Ripasudil zur antiangiogenen Therapie bei Keratoplastik

2021 ◽  
pp. 1-2
Author(s):  
Björn Bachmann
Keyword(s):  
Graft Survival ◽  
Immune Cell ◽  
Kinase Inhibitor ◽  
Healing Process ◽  
Corneal Transplantation ◽  
Inflammatory Factors ◽  
Wound Healing Process ◽  
Mrna Levels ◽  
Allograft Survival ◽  
Corneal Allograft

Corneal allograft survival is mediated by the variety of immunological reactions and wound healing process. Our aim was to explore the effects of topical administration of ripasudil, a selective Rho-associated coiled-coil protein kinase inhibitor, on corneal allograft survival. Ripasudil was administered to mice thrice a day after allogeneic corneal transplantation. Corneal graft survival, opacity, neovascularization, re-epithelization, immune cell infiltration, and mRNA levels of angiogenic and pro-inflammatory factors in the grafted cornea and draining lymph nodes (dLNs) were evaluated with slit-lamp microscopy, immunohistochemistry, flow cytometry, and polymerase chain reaction. Graft survival was significantly prolonged with lower graft opacity and neovascularization scores in 0.4% and 2.0% ripasudil-treated groups, and mRNA levels of angiogenic and pro-inflammatory factors in ripasudil-treated grafted corneas were reduced. Moreover, 0.4% and 2.0% ripasudil reduced CD45<sup>+</sup>-infiltrated leukocyte frequency, <i>Cd11b</i> and <i>Cd11c</i> mRNA levels, and the frequencies of mature dendritic cells, IFNγ-, and IL-17- producing CD4<sup>+</sup>T cells in the dLNs of recipients. Re-epithelization rate of the grafted cornea was significantly higher in the 0.4% and 2.0% ripasudil groups than in the control. Topically applied ripasudil prolonged graft survival by downregulating neovascularization and inflammation factors, while promoting corneal re-epithelization, suggesting that ripasudil may be useful for suppressing immunological rejection in corneal transplantation.

scholarly journals Novel immunotherapeutic effects of topically administered ripasudil (K-115) on corneal allograft survival

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Takenori Inomata ◽  
Keiichi Fujimoto ◽  
Yuichi Okumura ◽  
Jun Zhu ◽  
Kenta Fujio ◽  
...  
Keyword(s):  
Graft Survival ◽  
Immune Cell ◽  
Kinase Inhibitor ◽  
Healing Process ◽  
Corneal Transplantation ◽  
Inflammatory Factors ◽  
Wound Healing Process ◽  
Mrna Levels ◽  
Allograft Survival ◽  
Corneal Allograft

AbstractCorneal allograft survival is mediated by the variety of immunological reactions and wound healing process. Our aim was to explore the effects of topical administration of ripasudil, a selective Rho-associated coiled-coil protein kinase inhibitor, on corneal allograft survival. Ripasudil was administered to mice thrice a day after allogeneic corneal transplantation. Corneal graft survival, opacity, neovascularization, re-epithelization, immune cell infiltration, and mRNA levels of angiogenic and pro-inflammatory factors in the grafted cornea and draining lymph nodes (dLNs) were evaluated with slit-lamp microscopy, immunohistochemistry, flow cytometry, and polymerase chain reaction. Graft survival was significantly prolonged with lower graft opacity and neovascularization scores in 0.4% and 2.0% ripasudil-treated groups, and mRNA levels of angiogenic and pro-inflammatory factors in ripasudil-treated grafted corneas were reduced. Moreover, 0.4% and 2.0% ripasudil reduced CD45+-infiltrated leukocyte frequency, Cd11b and Cd11c mRNA levels, and the frequencies of mature dendritic cells, IFNγ-, and IL-17- producing CD4+T cells in the dLNs of recipients. Re-epithelization rate of the grafted cornea was significantly higher in the 0.4% and 2.0% ripasudil groups than in the control. Topically applied ripasudil prolonged graft survival by downregulating neovascularization and inflammation factors, while promoting corneal re-epithelization, suggesting that ripasudil may be useful for suppressing immunological rejection in corneal transplantation.

scholarly journals Subconjunctival administration of low-dose murine allogeneic mesenchymal stromal cells promotes corneal allograft survival in mice

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Oliver Treacy ◽  
Kevin Lynch ◽  
Nick Murphy ◽  
Xizhe Chen ◽  
Ellen Donohoe ◽  
...  
Keyword(s):  
Graft Survival ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Low Dose ◽  
Corneal Transplantation ◽  
Corneal Transplant ◽  
Subconjunctival Injection ◽  
Allograft Survival ◽  
Corneal Allograft ◽  
Allogeneic Mscs

Abstract Background Systemic administration of mesenchymal stromal cells (MSCs) has been efficacious in many inflammatory disease settings; however, little data are available on the potential immunomodulatory effects following local MSC administration in the context of corneal transplantation. The purpose of this study was to assess the potential of subconjunctival injection of MSCs to promote corneal allograft survival. Methods MSCs were isolated from female C57BL/6 (H-2k) or Balb/c (H-2d) mice and extensively characterized. An allogeneic mouse corneal transplant model was used with Balb/c mice as recipients of C57BL/6 grafts. A dose-finding study starting with 5 × 105 MSCs injected subconjunctivally at day − 7 was tested first followed by a more clinically translatable low-dose single or dual injection strategy on day − 1 and day + 1 before/after transplantation. Graft transparency served as the primary indicator of transplant rejection while neovascularization was also recorded. Lymphocytes (from draining lymph nodes) and splenocytes were isolated from treatment groups on day 2 post-transplantation and characterized by flow cytometry and qRT-PCR. Results Both high- and low-dose injection of allogeneic MSCs on day − 7 led to 100% graft survival over the observation period. Moreover, low-dose dual subconjunctival injection of 5 × 104 allogeneic MSCs on day − 1 or day + 1 led to 100% allograft survival in transplant recipients (n = 7). We also demonstrate that single administration of allogeneic MSCs on either day − 1 or day + 1 promotes rejection-free graft survival in 100% (n = 8) and 86% (n = 7) of transplanted mice, respectively. Early time point ex vivo analysis suggests modulation of innate immune responses towards anti-inflammatory, pro-repair responses by local MSC administration. Conclusion This work demonstrates that low-dose subconjunctival injection of allogeneic MSCs successfully promotes corneal allograft survival and may contribute to refining future MSC immunotherapies for prevention of corneal allograft rejection.

Biological activities of novel lipid mediator sphingosine 1-phosphate in rat hepatic stellate cells

2000 ◽  
Vol 279 (2) ◽  
pp. G304-G310 ◽  
Author(s):  
Hitoshi Ikeda ◽  
Yutaka Yatomi ◽  
Mikio Yanase ◽  
Hiroaki Satoh ◽  
Hisato Maekawa ◽  
...  
Keyword(s):  
Wound Healing ◽  
Hepatic Stellate Cells ◽  
Biological Activities ◽  
Healing Process ◽  
Stellate Cells ◽  
Mitogen Activated Protein Kinase ◽  
Lipid Mediator ◽  
Wound Healing Process ◽  
Mrna Levels ◽  
Sphingosine 1 Phosphate

Sphingosine 1-phosphate (S-1-P), a lipid mediator shown to be a ligand for G protein-coupled receptors (AGRs), endothelial differentiation gene (EDG)1, EDG3, and AGR16/EDG5, is stored in platelets and released on their activation. Platelet consumption occurs in acute liver injury. Hepatic stellate cells (HSCs) play an important role in wound healing. Effects of S-1-P on HSCs were investigated. S-1-P enhanced proliferation of culture-activated HSCs. The mitogenic effect was pertussis toxin sensitive, mitogen-activated protein kinase dependent, and more prominent at lower cell density. S-1-P increased contraction of collagen lattices containing HSCs, irrespective of activation state, in a C3 exotoxin-sensitive manner. mRNAs of EDG1 and AGR16, but not of EDG3, were detected in HSCs. In HSC activation, EDG1 mRNA levels were downregulated, whereas AGR16 mRNA levels were unchanged. Considering that HSCs are capable of production of extracellular matrices and modulation of blood flow in sinusoids, our results suggest that S-1-P may play a role in wound healing process in the liver.

scholarly journals Immunology of Acute and Chronic Wound Healing

Biomolecules ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 700
Author(s):  
Kamila Raziyeva ◽  
Yevgeniy Kim ◽  
Zharylkasyn Zharkinbekov ◽  
Kuat Kassymbek ◽  
Shiro Jimi ◽  
...  
Keyword(s):  
Wound Healing ◽  
Chronic Wounds ◽  
Chronic Wound ◽  
Healing Process ◽  
Inflammatory Factors ◽  
Wound Healing Process ◽  
Macrophage Phenotype ◽  
Microbial Biofilms ◽  
Inflammatory Phase ◽  
Persistent Inflammation

Skin wounds greatly affect the global healthcare system, creating a substantial burden on the economy and society. Moreover, the situation is exacerbated by low healing rates, which in fact are overestimated in reports. Cutaneous wounds are generally classified into acute and chronic. The immune response plays an important role during acute wound healing. The activation of immune cells and factors initiate the inflammatory process, facilitate wound cleansing and promote subsequent tissue healing. However, dysregulation of the immune system during the wound healing process leads to persistent inflammation and delayed healing, which ultimately result in chronic wounds. The microenvironment of a chronic wound is characterized by high quantities of pro-inflammatory macrophages, overexpression of inflammatory mediators such as TNF-α and IL-1β, increased activity of matrix metalloproteinases and abundance of reactive oxygen species. Moreover, chronic wounds are frequently complicated by bacterial biofilms, which perpetuate the inflammatory phase. Continuous inflammation and microbial biofilms make it very difficult for the chronic wounds to heal. In this review, we discuss the role of innate and adaptive immunity in the pathogenesis of acute and chronic wounds. Furthermore, we review the latest immunomodulatory therapeutic strategies, including modifying macrophage phenotype, regulating miRNA expression and targeting pro- and anti-inflammatory factors to improve wound healing.

Denatured Collagen Could Increase the Autophagy Level and Inhibit Apoptosis of Fibroblasts to Help Cell Survival and Influence Wound Healing

2020 ◽  
pp. 153473462092594
Author(s):  
Yidan Su ◽  
Min Li ◽  
Xiqiao Wang ◽  
Zhiyong Wang ◽  
Lei Yi
Keyword(s):  
Wound Healing ◽  
Cell Survival ◽  
Quantitative Polymerase Chain Reaction ◽  
Healing Process ◽  
Biological Behavior ◽  
Wound Healing Process ◽  
Mrna Levels ◽  
Promote Cell Survival ◽  
Vitro Model

When exposed to thermal factors, collagen in the dermis denatures, which could affect the biological behavior of cells. Previous studies have demonstrated that denatured collagen could influence the activity of fibroblasts and induce fibroblasts differentiate into myofibroblasts. However, information on the regulation of fibroblasts by denatured collagen-modulated autophagy and apoptosis during the wound healing process is limited. In this article, we researched the effect of denatured collagen-modulated autophagy and apoptosis on fibroblasts. An in vitro model comprising fibroblasts and denatured collagen was established to identify the potential ability of denatured collagen to modulate autophagy and apoptosis. Western blotting, quantitative polymerase chain reaction, transmission electron microscopy, TUNEL assay, and immunofluorescence staining were used to examine the changes induced by denatured collage. Protein and mRNA levels of LC3 and beclin-1 were significantly increased after stimulated by denatured collagen, while those of caspase-3 were reduced. Unlike stimulation with normal collagen, denatured collagen enhanced autophagy and inhibited apoptosis of fibroblasts. After blocking autophagy using 3-methyladenine, the apoptotic function was increased. Denatured collagen could increase autophagy and inhibit apoptosis of the fibroblasts to promote cell survival and influence wound healing.

scholarly journals The Balance of Th1/Th2 and LAP+Tregs/Th17 Cells Is Crucial for Graft Survival in Allogeneic Corneal Transplantation

2018 ◽  
Vol 2018 ◽  
pp. 1-10 ◽  
Author(s):  
Shang Li ◽  
Jing Yu ◽  
Chungang Guo ◽  
Ying Jie ◽  
Zhiqiang Pan
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Aqueous Humor ◽  
Allograft Rejection ◽  
Corneal Transplantation ◽  
Allograft Survival ◽  
Survival Group ◽  
Corneal Allograft ◽  
Group 3 ◽  
Corneal Allograft Rejection

Purpose. CD4+LAP+ T cells are newly discovered regulatory T cells (Tregs). The aim of this study is to investigate the balance of Th1/Th2 and LAP+Tregs/Th17 in mice after allogeneic corneal transplantation. Methods. A total of 65 mice received orthotopic penetrating transplantation. According to the survival scores of the grafts, the mice were divided into the rejection group and the survival group 3 weeks after transplantation. Th1, Th2, Th17, and regulatory T cells in the ipsilateral drainage lymph nodes and spleens were measured with flow cytometry. The related cytokines in aqueous humor were also analyzed. Results. The frequencies of Foxp3+Tregs, GARP+Tregs, and LAP+Tregs in the survival group were significantly higher than those in the rejection group. And the expression trend of CD4+LAP+ T cells and CD4+GARP+ T cells was consistent. The level of IFN-γ, TNF, IL-6, and IL-17A markedly increased in aqueous humor during corneal allograft rejection. The ratio of Th1/Th2 and Th17/LAP+Tregs significantly increased in the rejection group at the 3rd week after corneal transplantation. Conclusion. LAP+Tregs might be regarded as substitute for Foxp3+Tregs. The balance of Th1/Th2 and LAP+Tregs/Th17 is crucial for corneal allograft survival.

scholarly journals hucMSC Exosome Ameliorates Pressure Ulcers Through Inhibition of HMGB1.

2021 ◽  
Author(s):  
Fei Yan ◽  
Meihua Gong ◽  
Furong Li ◽  
Li Yu
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Pressure Ulcers ◽  
Healing Process ◽  
Wound Healing Process ◽  
Human Umbilical Cord ◽  
Mrna Levels ◽  
Tissue Samples ◽  
New Perspective

Abstract Background: Pressure ulcers (PU) are a chronic wound for elderly populations. Previous works have shown that exosomes from stem cells contain cytokines and growth factors that play a role in tissue repair and can represent a therapeutic strategy for wound healing. Thus, as a new cell-free treatment model, fully understanding the extraction of exosomes and its mechanism of action can help promote the management of clinically chronic refractory wound healing. Material and Methods: In this study, we initially isolated exosomes from human umbilical cord mesenchymal stem cells (hucMSC-Exo) and examined their roles in wound healing. Different time points were evaluated for 15 mice which were randomly divided into three groups to serve three I-R circles and took different dose of hucMSC-Exo. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to analyze collagen mRNA levels in tissue samples. HMGB1 content was explored by western blot and immunohistochemistry. Comparing α-SAM, CD34, HMGB1 were used to investigate the potential mechanisms.Results: We found that hucMSC-Exo could be taken up by fibroblasts and significantly regulate and improve fibroblast fibrotic status and in-vivo PU wound healing. Further, we observed that hucMSC-Exo treatment of PU wound was able to downregulate the expression of HMGB1 previously shown to have a deleterious role in the wound healing process. Conclusion: Our study indicates that hucMSC-Exo regulates the repair of pressure ulcer wounds in part by inhibiting HMGB1. Exosome treatment has opened up a new perspective in regenerative medicine and trauma management.

scholarly journals Von Hippel-Lindau (VHL) Protein Antagonist VH298 Improves Wound Healing in Streptozotocin-Induced Hyperglycaemic Rats by Activating Hypoxia-Inducible Factor- (HIF-) 1 Signalling

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Shuo Qiu ◽  
Yachao Jia ◽  
Yunchu Sun ◽  
Pei Han ◽  
Jia Xu ◽  
...  
Keyword(s):  
Wound Healing ◽  
Growth Factor ◽  
Rat Model ◽  
Healing Process ◽  
Wound Healing Process ◽  
Type I ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Vascular Endothelial ◽  
Vhl Protein

Aims. The purpose of the present research is to investigate the effects of the VHL protein antagonist, VH298, on functional activities of fibroblasts and vascular endothelial cells and the effects on the wound healing process in a streptozotocin-induced hyperglycaemic rat model. Methods. HIF-1α and hydroxy-HIF-1α protein levels in VH298-treated rat fibroblasts (rFb) were measured by immunoblotting, rFb proliferation was detected by the CCK-8 assay, and mRNA levels of related genes were measured by quantitative RT-PCR. In vitro wound healing was simulated by the scratch test; angiogenesis was measured by the human umbilical vein endothelial cell (hUVEC) tube formation assay. VH298 or PBS was locally injected into wounds in rat models with streptozotocin- (STZ-) induced hyperglycaemia, the wound tissues were harvested, and haematoxylin-eosin (HE) and Masson trichrome staining and immunohistochemical processes were conducted. Results. HIF-1α and hydroxy-HIF-1α levels increased in VH298-treated rFb, in a time- and dose-dependent manner. Thirty micromolar VH298 could significantly increase cell proliferation, angiogenesis, and gene expression of type I collagen-α1 (Col1-α1), vascular endothelial growth factor A (VEGF-A), and insulin-like growth factor 1 (IGF-1). The VH298-treated wound had a better healing pattern, activation of HIF-1 signalling, and vascularization. Conclusions. Taken together, VH298 activated the HIF-1 signalling pathway by stabilizing both HIF-1α and hydroxy-HIF-1α. VH298 enhanced rFb functions, promoted hUVEC angiogenesis, and accelerated wound healing in the rat model mimicking diabetes mellitus.

scholarly journals Immunomodulatory Strategies Targeting Dendritic Cells to Improve Corneal Graft Survival

2020 ◽  
Vol 9 (5) ◽  
pp. 1280
Author(s):  
Alfrun Schönberg ◽  
Matthias Hamdorf ◽  
Felix Bock
Keyword(s):  
Dendritic Cells ◽  
Risk Factor ◽  
Common Sense ◽  
Gold Standard ◽  
Corneal Transplantation ◽  
Lymphatic Vessels ◽  
Therapeutic Strategies ◽  
Allograft Survival ◽  
Corneal Allograft ◽  
Vessel Regression

Even though the cornea is regarded as an immune-privileged tissue, transplantation always comes with the risk of rejection due to mismatches between donor and recipient. It is common sense that an alternative to corticosteroids as the current gold standard for treatment of corneal transplantation is needed. Since blood and lymphatic vessels have been identified as a severe risk factor for corneal allograft survival, much research has focused on vessel regression or inhibition of hem- and lymphangiogenesis in general. However, lymphatic vessels have been identified as required for the inflammation’s resolution. Therefore, targeting other players of corneal engraftment could reveal new therapeutic strategies. The establishment of a tolerogenic microenvironment at the graft site would leave the recipient with the ability to manage pathogenic conditions independent from transplantation. Dendritic cells (DCs) as the central player of the immune system represent a target that allows the induction of tolerogenic mechanisms by many different strategies. These strategies are reviewed in this article with regard to their success in corneal transplantation.

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