Increased vascular endothelial growth factor may account for elevated level of plasminogen activator inhibitor-1 via activating ERK1/2 in keloid fibroblasts (original) (raw)
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The American Journal of Pathology, 2003
Proteolytic degradation of the provisional fibrin matrix and subsequent substitution by fibroblast-produced collagen are essential features of injury repair. Immunohistochemical studies revealed that although dermal fibroblasts of normal scars and keloids expressed both urokinase type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1), keloid fibroblasts had a much higher PAI-1 expression. In long-term three-dimensional fibrin gel cultures (the in vitro fibroplasia model), normal fibroblasts expressed moderate and modulated activity levels of uPA and PAI-1. In contrast, keloid fibroblasts expressed a persistently high level of PAI-1 and a low level of uPA. The high PAI-1 activity of keloid fibroblasts correlated with their elevated collagen accumulation in fibrin gel cultures. Substituting collagen for fibrin in the gel matrix resulted in increased uPA activity and reduced collagen accumulation of keloid fibroblasts. Furthermore, decreasing PAI-1 activity of keloid fibroblasts in fibrin gel cultures with anti-PAI-1-neutralizing antibodies also resulted in a reduction in collagen accumulation by keloid fibroblasts. Cumulatively, these results suggest that PAI-1 overexpression is a consistent feature of keloid fibroblasts both in vitro and in vivo, and PAI-1 may play a causative role in elevated collagen accumulation of keloid fibroblasts.
Journal of Investigative Dermatology, 1996
Using a 3-dimensional fibrin gel model system simulating fibroplasia of wound repair, we investigated the interaction between keloid fibroblasts and fibrin matrix and compared it with that of normal fibroblasts. Normal skin fibroblasts caused fibrin gel degradation under serum-free conditions, whereas keloid fibroblasts did not cause microscopically detectable gel degradation. Fibrin gel degradation occurred through plasmin-mediated fibrinolysis, which was initiated by fibroblasts exhibited high uPA but low plasminogen activator inhibitor-1 (PAI-1) activities, and transforming growth factor-beta 1 prevented fibrinolysis of normal fibroblasts by upregulating PAI-1 while downregulating uPA activities. In contrast, keloid fibroblasts exhibited an intrinsically high level of PAI-1 and a low level of uPA. This change in the ratio of activator and inhibitor activities was attributed to altered fibrin degradation by keloid fibroblasts. The PAI-1 increase was also demonstrated at the RNA level by Northern analysis. In terms of the pivotal role of the plasmin/plasminogen activator system in matrix remodeling, the elevated PAI-1 level exhibited by keloid fibroblasts may have significant consequences not only in altered fibrin degradation, but also in subsequent repair steps that lead to keloids and fibrosis.
Wound Repair and Regeneration, 2004
Keloids, which overgrow the boundaries of the original injury, represent aberrations in the fundamental process of wound healing that include over-abundant cell in-migration, cell proliferation, and inflammation, as well as increased extracellular matrix synthesis and defective remodeling. To understand the key events that result in the formation of these abnormal scars would open new avenues for better understanding of excessive repair, and might provide new therapeutic options. We examined epidermal growth factor receptor (EGFR)-induced cell motility in keloid fibroblasts, as this receptor initiates cell migration during normal wound repair. We show that keloid fibroblasts respond to EGF-induced cell migration but the response is somewhat diminished compared to normal adult fibroblasts (~30% reduced); the mitogenic response was similarly blunted (~5% reduced). Keloid fibroblasts express near normal levels of EGFR (82%), but show a much more attenuated activation of EGFR itself and the motility-associated phospholipase C-g. This was reflected in part by rapid loss of EGFR upon exposure to EGF. Interestingly, while extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK-MAPK) activation was relatively robust in keloid fibroblasts, the downstream triggering of the motility-associated calpain activity was blunted. This was reflected by high cell-substratum adhesiveness in the keloid fibroblasts. Thus, the blunted migratory response to EGF noted in keloid fibroblasts appears due to limited activation of two important biochemical switches for cell motility.(WOUND REP REG 2004;12:183-192) Keloids are neoplastic growths resulting from excessive response to cutaneous injury. Keloids are raised pathological scars that, over extended time periods, overgrow the margins of the original wound and cause damage to the healthy dermal tissue. 1,2 Keloids persist for years and cause pain, persistent itching, disfiguration, and limit function; unfortunately, they rarely regress. These disorders represent aberrations in the fundamental processes of wound healing, which include cell migration, cell proliferation, inflammation, increased synthesis of cytokines and extracellular matrix proteins, and defective remodeling. Probing the intracellular and molecular signaling events of cells derived from the keloid lesions would open new avenues for better understanding of excessive repair, which might provide new therapeutic considerations. Several hypotheses have been put forward regarding the biochemical, immunological, and molecular BSA Bovine serum albumin DMEM Dulbecco'
Role of vascular endothelial growth factor in keloids: a clinicopathologic study
International Journal of Dermatology, 2009
Background Despite their benign nature, keloids are usually associated with considerable cosmetic effects and may lead to functional problems. Recently, it has been reported that vascular endothelial growth factor (VEGF), a potent angiogenic factor, is overexpressed in keloid tissue and may have a potential role in its evolution.Methods Twenty patients with keloids were included in this study and classified into two groups according to the treatment received: intralesional triamcinolone acetonide 20 mg/mL (group 1) and cryotherapy spray technique (group 2). Treatment was continued until clearance or for a maximum of six sessions, and the follow-up period was 1 year. Skin biopsies were taken from patients before and after treatment to evaluate keloid pathology and from patients and 10 healthy controls to detect the immunohistochemical expression of VEGF.Results Histopathologic examination revealed a remarkable resolution of the nodular arrangement of collagen after therapy, particularly in group 1. A statistically significant difference in VEGF expression was found between patients before therapy and controls, and between patients before and after therapy in each group. There was no significant difference in the treatment outcome between intralesional steroids and cryotherapy. No significant correlation was observed between the clinical variables of keloids and both VEGF expression and clinical response to therapy.Conclusion VEGF seems to play an important role in the pathogenesis of keloids and may be a useful guide in the evaluation of keloid therapeutics. Modulation of its production may provide a valuable treatment for keloids.
The American Journal of Pathology, 2008
Keloids are tumor-like skin scars that grow as a result of the aberrant healing of skin injuries, with no effective treatment. We provide new evidence that both overexpression of plasminogen activator inhibitor-1 (PAI-1) and elevated collagen accumulation are intrinsic features of keloid fibroblasts and that these characteristics are causally linked. Using seven strains each of early passage normal and keloid fibroblasts, the keloid strains exhibited inherently elevated collagen accumulation and PAI-1 expression in serumfree, 0.1% ITS؉ culture; larger increases in these parameters occurred when cells were cultured in 3% serum. To demonstrate a causal relationship between PAI-1 overexpression and collagen accumulation, normal fibroblasts were infected with PAI-1-expressing adenovirus. Such cells exhibited a two-to fourfold increase in the accumulation of newly synthesized collagen in a viral dose-dependent fashion in both monolayers and fibrin gel, provisional matrix-like cultures. Three different PAI-1-targeted small interfering RNAs, alone or in combination, produced greater than an 80% PAI-1 knockdown and reduced collagen accumulation in PAI-1-overexpressing normal or keloid fibroblasts. A vitronectin-binding mutant of PAI-1 was equipotent with wild-type PAI-1 in inducing collagen accumulation, whereas a complete protease in-hibitor mutant retained approximately 50% activity. Thus, PAI-1 may use more than its protease inhibitory activity to control keloid collagen accumulation. PAI-1-targeted interventions , such as small interfering RNA and lentiviral short hairpin RNA-containing microRNA sequence suppression reported here , may have therapeutic utility in the prevention of keloid scarring.
Journal of Investigative Dermatology, 2003
Keloids are an excessive accumulation of extracellular matrix. Although numerous studies have shown elevated plasminogen activator inhibitor-1 (PAI-1) levels in keloid ¢broblasts compared with those of normal skin. Their speci¢c mechanisms involved in the di¡erential expression of PAI-1 in these cell types. In this study, the upregulation of PAI-1 expression is demonstrated in keloid tissues and their derived dermal ¢broblasts, attesting to the persistence, if any, of fundamental di¡erences between in vivo and in vitro paradigms. We further examined the mechanisms involved in hypoxia-induced regulation of PAI-1 gene in dermal ¢broblast derived from keloid lesions and associated clinically normal peripheral skins from the same patient. Primary cultures were exposed to an environ-
Keloids - A fibroproliferative disease
2008
Keloids are a fibroproliferative disorder of unknown etiology developing in the skin after injury or spontaneously. The aim of this thesis is to gain deeper insight into the role of TGF-β and its signaling pathway proteins, SMADs, in the pathogenesis of keloids and describe the gene expression profile in different keloid sites in the search for potential target genes for future treatment. Further aim is to develop an instrument to describe the quality of life of patients with keloids. We find cultured keloid fibroblasts to express an increased level of TGF-β1 mRNA and a decreased level of TGF-β3 mRNA compared to control skin. Keloid derived fibroblasts exhibit significantly decreased mRNA levels of TGF-β receptor type II (TβRII) and the ratio of TβRI/TβRII mRNA expression is increased. This suggests that a certain expression pattern of TGF-β subtypes and receptors may be important in keloid pathogenesis. Analysis of keloid derived fibroblasts reveal decreased SMAD3 mRNA expression and decreased ratio of SMAD2/SMAD3 mRNA implicating a disturbed SMAD signaling. Keloid fibroblasts up-regulate SMAD4 protein after stimulation with TGF-β1 and display diminished levels of the inhibitory proteins SMAD6 and 7. This may contribute to unlimited and deregulated TGF-β signaling leading to increased extracellular matrix production (ECM).
Accelerated skin wound healing in plasminogen activator inhibitor-1-deficient mice
American Journal of …, 2001
Components of the fibrinolytic system have been implicated in cell migratory events associated with tissue remodeling. Studies in plasminogen-deficient mice (PG ؊/؊ ) indicated that skin wound healing is impaired, but is resolved with an additional fibrinogen deficiency. Plasminogen activator inhibitor-1 (PAI-1) expression by keratinocytes has been identified shortly after wound injury. PAI-1 expression could affect wound healing by regulating the fibrinolytic environment of the wounded area, as well as influencing events associated with cell attachment and detachment through interactions with matrix proteins. The present study directly assesses PAI-1 involvement in skin wound healing through analyses of a dermal biopsy punch model in PAI-1-deficient (PAI-1 ؊/؊ ) mice. While the cellular events associated with the healing process are similar between wildtype (WT) and PAI-1 ؊/؊ mice, the rate of wound closure is significantly accelerated in PAI-1 ؊/؊ mice.
Altered angiogenic balance in keloids: a key to therapeutic intervention
Translational Research, 2012
Keloids are manifestations of abnormal wound repair with unresolved clinical complications. An effective therapeutic regimen has not been established for keloids, and current strategies are plagued by problems such as recurrence and side effects. Keloids, being a human-specific dermal fibroproliferative disorder are characterized by an excessive accumulation of extracellular matrix (ECM), thickened basement membrane, unregulated expression of matrix metalloproteases, growth factors, and cytokines. The internal milieu in a keloid bears a strong resemblance to a tumor with both exhibiting striking similarities with respect to tissue environment and unregulated vasculature. Abnormal angiogenesis manifested by an imbalance between proangiogenic and antiangiogenic factors has been recognized as a ''common denominator'' underlying many pathological conditions. However, such an imbalance has not been investigated in keloids. In this study, the angiogenic imbalance in keloids was explored with reference to circulating and tissue level expression of vascular endothelial growth factor (VEGF) and endostatin/collagen XVIII. It was observed that VEGF levels were upregulated and endostatin levels were downregulated in keloid patients in comparison to normal controls in both sera and tissue. Hence, antiangiogenic therapeutics based on endostatin in combination with current curative strategies as in tumors would present a scope for the effective management of keloids.