Keloid Fibroblasts Research Articles (Page 1)

Asiatic acid inhibits keloid fibroblast migration and collagen deposition via suppression of STAT3 activation.

NNMT enhances invasive and migratory capacity of keloid fibroblasts and M2 macrophage polarization.

ACSS3 protein macromolecule regulates glycolysis in keloid through Wnt/β-catenin signaling pathway: Bioinformatics, machine learning, and experimental validation.

Keloids, characterized by excessive collagen deposition and recurrence, pose significant therapeutic challenges due to limited mechanistic understanding. Mesenchymal stem cells (MSCs) exhibit potential for keloid management, but their precise mechanisms remain unclear. This study investigated how MSCs modulate extracellular matrix (ECM) remodeling in keloid pathogenesis. Using a co-culture system of human umbilical cord MSCs (UC-MSCs) and immortalized keloid fibroblasts (HDIKFs), we demonstrated that UC-MSCs significantly suppressed HDIKF proliferation (via CCK8 assay) and migration (via wound healing assay). Interestingly, UC-MSCs did not alter keloid xenograft growth in vivo. Mechanistically, quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) revealed selective downregulation of matrix metalloproteinases 9 (MMP9) in HDIKFs co-cultured with UC-MSCs, while MMP1, MMP2, and MMP3 remained unaffected. This suppression was linked to inhibition of the transforming growth factor-β1/SMAD (TGF-β1/SMAD) pathway, evidenced by reduced hypoxia‐inducible factor-1α (HIF-1α) and SMAD2 expression, alongside upregulated interleukin-10 receptor alpha (IL-10RA). Additionally, UC-MSCs did not alter collagen I/III (COL I/III) ratios or phosphatidylinositol‐3‐kinase (PI3K)/protein kinase B (AKT) signaling. These findings highlight that MSCs attenuate keloid fibroblast activity through TGF-β1/SMAD-driven MMP9 suppression and IL-10RA enhancement, offering novel insights into MSC-based strategies for ECM homeostasis. This study underscores MMP9 as a therapeutic target and provides a foundation for refining MSC efficacy in keloid treatment.

Background Mechanical stresses affect biological processes, cellular homeostasis, and skin wound healing. Objective We investigated the effects of mechanical stretching on epidermal keratinocytes, focusing on its role in wound healing and keloid formation. Methods Epidermal keratinocytes were stretched using a biochemical stretch device with or without a cyclooxygenase-2 inhibitor. Cells and supernatants were collected after 1−24 h of stimulation. Stretched keratinocytes were co-cultured with normal dermal or keloid-derived fibroblasts. Prostaglandin E2 and 12-hydroxyeicosatetraenoic acid were measured via enzyme-linked immunosorbent assay. Prostaglandin E2 receptor EP3 and transforming growth factor-β1 (TGF-β1) were analyzed using qRT-PCR. TGF-β1 was also examined using western blotting. Prostaglandin E2 synthase and TGF-β1 were also stained and analyzed using immunofluorescence microscopy. Results Mechanical stretching of keratinocytes increased the levels of prostaglandin E2, which stimulates keratinocyte proliferation and exerts antifibrotic effects, as well as those of prostaglandin E synthase 2 and prostaglandin E2 receptor 3. In contrast, mechanical stretching did not affect the levels of transforming growth factor-β1 in affected keratinocytes. However, co-culture with keloid-derived fibroblasts markedly increased the levels of transforming growth factor-β1 in mechanically stretched keratinocytes. Conclusion Mechanical stretching induced PGE2 production and increased the TGF-β1 levels in keratinocytes in the presence of keloid-derived fibroblasts. Our results suggest that stretch stimulation in the presence of keloid-derived fibroblasts promotes keloid development.

Keloids are an inflammatory cutaneous condition, which are characterized by fibroproliferative overgrowth of the skin. Although keloids are not life‑threatening, their incidence and recurrence are relatively high, thus decreasing the quality of life of patients due to pain, pruritus and cosmetic reasons. Additionally, the precise molecular mechanisms underlying the pathogenesis of keloids remain largely unexplored, thus limiting the development of therapeutic interventions. To screen the key molecules in keloids, microarray data were selected from three different datasets obtained from the Gene Expression Omnibus database, namely GSE145725, GSE7890 and GSE44270. One differentially expressed gene was identified, periostin (POSTN), which was upregulated in keloid fibroblasts (KFs) compared with normal fibroblasts. Its high expression was further validated in KFs using reverse transcription‑quantitative PCR (RT‑qPCR), western blotting and immunofluorescence staining. Its potential function were explored in keloids through loss-of-function assay. Notably, the EdU incorporation assay and cell cycle assay indicated that POSTN knockdown had limited effects on the proliferation of KFs; however, the RT‑qPCR, western blotting, and RNA sequencing results suggested that POSTN inhibition blocked the JAK‑STAT signaling pathway and decreased the expression levels of various proinflammatory factors in KFs. Additionally, the RT‑qPCR and western blotting results demonstrated that IL‑4 and IL‑13, two significant mediators of T helper 2 (Th2) signaling, could induce POSTN expression in KFs. Notably, IL‑4 receptor (IL‑4R), a receptor for both IL‑4 and IL‑13, could be positively modulated by POSTN through the Reactome enrichment, RT‑qPCR and western blotting analysis. Furthermore, IL‑4R was essential for IL‑4/IL‑13‑induced POSTN upregulation in KFs, thus indicating a positive feedback loop between POSTN and Th2 signaling. Overall, the current study uncovered a novel mechanism of POSTN, which could be associated with keloid inflammation, thus highlighting the POSTN/Th2 feedback loop as a potential therapeutic target for patients with keloids.

Secreted clusterin inhibits keloid formation by promoting fibroblast apoptosis.

Keloid is a benign form of dermis tumor formed due to imbalance of collagen deposition and degradation during wound healing. Preclinical studies showed Tithonia diversifolia (Hemsley) A. Gray (T. diversifolia) potency as keloid treatment by inhibiting keloid fibroblast proliferation, collagen deposition, TGFβ1 and VEGF expression. This study aims to examine the safety and efficacy of T. diversifolia extract gel for keloid treatment in keloid patient. Fourteen keloid patients were included in the study with ratio 1:1. The participants received either T. diversifolia extract gel 2% or triamcinolone acetate cream 0.025% for 12 weeks. Evaluation of efficacy was done every 4 weeks until week 12 using Vancouver Scar Score, the patient and observer Scar assessment. Vancouver scar total score was improved in week 12 from baseline in both groups. Improvement from baseline after 12 weeks of treatment also found in patient and observer scar assessment total scale. Those scores on week 12 were similar between those on T. diversifolia extract gel 2% and triamcinolone acetate cream 0.025% treatment. Conclusions: T. diversifolia extract gel 2% is efficacious and safe for keloid treatment.

Keloids are characterized by excessive extracellular matrix (ECM) accumulation and persistent inflammation, leading to disfiguring scars and poor therapeutic outcomes. The α7 nicotinic acetylcholine receptor (α7nAChR) has emerged as a key modulator of inflammatory and fibrotic signaling. This study evaluated the antifibrotic effects of tropisetron, a clinically available α7nAChR agonist, in keloid fibroblasts (KFs) and a rat incisional scar model. In vitro, KFs exhibited reduced α7nAChR expression, which was restored by tropisetron in a dose-dependent manner. Tropisetron treatment significantly decreased KF viability, downregulated pro-fibrotic genes (COL1A1, COL3A1, α-SMA), and upregulated matrix metalloproteinases (MMP1 and MMP3). Additionally, it suppressed phosphorylation of Smad2/3 and reduced expression of NF-κB and TNF-α, indicating inhibition of both TGF-β and inflammatory pathways. In vivo, tropisetron-treated rats showed a ~40% reduction in scar area, improved collagen organization, and increased α7nAChR expression in scar tissue. Western blot analysis confirmed decreased levels of collagen I, p-Smad2/3, α-SMA, NF-κB, and TNF-α. These results indicate that tropisetron exerts dual antifibrotic and anti-inflammatory effects through α7nAChR-mediated signaling and enhanced ECM remodeling. This study provides the first evidence supporting α7nAChR activation as a promising therapeutic strategy for managing keloids and other fibrotic skin disorders.

ABSTRACTBackground and AimsThe formation of keloids is influenced by mechanical stretch. Thrombospondin‐1 (TSP‐1) is identified as a tension‐sensitive protein. However, the relationship between TSP‐1 and keloid formation induced by mechanical stretch remains unknown.MethodsA simple customized mechanical stretch device was used for the application of homogeneous equibiaxial stretch (HES). Using Western blot and RT‐PCR, the optimal stretch strength and duration were determined. Regarding the functional changes induced by stretch in keloid fibroblasts (Kfbs), cell function assays were used. The relationship between TSP‐1 expression and stretch‐induced scar formation in human and animal models was investigated using immunohistochemistry. The knockdown of TSP‐1 in fibroblasts served as a reverse test.ResultsOptimal HES (oHES) could be achieved with four rotations of the screws on our mechanical stretch device, resulting in a significant increase in vimentin, Col I, and fibronectin expression in Kfbs on Day 5. Additionally, oHES significantly promoted cell proliferation and migration. oHES resulted in the upregulation of TSP‐1 expression in both in vitro and in vivo experimental settings. The inhibition of TSP‐1 may attenuate oHES‐induced keloid formation through the Akt and GSK‐3β/β‐catenin signaling pathways.ConclusionThe results confirmed that oHES promoted keloid formation by increasing Col I expression through TSP‐1‐mediated Akt and GSK‐3β/β‐catenin signaling pathways.

Aberrant extracellular matrix (ECM) production by dermal fibroblasts drives fibrotic skin diseases, which has an adverse impact on the lives of patients. Current treatments are limited; therefore, the development of new antifibrotic strategies is necessary. The aim of the present study was to investigate zinc finger 469 (ZNF469) as a potential ECM regulator in skin fibrosis. ZNF469 was knocked down in dermal fibroblasts using doxycycline-induced short hairpin RNA. ZNF469 knockdown was found to impair proliferation, migration, contraction and collagen production in dermal fibroblasts. In addition, RNA sequencing revealed that ZNF469 knockdown suppressed the expression of genes associated with the ECM and collagen biosynthesis, indicating that ZNF469 plays a role in ECM regulation. Analysis of publicly available RNA-sequencing data from hypertrophic scars and keloids revealed the upregulation of ZNF469 expression in these tissues and the positive correlation of ZNF469 expression with that of ECM-related genes. Single-cell analysis of keloids indicated that ZNF469 is localized in mesenchymal fibroblasts, a key collagen-producing fibroblast subpopulation. Pseudotime analysis suggested that ZNF469 plays a role in the establishment of this phenotype. Consistent with this notion, the knockdown of ZNF469 in dermal fibroblasts was shown to downregulate the expression of mesenchymal markers. In addition, it reduced the proliferation, migration, contraction and collagen production of keloid fibroblasts. These findings indicate that ZNF469 is a crucial regulator of ECM production and the mesenchymal fibroblast phenotype, suggesting it may be a potential therapeutic target for skin fibrosis.

Keloid is a trauma-induced fibroproliferative condition characterized by excessive extracellular matrix (ECM) deposition and aberrant keloid fibroblast activation, leading to physical, psychological, functional, and cosmetic impairments. This study investigates DNA methylation alterations at Long Interspersed Nuclear Element-1 (LINE-1) and Alu repetitive elements in keloid tissues compared to normal skin tissues. Methylation levels and patterns were analyzed in keloid (n = 38) and normal skin tissues (n = 32). The results revealed significantly lower LINE-1 (P < 0.0001) and Alu (P = 0.0147) methylation levels in keloids, with hypomethylation inversely correlated with chronological age. Younger individuals exhibited higher methylation levels compared to older individuals. These findings offer critical insights into the epigenetic mechanisms underlying keloid formation and progression, providing a foundation for developing epigenetic-based therapeutic strategies.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-17646-2.

PurposeTargeting the distinct genetic and protein expression profiles of keloids necessitates the identification of novel therapeutic targets. This study was aimed to elucidate the role of Bcl-2-associated athanogene 2 (BAG2) in keloid pathology and identify compounds with high-affinity to BAG2.Patients and MethodsCell migration, and cell proliferation assays, along with flow cytometry, were used to evaluate the effects of BAG2 on keloid fibroblasts (KFs) derived from tissue samples of patients with abdominal or chest keloids. Additionally, histological examinations and Western blotting were performed to investigate BAG2’s role in keloids. Surface plasmon resonance (SPR) was employed to identify compounds with high-affinity to BAG2, and the effects of these compounds on keloids was assessed.ResultsInhibition of BAG2 significantly decreased collagen deposition, cell proliferation and migration in keloid tissues. The modulatory effect of BAG2 on these processes appears to be mediated partly by the MEK signaling pathway. Among the tested compounds, Bazedoxifene acetate and Ponesimod showed high affinity for BAG2 and demonstrated a more pronounced inhibitory effect on collagen deposition of the keloid tissues than other candidates.ConclusionThis study revealed the pathogenic role of BAG2 in keloid and identified compounds with high-affinity to BAG2, Bazedoxifene acetate and Ponesimod. The therapeutic capabilities of these compounds demonstrated their potential to improve therapeutic strategies for localized, targeted treatment to keloids.

Purpose: To evaluate the effects of preoperative topical demethasone on CC chemokine receptor 3 (CCR3) expression in human pterygium. Methods: Forty cases of patients with pterygium were divided into a dexamethasone group and a control group, which were treated topically with 3 g/L tobramycin + dexamethasone or 3 g/L tobramycin eye drops in the surgical eyes 2 weeks before surgery. Pterygium samples were collected from the subjects after pterygium excision surgery andprocessed for real-time polymerase chain reaction and Western blot to examine CCR3 expression. The effects of CCR3 signaling on tube-forming properties and proliferation of human retinal vascular endothelial cells (HRECs) were investigated in vitro using a Matrigel assay and a cell counting kit (CCK)-8 assay. The effects of CCR3 signaling on the proliferation and migration of human keloid fibroblasts (HKFs) were investigated using the CCK-8 assay and wound-healing assay. Results: CCR3 mRNA and protein were detected in pterygium. CCR3 expression levels were reduced in the dexamethasone group compared with the control group (P < 0.05). CCR3 expression levels were associated with basic fibroblast growth factor and inducible nitric oxide synthase expression levels in pterygium. In vitro experiments showed that CCR3 signaling increased the number of intact capillary tubes and promoted the proliferation of HRECs. In addition, CCR3 signaling increased the proliferation and migration of HKFs. Conclusions: CCR3 signaling may play an important role in the pathogenesis of pterygium. The proangiogenic and profibrotic functions of CCR3 signaling may be by promoting tube formation and proliferation of vascular endothelial cells and promoting proliferation and migration of fibroblasts.

Keloids are a form of excessive fibrosis disease characterized by tumor‐like features, which are prone to recurrence. Circular RNAs play a role in various diseases, but its roles in keloids remain unclear. In this study, using high‐throughput RNA sequencing to compare keloids with normal scars, a novel circRNA, hsa_circ_0026782 is identified, whose expression is downregulated in keloids. The hsa_circ_0026782 inhibits the proliferation, migration, invasion, and apoptosis of primary human keloid fibroblasts, blocks the cell cycle in the S phase in vitro, and prevents keloid progression in vivo. Mechanistically, 1–90 nucleotides of hsa_circ_0026782 directly bind to the transcription factor CREB1. This interaction increases the exposure of the basic leucine zipper domain of CREB1 to enhance its dimerization. In addition, this interaction also promotes CREB1 phosphorylation at serine residue 142, inhibits the binding of CREB1 to the promoters or enhancers of its downstream target genes, and ultimately alters the transactivation of CREB1. The findings unveil that the hsa_circ_0026782/CREB1 axis acts as a transcriptional spatiotemporal “molecular break” in the formation of keloids, providing a new target for the therapy of keloids.

SYVN1 promotes keloid fbroblast viability, migration, and M2 macrophage polarization via downregulating SFRP1 protein expression through the mediation of ubiquitination

ABSTRACT Background Long non-coding RNAs (lncRNAs) are increasingly recognized in keloid pathogenesis. This study investigates the role and mechanisms of HOXA11-AS in keloid formation. Methods Expression levels of HOXA11-AS and related proteins were measured in keloid tissues and fibroblasts using qRT-PCR, Western blot, and ELISA. Functional assays assessed cell proliferation, migration, fibrosis, and oxidative stress. RIP, ChIP, Co-IP, FISH, and luciferase assays were used to explore interactions among HOXA11-AS, YY1, Nrf2, EZH2, and DNMT1. An in vivo mouse xenograft model validated the findings. Results HOXA11-AS was upregulated in keloids. Silencing HOXA11-AS reduced fibroblast proliferation, migration, fibrosis, and oxidative stress. Its overexpression had the opposite effect, which was reversed by Nrf2 pathway inhibition. HOXA11-AS promoted the methylation of the Nrf2 promoter via DNMT1 recruitment, mediated by EZH2. YY1 enhanced HOXA11-AS transcription by binding to its promoter. The YY1/HOXA11-AS axis was confirmed in vivo. Conclusion YY1-induced HOXA11-AS drives keloid formation by promoting oxidative stress and inflammation through epigenetic suppression of Nrf2 signaling.

Adhesive hydrogel based on Konjac Glucomannan (KGM) loaded with siACTC1-exosomes for enhanced post-surgical keloid treatment.

Dispel-Scar Ointment is used in Traditional Chinese Medicine to treat scarred tissue and increasing evidence has shown that DSO has potent therapeutic; however, its exact mechanism remains unexplored. This study explored the molecular mechanisms of action of DSO in scarring using network pharmacology, molecular docking, and experimental validation. Public databases were used to predict the bioactive ingredients and putative targets of DSO against scars. A compounds-targets network was constructed using the Cytoscape software. Enrichment analysis was performed using ClueGo and FunRich to specify the biological functions and associated pathways of hub targets. Molecular docking was used to verify the correlation between the major active components and hub targets, visualised using PyMol 2.3. Experimental validations were conducted to elucidate the influence of DSO on keloid fibroblast cells using the CCK-8, wound-scratch, cell reactive oxygen species, and western blot assays. Results：Network pharmacological analysis of DSO for scar treatment identified 146 ingredients and 1078 gene targets. Major targets included, prostaglandin-endoperoxide synthase 2 matrix metallopeptidases, and nitric oxide synthase 2. ClueGo analysis revealed 29 pathways (p&amp;amp;amp;lt;0.05) and FunRich 345 pathways (p&amp;amp;amp;lt;0.05), mainly toll-like receptor, TGF-β, interleukin-4/13, glypican, and tumour necrosis factor-related apoptosis-inducing ligand pathways. Molecular docking showed MMP2-flavoxanthin, MMP9-luteolin and MMP-9-kaempferol bound best to DSO. DSO could inhibit the proliferation and migration of scar fibroblasts and promote their apoptosis in a concentration-dependent manner. DSO also decreased TGF-β1, -βR2, pSMAD2, pSMAD3, SMAD4, CoL1a1, and MMP2 expression. Network pharmacology, molecular docking, and experimental validation showed DSO&amp;amp;#039;s potential in treating scars. It may inhibit scars via the TGF-β1/SMADs/MMPs signalling pathway, providing a basis for DSO&amp;amp;#039;s scar treatment application.

Keloid is a fibroproliferative disorder that poses a challenge in clinical management. This study aims to identify and functionally annotate differentially expressed genes (DEGs) in keloid and explore the potential role of SLC6A15. The data were obtained from GEO (GSE218922 and GSE7890), and the DEGs and module genes were obtained with Limma and WGCNA. KEGG and GO enrichment analysis, and machine learning algorithms (Random Forest, Boruta, and XGBoost) were conducted to explore the keloid-related key genes. Finally, qRT-PCR was carried out to detect SLC6A15 mRNA expression, and CCK-8 and flow cytometry were employed to assess cell proliferation and apoptosis. We obtained 147 DEGs between keloid fibroblasts and normal fibroblasts, and 193 DEGs between keloid stem cells and normal stem cells, followed by acquisition of 40 intersection DEGs. These intersection DEGs were mainly enriched in external encapsulating structure organization, extracellular matrix organization, and were closely related to cytoskeleton in muscle cells and arrhythmogenic right ventricular cardiomyopathy (ARVC). WGCNA analysis identified five modules, with the blue modules showing a significant negative correlation with keloid. Afterwards, three machine learning methods were applied to analyze DEGs in keloid, identifying SLC6A15 as the most important gene. Further validation demonstrated that SLC6A15 was lowly expressed in keloid tissues and fibroblasts, and SLC6A15 overexpression inhibited proliferation and facilitated apoptosis in keloid fibroblasts. This study identified SLC6A15 as a potential biomarker for keloid, providing new research clues for the treatment target of this disorder.