The Experts below are selected from a list of 318 Experts worldwide ranked by ideXlab platform
Sylvain L. Guérin - One of the best experts on this subject based on the ideXlab platform.
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contribution of the wnk1 kinase to Corneal Wound Healing using the tissue engineered human cornea as an in vitro model
Journal of Tissue Engineering and Regenerative Medicine, 2019Co-Authors: Pascale Desjardins, Lucie Germain, Camille Couture, Sylvain L. GuérinAbstract:Damage to the Corneal epithelium triggers important changes in the extracellular matrix (ECM) to which basal human Corneal epithelial cells (hCECs) attach. These changes are perceived by integrin receptors that activate different intracellular signalling pathways, ultimately leading to re-epithelialization of the injured epithelium. In this study, we investigated the impact of pharmacological inhibition of specific signal transduction mediators on Corneal Wound Healing using both monolayers of hCECs and the human tissue-engineered cornea (hTEC) as an in vitro 3D model. RNA and proteins were isolated from the Wounded and unWounded hTECs to conduct gene profiling analyses and protein kinase arrays. The impact of WNK1 inhibition was evaluated on the Wounded hTECs as well as on hCECs monolayers using a scratch Wound assay. Gene profiling and protein kinase arrays revealed that expression and activity of several mediators from the integrin-dependent signaling pathways were altered in response to the ECM changes occurring during Corneal Wound Healing. Phosphorylation of the WNK1 kinase turned out to be the most striking activation event going on during this process. The inhibition of WNK1 by WNK463 reduced the rate of Corneal Wound closure in both the hTEC and hCECs grown in monolayer compared with their respective negative controls. WNK463 also reduced phosphorylation of the WNK1 downstream targets SPAK/OSR1 in Wounded hTECs. These in vitro results allowed for a better understanding of the cellular and molecular mechanisms involved in Corneal Wound Healing and identified WNK1 as a kinase important to ensure proper Wound Healing of the cornea.
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Contribution of the WNK1 kinase to Corneal Wound Healing using the tissue‐engineered human cornea as an in vitro model
Journal of tissue engineering and regenerative medicine, 2019Co-Authors: Pascale Desjardins, Lucie Germain, Camille Couture, Sylvain L. GuérinAbstract:Damage to the Corneal epithelium triggers important changes in the extracellular matrix (ECM) to which basal human Corneal epithelial cells (hCECs) attach. These changes are perceived by integrin receptors that activate different intracellular signalling pathways, ultimately leading to re-epithelialization of the injured epithelium. In this study, we investigated the impact of pharmacological inhibition of specific signal transduction mediators on Corneal Wound Healing using both monolayers of hCECs and the human tissue-engineered cornea (hTEC) as an in vitro 3D model. RNA and proteins were isolated from the Wounded and unWounded hTECs to conduct gene profiling analyses and protein kinase arrays. The impact of WNK1 inhibition was evaluated on the Wounded hTECs as well as on hCECs monolayers using a scratch Wound assay. Gene profiling and protein kinase arrays revealed that expression and activity of several mediators from the integrin-dependent signaling pathways were altered in response to the ECM changes occurring during Corneal Wound Healing. Phosphorylation of the WNK1 kinase turned out to be the most striking activation event going on during this process. The inhibition of WNK1 by WNK463 reduced the rate of Corneal Wound closure in both the hTEC and hCECs grown in monolayer compared with their respective negative controls. WNK463 also reduced phosphorylation of the WNK1 downstream targets SPAK/OSR1 in Wounded hTECs. These in vitro results allowed for a better understanding of the cellular and molecular mechanisms involved in Corneal Wound Healing and identified WNK1 as a kinase important to ensure proper Wound Healing of the cornea.
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A tissue-engineered Corneal Wound Healing model for the characterization of reepithelialization.
Methods in molecular biology (Clifton N.J.), 2013Co-Authors: Karine Zaniolo, Patrick Carrier, Sylvain L. Guérin, François A. Auger, Lucie GermainAbstract:Progress in tissue engineering has led to the discovery of technologies allowing reconstruction of autologous tissues from the patient's own cells and the development of new in vitro models to study cellular and molecular mechanisms implicated in Wound Healing. The outer surface of the eye, the cornea, is involved in the sense of sight, thus an adequate reepithelialization process after Wounding is essential in order to maintain Corneal function. In this chapter, protocols to generate a new in vitro three-dimensional human Corneal Wound Healing model suitable for studying the different components that play important roles in Corneal reepithelialization are described in details. The methods include extraction and culture of human Corneal epithelial cells (HCECs), human Corneal fibroblasts, a complete description of the cornea reconstructed by tissue-engineering as well as the Corneal Wound Healing model.
Shizuya Saika - One of the best experts on this subject based on the ideXlab platform.
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TGF-β Signal Transduction in Corneal Wound Healing as a Therapeutic Target
Cornea, 2004Co-Authors: Shizuya SaikaAbstract:AbstractThis article reviews recent progress in research on the role of Smad signaling in Corneal Wound Healing. Smad2 and Smad3 are key signaling molecules downstream of the cell surface receptor of transforming growth factor-β (TGF-β) or activin. On ligand binding to the receptor, Smads2/3 undergo
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TGF-beta signal transduction in Corneal Wound Healing as a therapeutic target.
Cornea, 2004Co-Authors: Shizuya SaikaAbstract:This article reviews recent progress in research on the role of Smad signaling in Corneal Wound Healing. Smad2 and Smad3 are key signaling molecules downstream of the cell surface receptor of transforming growth factor-beta (TGF-beta) or activin. On ligand binding to the receptor, Smads2/3 undergo phosphorylation, form complexes with Smad4, and thence convey signaling. TGF-beta isoforms have been detected in Corneal epithelium and are also deposited in Wounded stroma, suggesting their participation in the Wound-Healing process in Corneal tissue. Human or mouse uninjured healthy Corneal epithelium shows nuclear accumulation of Smads3/4, indicating active Smad signaling in this tissue. Migrating Corneal epithelium lacks nuclear Smad accumulation with up-regulation of Smad7, but p38MAPK is activated. Organ-culture experiments show that p38MAPK activation depends on endogenous TGF-beta and that activation of p38MAPK results in cell proliferation cessation with a reduction of Erk activation and acceleration of cell migration in Healing Corneal epithelium. These findings indicate that during Healing of Corneal epithelial defects, endogenous TGF-beta activates p38MAPK for cell migration and suppression of cell proliferation and up-regulates Smad7 for inhibition of Smad2 and Smad3 signaling, resulting in rapid initial resurfacing of the epithelium. Such involvement of p38MAPK in cell migration has been reported in many cell types and observed in keratocyte culture. Possible benefits of preserving non-Smad cascades in treating problems in Corneal Wound Healing by manipulating TGF-beta signals have been suggested.
Lucie Germain - One of the best experts on this subject based on the ideXlab platform.
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contribution of the wnk1 kinase to Corneal Wound Healing using the tissue engineered human cornea as an in vitro model
Journal of Tissue Engineering and Regenerative Medicine, 2019Co-Authors: Pascale Desjardins, Lucie Germain, Camille Couture, Sylvain L. GuérinAbstract:Damage to the Corneal epithelium triggers important changes in the extracellular matrix (ECM) to which basal human Corneal epithelial cells (hCECs) attach. These changes are perceived by integrin receptors that activate different intracellular signalling pathways, ultimately leading to re-epithelialization of the injured epithelium. In this study, we investigated the impact of pharmacological inhibition of specific signal transduction mediators on Corneal Wound Healing using both monolayers of hCECs and the human tissue-engineered cornea (hTEC) as an in vitro 3D model. RNA and proteins were isolated from the Wounded and unWounded hTECs to conduct gene profiling analyses and protein kinase arrays. The impact of WNK1 inhibition was evaluated on the Wounded hTECs as well as on hCECs monolayers using a scratch Wound assay. Gene profiling and protein kinase arrays revealed that expression and activity of several mediators from the integrin-dependent signaling pathways were altered in response to the ECM changes occurring during Corneal Wound Healing. Phosphorylation of the WNK1 kinase turned out to be the most striking activation event going on during this process. The inhibition of WNK1 by WNK463 reduced the rate of Corneal Wound closure in both the hTEC and hCECs grown in monolayer compared with their respective negative controls. WNK463 also reduced phosphorylation of the WNK1 downstream targets SPAK/OSR1 in Wounded hTECs. These in vitro results allowed for a better understanding of the cellular and molecular mechanisms involved in Corneal Wound Healing and identified WNK1 as a kinase important to ensure proper Wound Healing of the cornea.
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Contribution of the WNK1 kinase to Corneal Wound Healing using the tissue‐engineered human cornea as an in vitro model
Journal of tissue engineering and regenerative medicine, 2019Co-Authors: Pascale Desjardins, Lucie Germain, Camille Couture, Sylvain L. GuérinAbstract:Damage to the Corneal epithelium triggers important changes in the extracellular matrix (ECM) to which basal human Corneal epithelial cells (hCECs) attach. These changes are perceived by integrin receptors that activate different intracellular signalling pathways, ultimately leading to re-epithelialization of the injured epithelium. In this study, we investigated the impact of pharmacological inhibition of specific signal transduction mediators on Corneal Wound Healing using both monolayers of hCECs and the human tissue-engineered cornea (hTEC) as an in vitro 3D model. RNA and proteins were isolated from the Wounded and unWounded hTECs to conduct gene profiling analyses and protein kinase arrays. The impact of WNK1 inhibition was evaluated on the Wounded hTECs as well as on hCECs monolayers using a scratch Wound assay. Gene profiling and protein kinase arrays revealed that expression and activity of several mediators from the integrin-dependent signaling pathways were altered in response to the ECM changes occurring during Corneal Wound Healing. Phosphorylation of the WNK1 kinase turned out to be the most striking activation event going on during this process. The inhibition of WNK1 by WNK463 reduced the rate of Corneal Wound closure in both the hTEC and hCECs grown in monolayer compared with their respective negative controls. WNK463 also reduced phosphorylation of the WNK1 downstream targets SPAK/OSR1 in Wounded hTECs. These in vitro results allowed for a better understanding of the cellular and molecular mechanisms involved in Corneal Wound Healing and identified WNK1 as a kinase important to ensure proper Wound Healing of the cornea.
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A tissue-engineered Corneal Wound Healing model for the characterization of reepithelialization.
Methods in molecular biology (Clifton N.J.), 2013Co-Authors: Karine Zaniolo, Patrick Carrier, Sylvain L. Guérin, François A. Auger, Lucie GermainAbstract:Progress in tissue engineering has led to the discovery of technologies allowing reconstruction of autologous tissues from the patient's own cells and the development of new in vitro models to study cellular and molecular mechanisms implicated in Wound Healing. The outer surface of the eye, the cornea, is involved in the sense of sight, thus an adequate reepithelialization process after Wounding is essential in order to maintain Corneal function. In this chapter, protocols to generate a new in vitro three-dimensional human Corneal Wound Healing model suitable for studying the different components that play important roles in Corneal reepithelialization are described in details. The methods include extraction and culture of human Corneal epithelial cells (HCECs), human Corneal fibroblasts, a complete description of the cornea reconstructed by tissue-engineering as well as the Corneal Wound Healing model.
Choun-ki Joo - One of the best experts on this subject based on the ideXlab platform.
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Wakayama Symposium: New Therapies for Modulation of Epithelialization in Corneal Wound Healing
The ocular surface, 2012Co-Authors: Jun-sub Choi, Choun-ki JooAbstract:Many factors are involved in the Corneal Wound Healing mechanism, including adhesion, migration, and proliferation of Corneal epithelial cells. Abnormal Corneal Wound Healing leads to Corneal edema, neovascularization, scar formation, and poor vision. Three agents, 17β-estradiol, nicergoline, and β-glucan, have demonstrated positive effects on the Wound Healing response in laboratory experiments and may be of help in controlling Wound Healing in corneas that have suffered epithelial damage or have undergone refractive surgery.
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Expression of Wnt and MMP in epithelial cells during Corneal Wound Healing.
Cornea, 2006Co-Authors: Jungmook Lyu, Choun-ki JooAbstract:Purpose To study the expression of Wnt during Corneal Wound Healing and to understand the signaling mechanism involved. Methods Rat cornea was demarcated on the central area by 4-mm trepine, and the epithelium within this area was removed by scalpel. The epithelium was scraped and isolated to extract RNA. To determine the proliferation of Corneal epithelial cells, corneoscleral rims from human donors were treated with dispase II for 15 minutes, and epithelial cells were isolated. Cells were plated on a 3T3 feeder cells layer. The proliferation of Corneal epithelial cells was evaluated by colony-forming efficiency. Results Wnt 5b and 7a were rapidly induced in Wounded cornea, and Wnt 7a promoted proliferation of Corneal epithelial cells. In addition, matrix metalloproteinase (MMP)-12 was expressed in Wounded rat Corneal epithelium. Transcription of MMP-12 was responsive to Wnt/beta-catenin signaling. Function blockade of MMP-12 delayed Wnt 7a-induced cell proliferation. Conclusion These results indicate that Wnt proteins and MMP-12 regulate the proliferation of Corneal epithelial cells and that Wnt signaling contributes to the resurfacing of defective areas during Corneal Wound Healing.
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Expression of Wnt and MMP in epithelial cells during Corneal Wound Healing.
Cornea, 2006Co-Authors: Jungmook Lyu, Choun-ki JooAbstract:To study the expression of Wnt during Corneal Wound Healing and to understand the signaling mechanism involved. Rat cornea was demarcated on the central area by 4-mm trepine, and the epithelium within this area was removed by scalpel. The epithelium was scraped and isolated to extract RNA. To determine the proliferation of Corneal epithelial cells, corneoscleral rims from human donors were treated with dispase II for 15 minutes, and epithelial cells were isolated. Cells were plated on a 3T3 feeder cells layer. The proliferation of Corneal epithelial cells was evaluated by colony-forming efficiency. Wnt 5b and 7a were rapidly induced in Wounded cornea, and Wnt 7a promoted proliferation of Corneal epithelial cells. In addition, matrix metalloproteinase (MMP)-12 was expressed in Wounded rat Corneal epithelium. Transcription of MMP-12 was responsive to Wnt/beta-catenin signaling. Function blockade of MMP-12 delayed Wnt 7a-induced cell proliferation. These results indicate that Wnt proteins and MMP-12 regulate the proliferation of Corneal epithelial cells and that Wnt signaling contributes to the resurfacing of defective areas during Corneal Wound Healing.
Mehrnoosh Saghizadeh - One of the best experts on this subject based on the ideXlab platform.
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Concise Review: Stem Cells for Corneal Wound Healing.
Stem cells (Dayton Ohio), 2017Co-Authors: Mehrnoosh Saghizadeh, Andrei A. Kramerov, Clive N. Svendsen, Alexander V. LjubimovAbstract:Corneal Wound Healing is a complex process that occurs in response to various injuries and commonly used refractive surgery. It is a significant clinical problem, which may lead to serious complications due to either incomplete (epithelial) or excessive (stromal) Healing. Epithelial stem cells clearly play a role in this process, whereas the contribution of stromal and endothelial progenitors is less well studied. The available evidence on stem cell participation in Corneal Wound Healing is reviewed, together with the data on the use of Corneal and non-Corneal stem cells to facilitate this process in diseased or postsurgical conditions. Important aspects of Corneal stem cell generation from alternative cell sources, including pluripotent stem cells, for possible transplantation upon Corneal injuries or in disease conditions are also presented. Stem Cells 2017;35:2105-2114.
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Progress in Corneal Wound Healing
Progress in retinal and eye research, 2015Co-Authors: Alexander V. Ljubimov, Mehrnoosh SaghizadehAbstract:Corneal Wound Healing is a complex process involving cell death, migration, proliferation, differentiation, and extracellular matrix remodeling. Many similarities are observed in the Healing processes of Corneal epithelial, stromal and endothelial cells, as well as cell-specific differences. Corneal epithelial Healing largely depends on limbal stem cells and remodeling of the basement membrane. During stromal Healing, keratocytes get transformed to motile and contractile myofibroblasts largely due to activation of transforming growth factor-β (TGF-β) system. Endothelial cells heal mostly by migration and spreading, with cell proliferation playing a secondary role. In the last decade, many aspects of Wound Healing process in different parts of the cornea have been elucidated, and some new therapeutic approaches have emerged. The concept of limbal stem cells received rigorous experimental corroboration, with new markers uncovered and new treatment options including gene and microRNA therapy tested in experimental systems. Transplantation of limbal stem cell-enriched cultures for efficient re-epithelialization in stem cell deficiency and Corneal injuries has become reality in clinical setting. Mediators and course of events during stromal Healing have been detailed, and new treatment regimens including gene (decorin) and stem cell therapy for excessive Healing have been designed. This is a very important advance given the popularity of various refractive surgeries entailing stromal Wound Healing. Successful surgical ways of replacing the diseased endothelium have been clinically tested, and new approaches to accelerate endothelial Healing and suppress endothelial-mesenchymal transformation have been proposed including Rho kinase (ROCK) inhibitor eye drops and gene therapy to activate TGF-β inhibitor SMAD7. Promising new technologies with potential for Corneal Wound Healing manipulation including microRNA, induced pluripotent stem cells to generate Corneal epithelium, and nanocarriers for Corneal drug delivery are discussed. Attention is also paid to problems in Wound Healing understanding and treatment, such as lack of specific epithelial stem cell markers, reliable identification of stem cells, efficient prevention of haze and stromal scar formation, lack of data on Wound regulating microRNAs in keratocytes and endothelial cells, as well as virtual lack of targeted systems for drug and gene delivery to select Corneal cells.