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Wu KY, Kearn N, Truong D, Choulakian MY, Tran SD. Advances in Regenerative Medicine, Cell Therapy, and 3D Bioprinting for Corneal, Oculoplastic, and Orbital Surgery. ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY 2025. [PMID: 40131704 DOI: 10.1007/5584_2025_855] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Subscribe] [Scholar Register] [Indexed: 03/27/2025]
Abstract
Advances in regenerative medicine, cell therapy, and 3D bioprinting are reshaping the landscape of ocular surgery, offering innovative approaches to address complex conditions affecting the cornea, ocular adnexal structures, and the orbit. These technologies hold the potential to enhance treatment precision, improve functional outcomes, and address limitations in traditional surgical and therapeutic interventions.The cornea, as the eye's primary refractive and protective barrier, is particularly well-suited for regenerative approaches due to its avascular and immune-privileged nature. Cell-based therapies, including limbal stem cell transplantation as well as stromal keratocyte and corneal endothelial cell regeneration, are being investigated for their potential to restore corneal clarity and function in conditions such as limbal stem cell deficiency, keratoconus, and endothelial dysfunction. Simultaneously, 3D bioprinting technologies are enabling the development of biomimetic corneal constructs, potentially addressing the global shortage of donor tissues and facilitating personalized surgical solutions.In oculoplastic and orbital surgery, regenerative strategies and cell therapies are emerging as possible alternatives to conventional approaches for conditions such as eyelid defects, meibomian gland dysfunction, and Graves' orbitopathy. Stem cell-based therapies and bioengineered scaffolds are showing potential in restoring lacrimal glands' function as well as reconstructing complex ocular adnexal and orbital structures. Moreover, 3D-printed orbital implants and scaffolds offer innovative solutions for repairing traumatic, post-tumor resection, and congenital defects, with the potential for improved biocompatibility and precision.Molecular and gene-based therapies, including exosome delivery systems, nanoparticle-based interventions, and gene-editing techniques, are expanding the therapeutic arsenal for ophthalmic disorders. These approaches aim to enhance the efficacy of regenerative treatments by addressing underlying pathophysiological mechanisms of diseases. This chapter provides an overview of these advancements and the challenges of translating laboratory discoveries into effective therapies in clinical practice.
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Affiliation(s)
- Kevin Y Wu
- Department of Surgery, Division of Ophthalmology, University of Sherbrooke, Sherbrooke, QC, Canada
| | - Natalie Kearn
- Department of Medicine, School of Medicine, Queen's University, Kingston, ON, Canada
| | - Doanh Truong
- College of Arts & Science, Case Western Reserve University, Cleveland, OH, USA
| | - Mazen Y Choulakian
- Department of Surgery, Division of Ophthalmology, University of Sherbrooke, Sherbrooke, QC, Canada
| | - Simon D Tran
- Faculty of Dental Medicine and Oral Health Sciences, McGill University, Montreal, QC, Canada.
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Bonnet C, Gonzalez S, Deng SX. Limbal stem cell therapy. Curr Opin Ophthalmol 2024; 35:309-314. [PMID: 38813737 DOI: 10.1097/icu.0000000000001061] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/31/2024]
Abstract
PURPOSE OF REVIEW To highlight the progress and future direction of limbal stem cell (LSC) therapies for the treatment of limbal stem cell deficiency (LSCD). RECENT FINDINGS Direct LSC transplantation have demonstrated good long-term outcomes. Cultivated limbal epithelial transplantation (CLET) has been an alternative to treat severe to total LSCD aiming to improve the safety and efficacy of the LSC transplant. A prospective early-stage uncontrolled clinical trial shows the feasibility and safety of CLET manufactured under xenobiotic free conditions. Other cell sources for repopulating of the corneal epithelium such as mesenchymal stem cells (MSCs) and induced pluripotent stem cells are being investigated. The first clinical trials of using MSCs showed short-term results, but long-term efficacy seems to be disappointing. A better understanding of the niche function and regulation of LSC survival and proliferation will lead to the development of medical therapies to rejuvenate the residual LSCs found in a majority of eyes with LSCD in vivo. Prior efforts have been largely focused on improving LSC transplantation. Additional effort should be placed on improving the accuracy of diagnosis and staging of LSCD, and implementing standardized outcome measures which enable comparison of efficacy of different LSCD treatments for different severity of LSCD. The choice of LSCD treatment will be customized based on the severity of LSCD in the future. SUMMARY New approaches for managing different stages of LSCD are being developed. This concise review summarizes the progresses in LSC therapies for LSCD, underlying mechanisms, limitations, and future areas of development.
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Affiliation(s)
- Clemence Bonnet
- Stein Eye Institute, University of California, Los Angeles, California, USA
- Centre de Recherche des Cordeliers, INSERM 1138, Paris Cité Université, AP-HP, Paris, France
| | - Sheyla Gonzalez
- Stein Eye Institute, University of California, Los Angeles, California, USA
| | - Sophie X Deng
- Stein Eye Institute, University of California, Los Angeles, California, USA
- Molecular Biology Institute, University of California, Los Angeles, California, USA
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Takayanagi H, Hayashi R. Status and prospects for the development of regenerative therapies for corneal and ocular diseases. Regen Ther 2024; 26:819-825. [PMID: 39329098 PMCID: PMC11424903 DOI: 10.1016/j.reth.2024.09.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/24/2024] [Revised: 08/13/2024] [Accepted: 09/05/2024] [Indexed: 09/28/2024] Open
Abstract
Among the regenerative therapies being put into clinical use, the field of corneal regenerative therapy is one of the most advanced, with several regulatory approved products. This article describes the progress from initial development through to clinical application in the eye field, with a particular focus on therapies for corneal epithelial and endothelial diseases that have already been regulatory approved as regenerative therapy products. The applications of regenerative therapy to the corneal epithelium were attempted and confirmed earlier than other parts of the cornea, following advancements in basic research on corneal epithelial stem cells. Based on these advances, four regenerative therapy products for corneal epithelial disease, each employing distinct cell sources and culture techniques, have been commercialized since the regulatory approval of Holoclar® in Italy as a regenerative therapy product for corneal epithelial disease in 2015. Corneal endothelial regenerative therapy was started by the development of an in vitro method to expand corneal endothelial cells which do not proliferate in adults. The product was approved in Japan as Vyznova® in 2023. The development of regenerative therapies for retinal and ocular surface diseases is actively being pursued, and these therapies use somatic stem cells and pluripotent stem cells (PSCs), especially induced pluripotent stem cells (iPSCs). Accordingly, the eye field is anticipated to play a pioneering role in regenerative therapy development going forward.
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Affiliation(s)
- Hiroshi Takayanagi
- Research, Development and Production Department of RAYMEI Incorporated, Osaka, Japan
- Department of Ophthalmology, Osaka University Graduate School of Medicine, Osaka, Japan
| | - Ryuhei Hayashi
- Department of Ophthalmology, Osaka University Graduate School of Medicine, Osaka, Japan
- Laboratory of Stem Cells and Applied Medicine, Osaka University Graduate School of Medicine, Osaka, Japan
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Shimmura S, Inagaki E, Hirayama M, Hatou S. The Cornea: An Ideal Tissue for Regenerative Medicine. Keio J Med 2024; 73:1-7. [PMID: 38369325 DOI: 10.2302/kjm.2023-0001-ir] [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] [Indexed: 02/20/2024]
Abstract
Regenerative medicine is a highly anticipated field with hopes to provide cures for previously uncurable diseases such as spinal cord injuries and retinal blindness. Most regenerative medical products use either autologous or allogeneic stem cells, which may or may not be genetically modified. The introduction of induced-pluripotent stem cells (iPSCs) has fueled research in the field, and several iPSC-derived cells/tissues are currently undergoing clinical trials. The cornea is one of the pioneering areas of regenerative medicine, and already four cell therapy products are approved for clinical use in Japan. There is one other government-approved cell therapy product approved in Europe, but none are approved in the USA at present. The cornea is transparent and avascular, making it unique as a target for stem cell therapy. This review discusses the unique properties of the cornea and ongoing research in the field.
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Affiliation(s)
- Shigeto Shimmura
- Department of Clinical Regenerative Medicine, Fujita Medical Innovation Center, Tokyo, Japan
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
| | - Emi Inagaki
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
| | - Masatoshi Hirayama
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
| | - Shin Hatou
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
- Cellusion Inc., Tokyo, Japan
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Wolosin JM. A Keratin 12 Expression-Based Analysis of Stem-Precursor Cells and Differentiation in the Limbal-Corneal Epithelium Using Single-Cell RNA-Seq Data. BIOLOGY 2024; 13:145. [PMID: 38534415 DOI: 10.3390/biology13030145] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/18/2024] [Revised: 02/18/2024] [Accepted: 02/22/2024] [Indexed: 03/28/2024]
Abstract
The corneal epithelium (CE) is spread between two domains, the outer vascularized limbus and the avascular cornea proper. Epithelial cells undergo constant migration from the limbus to the vision-critical central cornea. Coordinated with this migration, the cells undergo differentiation changes where a pool of unique stem/precursor cells at the limbus yields the mature cells that reach the corneal center. Differentiation is heralded by the expression of the corneal-specific Krt12. Processing data acquired by scRNA-Seq showed that the increase in Krt12 expression occurs in four distinct steps within the limbus, plus a single continuous increase in the cornea. Differential gene analysis demonstrated that these domains reflect discreet stages of CE differentiation and yielded extensive information of the genes undergoing down- or upregulation in the sequential transition from less to more differentiate conditions. The approach allowed the identification of multiple gene cohorts, including (a) the genes which have maximal expression in the most primitive, Krt12-negative cell cohort, which is likely to include the stem/precursor cells; (b) the sets of genes that undergo continuous increase or decrease along the whole differentiation path; and (c) the genes showing maximal positive or negative correlation with the changes in Krt12.
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Affiliation(s)
- J Mario Wolosin
- Department of Ophthalmology, Black Family Stem Cell Institute and Vision Research Institute, Icahn School of Medicine at Mount Sinai, One Gustave Levy Place, New York, NY 10029, USA
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Cabaña-Muñoz ME, Pelaz Fernández MJ, Parmigiani-Cabaña JM, Parmigiani-Izquierdo JM, Merino JJ. Adult Mesenchymal Stem Cells from Oral Cavity and Surrounding Areas: Types and Biomedical Applications. Pharmaceutics 2023; 15:2109. [PMID: 37631323 PMCID: PMC10459416 DOI: 10.3390/pharmaceutics15082109] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2023] [Revised: 07/28/2023] [Accepted: 08/02/2023] [Indexed: 08/27/2023] Open
Abstract
Adult mesenchymal stem cells are those obtained from the conformation of dental structures (DMSC), such as deciduous and permanent teeth and other surrounding tissues. Background: The self-renewal and differentiation capacities of these adult stem cells allow for great clinical potential. Because DMSC are cells of ectomesenchymal origin, they reveal a high capacity for complete regeneration of dental pulp, periodontal tissue, and other biomedical applications; their differentiation into other types of cells promotes repair in muscle tissue, cardiac, pancreatic, nervous, bone, cartilage, skin, and corneal tissues, among others, with a high predictability of success. Therefore, stem and progenitor cells, with their exosomes of dental origin and surrounding areas in the oral cavity due to their plasticity, are considered a fundamental pillar in medicine and regenerative dentistry. Tissue engineering (MSCs, scaffolds, and bioactive molecules) sustains and induces its multipotent and immunomodulatory effects. It is of vital importance to guarantee the safety and efficacy of the procedures designed for patients, and for this purpose, more clinical trials are needed to increase the efficacy of several pathologies. Conclusion: From a bioethical and transcendental anthropological point of view, the human person as a unique being facilitates better clinical and personalized therapy, given the higher prevalence of dental and chronic systemic diseases.
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Affiliation(s)
- María Eugenia Cabaña-Muñoz
- CIROM—Centro de Rehabilitación Oral Multidisciplinaria, 30001 Murcia, Spain; (M.E.C.-M.); (J.M.P.-C.); (J.M.P.-I.)
| | | | - José María Parmigiani-Cabaña
- CIROM—Centro de Rehabilitación Oral Multidisciplinaria, 30001 Murcia, Spain; (M.E.C.-M.); (J.M.P.-C.); (J.M.P.-I.)
| | | | - José Joaquín Merino
- Departamento de Farmacología, Farmacognosia y Botánica, Facultad de Farmacia, Universidad Complutense de Madrid (U.C.M), 28040 Madrid, Spain
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Tomczak W, Winkler-Lach W, Tomczyk-Socha M, Misiuk-Hojło M. Advancements in Ocular Regenerative Therapies. BIOLOGY 2023; 12:biology12050737. [PMID: 37237549 DOI: 10.3390/biology12050737] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/07/2023] [Revised: 05/12/2023] [Accepted: 05/16/2023] [Indexed: 05/28/2023]
Abstract
The use of stem cells (SCs) has emerged as a promising avenue in ophthalmology, offering potential therapeutic solutions for various vision impairments and degenerative eye diseases. SCs possess the unique ability to self-renew and differentiate into specialised cell types, making them valuable tools for repairing damaged tissues and restoring visual function. Stem cell-based therapies hold significant potential for addressing conditions such as age-related macular degeneration (AMD), retinitis pigmentosa (RP), corneal disorders, and optic nerve damage. Therefore, researchers have explored different sources of stem cells, including embryonic stem cells (ESC), induced pluripotent stem cells (iPSCs), and adult stem cells, for ocular tissue regeneration. Preclinical studies and early-phase clinical trials have demonstrated promising outcomes, with some patients experiencing improved vision following stem cell-based interventions. However, several challenges remain, including optimising the differentiation protocols, ensuring transplanted cells' safety and long-term viability, and developing effective delivery methods. The field of stem cell research in ophthalmology witnesses a constant influx of new reports and discoveries. To effectively navigate these tons of information, it becomes crucial to summarise and systematise these findings periodically. In light of recent discoveries, this paper demonstrates the potential applications of stem cells in ophthalmology, focusing on their use in various eye tissues, including the cornea, retina, conjunctiva, iris, trabecular meshwork, lens, ciliary body, sclera, and orbital fat.
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Affiliation(s)
| | | | | | - Marta Misiuk-Hojło
- Department of Ophthalmology, Wroclaw Medical University, 50556 Wroclaw, Poland
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Toshida H, Kasahara T, Kiriyama M, Iwasaki Y, Sugita J, Ichikawa K, Ohta T, Miyahara K. Early Clinical Outcomes of the First Commercialized Human Autologous Ex Vivo Cultivated Oral Mucosal Epithelial Cell Transplantation for Limbal Stem Cell Deficiency: Two Case Reports and Literature Review. Int J Mol Sci 2023; 24:ijms24108926. [PMID: 37240280 DOI: 10.3390/ijms24108926] [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: 04/30/2023] [Revised: 05/14/2023] [Accepted: 05/16/2023] [Indexed: 05/28/2023] Open
Abstract
The first product in the world for ex vivo cultivated oral mucosal epithelial cell transplantation (COMET) to treat limbal stem cell deficiency (LSCD), named Ocural®, was launched in June 2021 in Japan. COMET was performed on two patients, including the first case in the post-marketing phase of Ocural®. Pathological and immunohistochemical examinations were also carried out using specimens obtained before and after COMET and the spare cell sheet. In case 1, the ocular surface remained free from epithelial defects for approximately six months. In case 2, although defect of the cornea-like epithelia was observed after COMET for one month, it was resolved after the insertion of lacrimal punctal plugs. In case 1, adjuvant treatment was interrupted due to an accident during the second month after COMET, resulting in conjunctival ingrowth and corneal opacity. Eventually, a lamellar keratoplasty was required at six months after COMET. Immunohistochemistry revealed the presence of markers for stem cells (p63, p75), proliferation (Ki-67), and differentiation (Keratin-3, -4, and -13) in both the cornea-like tissue after COMET and a cultivated oral mucosal epithelial cell sheet. In conclusion, Ocural® can be accomplished without major complications, and the stem cells derived from oral mucosa might be successfully engrafted.
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Affiliation(s)
- Hiroshi Toshida
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Tomoto Kasahara
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Masamichi Kiriyama
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Yuma Iwasaki
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Jobu Sugita
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Kohei Ichikawa
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Toshihiko Ohta
- Department of Ophthalmology, Juntendo University Shizuoka Hospital, Shizuoka 410-2211, Japan
| | - Katsumi Miyahara
- Laboratory of Morphology and Image Analysis, Biomedical Research Core Facilities, Juntendo University Graduate School of Medicine, Tokyo 113-0033, Japan
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Aizawa S, Yoshida H, Umeshita K, Watanabe S, Takahashi Y, Sakane S, Sakaguchi H, Kataoka S. Development of an oral mucosal irritation test using a three-dimensional human buccal oral mucosal model. Toxicol In Vitro 2023; 87:105519. [PMID: 36403724 DOI: 10.1016/j.tiv.2022.105519] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2022] [Revised: 11/12/2022] [Accepted: 11/14/2022] [Indexed: 11/18/2022]
Abstract
The oral mucosa can become irritated by oral care products and lip cosmetics. Therefore, it is important to determine the irritation potential of their ingredients and products during safety evaluations. We developed a method for oral mucosal irritation test using EpiOral, which is a three-dimensional cultured model. Exposure of sodium lauryl sulphate (SLS) to EpiOral showed a dose-dependent decrease in cell viability. Under 120 min exposure conditions, SLS irritation was detected when 60% cell viability was set as a criterion. Evaluation of the irritancy of SLS and four other raw materials used in oral products at three laboratories under the above conditions confirmed good transferability of the test. Focused on the similarity of the oral and eye mucous, 32 chemicals categorised by the UN-GHS eye-irritation classification were evaluated to ensure the reliability of our criteria at these laboratories. The concordance rate between the UN-GHS classification and our test results was 100% for irritants and 60% for non-irritants. The good intra-laboratory reproducibility of our test was confirmed from the evaluation results of negative and positive controls, and the good inter-laboratory reproducibility was confirmed from the results of 32 chemicals. These findings showed that oral mucosal irritation can be evaluated using EpiOral.
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Affiliation(s)
- Seiya Aizawa
- Safety Science Research Laboratories, LION Corporation, Kanagawa, Japan.
| | - Hidenori Yoshida
- Safety Science Research Laboratories, Kao Corporation, Tochigi, Japan
| | | | - Shinichi Watanabe
- Safety Science Research Laboratories, LION Corporation, Kanagawa, Japan
| | - Yutaka Takahashi
- Safety Science Research Laboratories, Kao Corporation, Tochigi, Japan
| | - Shinji Sakane
- Safety & Analysis, R&D Support, Sunstar Inc., Osaka, Japan
| | - Hitoshi Sakaguchi
- Safety Science Research Laboratories, Kao Corporation, Tochigi, Japan
| | - Shinsuke Kataoka
- Safety Science Research Laboratories, LION Corporation, Kanagawa, Japan
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Mohammad-Pour N, Moghimi V, Bidkhori HR, Momeni-Moghaddam M, Naderi-Meshkin H. Comparing the Effects of Two Cryoprotectant Protocols, Dimethyl-Sulfoxide (DMSO) and Glycerol, on the Recovery Rate of Cultured Keratinocytes on Amniotic Membrane. INT J LOW EXTR WOUND 2023:15347346231155751. [PMID: 36794512 DOI: 10.1177/15347346231155751] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/17/2023]
Abstract
Background: Off-the-shelf supply of viable engineered tissue is critical for effective and fast treatment of life-threatening injuries such as deep burns. An expanded keratinocyte sheet on the human amniotic membrane (KC sheet-HAM) is a beneficial tissue-engineering product for wound healing. To access an on-hand supply for the widespread application and overcome the time-consuming process, it is necessary to develop a cryopreservation protocol that guarantees the higher recovery of viable keratinocyte sheets after freeze-thawing. This research aimed to compare the recovery rate of KC sheet-HAM after cryopreservation by dimethyl-sulfoxide (DMSO) and glycerol. Methods: Amniotic membrane was decellularized with trypsin, and keratinocytes were cultured on it to form a multilayer, flexible, easy-to-handle KC sheet-HAM. The effects of 2 different cryoprotectants were investigated by histological analysis, live-dead staining, and proliferative capacity assessments before and after cryopreservation. Results: KCs well adhered and proliferated on the decellularized amniotic membrane and successfully represented 3 to 4 stratified layers of epithelialization after 2 to 3 weeks culture period; making it easy to cut, transfer, and cryopreserve. However, viability and proliferation assay indicated that both DMSO and glycerol cryosolutions have detrimental effects on KCs, and KCs-sheet HAM could not recover to the control level after 8 days of culture post-cryo. The KC sheet lost its stratified multilayer nature on AM, and sheet layers were reduced in both cryo-groups compared to the control. Conclusion: Expanding keratinocytes on the decellularized amniotic membrane as a multilayer sheet made a viable easy-to-handle sheet, nonetheless cryopreservation reduced viability and affected histological structure after thawing. Although some viable cells were detectable, our research highlighted the need for a better cryoprotectant protocol other than DMSO and glycerol, specific for the successful banking of viable tissue constructs.
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Affiliation(s)
- Najmeh Mohammad-Pour
- Department of Biology, Faculty of Science, Hakim Sabzevari University, Sabzevar, Iran
- Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
| | - Vahid Moghimi
- Department of Biology, Faculty of Science, Hakim Sabzevari University, Sabzevar, Iran
- Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
| | - Hamid Reza Bidkhori
- Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
- Blood Borne Infections Research Center, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
| | - Madjid Momeni-Moghaddam
- Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
| | - Hojjat Naderi-Meshkin
- Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran
- Wellcome-Wolfson Institute for Experimental Medicine, Belfast, UK
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11
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Wang B, Wang X, Kenneth A, Drena A, Pacheco A, Kalvin L, Ibrahim ES, Rossi PJ, Thatcher K, Lincoln J. Developing small-diameter vascular grafts with human amniotic membrane: long-term evaluation of transplantation outcomes in a small animal model. Biofabrication 2023; 15. [PMID: 36626826 DOI: 10.1088/1758-5090/acb1da] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2022] [Accepted: 01/10/2023] [Indexed: 01/11/2023]
Abstract
While current clinical utilization of large vascular grafts for vascular transplantation is encouraging, tissue engineering of small grafts still faces numerous challenges. This study aims to investigate the feasibility of constructing a small vascular graft from decellularized amniotic membranes (DAMs). DAMs were rolled around a catheter and each of the resulting grafts was crosslinked with (a) 0.1% glutaraldehyde; (b) 1-ethyl-3-(3-dimethylaminopropyl) crbodiimidehydro-chloride (20 mM)-N-hydroxy-succinimide (10 mM); (c) 0.5% genipin; and (d) no-crosslinking, respectively. Our results demonstrated the feasibility of using a rolling technique followed by lyophilization to transform DAM into a vessel-like structure. The genipin-crosslinked DAM graft showed an improved integrated structure, prolonged stability, proper mechanical property, and superior biocompatibility. After transplantation in rat abdominal aorta, the genipin-crosslinked DAM graft remained patent up to 16 months, with both endothelial and smooth muscle cell regeneration, which suggests that the genipin-crosslinked DAM graft has great potential to beimplementedas a small tissue engineered graft for futurevasculartransplantation.
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Affiliation(s)
- Bo Wang
- Joint Department of Biomedical Engineering, Marquette University and the Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Xiaolong Wang
- Joint Department of Biomedical Engineering, Marquette University and the Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Allen Kenneth
- Biomedical Resource Center, Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Alexander Drena
- Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, United States of America
| | - Arsenio Pacheco
- Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, United States of America
| | - Lindsey Kalvin
- Department of Medicine, Division of Cardiovascular Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Ei-Sayed Ibrahim
- Department of Radiology, Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Peter J Rossi
- Heart and Vascular Center, Froedtert Hospital, Milwaukee, WI 53226, United States of America
| | - Kaitlyn Thatcher
- Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
| | - Joy Lincoln
- Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, United States of America
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12
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Tavakkoli F, Eleiwa TK, Elhusseiny AM, Damala M, Rai AK, Cheraqpour K, Ansari MH, Doroudian M, H Keshel S, Soleimani M, Djalilian AR, Sangwan VS, Singh V. Corneal stem cells niche and homeostasis impacts in regenerative medicine; concise review. Eur J Ophthalmol 2023:11206721221150065. [PMID: 36604831 DOI: 10.1177/11206721221150065] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
The limbal stem cells niche (LSCN) is an optimal microenvironment that provides the limbal epithelial stem cells (LESCs) and strictly regulates their proliferation and differentiation. Disturbing the LSCN homeostasis can lead to limbal stem cell dysfunction (LSCD) and subsequent ocular surface aberrations, such as corneal stromal inflammation, persistent epithelial defects, corneal neovascularisation, lymphangiogenesis, corneal opacification, and conjunctivalization. As ocular surface disorders are considered the second main cause of blindness, it becomes crucial to explore different therapeutic strategies for restoring the functions of the LSCN. A major limitation of corneal transplantation is the current shortage of donor tissue to meet the requirements worldwide. In this context, it becomes mandatory to find an alternative regenerative medicine, such as using cultured limbal epithelial/stromal stem cells, inducing the production of corneal like cells by using other sources of stem cells, and using tissue engineering methods aiming to produce the three-dimensional (3D) printed cornea. Limbal epithelial stem cells have been considered the magic potion for eye treatment. Epithelial and stromal stem cells in the limbal niche hold the responsibility of replenishing the corneal epithelium. These stem cells are being used for transplantation to maintain corneal epithelial integrity and ultimately sustain optimal vision. In this review, we summarised the characteristics of the LSCN and their current and future roles in restoring corneal homeostasis in eyes with LSCD.
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Affiliation(s)
- Fatemeh Tavakkoli
- Department of Community Health, College of Health Technology, Cihan University, Erbil, Iraq.,SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India.,Centre for Genetic Disorders, Banaras Hindu University, Varanasi, India
| | - Taher K Eleiwa
- Department of Ophthalmology, Benha University, Benha, Egypt
| | - Abdelrahman M Elhusseiny
- Department of Ophthalmology, Harvey and Bernice Jones Eye Institute, University of Arkansas for Medical Sciences, Little Rock, AR, USA
| | - Mukesh Damala
- SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India.,School of Life Sciences, University of Hyderabad, Hyderabad, India
| | - Amit K Rai
- Centre for Genetic Disorders, Banaras Hindu University, Varanasi, India
| | - Kasra Cheraqpour
- Translational Eye Research Center, Farabi Eye Hospital, 48439Tehran University of Medical Sciences, Tehran, Iran
| | - Mohammad H Ansari
- Ophthalmic Research Center, Department of Ophthalmology, Labbafinejad Medical Center, 556492Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mohammad Doroudian
- Department of Cell and Molecular Biology, Faculty of Biological Sciences, 145440Kharazmi University, Tehran, Iran
| | - Saeed H Keshel
- Department of Tissue Engineering and Applied Cell Sciences, 556492Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mohammad Soleimani
- Department of Ophthalmology, 159636Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA
| | - Ali R Djalilian
- Department of Ophthalmology, 159636Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA
| | | | - Vivek Singh
- SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India
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13
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Zhurenkov KE, Alexander-Sinkler EI, Gavrilyik IO, Yartseva NM, Aleksandrova SA, Mashel TV, Khorolskaya JI, Blinova MI, Kulikov AN, Churashov SV, Chernysh VF, Mikhailova NA. Labial Mucosa Stem Cells: Isolation, Characterization, and Their Potential for Corneal Epithelial Reconstruction. Invest Ophthalmol Vis Sci 2022; 63:16. [PMID: 35848889 PMCID: PMC9308017 DOI: 10.1167/iovs.63.8.16] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/02/2022] Open
Abstract
Purpose The purpose of this study was to characterize labial mucosa stem cells (LMSCs) and to investigate their potential for corneal epithelial reconstruction in a rabbit model of total limbal stem cell deficiency (LSCD). Methods Rabbit LMSCs (rLMSCs) and human (hLMSCs) LMSCs were derived from labial mucosa and characterized in terms of their proliferation activity by the evaluation of proliferation index (PI) and colony forming efficiency (CFE), cell senescence, and differentiation abilities. The expression of various limbus-specific, stem cell-specific, and epithelial markers was assessed via immunocytochemistry. Flow cytometry was used to evaluate mesenchymal and hematopoietic cell surface markers expression. Chromosomal stability of the derived cells was examined using the conventional GTG-banding technique. To assess the impact of LMSCs on corneal epithelial reconstruction, rLMSCs were seeded onto a decellularized human amniotic membrane (dHAM), thereafter their regeneration potential was examined in the rabbit model of total LSCD. Results Both rLMSCs and hLMSCs showed high proliferation and differentiation abilities, entered senescence at later passages, and expressed different stem cell-specific (ABCB5, ALDH3A1, ABCG2, and p63α), mesenchymal (vimentin), and epithelial (CK3/12, CK15) markers. Cell surface antigen expression was similar to other described mesenchymal stem cells. No clonal structural chromosome abnormalities (CSCAs) and the low percentage of non-clonal structural chromosome abnormalities (NSCAs) were observed. Transplantation of rLMSCs promoted corneal epithelial reconstruction and enhanced corneal transparency. Conclusions LMSCs have significant proliferation and differentiation abilities, display no detrimental chromosome aberrations, and demonstrate considerable potential for corneal repair.
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Affiliation(s)
- Kirill E Zhurenkov
- Institute of Cytology Russian Academy of Science, St. Petersburg, Russia.,Department of Cytology and Histology, St. Petersburg State University, St. Petersburg, Russia
| | | | | | - Natalia M Yartseva
- Institute of Cytology Russian Academy of Science, St. Petersburg, Russia
| | | | - Tatiana V Mashel
- Institute of Cytology Russian Academy of Science, St. Petersburg, Russia
| | | | - Miralda I Blinova
- Institute of Cytology Russian Academy of Science, St. Petersburg, Russia
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14
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Tibrewal S, Ratna R, Gour A, Agarkar S, Dubey S, Ganesh S, Kekunnaya R, Sangwan V, Liu Y, Vanita V. Clinical and molecular aspects of congenital aniridia - A review of current concepts. Indian J Ophthalmol 2022; 70:2280-2292. [PMID: 35791108 PMCID: PMC9426064 DOI: 10.4103/ijo.ijo_2255_21] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023] Open
Abstract
Congenital aniridia is a pan ocular disorder characterized by partial or total loss of iris tissue as the defining feature. Classic aniridia, however, has a spectrum of ocular findings, including foveal hypoplasia, optic nerve hypoplasia, nystagmus, late-onset cataract, glaucoma, and keratopathy. The latter three are reasons for further visual compromise in such patients. This entity is often due to mutations in the PAX6 (Paired box protein Pax-6) gene. Recently, aniridia-like phenotypes have been reported due to non-PAX6 mutations as in PITX2, FOXC1, FOXD3, TRIM44, and CYP1B1 as well wherein there is an overlap of aniridia, such as iris defects with congenital glaucoma or anterior segment dysgenesis. In this review, we describe the various clinical features of classic aniridia, the comorbidities and their management, the mutation spectrum of the genes involved, genotype-phenotype correlation of PAX6 and non-PAX6 mutations, and the genetic testing plan. The various systemic associations and their implications in screening and genetic testing have been discussed. Finally, the future course of aniridia treatment in the form of drugs (such as ataluren) and targeted gene therapy has been discussed.
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Affiliation(s)
- Shailja Tibrewal
- Department of Ocular Genetics; Department of Pediatric Ophthalmology, Strabismus and Neuro-ophthalmology, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Ria Ratna
- Department of Ocular Genetics, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Abha Gour
- Department of Cornea and Anterior Segment, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Sumita Agarkar
- Department of Pediatric Ophthalmology and Strabismus, Medical Research Foundation, Sankara Netralaya, Chennai, Tamil Nadu, India
| | - Suneeta Dubey
- Department of Glaucoma, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Suma Ganesh
- Department of Pediatric Ophthalmology, Strabismus and Neuro-ophthalmology, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Ramesh Kekunnaya
- Child Sight Institute, Jasti V Ramanamma Children's Eye Care Centre, L V Prasad Eye Institute, KAR Campus, Hyderabad, Telangana, India
| | - Virender Sangwan
- Department of Cornea and Anterior Segment, Dr Shroff's Charity Eye Hospital, Daryaganj, New Delhi, India
| | - Yutao Liu
- Department of Cellular Biology and Anatomy, Center for Biotechnology and Genomic Medicine, Augusta University, Augusta, USA
| | - Vanita Vanita
- Department of Human Genetics, Guru Nanak Dev University, Amritsar, Punjab, India
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15
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Kitazawa K, Sotozono C, Kinoshita S. Current Advancements in Corneal Cell-Based Therapy. Asia Pac J Ophthalmol (Phila) 2022; 11:335-345. [PMID: 36041148 DOI: 10.1097/apo.0000000000000530] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2021] [Accepted: 03/10/2022] [Indexed: 12/13/2022] Open
Abstract
Corneal epithelial stem cells (CEpSCs) mostly reside at the limbal area and are responsible for tissue homeostasis throughout life. Once complete CEpSC deficiency occurs, regenerative medicine cell-based therapy using CEpSCs or their alternatives can provide successful clinical outcomes. Due to an improved understanding of CEpSCs and mucosal epithelial stem cells, major advancements have been made over the past few decades in in vivo and ex vivo cell-based ocular surface reconstruction therapies for the treatment of severe ocular surface diseases. New therapeutic concepts and clinical strategies are emerging for the treatment of corneal endothelial dysfunction. For example, unlike corneal epithelial cells, in vivo corneal endothelial cells (CECs) stop proliferating and are arrested in the G1 phase of the cell cycle due to cell-to-cell contact inhibition and exposure to a high concentration of transforming growth factor-beta in the aqueous humor. Thus, the production of CECs with good functionality in culture has consistently been difficult. To solve this problem, Rho-associated protein kinase inhibition has taken center stage, as it not only makes the production of human CECs in culture closely mimic the functional characteristics of in vivo healthy CECs possible but also helps sustain those biological properties. Thus, cultured human CEC injection therapy is now moving to the forefront for the treatment of corneal endothelial failure. Herein, we summarize key historical discoveries in corneal cell-based regenerative medicine and illustrate the concept of corneal cell therapy for the treatment of refractory corneal epithelial and endothelial diseases.
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Affiliation(s)
- Koji Kitazawa
- Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
| | - Chie Sotozono
- Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
| | - Shigeru Kinoshita
- Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
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16
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Nguyen HT, Theerakittayakorn K, Somredngan S, Ngernsoungnern A, Ngernsoungnern P, Sritangos P, Ketudat-Cairns M, Imsoonthornruksa S, Assawachananont J, Keeratibharat N, Wongsan R, Rungsiwiwut R, Laowtammathron C, Bui NX, Parnpai R. Signaling Pathways Impact on Induction of Corneal Epithelial-like Cells Derived from Human Wharton’s Jelly Mesenchymal Stem Cells. Int J Mol Sci 2022; 23:ijms23063078. [PMID: 35328499 PMCID: PMC8949174 DOI: 10.3390/ijms23063078] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/08/2022] [Revised: 03/07/2022] [Accepted: 03/10/2022] [Indexed: 02/05/2023] Open
Abstract
Corneal epithelium, the outmost layer of the cornea, comprises corneal epithelial cells (CECs) that are continuously renewed by limbal epithelial stem cells (LESCs). Loss or dysfunction of LESCs causes limbal stem cell deficiency (LSCD) which results in corneal epithelial integrity loss and visual impairment. To regenerate the ocular surface, transplantation of stem cell-derived CECs is necessary. Human Wharton’s jelly derived mesenchymal stem cells (WJ-MSCs) are a good candidate for cellular therapies in allogeneic transplantation. This study aimed to test the effects of treatments on three signaling pathways involved in CEC differentiation as well as examine the optimal protocol for inducing corneal epithelial differentiation of human WJ-MSCs. All-trans retinoic acid (RA, 5 or 10 µM) inhibited the Wnt signaling pathway via suppressing the translocation of β-catenin from the cytoplasm into the nucleus. SB505124 downregulated the TGF-β signaling pathway via reducing phosphorylation of Smad2. BMP4 did not increase phosphorylation of Smad1/5/8 that is involved in BMP signaling. The combination of RA, SB505124, BMP4, and EGF for the first 3 days of differentiation followed by supplementing hormonal epidermal medium for an additional 6 days could generate corneal epithelial-like cells that expressed a CEC specific marker CK12. This study reveals that WJ-MSCs have the potential to transdifferentiate into CECs which would be beneficial for further applications in LSCD treatment therapy.
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Affiliation(s)
- Hong Thi Nguyen
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (H.T.N.); (K.T.); (S.S.)
- Laboratory of Embryo Technology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi 100000, Vietnam
| | - Kasem Theerakittayakorn
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (H.T.N.); (K.T.); (S.S.)
| | - Sirilak Somredngan
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (H.T.N.); (K.T.); (S.S.)
| | - Apichart Ngernsoungnern
- School of Preclinical Sciences, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (A.N.); (P.N.); (P.S.)
| | - Piyada Ngernsoungnern
- School of Preclinical Sciences, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (A.N.); (P.N.); (P.S.)
| | - Pishyaporn Sritangos
- School of Preclinical Sciences, Institute of Science, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (A.N.); (P.N.); (P.S.)
| | - Mariena Ketudat-Cairns
- School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (M.K.-C.); (S.I.)
| | - Sumeth Imsoonthornruksa
- School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (M.K.-C.); (S.I.)
| | - Juthaporn Assawachananont
- School of Ophthalmology, Institute of Medicine, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;
| | - Nattawut Keeratibharat
- School of Surgery, Institute of Medicine, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;
| | - Rangsirat Wongsan
- The Center for Scientific and Technological Equipment, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand;
| | - Ruttachuk Rungsiwiwut
- Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok 10000, Thailand;
| | - Chuti Laowtammathron
- Siriraj Center of Excellence for Stem Cell Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10000, Thailand;
| | | | - Rangsun Parnpai
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (H.T.N.); (K.T.); (S.S.)
- Correspondence: ; Tel.: +66-442-242-34
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17
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Induction of salivary gland-like cells from epithelial tissues transdifferentiated from mouse embryonic fibroblasts. Biochem Biophys Res Commun 2022; 586:55-62. [PMID: 34826701 DOI: 10.1016/j.bbrc.2021.11.064] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2021] [Revised: 10/16/2021] [Accepted: 11/15/2021] [Indexed: 11/21/2022]
Abstract
Salivary gland hypofunction due to radiation therapy for head and neck cancer or Sjögren syndrome may cause various oral diseases, which can lead to a decline in the quality of life. Cell therapy using salivary gland stem cells is a promising method for restoring hypofunction. Herein, we show that salivary gland-like cells can be induced from epithelial tissues that were transdifferentiated from mouse embryonic fibroblasts (MEFs). We introduced four genes, Dnp63a, Tfap2a, Grhl2, and Myc (PTMG) that are known to transdifferentiate fibroblasts into oral mucosa-like epithelium in vivo into MEFs. MEFs overexpressing these genes showed epithelial cell characteristics, such as cobblestone appearance and E-cadherin positivity, and formed oral epithelial-like tissue under air-liquid interface culture conditions. The epithelial sheet detached from the culture dish was infected with adenoviruses encoding Sox9 and Foxc1, which we previously identified as essential factors to induce salivary gland formation. The cells detached from the cell sheet formed spheres 10 days after infection and showed a branching morphology. The spheres expressed genes encoding basal/myoepithelial markers, cytokeratin 5, cytokeratin 14, acinar cell marker, aquaporin 5, and the myoepithelial marker α-smooth muscle actin. The dissociated cells of these primary spheres had the ability to form secondary spheres. Taken together, our results provide a new strategy for cell therapy of salivary glands and hold implications in treating patients with dry mouth.
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18
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Álvarez de Toledo Elizalde J, López García S, Benítez Del Castillo JM, Durán de la Colina J, Gris Castejón O, Celis Sánchez J, Herreras Cantalapiedra JM. Aniridia and the ocular surface: Medical and surgical problems and solutions. ARCHIVOS DE LA SOCIEDAD ESPANOLA DE OFTALMOLOGIA 2021; 96 Suppl 1:15-37. [PMID: 34836585 DOI: 10.1016/j.oftale.2021.04.003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/09/2020] [Accepted: 04/12/2021] [Indexed: 06/13/2023]
Abstract
Congenital aniridia is a multisystemic genetic disease due to a mutation in PAX6 gene which severely affects the development and functionality of the human eyes. In patients affected by the mutation, aside from the absence or defects of iris tissue formation, abnormalities in position or opacities of the crystalline lens, macular hypoplasia, ocular surface disease is the main cause of visual loss and the deterioration of the quality of life of most patients. Limbal stem cell deficiency combined with tear film instability and secondary dry eye cause aniridic keratopathy which, in advanced stages, ends up in corneal opacification. In this paper, the actual knowledge about congenital aniridia keratopathy physiopathology and medical and surgical treatment options and their efficacy are discussed. Indications and results of topical treatments with artificial tears and blood-derivatives in its initial stages, and different surgical techniques as limbal stem cell transplantation, keratoplasty and keratoprostheses are reviewed. Finally, recent advances and results in regenerative medicine techniques with ex vivo stem cell cultivation or other types of cultivated cells are presented.
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Affiliation(s)
| | - S López García
- Hospital Universitario Severo Ochoa, Leganés, Madrid, Spain
| | - J M Benítez Del Castillo
- Cátedra de Oftalmología, Hospital Clínico San Carlos, Universidad Complutense, Clínica Rementería, Madrid, Spain
| | - J Durán de la Colina
- Cátedra de Oftalmología, Universidad del País Vasco, Instituto Clínico-Quirúrgico de Oftalmología, Bilbao, Spain
| | - O Gris Castejón
- Departamento de Córnea y Superficie Ocular, Instituto de Microcirugía Ocular de Barcelona (IMO), Barcelona, Spain
| | - J Celis Sánchez
- Unidad de Córnea y Superficie ocular, Hospital La Mancha-Centro, Alcázar de San Juan, Spain
| | - J M Herreras Cantalapiedra
- Instituto Universitario de Oftalmobiología Aplicada (IOBA) de la Universidad de Valladolid, Servicio de Oftalmología del Hospital Clínico Universitario de Valladolid, Valladolid, Spain
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19
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Oral Mucosal Epithelial Transplantation and Limbal-Rigid Contact Lens: A Therapeutic Modality for the Treatment of Severe Ocular Surface Disorders. Cornea 2021; 39 Suppl 1:S19-S27. [PMID: 33181721 DOI: 10.1097/ico.0000000000002566] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Stevens-Johnson syndrome, ocular cicatricial pemphigoid, and severe thermal or chemical injury are considered severe ocular surface disorders (OSDs) because they affect the entire ocular surface, including corneal and conjunctival epithelial stem cells. In patients with severe OSDs, the long-term prognosis for limbal transplantation is poor, and the related corneal opacity and cicatrization lead to devastating visual impairment. To date, there is no standardized treatment to improve vision in cases with severe OSD. Investigating novel treatment methods for severe OSDs, our group began cultivated oral mucosal epithelial transplantation in 2002 and developed a limbal-supported rigid-type contact lens that can be applied as a nonsurgical treatment. When used in combination, these treatment methods make it possible to successfully restore vision in cases with severe OSDs.
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20
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Goals and Challenges of Stem Cell-Based Therapy for Corneal Blindness Due to Limbal Deficiency. Pharmaceutics 2021; 13:pharmaceutics13091483. [PMID: 34575560 PMCID: PMC8466237 DOI: 10.3390/pharmaceutics13091483] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2021] [Revised: 09/09/2021] [Accepted: 09/13/2021] [Indexed: 12/13/2022] Open
Abstract
Corneal failure is a highly prevalent cause of blindness. One special cause of corneal failure occurs due to malfunction or destruction of the limbal stem cell niche, upon which the superficial cornea depends for homeostatic maintenance and wound healing. Failure of the limbal niche is referred to as limbal stem cell deficiency. As the corneal epithelial stem cell niche is easily accessible, limbal stem cell-based therapy and regenerative medicine applied to the ocular surface are among the most highly advanced forms of this novel approach to disease therapy. However, the challenges are still great, including the development of cell-based products and understanding how they work in the patient's eye. Advances are being made at the molecular, cellular, and tissue levels to alter disease processes and to reduce or eliminate blindness. Efforts must be coordinated from the most basic research to the most clinically oriented projects so that cell-based therapies can become an integrated part of the therapeutic armamentarium to fight corneal blindness. We undoubtedly are progressing along the right path because cell-based therapy for eye diseases is one of the most successful examples of global regenerative medicine.
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21
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Corneal Epithelial Stem Cells-Physiology, Pathophysiology and Therapeutic Options. Cells 2021; 10:cells10092302. [PMID: 34571952 PMCID: PMC8465583 DOI: 10.3390/cells10092302] [Citation(s) in RCA: 32] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/01/2021] [Revised: 08/27/2021] [Accepted: 08/28/2021] [Indexed: 12/12/2022] Open
Abstract
In the human cornea, regeneration of the epithelium is regulated by the stem cell reservoir of the limbus, which is the marginal region of the cornea representing the anatomical and functional border between the corneal and conjunctival epithelium. In support of this concept, extensive limbal damage, e.g., by chemical or thermal injury, inflammation, or surgery, may induce limbal stem cell deficiency (LSCD) leading to vascularization and opacification of the cornea and eventually vision loss. These acquired forms of limbal stem cell deficiency may occur uni- or bilaterally, which is important for the choice of treatment. Moreover, a variety of inherited diseases, such as congenital aniridia or dyskeratosis congenita, are characterized by LSCD typically occurring bilaterally. Several techniques of autologous and allogenic stem cell transplantation have been established. The limbus can be restored by transplantation of whole limbal grafts, small limbal biopsies or by ex vivo-expanded limbal cells. In this review, the physiology of the corneal epithelium, the pathophysiology of LSCD, and the therapeutic options will be presented.
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22
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Sotozono C, Ueta M, Kinoshita S. Japan: Diagnosis and Management of Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis With Severe Ocular Complications. Front Med (Lausanne) 2021; 8:657327. [PMID: 34395463 PMCID: PMC8355416 DOI: 10.3389/fmed.2021.657327] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2021] [Accepted: 06/17/2021] [Indexed: 11/30/2022] Open
Abstract
In 2005, the “Japanese Research Committee on Severe Cutaneous Adverse Reaction” (J-SCAR) presented the official “Diagnostic Criteria” for SJS/TEN, and the specific ocular findings are included in these very important criteria. In SJS/TEN cases involving ocular disorder, conjunctivitis often occurs prior to the onset of the high fever. In a Japanese survey, ocular involvement was observed in 77% of the cases, and the incidence of ocular sequelae increased depending on the score of the acute ocular severity findings. Pseudo-membrane formation and epithelial defects are considered to be high-risk signs of ocular sequelae. At the chronic stage, limbal stem cell deficiency, visual disturbance, and severe dryness of the ocular surface are the primary disease characteristics. In 2002, we started performing Cultivated Oral Mucosal Epithelial Transplantation (COMET) for the treatment of severe ocular disorders, including SJS/TEN. As an additional treatment method, we developed a new type of rigid contact lens (CL) that is 13 to 14.0-mm in diameter, known as the “Limbal Rigid Contact Lens (Limbal CL).” Our Limbal Rigid CL greatly enhances the postoperative outcome of COMET. The detection rate of ocular surface bacteria is high in SJS/TEN cases. Thus, appropriate use of topical antibiotics reduces the risk of ocular surface inflammation. Moreover, rebamipide is an ophthalmic solution for dry eye that was developed in Japan, and it also has the effect of suppressing ocular surface inflammation. From disease onset until the chronic stage, the control of inflammation and stem cell loss is key to successfully treating eyes afflicted with SJS/TEN.
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Affiliation(s)
- Chie Sotozono
- Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
| | - Mayumi Ueta
- Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
| | - Shigeru Kinoshita
- Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
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cG-CAOMECS-clinical-grade cultured autologous oral mucosal epithelial cell sheet. Cell Tissue Res 2021; 386:47-57. [PMID: 34302219 DOI: 10.1007/s00441-021-03507-7] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2020] [Accepted: 07/10/2021] [Indexed: 10/20/2022]
Abstract
The present study reports the feasibility and successful production of rabbit cG-CAOMECS, designed to reconstruct corneal epithelium of patients with bilateral limbal stem cell deficiency. To produce a safe, chemically defined and FDA compliant cG-CAOMECS, oral mucosal epithelial cells were isolated from a biopsy of rabbit buccal tissue and seeded on a cGMP-certified cell culture surface coated with GMP-grade extracellular matrix. A newly designed clinical-grade medium (KaFa™ medium) was utilized to carry out cell expansion. Detachment and harvesting of the produced cell sheet was accomplished using collagenase treatment. Live cell imaging and morphological analysis techniques were used to examine cell growth. Cells attached onto the surface and self-assembled into colony-forming units (CFUs). Microscopic examination showed that CFUs formed during the first 5 days, and basal monolayer cell sheet formed in less than 10 days. Cells expanded to form a multilayered epithelial cell sheet that was harvested after 17-19 days in culture. Immunostaining and Western blot analyses showed that deltaNp63 was expressed in the basal cells and K3/K12 was expressed in the apical cells, indicating the presence of corneal epithelial-like cells in the produced cell sheet. Adhesion molecules, E-cadherin, beta-catenin, and Cnx43 were also expressed and exhibited the epithelial integrity of the cell sheet. The expression of integrin-beta1 and beta4 confirmed that the collagenase treatment used for detaching and harvesting the cell sheet did not have adverse effects. Our results showed that the utilization of clinical-grade and FDA-approved reagents successfully supported the production of cG-CAMECS.
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Maurizi E, Adamo D, Magrelli FM, Galaverni G, Attico E, Merra A, Maffezzoni MBR, Losi L, Genna VG, Sceberras V, Pellegrini G. Regenerative Medicine of Epithelia: Lessons From the Past and Future Goals. Front Bioeng Biotechnol 2021; 9:652214. [PMID: 33842447 PMCID: PMC8026866 DOI: 10.3389/fbioe.2021.652214] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/11/2021] [Accepted: 02/23/2021] [Indexed: 12/13/2022] Open
Abstract
This article explores examples of successful and unsuccessful regenerative medicine on human epithelia. To evaluate the applications of the first regenerated tissues, the analysis of the past successes and failures addresses some pending issues and lay the groundwork for developing new therapies. Research should still be encouraged to fill the gap between pathologies, clinical applications and what regenerative medicine can attain with current knowledge.
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Affiliation(s)
| | - Davide Adamo
- Interdepartmental Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
| | | | - Giulia Galaverni
- Interdepartmental Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
| | - Eustachio Attico
- Interdepartmental Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
| | | | | | - Lorena Losi
- Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy
| | | | | | - Graziella Pellegrini
- Holostem Terapie Avanzate S.r.l., Modena, Italy
- Interdepartmental Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
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Landsend ECS, Lagali N, Utheim TP. Congenital aniridia - A comprehensive review of clinical features and therapeutic approaches. Surv Ophthalmol 2021; 66:1031-1050. [PMID: 33675823 DOI: 10.1016/j.survophthal.2021.02.011] [Citation(s) in RCA: 64] [Impact Index Per Article: 16.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/18/2020] [Revised: 02/16/2021] [Accepted: 02/23/2021] [Indexed: 12/13/2022]
Abstract
Congenital aniridia is a rare genetic eye disorder with total or partial absence of the iris from birth. In most cases the genetic origin of aniridia is a mutation in the PAX6 gene, leading to involvement of most eye structures. Hypoplasia of the fovea is usually present and is associated with reduced visual acuity and nystagmus. Aniridia-associated keratopathy, glaucoma, and cataract are serious and progressive complications that can further reduce visual function. Treatment of the ocular complications of aniridia is challenging and has a high risk of side effects. New approaches such as stem cell therapy may, however, offer better prognoses. We describe the various ocular manifestations of aniridia, with a special focus on conditions that commonly require treatment. We also review the growing literature reporting systemic manifestations of the disease.
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Affiliation(s)
| | - Neil Lagali
- Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
| | - Tor P Utheim
- Department of Ophthalmology, Oslo University Hospital, Oslo, Norway; Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway
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Yazdani M, Shahdadfar A, Reppe S, Sapkota D, Vallenari EM, Lako M, Connon CJ, Figueiredo FC, Utheim TP. Response of human oral mucosal epithelial cells to different storage temperatures: A structural and transcriptional study. PLoS One 2020; 15:e0243914. [PMID: 33326470 PMCID: PMC7744058 DOI: 10.1371/journal.pone.0243914] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/08/2020] [Accepted: 11/30/2020] [Indexed: 11/26/2022] Open
Abstract
PURPOSE Seeking to improve the access to regenerative medicine, this study investigated the structural and transcriptional effects of storage temperature on human oral mucosal epithelial cells (OMECs). METHODS Cells were stored at four different temperatures (4°C, 12°C, 24°C and 37°C) for two weeks. Then, the morphology, cell viability and differential gene expression were examined using light and scanning electron microscopy, trypan blue exclusion test and TaqMan gene expression array cards, respectively. RESULTS Cells stored at 4°C had the most similar morphology to non-stored controls with the highest viability rate (58%), whereas the 37°C group was most dissimilar with no living cells. The genes involved in stress-induced growth arrest (GADD45B) and cell proliferation inhibition (TGFB2) were upregulated at 12°C and 24°C. Upregulation was also observed in multifunctional genes responsible for morphology, growth, adhesion and motility such as EFEMP1 (12°C) and EPHA4 (4°C-24°C). Among genes used as differentiation markers, PPARA and TP53 (along with its associated gene CDKN1A) were downregulated in all temperature conditions, whereas KRT1 and KRT10 were either unchanged (4°C) or downregulated (24°C and 12°C; and 24°C, respectively), except for upregulation at 12°C for KRT1. CONCLUSIONS Cells stored at 12°C and 24°C were stressed, although the expression levels of some adhesion-, growth- and apoptosis-related genes were favourable. Collectively, this study suggests that 4°C is the optimal storage temperature for maintenance of structure, viability and function of OMECs after two weeks.
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Affiliation(s)
- Mazyar Yazdani
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, Oslo, Norway
- Center for Eye Research, Department of Ophthalmology, Oslo University Hospital, Ullevål, Oslo, Norway
| | - Aboulghassem Shahdadfar
- Center for Eye Research, Department of Ophthalmology, Oslo University Hospital, Ullevål, Oslo, Norway
| | - Sjur Reppe
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, Oslo, Norway
- Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Oslo, Norway
- Lovisenberg Diaconal Hospital, Unger-Vetlesen Institute, Oslo, Norway
| | - Dipak Sapkota
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
| | - Evan M. Vallenari
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
| | - Majlinda Lako
- Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Life, Bioscience West Building, Newcastle upon Tyne, United Kingdom
| | - Che J. Connon
- Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Life, Bioscience West Building, Newcastle upon Tyne, United Kingdom
| | - Francisco C. Figueiredo
- Biosciences Institute, Faculty of Medical Sciences, Newcastle University, International Centre for Life, Bioscience West Building, Newcastle upon Tyne, United Kingdom
- Department of Ophthalmology, Royal Victoria Infirmary & Newcastle University, Newcastle upon Tyne, United Kingdom
| | - Tor Paaske Utheim
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, Oslo, Norway
- Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Oslo, Norway
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
- Department of Ophthalmology, Stavanger University Hospital, Stavanger, Norway
- Department of Ophthalmology, Sørlandet Hospital Arendal, Arendal, Norway
- Department of Computer Science, Oslo Metropolitan University, Oslo, Norway
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Nosrati H, Alizadeh Z, Nosrati A, Ashrafi-Dehkordi K, Banitalebi-Dehkordi M, Sanami S, Khodaei M. Stem cell-based therapeutic strategies for corneal epithelium regeneration. Tissue Cell 2020; 68:101470. [PMID: 33248403 DOI: 10.1016/j.tice.2020.101470] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2020] [Revised: 11/11/2020] [Accepted: 11/11/2020] [Indexed: 12/13/2022]
Abstract
Any significant loss of vision or blindness caused by corneal damages is referred to as corneal blindness. Corneal blindness is the fourth most common cause of blindness worldwide, representing more than 5% of the total blind population. Currently, corneal transplantation is used to treat many corneal diseases. In some cases, implantation of artificial cornea (keratoprosthesis) is suggested after a patient has had a donor corneal transplant failure. The shortage of donors and the side effects of keratoprosthesis are limiting these approaches. Recently, researchers have been actively pursuing new approaches for corneal regeneration because of these limitations. Nowadays, tissue engineering of different corneal layers (epithelium, stroma, endothelium, or full thickness tissue) is a promising approach that has attracted a great deal of interest from researchers and focuses on regenerative strategies using different cell sources and biomaterials. Various sources of corneal and non-corneal stem cells have shown significant advantages for corneal epithelium regeneration applications. Pluripotent stem cells (embryonic stem cells and iPS cells), epithelial stem cells (derived from oral mucus, amniotic membrane, epidermis and hair follicle), mesenchymal stem cells (bone marrow, adipose-derived, amniotic membrane, placenta, umbilical cord), and neural crest origin stem cells (dental pulp stem cells) are the most promising sources in this regard. These cells could also be used in combination with natural or synthetic scaffolds to improve the efficacy of the therapeutic approach. As the ocular surface is exposed to external damage, the number of studies on regeneration of the corneal epithelium is rising. In this paper, we reviewed the stem cell-based strategies for corneal epithelium regeneration.
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Affiliation(s)
- Hamed Nosrati
- Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran.
| | - Zohreh Alizadeh
- Endometrium and Endometriosis Research Center, Hamadan University of Medical Sciences, Hamadan, Iran; Department of Anatomical Sciences, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
| | - Ali Nosrati
- School of Mechanical Engineering, College of Engineering, University of Tehran, Tehran, Iran
| | - Korosh Ashrafi-Dehkordi
- Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Mehdi Banitalebi-Dehkordi
- Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Samira Sanami
- Department of Medical Biotechnology, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Mohammad Khodaei
- Department of Materials Science and Engineering, Golpayegan University of Technology, Golpayegan, Iran
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Xeno-free approach for the expansion of human adipose derived mesenchymal stem cells for ocular therapies. Exp Eye Res 2020; 202:108358. [PMID: 33207223 DOI: 10.1016/j.exer.2020.108358] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2020] [Revised: 11/05/2020] [Accepted: 11/09/2020] [Indexed: 11/21/2022]
Abstract
To restore corneal transparency and vision loss after an injury on the ocular surface, the use of human stem cells from different origins has been recently proposed. Mesenchymal stem cells (MSCs) seem to be an appropriate adult source of autologous stem cells due to their accessibility, high proliferation rate, and multipotent capacity. In this work, we developed a simple culture system to prepare a graft based on a fibrin membrane seeded with human MSCs. A commercial kit, PRGF Endoret®, was used to prepare both, the growth factors used as culture media supplement and the fibrin membrane grafts. Adipose-derived MSCs (Ad-MSCs) were expanded, characterised by flow cytometry and their multilineage differentiation potential confirmed by inducing adipogenesis, osteogenesis and chondrogenesis. Ad-MSCs seeded on the fibrin membranes were grafted onto athymic mice showing good biocompatibility with no adverse reactions observed during the follow up period. These findings support the assumption that a system in which all the biological components (cells, grow factors and carrier) are autologous, could potentially be used for future ex vivo expansion of Ad-MSCs to treat ocular conditions such as an inflammatory milieu, traumatic scars and loss of the regenerative capacity of the corneal epithelium that compromise the quality of vision.
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Nosrati H, Abpeikar Z, Mahmoudian ZG, Zafari M, Majidi J, Alizadeh A, Moradi L, Asadpour S. Corneal epithelium tissue engineering: recent advances in regeneration and replacement of corneal surface. Regen Med 2020; 15:2029-2044. [PMID: 33169642 DOI: 10.2217/rme-2019-0055] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
Currently, many corneal diseases are treated by corneal transplantation, artificial corneal implantation or, in severe cases, keratoprosthesis. Owing to the shortage of cornea donors and the risks involved with artificial corneal implants, such as infection transmission, researchers continually seek new approaches for corneal regeneration. Corneal tissue engineering is a promising approach that has attracted much attention from researchers and is focused on regenerative strategies using various biomaterials in combination with different cell types. These constructs should have the ability to mimic the native tissue microenvironment and present suitable optical, mechanical and biological properties. In this article, we review studies that have focused on the current clinical techniques for corneal replacement. We also describe tissue-engineering and cell-based approaches for corneal regeneration.
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Affiliation(s)
- Hamed Nosrati
- Department of Tissue Engineering & Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Zahra Abpeikar
- Department of Tissue Engineering & Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Zahra Gholami Mahmoudian
- Department of Anatomical Sciences, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran
| | - Mahdi Zafari
- Department of Bacteriology, Pasteur Institute of Iran, Tehran, Iran
| | - Jafar Majidi
- Cellular & Molecular Research Center, Basic Health Science Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
| | - Akram Alizadeh
- Department of Tissue Engineering & Applied Cell Sciences, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran
| | - Lida Moradi
- The Ronald O Perelman Department of Dermatology, New York University, School of Medicine, New York, NY 10016, USA.,Department of Cell Biology, New York University, School of Medicine, New York, NY, 10016 USA
| | - Shiva Asadpour
- Department of Tissue Engineering & Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran.,Cellular & Molecular Research Center, Basic Health Science Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
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Theerakittayakorn K, Thi Nguyen H, Musika J, Kunkanjanawan H, Imsoonthornruksa S, Somredngan S, Ketudat-Cairns M, Parnpai R. Differentiation Induction of Human Stem Cells for Corneal Epithelial Regeneration. Int J Mol Sci 2020; 21:E7834. [PMID: 33105778 PMCID: PMC7660084 DOI: 10.3390/ijms21217834] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/25/2020] [Revised: 10/16/2020] [Accepted: 10/17/2020] [Indexed: 12/13/2022] Open
Abstract
Deficiency of corneal epithelium causes vision impairment or blindness in severe cases. Transplantation of corneal epithelial cells is an effective treatment but the availability of the tissue source for those cells is inadequate. Stem cells can be induced to differentiate to corneal epithelial cells and used in the treatment. Multipotent stem cells (mesenchymal stem cells) and pluripotent stem cells (embryonic stem cells and induced pluripotent stem cells) are promising cells to address the problem. Various protocols have been developed to induce differentiation of the stem cells into corneal epithelial cells. The feasibility and efficacy of both human stem cells and animal stem cells have been investigated for corneal epithelium regeneration. However, some physiological aspects of animal stem cells are different from those of human stem cells, the protocols suited for animal stem cells might not be suitable for human stem cells. Therefore, in this review, only the investigations of corneal epithelial differentiation of human stem cells are taken into account. The available protocols for inducing the differentiation of human stem cells into corneal epithelial cells are gathered and compared. Also, the pathways involving in the differentiation are provided to elucidate the relevant mechanisms.
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Affiliation(s)
- Kasem Theerakittayakorn
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Hong Thi Nguyen
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Jidapa Musika
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Hataiwan Kunkanjanawan
- Medeze Research and Development Co., Ltd. 28/9 Moo 8, Phutthamonthon Sai 4 Rd., Krathum Lom, Sam Phran, Nakhon Pathom 73220, Thailand;
| | - Sumeth Imsoonthornruksa
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Sirilak Somredngan
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Mariena Ketudat-Cairns
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
| | - Rangsun Parnpai
- Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand; (K.T.); (H.T.N.); (J.M.); (S.I.); (S.S.); (M.K.-C.)
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Wang B, Li W, Harrison J. An Evaluation of Wound Healing Efficacy of a Film Dressing Made from Polymer-integrated Amnion Membrane. Organogenesis 2020; 16:126-136. [PMID: 33164697 PMCID: PMC7714469 DOI: 10.1080/15476278.2020.1844507] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/25/2020] [Revised: 09/01/2020] [Accepted: 09/09/2020] [Indexed: 10/23/2022] Open
Abstract
A film dressing is an easy and common wound management, which is flexible to cover many types of superficial injuries. In a recent study, we developed a scaffold from poly (1,8-octanediolco-citrate) incorporated decellularized amnion membrane (DAM-POC). The DAM-POC scaffold was biocompatible and could enhance soft and hard tissue regeneration when applied to repair the cleft palate in rat. The efficacy of the DAM-POC scaffold in oral repair had led us to hypothesize that it could be employed extensively in the medical field as a wound dressing. This study aimed to investigate the feasibility and efficacy of the DAM-POC scaffold as a film dressing in accelerating wound healing when applied in multiple tissue injuries. Our results demonstrated that both the DAM and DAM-POC scaffolds were biocompatible and anti-adhesive without causing severe foreign body reactions when covering wounds in abdominal wall, back muscle, tibia bone, and liver. In addition, the DAM-POC scaffold was superior to the DAM scaffold in reducing inflammation, preventing fibrosis, and regenerating tissues. In conclusion, the DAM-POC scaffold might potentially be adopted as a film dressing in a wide range of therapeutic applications and healing situations to protect the damaged tissues from the external environment and prevent infections.
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Affiliation(s)
- Bo Wang
- Joint Department of Biomedical Engineering, Marquette University and Medical College of Wisconsin, Milwaukee, WI, USA
| | - Wuwei Li
- Department of Oral and Maxillofacial Surgery, School of Stomatology, Dalian Medical University, Dalian, China
| | - Justin Harrison
- Department of Biological Sciences, Alabama State University, Montgomery, AL, USA
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He J, Ou S, Ren J, Sun H, He X, Zhao Z, Wu H, Qu Y, Liu T, Jeyalatha V, Zhang L, Li Q, Reinach PS, Quantock A, Hao J, Liu Z, Li W. Tissue engineered corneal epithelium derived from clinical-grade human embryonic stem cells. Ocul Surf 2020; 18:672-680. [PMID: 32710961 DOI: 10.1016/j.jtos.2020.07.009] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/14/2020] [Revised: 07/10/2020] [Accepted: 07/12/2020] [Indexed: 02/07/2023]
Abstract
PURPOSE To construct tissue engineered corneal epithelium from a clinical-grade human embryonic stem cells (hESCs) and investigate the dynamic gene profile and phenotypic transition in the process of differentiation. METHODS A stepwise protocol was applied to induce differentiation of clinical-grade hESCs Q-CTS-hESC-1 and construct tissue engineered corneal epithelium. Single cell RNA sequencing (scRNA-seq) analysis was performed to monitor gene expression and phenotypic changes at different differentiation stages. Immunostaining, real-time quantitative PCR and Western blot analysis were conducted to detect gene and protein expressions. After subcutaneous transplantation into nude mice to test the biosafety, the epithelial construct was transplanted in a rabbit corneal limbal stem cell deficiency (LSCD) model and followed up for eight weeks. RESULTS The hESCs were successfully induced into epithelial cells. scRNA-seq analysis revealed upregulation of ocular surface epithelial cell lineage related genes such as TP63, Pax6, KRT14, and activation of Wnt, Notch, Hippo, and Hedgehog signaling pathways during the differentiation process. Tissue engineered epithelial cell sheet derived from hESCs showed stratified structure and normal corneal epithelial phenotype with presence of clonogenic progenitor cells. Eight weeks after grafting the cell sheet onto the ocular surface of LSCD rabbit model, a full-thickness continuous corneal epithelium developed to fully cover the damaged areas with normal limbal and corneal epithelial phenotype. CONCLUSION The tissue engineered corneal epithelium generated from a clinical-grade hESCs may be feasible in the treatment of limbal stem cell deficiency.
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Affiliation(s)
- Jia He
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Shangkun Ou
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Jun Ren
- School of Informatics, Xiamen University, Xiamen, Fujian, China; National Institute for Data Science in Health and Medicine, Xiamen University, Xiamen, Fujian, China
| | - Huimin Sun
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Xin He
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Zhongyang Zhao
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Han Wu
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Yangluowa Qu
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Tingting Liu
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Vimalin Jeyalatha
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Liying Zhang
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Qiyuan Li
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China
| | - Peter Sol Reinach
- Wenzhou Medical University, Department of Ophthalmology, Wenzhou, Zhejiang, China; Wenzhou Medical University, Department of Optometry, Wenzhou, Zhejiang, China
| | - Andrew Quantock
- Structural Biophysics Group, School of Optometry and Vision Sciences, Cardiff University, Cardiff, Wales, United Kingdom
| | - Jie Hao
- State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China; National Stem Cell Resource Center, Chinese Academy of Sciences, Beijing, China.
| | - Zuguo Liu
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China.; Department of Ophthalmology, Xiang'an Hospital of Xiamen University, Xiamen, Fujian, China; Xiamen University Affiliated Xiamen Eye Center, Xiamen, Fujian, China.
| | - Wei Li
- Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, China.; Department of Ophthalmology, Xiang'an Hospital of Xiamen University, Xiamen, Fujian, China; Xiamen University Affiliated Xiamen Eye Center, Xiamen, Fujian, China.
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Tighe S, Mead OG, Lee A, Tseng SCG. Basic science review of birth tissue uses in ophthalmology. Taiwan J Ophthalmol 2020; 10:3-12. [PMID: 32309118 PMCID: PMC7158924 DOI: 10.4103/tjo.tjo_4_20] [Citation(s) in RCA: 31] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/25/2019] [Accepted: 01/12/2020] [Indexed: 12/31/2022] Open
Abstract
The birth tissue is predominantly comprised of amniotic membrane (AM) and umbilical cord (UC), which share the same cell origin as the fetus. These versatile biological tissues have been used to treat a wide range of conjunctival and corneal conditions since 1940. The therapeutic benefits of the birth tissue stem from its anti-inflammatory and anti-scarring properties that orchestrate regenerative healing. Although the birth tissue also contains many cytokines, growth factors, and proteins, the heavy chain 1-hyaluronic acid/pentraxin 3 (HC-HA/PTX3) matrix has been identified to be a major active tissue component responsible for AM/UC's multifactorial therapeutic actions. HC-HA/PTX3 complex is abundantly present in fresh and cryopreserved AM/UC, but not in dehydrated tissue. In this review, we discuss the tissue anatomy, the molecular mechanism of action based on HC-HA/ PTX3 to explain their therapeutic potentials, and the various forms available in ophthalmology.
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Affiliation(s)
- Sean Tighe
- R&D Department, TissueTech Inc., Miami, Florida, USA
- Department of Ophthalmology, Herbert Wertheim College of Medicine, Florida International University, Miami, Florida, USA
- Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, FL, USA
| | | | - Amy Lee
- R&D Department, TissueTech Inc., Miami, Florida, USA
| | - Scheffer C. G. Tseng
- R&D Department, TissueTech Inc., Miami, Florida, USA
- Ocular Surface Center and Ocular Surface Research Education Foundation, Miami, FL, USA
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Hatano H, Goda T, Matsumoto A, Miyahara Y. Induced Proton Dynamics on Semiconductor Surfaces for Sensing Tight Junction Formation Enhanced by an Extracellular Matrix and Drug. ACS Sens 2019; 4:3195-3202. [PMID: 31763825 DOI: 10.1021/acssensors.9b01635] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
Abstract
In the fields of tissue engineering and drug discovery, confirming the formation and maturation of epithelial cell tight junctions (TJs), which are necessary for blocking pathogenic invasion and absorption of nutrients and ions, at a high spatiotemporal resolution is essential. We previously developed a system of monitoring pH perturbation induced by weak acid exposure to cells cultured on an ion-sensitive field-effect transistor that enables a sensitive and specific detection of biomembrane injuries and TJ breakdowns caused by external stimuli such as nanomaterials and cytotoxins. In this study, we monitor time-lapse changes in the paracellular diffusion of growing epithelial cell monolayers using the pH perturbation assay as well as conventional permeability and trans-epithelial electrical resistance assays. The effects of the extracellular matrix and a TJ potentiator (KN-93) on epithelial TJ formation are evaluated. TJ formations were promoted on the substrate coated with Matrigel more than on the one coated with poly(l-lysine). KN-93 accelerated TJ formations in a dose-dependent manner. The pH perturbation assay denoted a longer incubation time for the completion of TJ formation compared with the conventional assays under the same conditions. Importantly, the pH perturbation assay is able to rigorously evaluate TJ formation, as the assay uses protons as the smallest indicator for detecting paracellular gaps, and the pH perturbation is specific to TJ alterations. These features for in vitro TJ evaluation using proton dynamics are advantageous for applications in tissue engineering and drug development.
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Affiliation(s)
- Hiroaki Hatano
- Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
| | - Tatsuro Goda
- Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
- Nano Innovation Institute, Inner Mongolia University for Nationalities, No. 22 Huoline Street, Tongliao, Inner Mongolia 028000, P. R. China
| | - Akira Matsumoto
- Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
- Kanagawa Institute of Industrial Science and Technology, 705-1 Shimoimaizumi, Ebina, Kanagawa 243-0435, Japan
| | - Yuji Miyahara
- Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
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Simple oral mucosal epithelial transplantation in a rabbit model. Sci Rep 2019; 9:18088. [PMID: 31792300 PMCID: PMC6889162 DOI: 10.1038/s41598-019-54571-7] [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: 03/06/2019] [Accepted: 11/12/2019] [Indexed: 01/22/2023] Open
Abstract
This study investigated a rabbit model of autologous simple oral mucosal epithelium transplantation (SOMET) for limbal stem cell deficiency (LSCD). LSCD was created in the SOMET group and the Control group. In the SOMET group, oral mucosa harvested from the buccal region was treated with dispase, cut into small pieces, and placed on the exposed corneal stroma without using graft sutures, amniotic membrane, and/or glue. A soft contact lens was positioned and tarsorrhaphy was performed in both groups. Postoperative corneal neovascularization and fluorescein staining scores were evaluated by slit lamp microscopy in both groups. At 2 weeks postoperatively, eyes were excised and subjected to immunohistochemical staining for CK3, CK13, CK15, and p63. In the SOMET group, transplantation of oral mucosa led to complete recovery of LSCD, as indicated by low neovascularization scores, low fluorescein staining scores, and detection of stratified K3/K13-positive cells on the stroma at 2 weeks after surgery. In contrast, corneal epithelial defects persisted in the Control group at 2 weeks. SOMET achieved re-epithelialization of the corneal surface in this rabbit LSCD model. It is a simple technique that does not require culture and could be a promising option for ocular surface reconstruction in bilateral LSCD.
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Yazdani M, Shahdadfar A, Jackson CJ, Utheim TP. A Hyaluronan Hydrogel Scaffold for Culture of Human Oral Mucosal Epithelial Cells in Limbal Stem-Cell Therapy. Bioengineering (Basel) 2019; 6:E97. [PMID: 31652804 PMCID: PMC6955856 DOI: 10.3390/bioengineering6040097] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/07/2019] [Revised: 10/20/2019] [Accepted: 10/22/2019] [Indexed: 12/26/2022] Open
Abstract
Hyaluronan (HA), a major component of the extracellular matrix, plays a key role in cell proliferation, growth, survival, polarization and differentiation. We investigated the optimization of a HA hydrogel scaffold for culture of human oral mucosal epithelial cells (OMECs) for potential application in limbal stem cell therapy. The effect of the optimized scaffold on OMEC cell sheet morphology, cell metabolic activity and expression of genes associated with stemness, adherence and cell damage was studied. The results indicate that HA hydrogels crosslinked with polyethylene glycol diacrylate (PEGDA) failed to support OMEC attachment and growth. However, HA hydrogel scaffolds dried for three days and coated with 1 mg/mL collagen IV produced a full OMEC sheet. Cell morphology was comparable to control after three weeks culture, maintaining 76% metabolic activity. Of apoptosis-related genes, the pro-apoptotic markers CASP3 and BAX2 were upregulated and downregulated, respectively, compared to control whereas the anti-apoptotic marker BCL2 was downregulated. The expression level of stemness genes ΔNp63α and ABCG2 was significantly higher than control. Genes associated with improved scar-less wound healing (integrin-V) and protection of the ocular surface (cadherin-1) had ~3-fold increased expression. These data suggest that our optimized HA-hydrogel scaffold could enhance culture of OMEC cell sheets for use in ocular reconstruction.
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Affiliation(s)
- Mazyar Yazdani
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, 0450 Oslo, Norway.
- Center for Eye Research, Department of Ophthalmology, Oslo University Hospital, Ullevål, 0450 Oslo, Norway.
- The Norwegian Dry Eye Clinic, 0366 Oslo, Norway.
| | - Aboulghassem Shahdadfar
- Center for Eye Research, Department of Ophthalmology, Oslo University Hospital, Ullevål, 0450 Oslo, Norway.
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, 0318 Oslo, Norway.
| | - Catherine Joan Jackson
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, 0450 Oslo, Norway.
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, 0318 Oslo, Norway.
- Department of Plastic and Reconstructive Surgery, Oslo University Hospital, 0450 Oslo, Norway.
| | - Tor Paaske Utheim
- Department of Medical Biochemistry, Oslo University Hospital, Ullevål, 0450 Oslo, Norway.
- The Norwegian Dry Eye Clinic, 0366 Oslo, Norway.
- Institute of Oral Biology, Faculty of Dentistry, University of Oslo, 0318 Oslo, Norway.
- Department of Plastic and Reconstructive Surgery, Oslo University Hospital, 0450 Oslo, Norway.
- Department of Ophthalmology, Stavanger University Hospital, 4011 Stavanger, Norway.
- Department of Ophthalmology, Sørlandet Hospital Arendal, 4604 Arendal, Norway.
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Gericke A, Wang X, Ackermann M, Neufurth M, Wiens M, Schröder HC, Pfeiffer N, Müller WEG. Utilization of metabolic energy in treatment of ocular surface disorders: polyphosphate as an energy source for corneal epithelial cell proliferation. RSC Adv 2019; 9:22531-22539. [PMID: 35519495 PMCID: PMC9066647 DOI: 10.1039/c9ra04409d] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2019] [Accepted: 07/16/2019] [Indexed: 12/18/2022] Open
Abstract
Impaired regeneration of the corneal epithelium, as found in many ocular surface diseases, is a major clinical problem in ophthalmology. We hypothesized that corneal epithelial regeneration can be promoted by the physiological, energy-delivering as well as "morphogenetically active" polymer, inorganic polyphosphate (polyP). Corneal limbal explants (diameter, 4 mm) were cultivated on collagen-coated well plates in the absence or presence of polyP (chain length, ∼40 Pi units; 50 μg ml-1) or human platelet lysate (hp-lysate; 5% v/v). Cell outgrowth and differentiation were analyzed after staining with DRAQ5 (nuclei) and rhodamine phalloidin (cytoskeleton), as well as by environmental scanning electron microscopy (ESEM). Cell growth/viability of hCECs was assessed by XTT assay. The expression of SDF-1 was quantitated by qRT-PCR. Exposure to hp-lysate (also containing polyP) increased cell migration already at day 1. Even stronger was the effect of polyP. This effect was blocked by a mast cell serine protease. The formation of cell multilayers was enhanced by hp-lysate or even more by polyP. ESEM revealed continuous cell junctions and prominent microvilli on the surface of adjacent cells exposed to polyP; those structures were only rarely seen in the controls. The hp-lysate and, more potently, polyP increased the proliferation of hCECs, as well as SDF-1 expression. The findings indicate the potential usefulness of the natural polymer, polyP, for topical treatment of corneal epithelial defects. Future studies are directed to develop suitable formulations of polyP, such as biomimetic polyP nano/microparticles showing an adjustable release kinetics.
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Affiliation(s)
- Adrian Gericke
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz Langenbeckstrasse 1 55131 Mainz Germany
| | - Xiaohong Wang
- ERC Advanced Investigator Grant Research Group at the Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Duesbergweg 6 D-55128 Mainz Germany +49-6131-39-25243 +49-6131-39-25910
| | - Maximilian Ackermann
- Institute of Functional and Clinical Anatomy, University Medical Center of the Johannes Gutenberg University Johann Joachim Becher Weg 13 55099 Mainz Germany
| | - Meik Neufurth
- ERC Advanced Investigator Grant Research Group at the Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Duesbergweg 6 D-55128 Mainz Germany +49-6131-39-25243 +49-6131-39-25910
| | - Matthias Wiens
- ERC Advanced Investigator Grant Research Group at the Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Duesbergweg 6 D-55128 Mainz Germany +49-6131-39-25243 +49-6131-39-25910
| | - Heinz C Schröder
- ERC Advanced Investigator Grant Research Group at the Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Duesbergweg 6 D-55128 Mainz Germany +49-6131-39-25243 +49-6131-39-25910
| | - Norbert Pfeiffer
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz Langenbeckstrasse 1 55131 Mainz Germany
| | - Werner E G Müller
- ERC Advanced Investigator Grant Research Group at the Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Duesbergweg 6 D-55128 Mainz Germany +49-6131-39-25243 +49-6131-39-25910
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Li W, Fu Y, Jiang B, Lo AY, Ameer GA, Barnett C, Wang B. Polymer-integrated amnion scaffold significantly improves cleft palate repair. Acta Biomater 2019; 92:104-114. [PMID: 31102764 DOI: 10.1016/j.actbio.2019.05.035] [Citation(s) in RCA: 23] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2018] [Revised: 04/08/2019] [Accepted: 05/13/2019] [Indexed: 11/17/2022]
Abstract
Cleft palate is a common oral and craniomaxillofacial birth defect. As the ideal surgery time is shortly after birth, clinical treatments should result in minimal disruption of theskeleton to allow tissue growth in children. A tissue-engineered graft was created in this study for cleft palate repair by integrating poly(1,8-octamethylene-citrate) (POC) with a decellularized amnion membrane (DAM-POC) to incorporate the advantages of both the synthetic polymer and the native tissue. The success of POC incorporation was confirmed by laser-induced breakdown spectroscopy and fluorescence detection. The DAM-POC scaffold showed a certain level of structure collapse and lower stiffness but better resistance to enzyme digestion than the native amnion and DAM scaffold. The DAM-POC scaffold is cell compatible when seeded with mesenchymal stem cells, as evidenced by adequate cell viability and improved alkaline phosphatase (ALP) activity and calcium deposit. A large palate defect was first surgically created in a young rat model and then repaired with the DAM-POC scaffold. Eight weeks postsurgery, histological study and CT scans showed nearly complete healing of both soft and hard tissues. In conclusion, we developed a cell-free, resorbable graft by incorporating and integrating a synthetic polymer with a human DAM. When the DAM-POC scaffold was applied to repair a large palate defect in young rats, it showed adequate biocompatibility as evidenced by its effectiveness in guiding hard and soft tissue regeneration and minimum interference with natural growth and palate development of rats. STATEMENT OF SIGNIFICANCE: Proper restoration of severe cleft palate remains a major challenge because of insufficient autologous soft tissues to close the open wounds, thereby causing high tension at the surgical junction, secondary palatal fistulas, wound contraction, scar tissue formation, and facial growth disturbances. In this study, we have developed a tissue-engineered graft through incorporating and integrating a synthetic polymer with the human amnion membrane for cleft palate repair. The significance of this study lies in our ability to develop a cell-free, resorbable graft that can provide a less surgically invasive option to cover the open defect and support palate regeneration and tissue growth. This technique could potentially advance soft and hard tissue regeneration in children with birth craniomaxillofacial defects.
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Affiliation(s)
- Wuwei Li
- Department of Oral and Maxillofacial Surgery, School of Stomatology, Dalian Medical University, Liaoning 116001, China
| | - Yuqian Fu
- Department of Oral and Maxillofacial Surgery, School of Stomatology, Dalian Medical University, Liaoning 116001, China
| | - Bin Jiang
- Biomedical Engineering Department, Northwestern University, Evanston, IL 60201, United States; Center for Advanced Regenerative Engineering, Northwestern University, Evanston, IL 60201, United States
| | - Aaron Y Lo
- Biomedical Engineering Department, Northwestern University, Evanston, IL 60201, United States; Center for Advanced Regenerative Engineering, Northwestern University, Evanston, IL 60201, United States
| | - Guillermo A Ameer
- Biomedical Engineering Department, Northwestern University, Evanston, IL 60201, United States; Center for Advanced Regenerative Engineering, Northwestern University, Evanston, IL 60201, United States
| | - Cleon Barnett
- Department of Physical Sciences, Alabama State University, Montgomery, AL 36104, United States
| | - Bo Wang
- Joint Department of Biomedical Engineering, Marquette University and the Medical College of Wisconsin, Milwaukee, WI 53226, United States.
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Choe HR, Yoon CH, Kim MK. Ocular Surface Reconstruction Using Circumferentially-trephined Autologous Oral Mucosal Graft Transplantation in Limbal Stem Cell Deficiency. KOREAN JOURNAL OF OPHTHALMOLOGY 2019; 33:16-25. [PMID: 30746908 PMCID: PMC6372385 DOI: 10.3341/kjo.2018.0111] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2018] [Accepted: 10/25/2018] [Indexed: 01/12/2023] Open
Abstract
PURPOSE To investigate the effects of transplantation of a circumferentially-trephined autologous oral mucosal graft using a vacuum trephine on ocular surface reconstruction in patients with limbal stem cell deficiency. METHODS Patients with a limbal stem cell deficiency who underwent transplantation of autologous oral mucosal graft performed by a particular surgeon in Seoul National University Hospital were included. The medical records of these five patients were retrospectively reviewed. The lower labial mucosal graft inside the inferior lip was trephined to a depth of 250 μm using a donor vacuum trephine with a 9-mm diameter. Outside markings were made using a 14-mm intraoperative keratometer. The oral mucosal graft was dissected under a microscope using a Beaver mini-blade as either a ring or a crescent-shaped strip with a 5-mm width. The mucosal graft was transplanted onto the limbus in the limbal-deficient eye. Best-corrected visual acuity and corneal status were measured during the follow-up period. RESULTS Four patients were diagnosed with Stevens-Johnson syndrome and one was diagnosed with atopy-associated immune keratitis. The mean follow-up period was 10.4 ± 2.9 months. After 4 months, visual acuity improved in all patients, and the mean improvement in logarithm of the minimum angle of resolution visual acuity was 0.526 ± 0.470 (range, 0.15 to 1.10). Corneal surface erosion and neovascularization decreased in four patients, and stromal opacity decreased in two patients. The engraftments maintained ocular surface stabilization in four of the five patients at the last follow-up. CONCLUSIONS Transplantation of circumferential autologous oral mucosal grafts may be effective for the treatment of limbal stem cell deficiency.
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Affiliation(s)
- Hye Rim Choe
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea
| | - Chang Ho Yoon
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.,Laboratory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Center, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea
| | - Mee Kum Kim
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.,Laboratory of Ocular Regenerative Medicine and Immunology, Seoul Artificial Eye Center, Seoul National University Hospital Biomedical Research Institute, Seoul, Korea.
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Duan CY, Xie HT, Zhao XY, Xu WH, Zhang MC. Limbal niche cells can reduce the angiogenic potential of cultivated oral mucosal epithelial cells. Cell Mol Biol Lett 2019; 24:3. [PMID: 30988673 PMCID: PMC6448320 DOI: 10.1186/s11658-018-0133-x] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/28/2018] [Accepted: 12/11/2018] [Indexed: 12/16/2022] Open
Abstract
Background Autologous cultivated oral mucosal epithelial transplantation (COMET) is an important treatment for limbal stem cell deficiency. However, peripheral corneal neovascularization after surgery hinders its application. This study aims to employ a culture system using allogenic limbal niche cells (LNCs) instead of mouse-derived 3T3 cells as a feeder layer that could relieve postoperative neovascularization. Methods Rat oral mucosal epithelial cells (OMECs) were co-cultured with rat LNCs or 3T3 cells. Cultivated oral mucosal epithelial cells (COMECs) of different culture systems were identified by hematoxylin and eosin staining and immunocytochemistry. The expression levels of the angiogenesis-related factors were analyzed by RT-qPCR and western blotting/ELISA. Angiogenic potential was reconfirmed by cell viability and tube formation assays with human umbilical vein endothelial cells (HUVECs). Results COMECs were obtained from both culture systems successfully. Immunocytochemistry showed approximately equal percentages of positive staining cells for p63α (p = 0.9177), ABCG2 (p = 0.526), Ki67 (p = 0.0987), and CK3 (p = 0.4000) in COMECs of different groups. RT-qPCR and western blotting/ELISA showed that COMECs of the LNC group expressed a significantly lower amount of basic fibroblast growth factor (bFGF) (p = 0.0038 for RT-qPCR, p = 0.0026 for western blotting) but more pigment epithelium-derived factor (PEDF) (p = 0.0172 for RT-qPCR, p = 0.0253 for western blotting) and soluble fms-like tyrosine kinase-1 (sFlt-1) (p < 0.0001 for RT-qPCR, p = 0.0064 for ELISA) than the COMECs of the 3T3 group. Furthermore, compared with COMECs of the 3T3 group, COMECs of the LNC group could reduce the viability (p = 0.0002) and tube formation (p = 0.0002) of HUVECs. Conclusions LNCs could substitute 3T3 cells for expanding OMECs in vitro, and the COMECs obtained in this system are less likely to induce postsurgical neovascularization, which provides an alternative option for an ex vivo culture system and promotes the application of COMET. Electronic supplementary material The online version of this article (10.1186/s11658-018-0133-x) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Chao-Ye Duan
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China
| | - Hua-Tao Xie
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China
| | - Xin-Yue Zhao
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China
| | - Wen-Han Xu
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China
| | - Ming-Chang Zhang
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China
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John S, Kesting MR, Paulitschke P, Stöckelhuber M, von Bomhard A. Development of a tissue-engineered skin substitute on a base of human amniotic membrane. J Tissue Eng 2019; 10:2041731418825378. [PMID: 30746119 PMCID: PMC6360641 DOI: 10.1177/2041731418825378] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2018] [Accepted: 12/28/2018] [Indexed: 12/03/2022] Open
Abstract
Allogenic graft material and tissue engineering have recently shown promising results for the improvement of both esthetic and functional outcomes in the treatment of large skin defects. We chose human amniotic membrane as a cellular scaffold in order to develop a skin substitute for later in vivo uses. Various methods of de-epithelialization of the human amniotic membrane were evaluated by histological analysis including hematoxylin–eosin and laminin staining, optic coherence tomography, and scanning electron microscopy with 0.25/0.02% trypsin/ethylenediaminetetraacetic acid treatment and mechanical cell removal showing an almost complete loss of the epithelium and a mainly intact basement membrane. Novel examination of human amniotic membrane by optic coherence tomography was feasible, but difficulties were experienced in handling and interpretation of the tissue as no comparable data exist. Subsequently, we developed an air–liquid interface cell culture to cultivate keratinocytes and fibroblasts on the de-epithelialized human amniotic membrane. We achieved a mostly keratinized surface on the epidermal side with a confluent fibroblast network on the chorion side.
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Affiliation(s)
| | - Marco Rainer Kesting
- Department of Oral and Maxillofacial Surgery, Technical University of Munich, Munich, Germany
| | | | - Mechthild Stöckelhuber
- Department of Oral and Maxillofacial Surgery, Technical University of Munich, Munich, Germany
| | - Achim von Bomhard
- Department of Oral and Maxillofacial Surgery, Technical University of Munich, Munich, Germany
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Zhang Q, Qian C, Xiao W, Zhu H, Guo J, Ge Z, Cui W. Development of a visible light, cross-linked GelMA hydrogel containing decellularized human amniotic particles as a soft tissue replacement for oral mucosa repair. RSC Adv 2019; 9:18344-18352. [PMID: 35547651 PMCID: PMC9087906 DOI: 10.1039/c9ra03009c] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2019] [Accepted: 05/17/2019] [Indexed: 01/07/2023] Open
Abstract
Early effective treatment of oral mucosal defects is the key to ensuring defect healing and functional recovery. The application of human amniotic membrane (HAM) in promoting wound healing has been shown to be safe and effective. However, amniotic membrane is thin, easy to tear and difficult to handle. Combined with the natural forces at play in the oral cavity, this has restricted the clinical applications of HAM for healing of mucosal defects. Methacrylated gelatin (GelMA) has good mechanical strength and adhesion, and can be used as a bionic repair film to attach to the damaged surface of oral mucosa, but GelMA lacks bioactive substances and cannot promote the rapid repair of oral mucosal defects. The aim of this study was to design a type of composite GelMA hydrogel mixed with decellularized human amniotic particles (dHAP) as an oral mucosa substitute, to promote regeneration of defective mucosa by stimulating rapid angiogenesis. The composite substitute GelMA–dHAP was easy to synthesize and store, and easy to operate for repair of oral mucosal defects. We show the angiogenic potential of GelMA–dHAP on chick chorioallontoic membrane and the curative effect of GelMA–dHAP as a treatment in the rabbit oral mucosa defect model. In conclusion, this study confirms the effectiveness of GelMA–dHAP as an ideal soft tissue substitute for the repair of oral mucosal defects, overcoming the shortcomings of using HAM or GelMA alone. Early effective treatment of oral mucosal defects is the key to ensuring defect healing and functional recovery.![]()
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Affiliation(s)
- Qiang Zhang
- Department of Oral and Maxillofacial Surgery
- The First Affiliated Hospital of Soochow University
- Soochow University
- Suzhou
- P. R. China
| | - Chunyu Qian
- Department of Oral and Maxillofacial Surgery
- The First Affiliated Hospital of Soochow University
- Soochow University
- Suzhou
- P. R. China
| | - Wanshu Xiao
- Department of Oral and Maxillofacial Surgery
- The First Affiliated Hospital of Soochow University
- Soochow University
- Suzhou
- P. R. China
| | - Huajun Zhu
- Department of Oral and Maxillofacial Surgery
- The First Affiliated Hospital of Soochow University
- Soochow University
- Suzhou
- P. R. China
| | - Jun Guo
- Department of Maxillofacial Surgery
- The Affiliated Hospital of Yangzhou University
- Yangzhou University
- Yangzhou
- P. R. China
| | - Zili Ge
- Department of Oral and Maxillofacial Surgery
- The First Affiliated Hospital of Soochow University
- Soochow University
- Suzhou
- P. R. China
| | - Wenguo Cui
- Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases
- Shanghai Institute of Traumatology and Orthopaedics
- Ruijin Hospital
- Shanghai Jiao Tong University School of Medicine
- P. R. China
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Duan CY, Xie HT, Zhao XY, Zhang MC. Limbal niche cells: a novel feeder cell for autologous cultivated oral mucosal epithelial transplantation. Regen Med 2018; 14:49-62. [PMID: 30572808 DOI: 10.2217/rme-2018-0122] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/17/2023] Open
Abstract
AIM To establish a culture system using conspecific limbal niche cells (LNCs) as feeders for autologous cultivated oral mucosal epithelial transplantation (COMET). MATERIALS & METHODS Rabbit oral epithelial sheets, harvested from culture systems containing LNCs or 3T3 cells, were transplanted onto limbal stem cell-deficient rabbit eyes (COMET-3T3 or COMET-LNCs). RESULTS After COMET, corneas were relatively restored, with the exception of mild neovascularization in one cornea of the COMET-3T3 group. CD34 was detected in COMET-3T3 group corneas. Corneas of the COMET-LNCs group expressed high levels of PEDF and sFlt-1, but low levels of bFGF, compared with expression in COMET-3T3 corneas. CONCLUSION The culture system containing conspecific LNC feeders could substitute for the 3T3 cell system and decrease the risk of neovascularization after COMET.
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Affiliation(s)
- Chao-Ye Duan
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
| | - Hua-Tao Xie
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
| | - Xin-Yue Zhao
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
| | - Ming-Chang Zhang
- Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
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Limbal Stem Cell Transplantation: Clinical Results, Limits, and Perspectives. Stem Cells Int 2018; 2018:8086269. [PMID: 30405723 PMCID: PMC6201383 DOI: 10.1155/2018/8086269] [Citation(s) in RCA: 57] [Impact Index Per Article: 8.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2018] [Revised: 07/10/2018] [Accepted: 08/27/2018] [Indexed: 12/13/2022] Open
Abstract
Limbal stem cell deficiency (LSCD) is a clinical condition characterized by damage of cornea limbal stem cells, which results in an impairment of corneal epithelium turnover and in an invasion of the cornea by the conjunctival epithelium. In these patients, the conjunctivalization of the cornea is associated with visual impairment and cornea transplantation has poor prognosis for recurrence of the conjunctivalization. Current treatments of LSCD are aimed at replacing the damaged corneal stem cells in order to restore a healthy corneal epithelium. The autotransplantation of limbal tissue from the healthy, fellow eye is effective in unilateral LSCD but leads to depauperation of the stem cell reservoir. In the last decades, novel techniques such as cultivated limbal epithelial transplantation (CLET) have been proposed in order to reduce the damage of the healthy fellow eye. Clinical and experimental evidence showed that CLET is effective in inducing long-term regeneration of a healthy corneal epithelium in patients with LSCD with a success rate of 70%–80%. Current limitations for the treatment of LSCD are represented by the lack of a marker able to unequivocally identify limbal stem cells and the treatment of total, bilateral LSCD which requires other sources of stem cells for ocular surface reconstruction.
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Al Shehadat S, Gorduysus MO, Abdul Hamid SS, Abdullah NA, Samsudin AR, Ahmad A. Optimization of scanning electron microscope technique for amniotic membrane investigation: A preliminary study. Eur J Dent 2018; 12:574-578. [PMID: 30369805 PMCID: PMC6178683 DOI: 10.4103/ejd.ejd_401_17] [Citation(s) in RCA: 24] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
Abstract
OBJECTIVE The objective of this study was to evaluate and compare the two scanning electron microscope (SEM) preparation protocols and determine the better SEM preparation technique to study stem cells on human amniotic membrane (hAM) scaffold. MATERIALS AND METHODS Formaldehyde-based protocol and glutaraldehyde-based protocol were compared to evaluate the quality of SEM images for stem cells cultured on hAM scaffold. RESULTS The results suggested that formaldehyde-based protocol is better than glutaraldehyde-based protocol in terms of showing clearer topography of the membrane as well as the boarders of the cells. To provide intact surface of the SEM sample and avoid possible ruptures of the hAM or the thin cell layer, it is recommended to perform the dehydration step using graded alcohol concentrations of 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90%, one time for each and twice in 100% for 10 min each. Gold sputter-coating step is not recommended as it does not improve the image quality. CONCLUSIONS To obtain clear SEM images, it is recommended to run a preliminary study to determine the better chemicals and conditions of sample preparation even when following preexisting protocols.
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Affiliation(s)
- Saaid Al Shehadat
- Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, UAE
| | - Mehmet Omer Gorduysus
- Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, UAE
| | | | - Nurul Asma Abdullah
- Department of Biomedicine, School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
| | - Abdul Rani Samsudin
- Department of Oral and Craniofacial Health Sciences, College of Dental Medicine, University of Sharjah, Sharjah, UAE
| | - Azlina Ahmad
- Department of Craniofacial Sciences and Oral Biology, School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Malaysia
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Fernandez-Buenaga R, Aiello F, Zaher SS, Grixti A, Ahmad S. Twenty years of limbal epithelial therapy: an update on managing limbal stem cell deficiency. BMJ Open Ophthalmol 2018; 3:e000164. [PMID: 30123847 PMCID: PMC6093197 DOI: 10.1136/bmjophth-2018-000164] [Citation(s) in RCA: 31] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2018] [Revised: 07/06/2018] [Accepted: 07/20/2018] [Indexed: 01/19/2023] Open
Abstract
Limbal stem cell damage after chemical injury, autoimmune disorders or iatrogenic trauma leads to corneal conjunctivalisation with new vessel formation, epithelium instability and visual loss. Limbal stem cell transplantation includes reconstructive surgical procedures to restore a corneal epithelium. The recognised options are: conjunctival limbal autograft, in which stem cells are taken from the patient's healthy eye; conjunctival limbal allograft, in which stem cells are taken from a living, related or dead donor and the keratolimbal allograft. Each of these procedures has some drawbacks; in particular, the conjunctival limbal autograft needs a relatively healthy fellow eye and needs a relatively large amount of donor tissue from the healthy eye (about one-third of the healthy limbal stem cell tissue) with potential risks to the donor eye. In the case of keratolimbal allograft transplants, the recipient needs an immunosuppressive treatment to reduce the risk of rejection with the associate possible side effects. More modern treatment options are reviewed. Cultivated oral mucosa epithelial transplantation success rate can vary between 50% and 70% at 3-4 years of follow-up. Simple limbal epithelial transplantation results show a success rate from 75.2% to 83.8% after 1 year of follow-up. Inclusion criteria for autologous cultivated limbal epithelial transplantation as approved by the National Institute of Health and Care Excellence are also shown in this paper. On the basis of these more contemporary treatment options, a stepladder approach to evaluate which procedure is most appropriate and personalised to the patient's conditions is proposed.
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Affiliation(s)
- Roberto Fernandez-Buenaga
- Department of Cornea and External Diseases, Moorfields Eye Hospital-NHS Foundation Trust, London, UK
| | - Francesco Aiello
- Department of Cornea and External Diseases, Moorfields Eye Hospital-NHS Foundation Trust, London, UK
- Department of Experimental Medicine, University of Rome Tor Vergata, Rome, Italy
| | - Sarah S Zaher
- Department of Cornea and External Diseases, Moorfields Eye Hospital-NHS Foundation Trust, London, UK
| | - Andre Grixti
- Department of Cornea and External Diseases, Moorfields Eye Hospital-NHS Foundation Trust, London, UK
| | - Sajjad Ahmad
- Department of Cornea and External Diseases, Moorfields Eye Hospital-NHS Foundation Trust, London, UK
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Attico E, Sceberras V, Pellegrini G. Approaches for Effective Clinical Application of Stem Cell Transplantation. CURRENT TRANSPLANTATION REPORTS 2018; 5:244-250. [PMID: 30221121 PMCID: PMC6132451 DOI: 10.1007/s40472-018-0202-0] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/01/2022]
Abstract
PURPOSE OF REVIEW This review highlights problems related to translation of advanced therapy medicinal products (ATMPs) from bench to bedsite. Regenerative medicine within the current regulatory frame reveals common hitches in the course of development, translation, and clinical application. This paper suggests outlining a path from the few examples of successfully approved vs unsuccessful advanced therapies. RECENT FINDINGS In the multitude of ongoing studies, few of them achieved positive results with a final treatment available to patients; this result was possible due to multidisciplinary teams working together from the beginning of the development and during the hard route to standardization and clinical application. SUMMARY The root of success of an advanced therapy requires not only the inescapable scientific and biological knowledge but also requires several contributions as regulatory, ethical, medical, and bio-engineering expertise, from the real beginning. A strong scientific rationale and an integrated network of expertises would contribute to a successful investment of available resources in advanced therapy medicinal products and to a greater confidence in future medicine.
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Affiliation(s)
- E. Attico
- Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
| | | | - G. Pellegrini
- Centre for Regenerative Medicine “Stefano Ferrari”, University of Modena and Reggio Emilia, Modena, Italy
- Holostem Terapie Avanzate, Modena, Italy
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Sasamoto Y, Ksander BR, Frank MH, Frank NY. Repairing the corneal epithelium using limbal stem cells or alternative cell-based therapies. Expert Opin Biol Ther 2018; 18:505-513. [PMID: 29471701 PMCID: PMC6317528 DOI: 10.1080/14712598.2018.1443442] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
INTRODUCTION The corneal epithelium is maintained by limbal stem cells (LSCs) that reside in the basal epithelial layer of the tissue surrounding the cornea termed the limbus. Loss of LSCs results in limbal stem cell deficiency (LSCD) that can cause severe visual impairment. Patients with partial LSCD may respond to conservative therapies designed to rehabilitate the remaining LSCs. However, if these conservative approaches fail or, if complete loss of LSCs occurs, transplantation of LSCs or their alternatives is the only option. While a number of clinical studies utilizing diverse surgical and cell culture techniques have shown favorable results, a universal cure for LSCD is still not available. Knowledge of the potential risks and benefits of current approaches, and development of new technologies, is essential for further improvement of LSCD therapies. AREAS COVERED This review focuses on cell-based LSCD treatment approaches ranging from current available clinical therapies to preclinical studies of novel promising applications. EXPERT OPINION Improved understanding of LSC identity and development of LSC expansion methods will influence the evolution of successful LSCD therapies. Ultimately, future controlled clinical studies enabling direct comparison of the diverse employed approaches will help to identify the most effective treatment strategies.
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Affiliation(s)
- Yuzuru Sasamoto
- Division of Genetics, Brigham & Women’s Hospital, Harvard Medical School, Boston, MA, USA
| | - Bruce R. Ksander
- Mass Eye & Ear, Schepens Eye Research Institute, Harvard Medical School, Boston, MA
| | - Markus H. Frank
- Transplant Research Program, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA
- Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA
- Western School of Medical Sciences, Edith Cowan University, Joondalup, WA, Australia
| | - Natasha Y. Frank
- Division of Genetics, Brigham & Women’s Hospital, Harvard Medical School, Boston, MA, USA
- Department of Medicine, VA Boston Healthcare System, Boston, MA, USA
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Jie J, Yang J, He H, Zheng J, Wang W, Zhang L, Li Z, Chen J, Vimalin Jeyalatha M, Dong N, Wu H, Liu Z, Li W. Tissue remodeling after ocular surface reconstruction with denuded amniotic membrane. Sci Rep 2018; 8:6400. [PMID: 29686390 PMCID: PMC5913251 DOI: 10.1038/s41598-018-24694-4] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/15/2018] [Accepted: 04/05/2018] [Indexed: 12/16/2022] Open
Abstract
Amniotic membrane (AM) has been widely used as a temporary or permanent graft in the treatment of various ocular surface diseases. In this study, we compared the epithelial wound healing and tissue remodeling after ocular surface reconstruction with intact amniotic membrane (iAM) or denuded amniotic membrane (dAM). Partial limbal and bulbar conjunctival removal was performed on New Zealand rabbits followed by transplantation of cryo-preserved human iAM or dAM. In vivo observation showed that the epithelial ingrowth was faster on dAM compared to iAM after AM transplantation. Histological observation showed prominent epithelial stratification and increased goblet cell number on dAM after 2 weeks of follow up. Collagen VII degraded in dAM within 2 weeks, while remained in iAM even after 3 weeks. The number of macrophages and α-SMA positive cells in the stroma of remodelized conjunctiva in the dAM transplantation group was considerably less. In conclusion, dAM facilitates epithelial repopulation and goblet cell differentiation, further reduces inflammation and scar formation during conjunctival and corneal limbal reconstruction.
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Affiliation(s)
- Jing Jie
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
- Guilin Women and Children's Hospital, Guilin, Guangxi, China
| | - Jie Yang
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
- Zhengzhou Second Hospital, Zhengzhou, Henan, China
| | - Hui He
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
| | - Jianlan Zheng
- Xiamen University affiliated Chenggong Hospital, Xiamen, Fujian, China
| | - Wenyan Wang
- Xiamen University affiliated Chenggong Hospital, Xiamen, Fujian, China
| | - Liying Zhang
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
- Xiamen University affiliated Xiamen Eye Center, Xiamen, Fujian, China
| | - Zhiyuan Li
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
| | - Jingyao Chen
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
| | - M Vimalin Jeyalatha
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
| | - Nuo Dong
- Xiamen University affiliated Xiamen Eye Center, Xiamen, Fujian, China
| | - Huping Wu
- Xiamen University affiliated Xiamen Eye Center, Xiamen, Fujian, China
| | - Zuguo Liu
- Eye Institute of Xiamen University, Xiamen, Fujian, China
- Medical College of Xiamen University, Xiamen, Fujian, China
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China
- Xiamen University affiliated Xiamen Eye Center, Xiamen, Fujian, China
| | - Wei Li
- Eye Institute of Xiamen University, Xiamen, Fujian, China.
- Medical College of Xiamen University, Xiamen, Fujian, China.
- Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, Xiamen, Fujian, China.
- The Affiliated Xiang'an Hospital of Xiamen University, Xiamen, Fujian, China.
- Xiamen University affiliated Xiamen Eye Center, Xiamen, Fujian, China.
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Feeder Cells Free Rabbit Oral Mucosa Epithelial Cell Sheet Engineering. Tissue Eng Regen Med 2018; 15:321-332. [PMID: 30603557 DOI: 10.1007/s13770-017-0108-4] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/06/2017] [Revised: 12/11/2017] [Accepted: 12/15/2017] [Indexed: 12/24/2022] Open
Abstract
The optimal cell culture method of autologous oral mucosal epithelial cell sheet is not well established for a safe transplantation on to the patients' ocular surface. Animal serum and 3T3 mouse feeder cells are currently being used to stimulate the growth of the epithelial cells. However, the use of animal compounds can have potential side effects for the patient after transplantation of the engineered cell sheet. In the present study, we focused on engineering a rabbit oral mucosal epithelial cell sheet without 3T3 mouse feeder cells using a mix of Dulbecco's Modified Eagle Medium/Bronchial Epithelial Cell Growth Medium culture media (DMEM/BEGM). Autologous oral mucosal epithelial cell sheets, engineered with DMEM/BEGM feeder cell free culture media, were compared to those cultured in presence of serum and feeder cells. Using a DMEM/BEGM mix culture media, feeder cell free culture condition, autologous oral mucosal epithelial cells reached confluence and formed a multilayered sheet. The phenotype of engineered cell sheets cultured with DMEM/BEGM were characterized and compared to those cultured with serum and feeder. Hematoxylin and eosin staining showed the formation of a similar stratified multilayer cell sheets, in both culture conditions. The expression of deltaN-p63, ABCG2, PCNA, E-cadherin, Beta-catenin, CK3, CK4, CK13, Muc5AC, was similar in both culture conditions. We demonstrated that rabbit autologous oral mucosal epithelial cell sheet can be engineered, in feeder cell free conditions. The use of the DMEM/BEGM culture media to engineer culture autologous oral mucosa epithelial cell sheet will help to identify key factors involved in the growth and differentiation of oral mucosal epithelial cells.
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