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In vivo assay of BrdU incorporation during corneal epithelial wound healing. BrdU-positive corneal epithelial cells were counted in sections of WBB6F1+/+ and c-kit mutant corneas. Although the number of BrdU incorporated cells gradually increased after the injury, there were no significant differences between WBB6F1+/+ and c-kit mutant mice at each time point (unpaired t tests). +/+=WBB6F1+/+ mice, W/Wv=c-kit mutant mice. Error bars represent the standard errors (n=6).
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To study the roles played by stem cell factor (SCF) and SCF receptor c-kit in wound healing of corneal epithelial cells.
A 2 mm corneal epithelial wound was made in control (WBB6F1(+/+)), SCF (Sl/Sl(d))-, and c-kit (W/W(v)) mutant mice, and the speed of wound healing, 5-bromo-2'-deoxyuridine (BrdU) incorporation, and scanning electron microscopic (...
Citations
... SCF/C-Kit is another biomolecule that is present in normal mouse cornea and plays an important role in promoting corneal wound healing. According to Miyamoto et al. (2012) [89], SCF/C-kit enhanced cell attachment to FN, laminin, and collagen type IV during corneal wound healing via the induction of the avidity and affinity of integrin members in vitro. ...
... SCF/C-Kit is another biomolecule that is present in normal mouse cornea and plays an important role in promoting corneal wound healing. According to Miyamoto et al. (2012) [89], SCF/C-kit enhanced cell attachment to FN, laminin, and collagen type IV during corneal wound healing via the induction of the avidity and affinity of integrin members in vitro. ...
Over the last several decades, numerous modifications and advancements have been made to design the optimal corneal biomatrix for corneal epithelial cell (CECs) or limbal epithelial stem cell (LESC) carriers. However, researchers have yet to discover the ideal optimization strategies for corneal biomatrix design and its effects on cultured CECs or LESCs. This review discusses and summarizes recent optimization strategies for developing an ideal collagen biomatrix and its interactions with CECs and LESCs. Using PRISMA guidelines, articles published from June 2012 to June 2022 were systematically searched using Web of Science (WoS), Scopus, PubMed, Wiley, and EBSCOhost databases. The literature search identified 444 potential relevant published articles, with 29 relevant articles selected based on inclusion and exclusion criteria following screening and appraising processes. Physicochemical and biocompatibility (in vitro and in vivo) characterization methods are highlighted, which are inconsistent throughout various studies. Despite the variability in the methodology approach, it is postulated that the modification of the collagen biomatrix improves its mechanical and biocompatibility properties toward CECs and LESCs. All findings are discussed in this review, which provides a general view of recent trends in this field.
... While the pathways specific to DFUs may be compensatory, the absence of KIT and TGFb signaling in DFUs could contribute to defects in healing. Kit signaling promotes myocardial healing after infarction (Cimini et al., 2007), corneal wound repair (Miyamoto et al., 2012), and may improve healing in skin wounds (Zgheib et al., 2015). Furthermore, TGFb signaling contributes to tissue repair in a context and isoform specific manner (Divoux et al., 2010), and may impair fibroblast ECM regulation, driving fibrosis (Shi-wen et al., 2010) or generating aberrant fibroblast populations identified here. ...
Wound repair requires the coordination of multiple cell types including immune cells and tissue resident cells to coordinate healing and return of tissue function. Diabetic foot ulceration is a type of chronic wound that impacts over 4 million patients in the US and over 7 million worldwide (Edmonds et al., 2021). Yet, the cellular and molecular mechanisms that go awry in these wounds are not fully understood. Here, by profiling chronic foot ulcers from non-diabetic (NDFUs) and diabetic (DFUs) patients using single-cell RNA sequencing, we find that DFUs display transcription changes that implicate reduced keratinocyte differentiation, altered fibroblast function and lineages, and defects in macrophage metabolism, inflammation, and ECM production compared to NDFUs. Furthermore, analysis of cellular interactions reveals major alterations in several signaling pathways that are altered in DFUs. These data provide a view of the mechanisms by which diabetes alters healing of foot ulcers and may provide therapeutic avenues for DFU treatments.
... SCF/C-Kit are another biomolecule that is present in normal mouse cornea and plays an important role in promoting corneal wound healing. According to Miyamoto and colleagues [89], SCF/C-kit enhanced cell attachment to FN, laminin and collagen type IV during corneal wound healing via the induction of the avidity and affinity of integrin members in vitro. ...
Since the past few decades, numerous modifications and innovations have been done to design the optimal corneal biomatrix for corneal epithelial cells (CECs) or limbal epithelial stem cells (LESCs) carriers. However, researchers have yet to discover the ideal optimization strategies in the development of corneal biomatrix design and its effects on cultured CECs or LESCs. This review further discusses and summarizes recent optimization strategies to develop an ideal collagen biomatrix and its interaction with CECs and LESCs. Using PRISMA guidelines, the articles published from June 2012 to June 2022 were systematically searched using Web of Science (WoS), Scopus, PubMed, Wiley, and EBSCOhost databases. The literature search identified 444 potential relevant published articles, with 29 relevant articles selected based on the inclusion and exclusion criteria after the screening and appraising processes. The current paper highlights the physicochemical and biocompatibility (in vitro and in vivo) characterization methods, which were inconsistent throughout the different studies. Despite the variability in the methodology approach, the reviewer postulated that the modification of the collagen biomatrix improves its mechanical and biocompatibility properties toward CECs and LESCs. All findings were discussed in this review; thus, it provides a general view of up-to-date trends in this field.
... Kit is also expressed by corneal epithelial cells. Using an in vitro corneal epithelial adhesion assay, Miyamoto and colleagues suggested that the binding of stem cell factor (SCF) ligand to the epithelial c-kit receptor positively influences epithelial cell attachment [59]. Indeed, the SCF/c-kit system is known to be important for epithelial cell maintenance [60]. ...
Platelet extravasation during inflammation is under-appreciated. In wild-type (WT) mice, a central corneal epithelial abrasion initiates neutrophil (PMN) and platelet extravasation from peripheral limbal venules. The same injury in mice expressing low levels of the β2-integrin, CD18 (CD18hypo mice) shows reduced platelet extravasation with PMN extravasation apparently unaffected. To better define the role of CD18 on platelet extravasation, we focused on two relevant cell types expressing CD18: PMNs and mast cells. Following corneal abrasion in WT mice, we observed not only extravasated PMNs and platelets but also extravasated erythrocytes (RBCs). Ultrastructural observations of engorged limbal venules showed platelets and RBCs passing through endothelial pores. In contrast, injured CD18hypo mice showed significantly less venule engorgement and markedly reduced platelet and RBC extravasation; mast cell degranulation was also reduced compared to WT mice. Corneal abrasion in mast cell-deficient (KitW-sh/W-sh) mice showed less venule engorgement, delayed PMN extravasation, reduced platelet and RBC extravasation and delayed wound healing compared to WT mice. Finally, antibody-induced depletion of circulating PMNs prior to corneal abrasion reduced mast cell degranulation, venule engorgement, and extravasation of PMNs, platelets, and RBCs. In summary, in the injured cornea, platelet and RBC extravasation depends on CD18, PMNs, and mast cell degranulation.
... KIT signalling is required for glycogen metabolism of the corneal epithelium, 54 and loss of either KITL or KIT significantly impairs corneal wound healing. 55 Interestingly, KIT is not only expressed in LESCs but also in photoreceptors, lacrimal canaliculus epithelial stem cells and eye wall cells. [56][57][58] These cells are all essential for sustaining the physiological functions of the eye and are located in close proximity to SOX10-positive NCCs or their derivatives. ...
Homeostasis and function of limbal epithelial stem cells (LESCs) rely on the limbal niche, which, if dysfunctional, leads to limbal epithelial stem cell deficiency (LSCD) and impaired vision. Hence, recovery of niche function is a principal therapeutic goal in LSCD, but the molecular mechanisms of limbal niche homeostasis are still largely unknown. Here, we report that the neural crest transcription factor SOX10, which is expressed in neural crest-derived limbal niche cells (LNCs), is required for LNCs to promote survival of LESCs both in vivo and in vitro. In fact, using mice with a Sox10 mutation and in vitro coculture experiments, we show that SOX10 in LNCs stimulates the production of KIT ligand (KITL), which in turn activates in LESCs the KIT-AKT signalling pathway that protects the cells against activated CASPASE 3-associated cell death. These results suggest that SOX10 and the KITL/KIT-AKT pathway play key roles in limbal niche homeostasis and LESC survival. These findings provide molecular insights into limbal niche function and may point to rational approaches for therapeutic interventions in LSCD.
... Accordingly, corneal wound healing should elicit a tissue response in which limbal stem cells undergo few cell cycles and give rise to numerous transient amplifying cells that constitute the migratory/proliferative edge of the wound. The size of the transient amplification of early precursors and committed cells, would then be modulated by changes in the ECM composition and its ECM receptors during corneal wound healing [208][209][210], and by changes in the expression of growth factors such as IGF-1 [183], Epiregulin [211] or Stem Cell Factor (c-kit ligand) [212]. ...
Stem cell niche may be described as an anatomically defined and protected location that provides housing, positioning information and signaling inputs necessary to support normal stem cell activity. Based on the distribution of the differentiation-linked keratins, proliferative potential and wound healing abilities in ocular surface, it was proposed that an anatomical structure, the limbus, was the presumptive site of residence of corneal stem cells. Further analysis determined that the limbus contains a specific anatomical structure that probably provides the microenvironmental characteristics that correspond to the stem cell niche. This structure was termed as the Limbal Epithelial Crypt (LEC). Accumulated evidences show that the LEC is the site where stem cells interact directly and/or indirectly with at least six different cell types: epithelial, stromal, Langerhans cells, melanocytes, and telocytes. In addition, a rich and distinctive vasculature as well as an extensive neural network exist at limbal niches. These cell types, together with growth factors, cytokines and specific components of the Extracellular Matrix establish the conditions for the regulated growth, migration and delayed differentiation of the corneal stem cells. In spite of the wide variety of molecular markers described for limbal epithelial cells, it has been extremely difficult to isolate stem cells. This is explained by the persistence of stem cell markers in the transient amplifying cell population and in the early differentiating cells. Consequently, the use of stem cells in ophthalmic therapy shows variable outcomes and research must be increased before a formal clinical use. Nevertheless, the analysis of the different cells and factors involved in stem cell regulation, may help the development of new therapies based on the interference or stimulation of the signaling pathways and microenvironmental components that control limbal stem cells.
... A recent study investigated the effects of stem cell factor (SCF) and c-kit in corneal wound healing in mice [20]. In vivo and in vitro experiments indicated that the SCF/c-kit system is most likely implicated in cell migration and cell attachment, not cell proliferation, during corneal epithelial wound healing [20]. ...
... A recent study investigated the effects of stem cell factor (SCF) and c-kit in corneal wound healing in mice [20]. In vivo and in vitro experiments indicated that the SCF/c-kit system is most likely implicated in cell migration and cell attachment, not cell proliferation, during corneal epithelial wound healing [20]. These results are in accordance with those obtained in the present study. ...
... However, we found no specific adhesion molecules related to GM-CSF. Although Miyamoto et al. attempted to identify adhesion molecules regulated by SCF, none of the molecules changed significantly [20]. However, they suggested that the SCF/c-kit system might be involved in cell attachment through enhancement of the avidity or affinity of integrins to fibronectin, laminin, and type IV collagen. ...
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pleiotropic cytokine that activates granulocyte and macrophage cell lineages. It is also known to have an important function in wound healing. This study investigated the effect of GM-CSF in wound healing of human corneal epithelial cells (HCECs). We used human GM-CSF derived from rice cells (rice cell-derived recombinant human GM-CSF; rhGM-CSF). An in vitro migration assay was performed to investigate the migration rate of HCECs treated with various concentrations of rhGM-CSF (0.1, 1.0, and 10.0 μg/ml). MTT assay and flow cytometric analysis were used to evaluate the proliferative effect of rhGM-CSF. The protein level of p38MAPK was analyzed by western blotting. For in vivo analysis, 100 golden Syrian hamsters were divided into four groups, and their corneas were de-epithelialized with alcohol and a blade. The experimental groups were treated with 10, 20, or 50 μg/ml rhGM-CSF four times daily, and the control group was treated with phosphate-buffered saline. The corneal wound-healing rate was evaluated by fluorescein staining at the initial wounding and 12, 24, 36, and 48 hours after epithelial debridement. rhGM-CSF accelerated corneal epithelial wound healing both in vitro and in vivo. MTT assay and flow cytometric analysis revealed that rhGM-CSF treatment had no effects on HCEC proliferation. Western blot analysis demonstrated that the expression level of phosphorylated p38MAPK increased with rhGM-CSF treatment. These findings indicate that rhGM-CSF enhances corneal wound healing by accelerating cell migration.
... They conclude that although extensive knowledge has been gained on the role of certain growth factors and their receptors in wound healing, there is still little information on the function of some other growth factors in this regard. Cazander et al. [101] have reviewed the evidence of the role of complement, whereas Miyamoto et al. [102] have highlighted a potential role of stem cell factor in wound healing. ...
Small-molecule tyrosine kinase inhibitors (TKIs) represent a major advance in the treatment of certain forms of cancer. Unexpectedly, however, their use is associated with serious toxic effects on many vital organs and functions. Some of these effects, such as venous thromboembolism, haemorrhage, gastric perforation and a potential for impaired tissue healing, have direct implications for the safety of surgery in cancer patients. A number of currently approved TKIs are suspected or have been reported to impair wound healing but, understandably, there have been no formal pre- or post-approval clinical trials to evaluate the extent of the risk. Consequently, drug labels typically recommend discontinuation of the TKI concerned prior to elective surgery. In patients with gastric perforation, permanent discontinuation is advised. These recommendations, which are based on a precautionary principle, raise a dilemma, especially in patients with TKI-responsive tumours. This review focuses on the labelled potential of these novel antineoplastic agents to impair tissue repair and wound healing, and the evidence concerning the likely mechanisms involved. At present, because of the lack of formal clinical data, there are no evidence-based guidelines on the management of surgery in patients treated with TKIs. There is a need for a central registry of clinical outcomes following emergency surgery in cancer patients receiving TKIs and TKI-naïve matched controls. Analysis of outcomes data from such registries will assist in formulating guidelines on the management of elective surgery in TKI-treated patients. If TKIs are shown to significantly impair wound healing, patients receiving TKI therapy will require special monitoring and a collaborative approach between oncologists and surgeons for individualized reappraisal of the risk/benefit of the TKI treatment.
... Более 90% нейральь ных стволовых и прогениторных клеток челоо века несут на своей поверхности рецептор ссkit [56]. Взаимодействие пары SCF\cckit активии рует многочисленные сигнальные каскады, включая RAS/ERK, PI33K, JAK/STAT и Srcc киназы, результатом чего становится направв ленная миграция, выживание и пролиферация стволовых клеток, как в области ишемического повреждения, так и в зоне неоплазии [57,58]. ...
Modern methods for the treatment of malignant brain tumors are insufficiently effective. One reason for this is that the existing technologies and methods are focused on removing all neoplastic cellsfrom the body. Understanding the mechanisms of systemic migration of stem cells provides a new view on the role of this phenomenon in the development of malignant tumors. Migration and homing of normal stem cells, being originally the regulatory process, ensuring revascularization and remodeling of ischemic or traumatic injury of brain, play a role of the axial conductor of neoplastic process in carcinogenesis. The use of the phenomenon of migration and homing of stem cells in the tumor center for therapeutic purposes opens the possibility of overcoming the blood-brain barrier, reducing the toxicity of chemotherapy and increasing the radiation therapy efficiency, makes possible the directed influence on the hypoxic zone of the tumor, can directly affect to the key life processes of tumor stem cells. These arguments allow to consider the mechanisms of systemic migration and homing of stem cells to neoplastic foci as a fundamental theoretical platform for the creation of a fundamentally new class of anti-cancer, cell personalized medicines.
... Accordingly, corneal wound healing should elicit a tissue response in which limbal stem cells undergo a few cell cycles and give rise to numerous transient amplifying cells that constitute the migratory/proliferative edge of the wound. The size of the transient amplification of early precursors and committed cells would then be modulated by changes in the ECM composition and ECM receptors during corneal wound healing148149150, and by changes in the expression of growth factors such as insulin-like growth factor 1 [128], epiregulin [151], or stem cell factor (c-kit ligand) [152].Figure 2. Schematic representation of the limbal epithelial crypt. The extracellular matrix composition and structure may regulate limbal stem cell fate providing information about their position. ...
Stem cells emerged as a concept during the second half of 19(th) century, first as a theoretical entity, but then became one of the most promising research fields in cell biology. This work describes the most important characteristics of adult stem cells, including the experimental criteria used to identify them, and discusses current knowledge that led to the proposal that stem cells existed in different parts of the eye, such as the retina, lens, conjunctiva, corneal stroma, Descemet's membrane, and the subject of this review: the corneal epithelium. Evidence includes results that support the presence of corneal epithelial stem cells at the limbus, as well as the major obstacles to isolating them as pure cell populations. Part of this review describes the variation in the basement membrane composition between the limbus and the central cornea, to show the importance of the corneal stem cell niche, its structure, and the participation of extracellular matrix (ECM) components in regulating corneal stem cell compartment. Results obtained by various laboratories suggest that the extracellular matrix plays a central role in regulating stem cell commitment, corneal differentiation, and participation in corneal wound healing, in addition to other environmental signals such as cytokines and growth factors. The niche could define cell division patterns in corneal stem cell populations, establishing whether stem cells divide asymmetrically or symmetrically. Characterization and understanding of the factors that regulate corneal epithelial stem cells should open up new paths for developing new therapies and strategies for accelerating and improving corneal wound healing.
