self-Retained Amniotic Membrane Combined With Antiviral Therapy for Herpetic Epithelial Keratitis. Cornea 2018; 36:1383-1386. [PMID: 28834819 DOI: 10.1097/ico.0000000000001316] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023]
Abstract
PURPOSE To evaluate the therapeutic benefit of self-retained cryopreserved amniotic membrane in conjunction with oral antiviral therapy in herpetic epithelial keratitis. METHODS Retrospective review of 4 patients with primary (1 eye) and recurrent (3 eyes) unilateral herpetic epithelial keratitis treated with cryopreserved amniotic membrane through the placement of the PROKERA Slim (PKS) (Bio-Tissue, Inc) in conjunction with oral acyclovir. Their symptoms, conjunctival inflammation, corneal staining, and visual acuity were compared before and after treatment. RESULTS Herpetic epithelial keratitis presented as dendritic (3 eyes) and geographic (1 eye) epithelial lesions. After epithelial debridement and placement of the PKS for 5 ± 3.7 days, all patients reported significant relief of symptoms, rapid corneal epithelialization, and reduction of ocular surface inflammation. The visual acuity was also improved in all eyes from 0.7 ± 0.7 to 0.4 ± 0.7 logarithm of the minimum angle of resolution (P = 0.2). They remained symptom-free during a follow-up period of 2.7 to 50.8 (20.3 ± 21.7) months. CONCLUSIONS The PKS in conjunction with oral acyclovir facilitates the ease of early intervention to accelerate restoration of a normal corneal epithelium in herpetic epithelial keratitis.
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Affiliation(s)
- Anny M S Cheng
- *Ocular Surface Center, Miami, FL; and †Department of Ophthalmology, Florida International University, Herbert Wertheim College of Medicine, Miami, FL
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Sheha H, Tighe S, Cheng AM, Tseng SC. A stepping stone in treating dendritic keratitis. Am J Ophthalmol Case Rep 2017; 7:55-58. [PMID: 29260079 PMCID: PMC5722148 DOI: 10.1016/j.ajoc.2017.06.002] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2016] [Revised: 05/17/2017] [Accepted: 06/02/2017] [Indexed: 11/11/2022] Open
Abstract
PURPOSE To report the outcome of self-retained amniotic membrane after debridement in recurrent dendritic keratitis. OBSERVATIONS A 70-year-old female with a recurrent dendritic corneal ulcer received debridement followed by placement of self-retained amniotic membrane. Five days after treatment, the patient experienced a complete resolution of symptoms, marked reduction of inflammation, complete re-epithelialization of the cornea and improvement of visual acuity. The corneal surface remained stable for 18 months despite noncompliance in using antiviral therapy. CONCLUSIONS AND IMPORTANCE Self-retained amniotic membrane after debridement appears effective in treating dendritic keratitis. While early debridement is crucial to remove the infected corneal epithelium, amniotic membrane was shown to enhance the healing without scarring or recurrence. Besides the known anti-inflammatory and anti-scarring effects of the amniotic membrane, it may have a potential topical antiviral effect that warrants further investigation.
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Affiliation(s)
- Hosam Sheha
- Ocular Surface Center, Miami, FL, USA
- Department of Ophthalmology, Florida International University, Herbert Wertheim College of Medicine, Miami, FL, USA
| | - Sean Tighe
- Ocular Surface Center, Miami, FL, USA
- Department of Ophthalmology, Florida International University, Herbert Wertheim College of Medicine, Miami, FL, USA
| | - Anny M.S. Cheng
- Ocular Surface Center, Miami, FL, USA
- Department of Ophthalmology, Florida International University, Herbert Wertheim College of Medicine, Miami, FL, USA
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18
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Corneal Nerve Regeneration after Self-Retained Cryopreserved Amniotic Membrane in Dry Eye Disease. J Ophthalmol 2017; 2017:6404918. [PMID: 28894606 PMCID: PMC5574308 DOI: 10.1155/2017/6404918] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/12/2017] [Accepted: 06/28/2017] [Indexed: 01/22/2023] Open
Abstract
Purpose To evaluate the efficacy of self-retained cryopreserved amniotic membrane (CAM) in promoting corneal nerve regeneration and improving corneal sensitivity in dry eye disease (DED). Methods In this prospective randomized clinical trial, subjects with DED were randomized to receive CAM (study group) or conventional maximum treatment (control). Changes in signs and symptoms, corneal sensitivity, topography, and in vivo confocal microscopy (IVCM) were evaluated at baseline, 1 month, and 3 months. Results Twenty subjects (age 66.9 ± 8.9) were enrolled and 17 completed all follow-up visits. Signs and symptoms were significantly improved in the study group yet remained constant in the control. IVCM showed a significant increase in corneal nerve density in the study group (12,241 ± 5083 μm/mm2 at baseline, 16,364 ± 3734 μm/mm2 at 1 month, and 18,827 ± 5453 μm/mm2 at 3 months, p = 0.015) but was unchanged in the control. This improvement was accompanied with a significant increase in corneal sensitivity (3.25 ± 0.6 cm at baseline, 5.2 ± 0.5 cm at 1 month, and 5.6 ± 0.4 cm at 3 months, p < 0.001) and corneal topography only in the study group. Conclusions Self-retained CAM is a promising therapy for corneal nerve regeneration and accelerated recovery of the ocular surface health in patients with DED. The study is registered at clinicaltrials.gov with trial identifier: NCT02764814.
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Ogawa Y, He H, Mukai S, Imada T, Nakamura S, Su CW, Mahabole M, Tseng SCG, Tsubota K. Heavy Chain-Hyaluronan/Pentraxin 3 from Amniotic Membrane Suppresses Inflammation and Scarring in Murine Lacrimal Gland and Conjunctiva of Chronic Graft-versus-Host Disease. Sci Rep 2017; 7:42195. [PMID: 28165063 PMCID: PMC5292704 DOI: 10.1038/srep42195] [Citation(s) in RCA: 36] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/02/2016] [Accepted: 01/06/2017] [Indexed: 12/21/2022] Open
Abstract
Chronic graft-versus-host disease (cGVHD) is a major complication of hematopoietic stem cell transplantation. Dry eye disease is the prominent ocular sequel of cGVHD and is caused by excessive inflammation and fibrosis in the lacrimal glands. Heavy chain-Hyaluronan/Pentraxin 3 (HC-HA/PTX3) is a complex purified from human amniotic membrane (AM) and known to exert anti-inflammatory and anti-scarring actions. In this study, we utilized a mouse model of cGVHD to examine whether HC-HA/PTX3 could attenuate dry eye disease elicited by cGVHD. Our results indicated that subconjunctival and subcutaneous injection of HC-HA/PTX3 preserved tear secretion and conjunctival goblet cell density and mitigated inflammation and scarring of the conjunctiva. Such therapeutic benefits were associated with suppression of scarring and infiltration of inflammatory/immune cells in the lacrimal glands. Furthermore, HC-HA/PTX3 significantly reduced the extent of infiltration of CD45+ CD4+ IL-17+ cells, CD45+ CD34+ collagen I+ CXCR4+ fibrocytes, and HSP47+ activated fibroblasts that were accompanied by upregulation of collagen type Iα1, collagen type IIIα1 and NF-kB in lacrimal glands. Collectively, these pre-clinical data help prove the concept that subcutaneous and subconjunctival injection of HC-HA/PTX3 is a novel approach to prevent dry eye disease caused by cGVHD and allow us to test its safety and efficacy in future human clinical trials.
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Affiliation(s)
- Yoko Ogawa
- Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo, 160-8582, Japan
| | - Hua He
- TissueTech, Inc., Miami, FL 33173, USA
| | - Shin Mukai
- Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo, 160-8582, Japan
| | - Toshihiro Imada
- Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo, 160-8582, Japan
| | - Shigeru Nakamura
- Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo, 160-8582, Japan
| | | | | | | | - Kazuo Tsubota
- Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo, 160-8582, Japan
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20
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Tseng SCG. HC-HA/PTX3 Purified From Amniotic Membrane as Novel Regenerative Matrix: Insight Into Relationship Between Inflammation and Regeneration. Invest Ophthalmol Vis Sci 2016; 57:ORSFh1-8. [PMID: 27116665 PMCID: PMC4855828 DOI: 10.1167/iovs.15-17637] [Citation(s) in RCA: 95] [Impact Index Per Article: 10.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/09/2015] [Accepted: 08/03/2015] [Indexed: 12/22/2022] Open
Abstract
PURPOSE Human limbal palisade of Vogt is an ideal model for studying and practicing regenerative medicine due to their accessibility. Nonresolving inflammation is a common manifestation of limbal stem cell deficiency, which is the major cause of corneal blindness, and presents as a threat to the success of transplanted limbal epithelial stem cells. Clinical studies have shown that the efficacy of transplantation of limbal epithelial stem cells can be augmented by transplantation of cryopreserved human amniotic membrane (AM), which exerts anti-inflammatory, antiscarring, and antiangiogenic action to promote wound healing. METHODS Review of published data to determine the molecular action mechanism explaining how AM exerts the aforementioned therapeutic actions. RESULTS From the water-soluble extract of cryopreserved AM, we have biochemically purified one novel matrix component termed heavy chain (HC)-hyaluronan (HA)/pentraxin 3 (PTX3) as the key relevant tissue characteristic responsible for the aforementioned AM's efficacy. Heavy chain-HA is a complex formed by a covalent linkage between HA and HC1 of inter-α-trypsin inhibitor (IαI) by tumor necrosis factor-stimulated gene-6 (TSG-6). This complex may then be tightly associated with PTX3 to form HC-HA/PTX3 complex. Besides exerting an anti-inflammatory, antiscarring, and antiangiogenic effects, HC-HA/PTX3 complex also uniquely maintains limbal niche cells to support the quiescence of limbal epithelial stem cells. CONCLUSIONS We envision that HC-HA/PTX3 purified from AM can be used as a unique substrate to refine ex vivo expansion of limbal epithelial stem cells by maintaining stem cell quiescence, self-renewal and fate decision. Furthermore, it can also be deployed as a platform to launch new therapeutics in regenerative medicine by mitigating nonresolving inflammation and reinforcing the well-being of stem cell niche.
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Affiliation(s)
- Scheffer C. G. Tseng
- The R&D Department of TissueTech Inc., Ocular Surface Center, and Ocular Surface Research and Education Foundation, Miami, Florida, United States
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21
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Induction of apoptosis, stimulation of cell-cycle arrest and inhibition of angiogenesis make human amnion-derived cells promising sources for cell therapy of cancer. Cell Tissue Res 2016; 363:599-608. [DOI: 10.1007/s00441-016-2364-3] [Citation(s) in RCA: 51] [Impact Index Per Article: 5.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2015] [Accepted: 01/14/2016] [Indexed: 12/11/2022]
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Tseng SCG, He H, Zhang S, Chen SY. Niche Regulation of Limbal Epithelial Stem Cells: Relationship between Inflammation and Regeneration. Ocul Surf 2016; 14:100-12. [PMID: 26769483 DOI: 10.1016/j.jtos.2015.12.002] [Citation(s) in RCA: 58] [Impact Index Per Article: 6.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/20/2015] [Revised: 12/17/2015] [Accepted: 12/24/2015] [Indexed: 02/07/2023]
Abstract
Human limbal palisades of Vogt are the ideal site for studying and practicing regenerative medicine due to their accessibility. Nonresolving inflammation in limbal stroma is common manifestation of limbal stem cell (SC) deficiency and presents as a threat to the success of transplanted limbal epithelial SCs. This pathologic process can be overcome by transplantation of cryopreserved human amniotic membrane (AM), which exerts anti-inflammatory, antiscarring and anti-angiogenic action to promote wound healing. To determine how AM might exert anti-inflammation and promote regeneration, we have purified a novel matrix, HC-HA/PTX3, responsible for the efficacy of AM efficacy. HC-HA complex is covalently formed by hyaluronan (HA) and heavy chain 1 (HC1) of inter-α-trypsin inhibitor by the catalytic action of tumor necrosis factor-stimulated gene-6 (TSG-6) and are tightly associated with pentraxin 3 (PTX3) to form HC-HA/PTX3. In vitro reconstitution of the limbal niche can be established by reunion between limbal epithelial progenitors and limbal niche cells on different substrates. In 3-dimensional Matrigel, clonal expansion indicative of SC renewal is correlated with activation of canonical Wnt signaling and suppression of canonical bone morphogenetic protein (BMP) signaling. In contrast, SC quiescence can be achieved in HC-HA/PTX3 by activation of canonical BMP signaling and non-canonical planar cell polarity (PCP) Wnt signaling, but suppression of canonical Wnt signaling. HC-HA/PTX3 is a novel matrix mitigating nonresolving inflammation and restoring SC quiescence in the niche for various applications in regenerative medicine.
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Affiliation(s)
- Scheffer C G Tseng
- R&D Department, TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research and Education Foundation, Miami, Florida, USA.
| | - Hua He
- R&D Department, TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research and Education Foundation, Miami, Florida, USA
| | - Suzhen Zhang
- R&D Department, TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research and Education Foundation, Miami, Florida, USA
| | - Szu-Yu Chen
- R&D Department, TissueTech, Inc., Ocular Surface Center, and Ocular Surface Research and Education Foundation, Miami, Florida, USA
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Cheng AMS, Zhao D, Chen R, Yin HY, Tighe S, Sheha H, Casas V, Tseng SCG. Accelerated Restoration of Ocular Surface Health in Dry Eye Disease by Self-Retained Cryopreserved Amniotic Membrane. Ocul Surf 2015; 14:56-63. [PMID: 26387870 DOI: 10.1016/j.jtos.2015.07.003] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/06/2015] [Revised: 06/18/2015] [Accepted: 07/10/2015] [Indexed: 10/23/2022]
Abstract
PURPOSE To evaluate the clinical efficacy of self-retained cryopreserved amniotic membrane in treating dry eye disease. METHODS Retrospective review of 10 patients treated with self-retained cryopreserved amniotic membrane (PROKERA® Slim [PKS], Bio-Tissue, Miami, FL) for moderate-to-severe dry eye refractory to conventional maximal medical treatments. Patients' symptoms, use of medications, conjunctival inflammation, corneal staining, and visual acuity were compared before and after treatment. RESULTS PKS was placed in 15 eyes of the 10 patients for 4.9 ± 1.5 days. All patients experienced symptomatic relief for a period of 4.2 ± 4.7 months (P<.001). Such improvement was accompanied by reduction of OSDI scores (P<.001), use of topical medications (P<.001), conjunctival hyperemia (P<.001), corneal staining (P<.001), and improvement of the visual acuity (P=.06). Linear regression analysis estimated that the optimal duration of PKS placement was 5 days to achieve an average symptom-free duration of 4 months in patients with dry eye. Surprisingly, PKS placement also generated improvement in the contralateral eyes. CONCLUSION This pilot study suggests that self-retained cryopreserved amniotic membrane via PKS can be used to treat moderate dry eye diseases and warrants further prospective controlled studies.
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Affiliation(s)
- Anny M S Cheng
- Ocular Surface Center and Tissuetech Inc., Miami FL, USA
| | - Dandan Zhao
- Department of Ophthalmology, Yan'An Hospital of Kunming City, Kunming, Yunnan, China
| | - Rendian Chen
- Department of Ophthalmology, Shenzhen Children's Hospital, Shenzhen, Guangzhou, China
| | - Han Y Yin
- Florida International University Herbert Wertheim College of Medicine, Miami, FL, USA
| | - Sean Tighe
- Ocular Surface Center and Tissuetech Inc., Miami FL, USA
| | - Hosam Sheha
- Ocular Surface Center and Tissuetech Inc., Miami FL, USA
| | - Victoria Casas
- Ocular Surface Center and Tissuetech Inc., Miami FL, USA
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Song TH, Jang J, Choi YJ, Shim JH, Cho DW. 3D-Printed Drug/Cell Carrier Enabling Effective Release of Cyclosporin A for Xenogeneic Cell-Based Therapy. Cell Transplant 2015; 24:2513-25. [PMID: 25608278 DOI: 10.3727/096368915x686779] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023] Open
Abstract
Systemic administration of the immunosuppressive drug cyclosporin A (CsA) is frequently associated with a number of side effects; therefore, sometimes it cannot be applied in sufficient dosage after allogeneic or xenogeneic cell transplantation. Local delivery is a possible solution to this problem. We used 3D printing to develop a CsA-loaded 3D drug carrier for the purpose of local and sustained delivery of CsA. The carrier is a hybrid of CsA-poly(lactic-co-glycolic acid) (PLGA) microsphere-loaded hydrogel and a polymeric framework so that external force can be endured under physiological conditions. The expression of cytokines, which are secreted by spleen cells activated by Con A, and which are related to immune rejection, was significantly decreased in vitro by the released CsA from the drug carrier. Drug carriers seeded with xenogeneic cells (human lung fibroblast) were subcutaneously implanted into the BALB/c mouse. As a result, T-cell-mediated rejection was also significantly suppressed for 4 weeks. These results show that the developed 3D drug carrier can be used as an effective xenogeneic cell delivery system with controllable immunosuppressive drugs for cell-based therapy.
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Affiliation(s)
- Tae-Ha Song
- Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), Hyoja-dong, Nam-gu, Pohang, Kyungbuk, Korea
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Dudok DV, Nagdee I, Cheung K, Liu H, Vedovelli L, Ghinelli E, Kenyon K, Parapuram S, Hutnik CM. Effects of amniotic membrane extract on primary human corneal epithelial and limbal cells. Clin Exp Ophthalmol 2015; 43:443-8. [PMID: 25495256 DOI: 10.1111/ceo.12480] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2014] [Accepted: 12/07/2014] [Indexed: 11/30/2022]
Abstract
BACKGROUND To assess the effects of amniotic membrane extract (AMX) on cellular activity of primary human corneal epithelial (HCE) cells under mechanical and oxidative stress, and on human limbal cells under oxidative stress. METHODS Corneal mechanical stress was simulated with a linear scratch in confluent HCE cell plates, then incubated with 0.1% AMX for 48 and 72 h. Subjecting HCE cultures to 0.5 mmol/L tertiary-butylhydroperoxide for 1 h simulated an oxidative stress. 0.1% AMX-treated cultures were compared with controls at 24 and 48 h using cellular viability assay, along with 12-h AMX pretreatment and human limbal cell comparisons. RESULTS Mechanical stress on HCE cultures revealed a statistically significant distance ratio at 48 and 72 h in favour of 0.1% AMX-treated cultures (P = 0.021 and 0.035, respectively). Oxidative stress did not reveal any significant difference in cellular viability of AMX-treated versus control cultures. Twelve hour AMX pre-treatment prior to oxidative stress revealed a significant difference after 24 h from oxidative injury (73.3% AMX vs. 66.0% control, P = 0.035), but not after 48 h. Human limbal cells demonstrated significantly improved oxidative viability compared with HCE cells, with (91.0% vs. 82.0% control, P = 0.017) and without 0.1% AMX pre-treatment (91.2% vs. 83.7% control, P = 0.019). CONCLUSIONS HCE cells treated with AMX healed faster after mechanical insult, suggesting a potential benefit in acute corneal injuries. Under oxidative stress, human limbal cells, a more proliferative cell type, showed superior viability compared with HCE cells.
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Affiliation(s)
- David V Dudok
- Ivey Eye Institute, Western University, London, Ontario, Canada
| | - Imraan Nagdee
- Ivey Eye Institute, Western University, London, Ontario, Canada
| | - Kevin Cheung
- Ivey Eye Institute, Western University, London, Ontario, Canada
| | - Hong Liu
- Ivey Eye Institute, Western University, London, Ontario, Canada
| | - Luca Vedovelli
- Division of Nephrology, Cell Biology, Human Biology and Physiology, Massachusetts General Hospital, Boston, Massachusetts, USA
| | - Emiliano Ghinelli
- Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts, USA
| | - Kenneth Kenyon
- Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts, USA.,New England Medical Center, Tufts University, Boston, Massachusetts, USA
| | - Sunil Parapuram
- Ivey Eye Institute, Western University, London, Ontario, Canada
| | - Cindy M Hutnik
- Ivey Eye Institute, Western University, London, Ontario, Canada
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Krawczyk A, Dirks M, Kasper M, Buch A, Dittmer U, Giebel B, Wildschütz L, Busch M, Goergens A, Schneweis KE, Eis-Hübinger AM, Sodeik B, Heiligenhaus A, Roggendorf M, Bauer D. Prevention of herpes simplex virus induced stromal keratitis by a glycoprotein B-specific monoclonal antibody. PLoS One 2015; 10:e0116800. [PMID: 25587898 PMCID: PMC4294644 DOI: 10.1371/journal.pone.0116800] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/17/2014] [Accepted: 12/15/2014] [Indexed: 10/24/2022] Open
Abstract
The increasing incidence of acyclovir (ACV) and multidrug-resistant strains in patients with corneal HSV-1 infections leading to Herpetic Stromal Keratitis (HSK) is a major health problem in industrialized countries and often results in blindness. To overcome this obstacle, we have previously developed an HSV-gB-specific monoclonal antibody (mAb 2c) that proved to be highly protective in immunodeficient NOD/SCID-mice towards genital infections. In the present study, we examined the effectivity of mAb 2c in preventing the immunopathological disease HSK in the HSK BALB/c mouse model. Therefore, mice were inoculated with HSV-1 strain KOS on the scarified cornea to induce HSK and subsequently either systemically or topically treated with mAb 2c. Systemic treatment was performed by intravenous administration of mAb 2c 24 h prior to infection (pre-exposure prophylaxis) or 24, 40, and 56 hours after infection (post-exposure immunotherapy). Topical treatment was performed by periodical inoculations (5 times per day) of antibody-containing eye drops as control, starting at 24 h post infection. Systemic antibody treatment markedly reduced viral loads at the site of infection and completely protected mice from developing HSK. The administration of the antiviral antibody prior or post infection was equally effective. Topical treatment had no improving effect on the severity of HSK. In conclusion, our data demonstrate that mAb 2c proved to be an excellent drug for the treatment of corneal HSV-infections and for prevention of HSK and blindness. Moreover, the humanized counterpart (mAb hu2c) was equally effective in protecting mice from HSV-induced HSK when compared to the parental mouse antibody. These results warrant the future development of this antibody as a novel approach for the treatment of corneal HSV-infections in humans.
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Affiliation(s)
- Adalbert Krawczyk
- Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
- * E-mail:
| | - Miriam Dirks
- Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
| | - Maren Kasper
- Ophtha-Lab, Department of Ophthalmology at St. Franziskus Hospital, Muenster, Germany
| | - Anna Buch
- Institute of Virology, Hannover Medical School, Hannover, Germany
| | - Ulf Dittmer
- Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
| | - Bernd Giebel
- Institute for Transfusion Medicine, University Hospital Essen, Essen, Germany
| | - Lena Wildschütz
- Ophtha-Lab, Department of Ophthalmology at St. Franziskus Hospital, Muenster, Germany
| | - Martin Busch
- Ophtha-Lab, Department of Ophthalmology at St. Franziskus Hospital, Muenster, Germany
| | - Andre Goergens
- Institute for Transfusion Medicine, University Hospital Essen, Essen, Germany
| | - Karl E. Schneweis
- Institute of Virology, University Medical Center Bonn, Bonn, Germany
| | | | - Beate Sodeik
- Institute of Virology, Hannover Medical School, Hannover, Germany
| | - Arnd Heiligenhaus
- Ophtha-Lab, Department of Ophthalmology at St. Franziskus Hospital, Muenster, Germany
| | - Michael Roggendorf
- Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany
| | - Dirk Bauer
- Ophtha-Lab, Department of Ophthalmology at St. Franziskus Hospital, Muenster, Germany
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Domínguez-López A, Bautista-de Lucio VM, Serafín-López J, Robles-Sánchez E, Garfias Y. Amniotic membrane modulates innate immune response inhibiting PRRs expression and NF-κB nuclear translocation on limbal myofibroblasts. Exp Eye Res 2014; 127:215-23. [PMID: 25117451 DOI: 10.1016/j.exer.2014.08.002] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2014] [Revised: 06/18/2014] [Accepted: 08/02/2014] [Indexed: 10/24/2022]
Abstract
Corneal damage observed in a viral infection such as herpetic stromal keratitis is mainly caused by proinflammatory molecules released by resident cells in the response to viral antigens. There are pattern recognition receptors like MDA5, RIG-1, and TLR3, that recognize viral dsRNA and after activation, the innate immune response is exacerbated inducing the synthesis and secretion of inflammatory cytokines through NF-κB activation. Amniotic membrane (AM) has demonstrated to reduce inflammation by several mechanisms, however the effect of AM on innate immune receptors such as MDA5, RIG-1, and TLR3 has not been reported. In this study, we have determined that the presence of AM significantly inhibited the synthesis and secretion of proinflammatory cytokines on human limbal myofibroblasts (HLM) stimulated with poly I:C. Similarly, the presence of AM reduced the protein expression of MDA5, RIG-1, and TLR3 on poly I:C stimulated HLM. Additionally, the presence of the AM significantly inhibited the NF-κB nuclear translocation when the HLM were poly I:C stimulated, and concomitantly, the AM was able to relocate cadherins affecting the myofibroblastic cellular morphology. These results suggest that AM generates an anti-inflammatory microenvironment, and specific inhibition of NFκB nuclear translocation on infected corneal tissue would reduce the inflammation undesirable effects, explaining in part the beneficial usefulness of transplanting AM on herpetic stromal keratitis.
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Affiliation(s)
- Alfredo Domínguez-López
- Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800 Mexico City, Mexico; Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Insurgentes Sur 3000, 04510 Mexico City, Mexico
| | | | - Janet Serafín-López
- Department of Immunology, National School of Biological Sciences, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala, 11340 Mexico City, Mexico
| | - Edson Robles-Sánchez
- Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800 Mexico City, Mexico; Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Insurgentes Sur 3000, 04510 Mexico City, Mexico
| | - Yonathan Garfias
- Research Unit, Institute of Ophthalmology Conde de Valenciana Foundation, Chimalpopoca 14, 06800 Mexico City, Mexico; Department of Biochemistry, Faculty of Medicine, Universidad Nacional Autónoma de México, Insurgentes Sur 3000, 04510 Mexico City, Mexico.
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Nguyen P, Rue K, Heur M, Yiu SC. Ocular surface rehabilitation: Application of human amniotic membrane in high-risk penetrating keratoplasties. Saudi J Ophthalmol 2014; 28:198-202. [PMID: 25278797 DOI: 10.1016/j.sjopt.2014.06.010] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2014] [Revised: 06/01/2014] [Accepted: 06/15/2014] [Indexed: 11/17/2022] Open
Abstract
BACKGROUND Human amniotic membrane is a versatile tool for management of ocular surface disorders. This study evaluates the effect of cryopreserved human amniotic membrane (hAM) on one-year survival of penetrating keratoplasties (PKP) in high-risk recipients. METHOD This is a retrospective noncomparative cohort study of 58 consecutive eyes undergoing PKP with concurrent placement of a self-retained cryopreserved hAM (PROKERA®) at a tertiary care center from January 2009 to July 2010. RESULTS Mean patient age was 66.7 ± 17.2 years and 30 (54%) were males. 51 eyes were pseudophakic and one aphakic. 27 eyes were glaucomatous; 24 had glaucoma drainage device and 2 had previous endocyclophotocoagulation. 12 patients had PKP for the first time and 46 had repeat PKP (average number of prior PKP = 1.63 ± 1.1, range: 1-5). Risk factors for graft failure included repeat PKP (79.3%), corneal neovascularization (51.7%), preexisting glaucoma (46.6%), and presence of anterior synechiae (37.9%). Both First Transplant and Repeat Transplant groups had similar survival rates until 6 months after transplant (75% vs 74%, odds ratio = 1.06, p = 1.00). At 12 months, First Transplant group showed a better survival rate (67% vs 43%, odds ratio = 2.60, p = 0.20). Eyes with >3 risk factors had a higher graft failure rate (odds ratio = 5.81, p = 0.003). CONCLUSION Survey of the literature suggests that high-risk PKP with concurrent hAM placement demonstrate comparable graft survival. Presence of multiple risk factors is associated with poor survival.
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Affiliation(s)
- Pho Nguyen
- Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA
| | - Kelly Rue
- Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA
| | - Martin Heur
- Department of Ophthalmology, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA
| | - Samuel C Yiu
- Department of Ophthalmology, The Wilmer Eye Institute, The John Hopkins University, Baltimore, MD, USA
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He H, Tan Y, Duffort S, Perez VL, Tseng SCG. In vivo downregulation of innate and adaptive immune responses in corneal allograft rejection by HC-HA/PTX3 complex purified from amniotic membrane. Invest Ophthalmol Vis Sci 2014; 55:1647-56. [PMID: 24519420 DOI: 10.1167/iovs.13-13094] [Citation(s) in RCA: 43] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/19/2023] Open
Abstract
PURPOSE Heavy chain-hyaluronic acid (HC-HA)/PTX3 purified from human amniotic membrane (AM) was previously observed to suppress inflammatory responses in vitro. We now examine whether HC-HA/PTX3 is able to exert a similar effect in vivo, using murine models for keratitis and corneal allograft rejection. METHODS The in vitro effect of HC-HA/PTX3 was tested using OTII ovalbumin (OVA) transgenic, purified CD4(+) T cells, or IFN-γ/lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Cytokine production was measured by ELISA, while cell surface markers and cell proliferation were determined by flow cytometry. In vivo effects of HC-HA/PTX3 were analyzed by quantifying the recruitment of enhanced green fluorescence-labeled macrophages and by measuring the expression of arginase 1 (Arg-1), IL-10, and IL-12 in LPS-induced keratitis in the macrophage Fas-induced apoptosis (Mafia) mouse. The effect of corneal allograft survival in a complete major histocompatibility complex (MHC) mismatched mouse model was assessed by grading corneal opacification. RESULTS In vitro studies demonstrated that HC-HA/PTX3 significantly enhanced the expansion of FOXP3 T cells and suppressed cell proliferation and protein expression of IFN-γ, IL-2, CD25, and CD69 in activated CD4(+) T cells. Furthermore, immobilized HC-HA/PTX3 significantly upregulated IL-10 gene expression but downregulated that of IL-12 and IL-23 in activated RAW264.7 cells. Finally, in vivo subconjunctival injection of HC-HA/PTX3 significantly prolonged corneal allograft survival, suppressed macrophage infiltration, and promoted M2 polarization by upregulating Arg-1 and IL-10 but downregulating IL-12. CONCLUSIONS HC-HA/PTX3 can suppress inflammatory responses in vivo by modulating both innate and adaptive immunity of macrophages and CD4(+) T cells.
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Affiliation(s)
- Hua He
- TissueTech, Inc., Miami, Florida
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Liu J, Sheha H, Fu Y, Liang L, Tseng SC. Update on amniotic membrane transplantation. EXPERT REVIEW OF OPHTHALMOLOGY 2014; 5:645-661. [PMID: 21436959 DOI: 10.1586/eop.10.63] [Citation(s) in RCA: 134] [Impact Index Per Article: 12.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Abstract
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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Affiliation(s)
- Jingbo Liu
- Ocular Surface Center, 7000 SW, 97 Avenue, Suite 213, Miami, FL 33173, USA
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31
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Cirman T, Beltram M, Schollmayer P, Rožman P, Kreft ME. Amniotic membrane properties and current practice of amniotic membrane use in ophthalmology in Slovenia. Cell Tissue Bank 2013; 15:177-92. [PMID: 24352631 DOI: 10.1007/s10561-013-9417-6] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/16/2013] [Accepted: 12/12/2013] [Indexed: 01/01/2023]
Abstract
Amniotic membrane (AM) is the innermost, multilayered part of the placenta. When harvested, processed and stored properly, its properties, stemming from AM biological composition, make it a useful tissue for ophthalmic surgery. AM was shown to have several beneficial effects: it promotes epithelization, has antimicrobial effects, decreases inflammation, fibrosis and neovascularization. Many case reports and case series as well as practical experience (e.g. reconstruction of conjunctival and corneal defects, treatment of corneal ulcers) demonstrated the beneficial effect of AM for different ophthalmological indications. The combination of the above mentioned beneficial effects and reasonable mechanical properties are also the reason why AM is used as a substrate for ex vivo expansion of epithelial progenitor cells. Recently, amnion-derived cells, which also have stem cell characteristics, have been proposed as potential contributors to cell-based treatment of ocular surface disease. However, the use of AM remains one of the least standardized methods in ophthalmic surgery. In this review, the various properties of AM and its current clinical use in ophthalmology in Slovenia are discussed.
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Affiliation(s)
- Tina Cirman
- Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia
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He H, Zhang S, Tighe S, Son J, Tseng SCG. Immobilized heavy chain-hyaluronic acid polarizes lipopolysaccharide-activated macrophages toward M2 phenotype. J Biol Chem 2013; 288:25792-25803. [PMID: 23878196 DOI: 10.1074/jbc.m113.479584] [Citation(s) in RCA: 100] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023] Open
Abstract
Despite the known anti-inflammatory effect of amniotic membrane, its action mechanism remains largely unknown. HC-HA complex (HC-HA) purified from human amniotic membrane consists of high molecular weight hyaluronic acid (HA) covalently linked to the heavy chain (HC) 1 of inter-α-trypsin inhibitor. In this study, we show that soluble HC-HA also contained pentraxin 3 and induced the apoptosis of both formyl-Met-Leu-Phe or LPS-activated neutrophils and LPS-activated macrophages while not affecting the resting cells. This enhanced apoptosis was caused by the inhibition of cell adhesion, spreading, and proliferation caused by HC-HA binding of LPS-activated macrophages and preventing adhesion to the plastic surface. Preferentially, soluble HC-HA promoted phagocytosis of apoptotic neutrophils in resting macrophages, whereas immobilized HC-HA promoted phagocytosis in LPS-activated macrophages. Upon concomitant LPS stimulation, immobilized HC-HA but not HA polarized macrophages toward the M2 phenotype by down-regulating IRF5 protein and preventing its nuclear localization and by down-regulating IL-12, TNF-α, and NO synthase 2. Additionally, IL-10, TGF-β1, peroxisome proliferator-activated receptor γ, LIGHT (TNF superfamily 14), and sphingosine kinase-1 were up-regulated, and such M2 polarization was dependent on TLR ligation. Collectively, these data suggest that HC-HA is a unique matrix component different from HA and uses multiple mechanisms to suppress M1 while promoting M2 phenotype. This anti-inflammatory action of HC-HA is highly desirable to promote wound healing in diseases heightened by unsuccessful transition from M1 to M2 phenotypes.
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Affiliation(s)
| | | | | | - Ji Son
- the Ocular Surface Research Education Foundation, and
| | - Scheffer C G Tseng
- From the TissueTech, Inc.,; the Ocular Surface Research Education Foundation, and; the Ocular Surface Center, Miami, Florida 33173.
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Labetoulle M, Colin J. Aspects actuels du traitement des kératites herpétiques. J Fr Ophtalmol 2012; 35:292-307. [DOI: 10.1016/j.jfo.2011.10.002] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/04/2011] [Revised: 09/26/2011] [Accepted: 10/05/2011] [Indexed: 01/18/2023]
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Of mice and not humans: how reliable are animal models for evaluation of herpes CD8(+)-T cell-epitopes-based immunotherapeutic vaccine candidates? Vaccine 2011; 29:5824-36. [PMID: 21718746 DOI: 10.1016/j.vaccine.2011.06.083] [Citation(s) in RCA: 53] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2011] [Revised: 06/09/2011] [Accepted: 06/14/2011] [Indexed: 11/23/2022]
Abstract
Herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2)-specific CD8(+) T cells that reside in sensory ganglia, appear to control recurrent herpetic disease by aborting or reducing spontaneous and sporadic reactivations of latent virus. A reliable animal model is the ultimate key factor to test the efficacy of therapeutic vaccines that boost the level and the quality of sensory ganglia-resident CD8(+) T cells against spontaneous herpes reactivation from sensory neurons, yet its relevance has been often overlooked. Herpes vaccinologists are hesitant about using mouse as a model in pre-clinical development of therapeutic vaccines because they do not adequately mimic spontaneous viral shedding or recurrent symptomatic diseases, as occurs in human. Alternatives to mouse models are rabbits and guinea pigs in which reactivation arise spontaneously with clinical herpetic features relevant to human disease. However, while rabbits and guinea pigs develop spontaneous HSV reactivation and recurrent ocular and genital disease none of them can mount CD8(+) T cell responses specific to Human Leukocyte Antigen- (HLA-)restricted epitopes. In this review, we discuss the advantages and limitations of these animal models and describe a novel "humanized" HLA transgenic rabbit, which shows spontaneous HSV-1 reactivation, recurrent ocular disease and mounts CD8(+) T cell responses to HLA-restricted epitopes. Adequate investments are needed to develop reliable preclinical animal models, such as HLA class I and class II double transgenic rabbits and guinea pigs to balance the ethical and financial concerns associated with the rising number of unsuccessful clinical trials for therapeutic vaccine formulations tested in unreliable mouse models.
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Lin JA, Chen JH, Lee YW, Lin CS, Hsieh MH, Chang CC, Wong CS, Chen JJY, Yeh GC, Lin FY, Chen TL. Biphasic effect of curcumin on morphine tolerance: a preliminary evidence from cytokine/chemokine protein array analysis. EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE 2011; 2011:452153. [PMID: 21826185 PMCID: PMC3150782 DOI: 10.1093/ecam/neq018] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 06/22/2009] [Accepted: 02/08/2010] [Indexed: 11/26/2022]
Abstract
The aim of this study was to evaluate the effect of curcumin on morphine tolerance and the corresponding cytokine/chemokine changes. Male ICR mice were made tolerant to morphine by daily subcutaneous injection for 7 days. Intraperitoneal injections of vehicle, low-dose or high-dose curcumin were administered 15 min after morphine injection, either acutely or chronically for 7 days to test the effect of curcumin on morphine-induced antinociception and development of morphine tolerance. On day 8, cumulative dose-response curves were generated and the 50% of maximal analgesic dose values were calculated and compared among groups. Corresponding set of mice were used for analyzing the cytokine responses by antibody-based cytokine protein array. Acute, high-dose curcumin enhanced morphine-induced antinociception. While morphine tolerance was attenuated by administration of low-dose curcumin following morphine injections for 7 days, it was aggravated by chronic high-dose curcumin following morphine injection, suggesting a biphasic effect of curcumin on morphine-induced tolerance. Of the 96 cytokine/chemokines analyzed by mouse cytokine protein array, 14 cytokines exhibited significant changes after the different 7-day treatments. Mechanisms for the modulatory effects of low-dose and high-dose curcumin on morphine tolerance were discussed. Even though curcumin itself is a neuroprotectant and low doses of the compound serve to attenuate morphine tolerance, high-doses of curcumin might cause neurotoxicity and aggravate morphine tolerance by inhibiting the expression of antiapoptotic cytokines and neuroprotective factors. Our results indicate that the effect of curcumin on morphine tolerance may be biphasic, and therefore curcumin should be used cautiously.
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Affiliation(s)
- Jui-An Lin
- Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taiwan
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36
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Ocular involvement in chronic graft-versus-host disease: therapeutic approaches to complicated courses. Cornea 2011; 30:107-13. [PMID: 20847653 DOI: 10.1097/ico.0b013e3181e2ecf0] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Chronic graft-versus-host disease (cGvHD) is a major concomitant phenomenon in recipients of allogeneic hematopoietic stem cell transplantations, affecting multiple organ systems including the eye. Ocular structures, such as lacrimal gland, conjunctiva, and eyelids with meibomian glands, are frequently involved with clinical features ranging from dry eyes and common inflammatory conjunctival disease to severe complications like corneal ulcerations or even perforations. We present 2 patients with complicated courses of ocular cGvHD. In both cases, keratoconjunctivitis sicca refractory to systemic immunosuppressive therapy and to conventional topical treatment resulted in progressive binocular corneal melting and finally repeated perforations. According to our 2 cases and to the current pathophysiological understanding, we discuss possible strategies for the treatment and prevention of ocular cGvHD complications.
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Gu HW, Bian DM, Hu N, Zhang JF. Effects of amniotic membrane transplantation on cytokines expression in chemically burned rat corneas. Int J Ophthalmol 2011; 4:33-6. [PMID: 22553604 DOI: 10.3980/j.issn.2222-3959.2011.01.07] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/24/2010] [Accepted: 01/15/2011] [Indexed: 11/02/2022] Open
Abstract
AIM To investigate the effect of amniotic membrane transplantation (AMT) on the expressions of inflammatory-related, angiogenic-related and growth-related cytokines in rat corneas after chemical injury. METHODS Alkali wounds were inflicted on the central corneas of rats by applying a round filter paper soaked in 1mol/L NaOH for 40 seconds. One week after alkali burn, 12 rats were randomly divided into 2 groups: the AMT group and the control group, and AMT was performed on the rats in the AMT group. Corneal opacity and neovascularization were observed by slit-lamp microscopy. The protein levels of interleukin (IL)-2, interferon (IFN)-γ, IL-10 and transforming growth factor (TGF)-β were determined by enzyme-linked immunosorbent assay 2 weeks after AMT. The mRNA levels of matrix metalloproteinase-2 (MMP-2), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) were evaluated by real-time quantitative PCR. RESULTS In the AMT group, the corneal opacity was improved (P=0.011) and the area of corneal neovascularization was significantly decreased (P=0.005) compared with the control group. The amount of IL-2 and IFN-γ secreted by Th1 cells were decreased after AMT, whereas the amount of IL-10 and TGF-β secreted by Th2 cells were increased (P<0.05). The level of MMP-2 was significantly down-regulated (P=0.013) at the mRNA level in the AMT group, while the expression of EGF was significantly higher (P=0.022) compared with controls. CONCLUSION [corrected] AMT may suppress corneal neovascularization after chemical injury by modulating the expressions of soluble factors.
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Affiliation(s)
- Hong-Wei Gu
- Department of Ophthalmology, the Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
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Garfias Y, Zaga-Clavellina V, Vadillo-Ortega F, Osorio M, Jimenez-Martinez MC. Amniotic membrane is an immunosuppressor of peripheral blood mononuclear cells. Immunol Invest 2010; 40:183-96. [PMID: 21080833 DOI: 10.3109/08820139.2010.532266] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
UNLABELLED Amniotic membrane (AM) is the inner layer of the placenta, which is in contact with the fetus; it has been used for transplantation in ocular surface diseases. It has been reported that amniotic membrane promotes epithelialization, inhibits angiogenesis and diminishes ocular inflammation. A persistent epithelial defect is the delay in epithelial wound healing caused by infiltrating inflammatory cells into the cornea and amniotic membrane transplantation has been successfully used in its treatment, however the mechanism of action in inhibiting inflammation it is not well understood. This study was aimed at determining whether denuded amniotic membrane (dAM) induces anti-inflammatory effects on peripheral blood mononuclear cells. METHODS Human peripheral blood mononuclear cells (PBMC) were cultured on dAM. Proliferation and apoptosis assays were performed on PBMC; and synthesis and secretion of pro-inflammatory cytokines by these cells was analyzed. RESULTS dAM induced apoptosis and inhibited cell proliferation of PBMC; and abolished the synthesis and the secretion of pro-inflammatory cytokines even when they were LPS stimulated. In contrast, when PBMC were cultured on hydrophilic membrane cell culture inserts, apoptosis was not significantly induced, cell proliferation was conserved, and synthesis and secretion of pro-inflammatory cytokines were not affected. CONCLUSIONS Taken together, these results could explain the anti-inflammatory in vivo effects observed when the amniotic membrane is used as a transplant.
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Affiliation(s)
- Yonathan Garfias
- Research Unit, Ophthalmology Institute, Conde de Valenciana Foundation, Mexico City, 06800.
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