Search Results (1,035)

Background: GDF15 (growth/differentiation factor-15) is part of the transforming growth factor-beta family and represents a cellular stress-induced gene. It might have a role in metaflammation and adipoflammation. We aimed to investigate the effects of Toll-like receptor (TLR) activation and hypoxia-related pathways together with metabolic factors on GDF15 regulation in adipocytes and adipose tissue (AT). Methods: GDF15 mRNA quantities in the human adipocyte cell line SGBS, in visceral (VAT) and subcutaneous adipose tissue (SAT) (resected from n = 96 obese and characterized patients), and in murine 3T3-L1 adipocytes were measured by real-time RT-PCR. GDF15 protein concentrations in cell supernatants and serum were quantified by ELISA. The following stimuli/pathways were investigated: insulin, glucose, TLR ligands (TLR2/6, TLR3, TLR4, TLR7, TLR9), bile acids, synthetic FXR/TGR5 activators, and HIF1α activators. Results: Basal GDF15 expression is low and only marginally induced in SGBS cells. In contrast, GDF15 is expressed in human SAT and VAT and correlates positively with the corresponding GDF15 protein concentration in peripheral blood serum of obese patients. Among metabolic factors, insulin and bile acids such as ursodeoxycholic acid upregulate GDF15 expression in 3T3-L1 adipocytes, the latter via FXR but not via TGR5. Among innate immune regulators, only TLR7 activation and hypoxic mediators upregulate whereas STAT3 signaling downregulates GDF15. Conclusion: GDF15 expression in human SAT and VAT is correlated to peripheral blood GDF15 concentrations and is regulated by metabolic and innate immune response pathways involved in AT inflammation and metaflammation. Full article

Sudden cardiac death (SCD) in athletes often represents the first manifestation of an underlying inherited cardiovascular disorder exposed by adrenergic stress, altered calcium cycling, mechanical loading, and metabolic demand during intense exercise. This review focuses on the molecular architecture that links genotype to arrhythmogenic phenotype in athletes, emphasizing sarcomeric force generation and energetic inefficiency in hypertrophic cardiomyopathy, desmosomal failure and Hippo/Wnt/transforming growth factor-beta (TGF-β) signaling in arrhythmogenic cardiomyopathy, and ion-channel and calcium/calmodulin-dependent protein kinase II (CaMKII)calcium handling abnormalities in inherited channelopathies. This review further examines how exercise-induced physiological remodeling intersects with these pathways through insulin-like growth factor-1 (IGF-1)/phosphoinositide 3-kinase (PI3K)/ protein kinase B (AKT) signaling, mitochondrial biogenesis, oxidative stress, inflammatory signaling, and epigenetic regulation. Attention is given to the molecular basis of genotype-positive/phenotype-negative states, variable penetrance, and exercise-mediated disease expression. Finally, the integration of molecular biology with genomic data, polygenic risk, and emerging digital phenotyping is discussed to refine mechanism-based risk stratification and identify future therapeutic targets for prevention of SCD in athletes. Full article

Wound healing remains a persistent health problem with no definitive solution. It is crucial to characterize the complex wound healing process and the various growth factors, cytokines, and polypeptides involved. Transforming growth factor beta1 (rhTGF-β1) stimulates different cell types, providing multifunctionality in the wound healing process. Since proteins are sensitive to proteases, drug delivery systems are needed. Developed polymeric carrier systems are as important as the active substance. The carrier systems used in our study aim to contribute to wound healing in addition to the rhTGF-β1. We hypothesized that PLGA nanoparticles embedded in PVA/Chitosan (PVA/Chi) hydrogels could enhance the therapeutic effect of rhTGF-β1. PVA/Chitosan hydrogels were prepared by the freezing/thawing method. Several characterization studies (Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), texture analysis, and cell culture) were performed to investigate the potential of the prepared formulations to enhance the therapeutic effect of rhTGF-β1. Hydrogel formulations reduced the inhibitory effect of rhTGF-β1 on keratinocytes. The H5 hydrogel exhibited a proliferative effect on fibroblast cells, which play a crucial role in wound healing, resulting in a 78.8% increase compared to the control. As the PVA content in the hydrogel formulations increased, bioadhesion and viscosity also increased. Although TGF-β1 inhibited keratinocytes, it induced migration of both NIH-3T3 and HACAT cell lines. The formulations developed exhibit the potential to improve the therapeutic efficacy of rhTGF-β1 in wound healing. A small amount of the protein can have the same therapeutic efficacy and fewer side effects because the developed polymeric carrier systems contribute to the therapeutic efficacy. Full article

Background/Objectives: Regenerative endodontic treatment (RET) depends on the release of dentin-derived bioactive molecules, which is commonly promoted by ethylenediaminetetraacetic acid (EDTA)-based dentin conditioning. However, whether adjunctive ozone delivery protocols can modify the measurable release of dentin-derived transforming growth factor beta 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1) remains unclear. This study evaluated the effects of two adjunctive ozone application protocols used with chelation on dentin-derived TGF-β1 and IGF-1 release, without directly assessing the in situ activation or functional bioactivity of TGF-β1. Methods: Sixty-four freshly extracted human mandibular premolars were randomly assigned to four groups (n = 16). The experimental protocols were as follows: 17% EDTA alone (Group A), 17% EDTA followed by ozonated distilled water and ozone gas (Group B), ozonated 17% EDTA followed by ozone gas (Group C), and a negative control group. Root segments were standardized. In the experimental groups, all external surfaces were coated with nail varnish, leaving only the intracanal dentin surface exposed. In the negative control group, all surfaces were sealed. After ultrasonic activation, the specimens were incubated in phosphate-buffered saline (PBS) at 37 °C. PBS samples were collected on day 1 to evaluate early measurable growth factor release and on day 7 to assess short-term changes in detectable growth factor levels. TGF-β1 and IGF-1 levels were measured by ELISA and normalized to internal dentin surface area derived from micro-computed tomography (micro-CT) analysis. Results: No detectable growth factor values were observed in the negative control group. For TGF-β1, no significant intergroup difference was observed on day 1, whereas a significant difference was found on day 7 (p = 0.022). On day 7, the ozonated EDTA followed by ozone gas group showed approximately threefold higher surface-area-normalized TGF-β1 values than the EDTA followed by ozonated distilled water and ozone gas group (p = 0.018). TGF-β1 values increased from day 1 to day 7 in Groups A and C, whereas no significant temporal change was observed in Group B. IGF-1 values showed no significant intergroup or intragroup differences. Conclusions: Adjunctive ozone application showed a protocol-dependent effect on dentin-derived growth factor values, mainly for TGF-β1, while IGF-1 remained unaffected. The highest TGF-β1 values were observed when ozonated EDTA was followed by ozone gas. However, these in vitro findings indicate measurable growth factor release and should not be interpreted as direct evidence of TGF-β1 activation or clinical regenerative efficacy. Full article

Background/Objectives: Growth differentiation factor-15 (GDF-15) is a protein that belongs to the transforming growth factor beta superfamily and has been found elevated in cases of organ injury such as liver, kidney, heart, and lung, as well as cardiovascular diseases and cancer. Soluble urokinase plasminogen activator receptor (suPAR) is a protein which is expressed mainly on immune cells and endothelial and smooth muscle cells, and is a marker of severity and intensity of inflammation in acute and chronic diseases. The aim of the present study was to compare GDF-15 serum levels between patients with acute cerebrovascular incidents and healthy controls and to investigate the possible correlation of GDF-15 serum levels and inflammatory markers, such as serum C-reactive protein (CRP) and plasma suPAR, in the above-mentioned groups. Methods: This is a retrospective study. Thirty-one patients were included in the study, with a mean age ± SD of 67 ± 13 years, compared to 18 age-matched healthy controls. Results: In the patient group a statistically significant positive correlation of serum levels of GDF15 values with suPAR and CRP emerged (rs = 0.516, p = 0.003) and (rs = 0.409, p = 0.022), respectively, and no significant correlation was found in the group of controls (rs = 0.271, p = 0.277) and (rs = 0.423, p = 0.080), respectively. Conclusions: These findings support the role of inflammation as a key underlying mechanism in acute cerebrovascular injury and suggest that GDF-15 may serve as a valuable adjunct biomarker for assessing disease severity and inflammatory burden. Full article

Circulating sex hormone-binding globulin (SHBG) concentrations are lower in individuals with metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH), reflecting its potential role in metabolic liver dysfunction. Our prior studies demonstrated that SHBG can attenuate MASLD by limiting hepatic lipid deposition, partly through suppression of lipogenic pathways, in both cellular and animal models. In the present work, we have examined whether SHBG could protect against development of liver fibrosis. For this purpose, in vitro and in vivo studies were performed. In vitro, we used co-cultures of human hepatocellular carcinoma cell line (HepG2) and human hepatic stellate cell line (LX-2) cells transfected using an SHBG expression vector vs. vehicle and treated with transforming growth factor beta 1 (TGF-β1). For in vivo studies we used wild-type and human SHBG transgenic mice developing liver fibrosis induced by carbon tetrachloride (CCl₄). Our results clearly showed that SHBG overexpression reduced the TGF-β1-induced expression in collagen in LX-2 cells. Moreover, SHBG overexpression reduced the CCl₄ induced liver fibrosis in both male and female mice. Histological examination revealed that SHBG transgenic mice had reduced NAS score and decreased collagen accumulation, assessed by Sirious Red staining. In addition, human SHBG transgenic mice treated with CCl₄ exhibited lower collagen 1A1 (Col1A1) protein levels when compared with wild-type CCl₄ treated mice. Mechanistically, SHBG attenuated fibrosis primarily through modulation of the TGF-β1/matrix metalloproteinases (MMPs)/tissue inhibitor metalloproteinases 1 (TIMP1) axis, characterized by reduced TGF-β1 levels, increased metalloprotease activity, and decreased TIMP1 levels compared with wild-type CCl₄ treated mice. Notably, female SHBG transgenic mice exhibited greater protection against fibrosis than males, indicating a sex-dependent effect likely mediated by differences in sex steroid signaling. Taken together, we demonstrate for the first time that SHBG protects against liver fibrosis by promoting collagen degradation via the TGF-β1/MMPs/TIMP1 pathway. Further research is needed to elucidate the role of sex steroids in the regulation of MMPs and the observed sexual dimorphism. Full article

Matrix metalloproteinases (MMPs) are zinc-dependent proteolytic enzymes involved in extracellular matrix remodelling in oral and dental tissues, including the periodontal ligament, alveolar bone, dentin, dental pulp, and periapical tissues. This narrative review summarises selected evidence on the role of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in orthodontic tooth movement, dental trauma and root resorption, restorative adhesive dentistry, and pulp/periapical disease. Particular attention is given to signalling pathways that regulate MMP/TIMP activity, including nuclear factor kappa B (NF-κB), mitogen-activated protein kinase (MAPK), Wnt/β-catenin, and transforming growth factor beta (TGF-β)/Smad-related mechanisms. The review also discusses the biomarker potential and translational status of MMP-targeted strategies. Across clinical contexts, MMP activity contributes to both matrix degradation and tissue repair, and its biological effect depends on local stimuli, TIMP-mediated regulation, pathway crosstalk, and the stage of disease or treatment. Full article

Crohn’s disease (CD) is a chronic immune-mediated inflammatory disorder characterized by transmural inflammation and a progressive course that frequently leads to structural complications such as intestinal fibrosis. Fibrostenosing disease represents a major clinical challenge, affecting up to 50% of patients over time and often requiring surgical intervention. Despite advances in anti-inflammatory therapies, no effective treatments currently exist to prevent or reverse established fibrosis. Intestinal fibrosis arises from a dysregulated tissue remodeling process driven by excessive extracellular matrix deposition and persistent activation of mesenchymal cells, particularly fibroblasts and myofibroblasts. This process is orchestrated through complex interactions between immune and non-immune cells and mediated by key signaling pathways, including transforming growth factor beta (TGF-β1) and the TL1A/DR3 axis. Genetic susceptibility, notably variants in NOD2 and other fibrosis-related genes, contributes not only to disease risk but also to phenotype progression. Epigenetic mechanisms, particularly microRNAs such as the miR-29 and miR-200 families, further modulate fibrogenesis and represent promising non-invasive biomarkers. Additionally, intestinal dysbiosis and specific microbial signatures, including reduced short-chain fatty acid-producing bacteria and the presence of adherent-invasive Escherichia coli, play a critical role in promoting fibrotic pathways. Mesenteric adipose tissue, especially creeping fat, also contributes to fibrosis through immune and metabolic signaling. Emerging biomarkers related to collagen metabolism and advances in molecular profiling are improving early detection strategies. Novel therapeutic approaches targeting fibrogenic pathways, including anti-TL1A agents, show promising preliminary results. A deeper understanding of these mechanisms is essential to develop effective antifibrotic therapies and improve long-term outcomes in CD. Full article

Pyrrolizidine alkaloids (PAs), a category of naturally occurring secondary metabolites, are commonly found in various botanical sources. Accumulating evidence indicates that PAs and their biologically active metabolites can interact with cellular components and trigger a variety of toxic effects in animals and humans. Notably, PAs exhibit significant hepatotoxic potential via nutritional supplements, environmental dissemination, food chain contamination, and broader ecological pollution. In this review, we summarize PA-induced hepatotoxicity in humans and animals and the underlying molecular mechanisms. It involves oxidative stress, mitochondrial dysfunction, apoptosis, ER stress, inflammation, autophagy, and ferroptosis. Several key signaling pathways, such as nuclear factor-erythroid 2 related factor 2 (Nrf2), mitogen-activated protein kinase (MAPK), protein kinase RNA-like endoplasmic reticulum kinase (PERK), toll like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB), transforming growth factor beta (TGF-β), p53, farnesoid X receptor (FXR), and pregnane X receptor (PXR), are also implicated. Furthermore, this review discusses diagnostic approaches, metabolic activation pathways, and detoxification strategies targeting PA-induced liver injury. Collectively, this review provides a comprehensive understanding of the molecular basis of PA hepatotoxicity and underscores the urgent need for improved risk assessment, early diagnosis, and effective detoxification interventions to mitigate PA-related liver diseases in humans and animals. Full article

Mesenchymal stromal cells (MSCs) are multipotent cells characterized by their regenerative capacity and strong immunomodulatory properties. In recent years, MSC-based therapy has attracted significant attention as a potential treatment for a wide range of immune-mediated and degenerative diseases. The therapeutic effects of MSCs are primarily mediated through paracrine signaling and secretion of cytokines that regulate immune responses and promote tissue repair. This review focuses on five key cytokines involved in MSC immunomodulation: interleukin-6 (IL-6), interleukin-10 (IL-10), transforming growth factor-beta (TGF-β), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β). These cytokines interact within a complex signaling network that allows MSCs to suppress excessive inflammation and restore immune balance. The role of MSC therapy is examined in several clinically relevant conditions, including systemic lupus erythematosus, systemic sclerosis, ischemic stroke, spinal cord injury, diabetes mellitus, and female infertility. Across these diseases, MSCs demonstrate the ability to inhibit pro-inflammatory immune cell activity, promote regulatory immune phenotypes, reduce oxidative stress, and stimulate regeneration through the secretion of growth factors and extracellular vesicles. Despite promising experimental and early clinical findings, several limitations remain, including variability in MSC sources, limited cell survival after transplantation, and the need for optimized dosing strategies. Overall, MSC therapy represents a multifunctional therapeutic approach combining immunomodulation, anti-inflammatory activity, and regenerative support. Further research is required to better understand cytokine interactions, improve standardization of MSC-based treatments, and enhance clinical efficacy across diverse pathological conditions. Full article

Systemic sclerosis (SSc) is a fibrotic disease with high mortality. SSc-associated pulmonary fibrosis (SSc-PF) is currently the leading cause of death. SSc shows a significant sex disparity, with an average sex ratio of 1:5 men to women, yet SSc-PF is more severe in men. We compared gene expression profiles of SSc-PF lung tissues from male and female donors. Whole lung tissues from healthy donors and SSc-PF patients of both sexes were analyzed by RNA sequencing. Selected genes were validated by quantitative polymerase chain reaction and Western blotting analyses. Our results show that genes related to extracellular matrix production were upregulated in females, while genes that are less explicitly related to fibrosis were upregulated in males. Additionally, recombinant transforming growth factor beta (TGFβ) elicited a different response in female and male normal lung fibroblasts. Further, the transcriptomic signatures in male and female lungs only overlapped by 9.19%, highlighting that SSc-PF progresses using different pathways in individuals of different sex. Furthermore, this sex-specific signature of SSc-PF highlights the importance of precision medicine when considering disease-modulating therapies. Full article

Platelet-rich gel supernatants (PRGS) are increasingly used in veterinary medicine due to their regenerative and immunomodulatory properties; however, most studies focus on individual mediators and provide limited insight into their coordinated biological behavior. This study aimed to characterize the integrated inflammatory–regenerative signatures of bovine PRGS stored under different temperature conditions using a multivariate approach. Concentrations of transforming growth factor beta-1 (TGF-β1), tumor necrosis factor alpha (TNF-α), interleukin-2 (IL-2), and interleukin-6 (IL-6) were evaluated in PRGS samples from six clinically healthy cows stored at −80, −20, 4, 21, and 37 °C for up to 326 h. Data were standardized and explored using hierarchical clustering and heatmaps, and principal component analysis (PCA) based on area under the concentration–time curve (AUC) was used to integrate temporal behavior. Temperature-dependent multivariate signatures were identified, with frozen PRGS clustering separately from samples stored at moderate temperatures. The first two principal components explained 43.0% and 28.9% of the variance and defined an inflammatory–regenerative gradient contrasting TGF-β1/IL-2 versus TNF-α/IL-6 profiles. Linear mixed-effects modeling showed that PC1 was significantly affected by temperature and time (p < 0.001), whereas PC2 was influenced by temperature, time, and their interaction (p ≤ 0.048). Differences among temperatures were minimal at early time points but became more pronounced from 48 to 96 h onward, following a temperature gradient with higher values at moderate temperatures and lower values under frozen conditions. These findings indicate that storage temperature reshapes the integrated biological profile of PRGS, rather than merely preserving mediator composition. Full article

Background: Stromal remodeling in the tumor microenvironment contributes to multiple myeloma (MM) progression and drug resistance, but the extracellular mediators that drive these changes remain incompletely defined. Extracellular enolase-1 (ENO1), including membrane-associated and secreted forms, has been implicated in tumor progression; however, its role in cancer-associated fibroblast (CAF)-associated stromal reprogramming in MM is unclear. Methods: The effects of extracellular ENO1 on stromal activation and tumor-supportive functions were examined in MM using MM–bone marrow stromal cell (BMSC) co-cultures, lactate production and viability assays, immunoblotting, cytokine analyses, and a subcutaneous xenograft model of bortezomib (BTZ)-resistant MM in male 6–7-week-old NOD.Cg-Prkdc^scid Il2rg^tm1Vst/Vst (NPG) mice. HuL001, an anti-ENO1 monoclonal antibody, was used to evaluate the therapeutic relevance of extracellular ENO1 targeting. Results: Extracellular ENO1 promoted fibroblast activation protein expression through plasmin-mediated transforming growth factor-β (TGF-β) activation and induced a CAF-associated stromal phenotype marked by enhanced glycolytic activity and increased secretion of tumor-promoting cytokines in MM-BMSC co-cultures. HuL001 suppressed these ENO1-driven effects. HuL001-pretreated stromal cells also exhibited reduced tumor-supportive activity in a BTZ-resistant MM xenograft model. In addition, HuL001 combined with lenalidomide overcame BTZ resistance in MM. Conclusions: Extracellular ENO1 drives CAF-associated stromal reprogramming in the MM microenvironment through the ENO1/plasminogen/plasmin/TGF-β axis. Therapeutic targeting of extracellular ENO1 with HuL001 may disrupt these tumor-supportive stromal activities and help overcome drug resistance in MM. Full article

Endometriosis is a chronic inflammatory disease in which transforming growth factor-beta (TGF-β) has been implicated in immune dysregulation, extracellular matrix remodeling, and fibrosis. Data on baseline secretion of TGF-β isoforms by systemic immune cells remain limited. This pilot study quantified unstimulated secretion of TGF-β1, TGF-β2, and TGF-β3 by peripheral blood mononuclear cell (PBMC) cultures from women with and without endometriosis and explored stage-related patterns. In this prospective case–control study, PBMCs from 50 women with surgically confirmed endometriosis and 30 controls were cultured for 24 h without exogenous stimulation. Supernatant concentrations were measured using a multiplex bead-based immunoassay (Bio-Plex, Bio-Rad) and expressed as pg/mL; between-group and stage-related differences were assessed using non-parametric tests. Median 24 h secretion was similar between groups (TGF-β1: 103,816 vs. 114,700 pg/mL, p = 0.25; TGF-β2: 3735 vs. 3732 pg/mL, p = 0.32; TGF-β3: 3280 vs. 3284 pg/mL, p = 0.70). Within the endometriosis cohort, TGF-β2 was significantly higher in moderate/advanced disease (rASRM stages III–IV) than in minimal/mild disease (stages I–II), whereas TGF-β1 and TGF-β3 did not reach statistical significance for a stage-dependent pattern in this pilot cohort (p = 0.42 and p = 0.41, respectively; Kruskal–Wallis), and a type II error cannot be excluded given the small sample size per rASRM (revised American Society of Reproductive Medicine)stage (n = 11–14). These findings suggest that TGF-β dysregulation is compartmentalized to the peritoneal environment rather than systemically imprinted in circulating immune cells. The stage-dependent elevation of TGF-β2 supports its role in progressive fibrogenesis and as a candidate severity biomarker, warranting confirmation in larger, stimulus-augmented studies. Full article

Mesenchymal stromal/stem cells (MSCs) are increasingly explored for immune-mediated diseases, yet standardized analytical readouts that capture coordinated immunomodulatory output across complementary secretory pathways remain limited. Here, we report the feasibility of an HPLC-based multi-analyte secretome characterization panel that quantifies two small-molecule outputs—adenosine and kynurenine—alongside two immunomodulatory proteins—interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β)—in conditioned media from canine adipose-derived MSCs (cAD-MSCs). Canine immune-mediated hemolytic anemia (IMHA) was used as a disease context to motivate the selection of these analytes, given the pro-inflammatory cytokine environment characteristic of this condition. Three independent cAD-MSC lines were evaluated under baseline conditions and following cytokine stimulation with recombinant interferon-gamma (IFN-γ; 100 ng/mL) and tumor necrosis factor-alpha (TNF-α; 50 ng/mL), referred to herein as inflammatory priming or licensing. Conditioned media were collected at 72 h for metabolite analysis and 48 h for protein analysis, and quantified by HPLC using external calibration and peak integration. Across all three lines, licensing produced directionally consistent increases: mean adenosine increased 2.3-fold, mean kynurenine increased 3.1-fold, mean IL-10 increased 1.6-fold, and mean TGF-β increased 1.7-fold compared with unlicensed controls. Metabolite measurements for adenosine and kynurenine are reported with full chromatographic selectivity data; IL-10 and TGF-β measurements by reversed-phase HPLC with UV detection are presented as exploratory/semi-quantitative outputs and will require orthogonal confirmation (e.g., immunoassay) in future work. These findings are preliminary, derived from three independent donor lines with no comparator group, and are intended to support feasibility of the analytical framework rather than establish definitive performance specifications. Collectively, the data support the potential of a multi-analyte HPLC-based characterization panel to capture licensing-responsive secretory shifts across mechanistically complementary pathways, providing a foundation for expanded development and validation. Full article