Search Results (121)

Field-cultivated roots of Panax quinquefolium represent the natural source of biologically active compounds, e.g., ginsenosides, while transformed roots provide a controlled alternative for their production. Ginsenoside levels from both the sources were determined with the use of the HPLC method. The extracts were tested for antimicrobial activity using the MIC and MBC/MFC methods, as well as for cytotoxic activity on the AGS (gastric cancer) cell line, Hs68 (human fibroblasts), and L929 (mouse fibroblasts) lines using the MTT assay. Additionally, the lack of pro-inflammatory activity of the plant materials was assessed using a monocyte activation test. The tested P. quinquefolium roots differed quantitatively and qualitatively in their ginsenoside profiles, and the highest amount was recorded in the transformed roots (204.62 ± 5.56 mg/g extract ± SE). The extracts exhibited the strongest antimicrobial activity against the Escherichia coli strain. Low activity of the tested extracts was observed against Candida species. In the tested cell lines (AGS, Hs68, L929), a dose-dependent decrease in cell viability was observed, with the field root extract exhibiting the highest cytotoxic activity in the concentration range of 2.5–10 mg/mL. All tested extracts proved to be safe and did not stimulate a pro-inflammatory response. Full article

The worldwide rise in antimicrobial resistance, along with the ongoing prevalence of cancer, underscores the pressing need for novel, safe, and multifunctional therapeutic candidates. Medicinal plants continue to serve as a valuable source of chemically diverse bioactive molecules that modulate multiple biological targets. In this investigation, the preliminary screening of the antibacterial and anticancer activities of an ethanolic extract of Alhagi maurorum (A. maurorum) was comprehensively evaluated using integrated chemical characterization, in vitro bioassays, and in silico approaches. A liquid chromatography–mass spectrometry (LC-MS) analysis demonstrated a rich phytochemical profile including glucosinolates, phenolic acids, gallotannins, fatty acids, alkaloids, carotenoid derivatives, and 2-hexyldecanoic acid-associated constituents. Antibacterial efficacy was assessed by disk diffusion and minimum inhibitory concentration (MIC) testing against Escherichia coli (E. coli) and Bacillus cereus (B. cereus), with the extract producing inhibition zones similar to those observed with streptomycin. Anticancer effects were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays with MCF-7 breast carcinoma cells and Hs27 normal fibroblasts over 24, 48, and 72 h, revealing a time-dependent, selective decrease in malignant cell viability with relatively limited toxicity towards normal cells. Induction of apoptosis was further verified by propidium iodide (PI) staining. A molecular docking analysis highlighted 2-hexyldecanoic acid as the principal active compound, with a strong binding affinity for critical bacterial targets (GyrA, GyrB, and RpoB). In silico toxicity and ADME (absorption, distribution, metabolism, and excretion) assessments indicated favorable drug-like properties, good gastrointestinal uptake, and acceptable safety profiles. Altogether, these results provide combined experimental and computational support for A. maurorum as a promising source of dual-purpose antibacterial and anticancer agents and lay a mechanistic foundation for subsequent preclinical studies. Full article

Background: Apoptotic vesicles (ApoVs) derived from adipose stem cells (ASCs) have recently emerged as important mediators of tissue repair and are implicated in pathways relevant to hypertrophic scar (HS). Although ASCs exhibit potential in scar modulation, the therapeutic value of their apoptotic clearance products remains largely unexplored. Methods: In this study, we investigated the efficacy and mechanism of staurosporine (STS)-induced adipose stem cell derived apoptotic vesicles (ASCs-ApoVs) in mitigating HS. Western blot, RT-qPCR, and immunofluorescence were used to assess fibrotic markers including α-SMA, COL1A1, and COL3A1 and so on in hypertrophic scar derived fibroblasts (HS-fibroblasts). Results: ASCs-ApoVs significantly reduced profibrotic marker expression in HS-fibroblasts without short-term cytotoxicity. CDC20 down-regulation was identified as a critical target, through which ASCs-ApoVs suppressed Wnt/β-catenin signaling, as evidenced by the downregulation of β-catenin, c-MYC, Cyclin D1, and AXIN2. The efficacy of ASCs-ApoVs in hypertrophic scar regulation was also confirmed by the rabbit ear scar model. Furthermore, ASCs-ApoVs demonstrated notable structural and functional stability. Conclusions: In summary, our results established STS-induced ASCs-ApoVs as a potent multi-target strategy for hypertrophic scar regulation. Besides, the scalable production, functional stability, and favorable safety profile of ASCs-ApoVs underscore a strong promise for clinical translation. Full article

Fisetin (FIS) is a bioactive flavonoid with antioxidant, anti-inflammatory, and anticancer activity, but its poor aqueous solubility and high lipophilicity limit its therapeutic use. In this study, three-component FIS-loaded mixed micelles based on Poloxamer 407 (P407) or Poloxamer 188 (P188), sodium deoxycholate, and Kolliphor HS15 or Kolliphor ELP were developed and comparatively evaluated. The formulations were prepared by the thin-film hydration method and characterized in terms of physicochemical properties, storage stability, solid-state properties, and in vitro biological activity. All freshly prepared formulations formed nanosized systems with high encapsulation efficiency. Although P188-based micelles showed smaller initial particle sizes, P407-based systems exhibited superior stability after lyophilization and rehydration. Formulations containing Kolliphor ELP showed the most favorable stability profile over 28 days of storage. FT-IR, TG, DSC, and XRPD analyses confirmed successful incorporation of FIS into the polymeric matrix and transformation of the drug into an amorphous or molecularly dispersed state. In vitro studies demonstrated that micellar encapsulation enhanced the cytotoxic activity of FIS against MICH-2 melanoma cells compared with the free compound, while P407-based systems showed a more favorable safety profile toward MRC-5 fibroblasts. These findings indicate that P407-based mixed micelles, particularly those containing Kolliphor ELP, may serve as promising nanocarriers for improving FIS delivery with potential relevance for dermal and anticancer applications. Full article

Inflammation is associated with metabolic alterations that can lead to the release of volatile organic compounds (VOCs) reflecting cellular biochemical activity. Profiling these volatile metabolites may provide insight into cellular responses to inflammatory stimuli, although their characterization in skin-derived cells remains limited. In this exploratory proof-of-concept study, we investigated the volatile metabolite profiles of human skin fibroblasts exposed to different inflammatory stimuli. Fibroblast cell lines were stimulated with polyinosinic:polycytidylic acid (Poly I:C), tumor necrosis factor-alpha (TNF-α), and lipopolysaccharide (LPS) to model viral-, cytokine-, and bacterial-associated stress conditions. Headspace solid-phase microextraction coupled with comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (HS-SPME-GC×GC-TOFMS) was applied to analyze volatile metabolites released from the cell cultures, enabling exploratory profiling of the fibroblast volatilome. A data-processing workflow including pairwise comparisons between experimental groups and statistical filtering was implemented to identify volatile features associated with the different conditions. Several VOCs were tentatively identified, mainly belonging to alcohol, ester, and hydrocarbon classes, and showed differential abundance patterns between stimulated and control samples. Multivariate analysis indicated a separation between stimulated and non-stimulated groups, suggesting stimulus-associated differences in the volatile profiles of fibroblast cultures. While these observations may reflect metabolic responses occurring under inflammatory stimulation, the chemical identity and biochemical origins of several detected features remain to be confirmed. All in all, this study demonstrates the feasibility of applying HS-SPME-GC×GC-TOFMS-based volatilome profiling to investigate stimulus-associated changes in fibroblast cultures. The detected VOC patterns should therefore be considered preliminary observations requiring further chemical characterization and independent validation. Future studies including larger sample numbers, complementary biological verification of the inflammatory response, and more physiologically relevant experimental models will be necessary to further assess the robustness and potential relevance of these volatile signatures in the context of inflammatory processes. Full article

Background/Objectives: Non-covalent nanocarrier-based systems have become a promising platform as they offer a strategy to improve the efficacy-safety profile of doxorubicin (DOX) without altering its chemical structure. Praised for biocompatibility and rich surface chemistry, nanodiamonds (NDs) have launched as nanocarriers of choice for advanced cancer therapy. By investigating DOX-ND physicochemical interactions, this work advances the structural understanding of a non-covalent potential anticancer system, which has not been quantitatively experimentally explored so far. Methods: To our knowledge, this is among the first studies combining ultraviolet–visible (UV–VIS) spectroscopy with spectral deconvolution to reveal the redistribution of different DOX species in the presence of NDs. Centrifugation-assisted analysis enabled differentiation between hypothetical labile and stable ND/DOX fractions. Adsorption kinetics was studied, and dynamic light scattering (DLS) measured particle size and zeta potential. In vitro screening was performed in non-malignant fibroblasts (MRC-5) and malignant melanoma (HS294T), glioblastoma (U251), and breast cancer (MCF-7) cells to evaluate ND/DOX combinations. Results: Centrifugation analysis revealed heterogeneous ND-DOX binding. Kinetic experiments showed fast multi-stage adsorption kinetics, best described by a bi-exponential decay function and the Weber–Morris model. DLS suggested stable systems with a particle size within 10–80 nm, predominantly around 20 nm, and positive zeta potential. Comparative in vitro screening demonstrated differential cellular responses across cell types, highlighting the relevance of ND/DOX interactions. Conclusions: The findings contribute to elucidating ND-DOX interactions relevant for the design and optimization of drug delivery systems, emphasizing the importance of spectroscopic insights for the design of nanodiamond-based drug delivery systems. Full article

Background/Objectives: Hidradenitis suppurativa (HS) is a chronic, immune-mediated inflammatory skin disease characterized by painful nodules, abscesses, sinus tracts, and progressive fibrosis. Vascular activation is becoming increasingly acknowledged as an important factor in HS pathogenesis; however, the effects of tumor necrosis factor alpha (TNF-α) blockade on vascular remodeling in HS remain poorly characterized. This study investigated the impact of TNF-α inhibition by adalimumab (ADA) on endothelial and fibroblast-associated markers in HS lesions. Methods: Formalin-fixed paraffin-embedded skin samples from 71 HS patients were analyzed, including treatment-naive (n = 38) and adalimumab-treated (n = 33) cases. Histopathology and immunofluorescence were performed using antibodies against CD31, von Willebrand factor (vWF), α-smooth muscle actin (αSMA), vimentin, Ki-67 (proliferation), and cleaved Caspase-3 (apoptosis). ImageJ software was used to determine the immunoexpression of selected markers and vascular density. Vascular density, assessed as vessel count per mm², was designated as the primary endpoint. Sex-related differences were analyzed as exploratory endpoints. Results: Adalimumab-treated tissue exhibited significantly reduced vascular density (p < 0.01) compared to the treatment-naive group. Conversely, vimentin immunoexpression was significantly higher (p < 0.01) in the adalimumab-treated group. No significant differences were found in endothelial Ki-67 or cleaved Caspase-3 expression between treatment groups, indicating that the observed reduction in vascular density is not associated with direct effects on endothelial cell proliferation or apoptosis, but rather may occur indirectly through attenuation of the pro-angiogenic inflammatory milieu. Exploratory sex-stratified analysis revealed that treatment-naive males had significantly higher endothelial proliferation (Ki-67; p = 0.031) and vimentin expression (p = 0.017) compared to treatment-naive females. In the ADA-treated group, males exhibited significantly lower vascular density (p = 0.036) and higher endothelial apoptosis (p = 0.039) compared to females, whereas females showed a significant increase in vimentin expression following treatment (p = 0.008), suggesting possible sex-dependent differences in vascular remodeling. Conclusions: TNF-α blockade is associated with reduced vascular density, consistent with indirect anti-angiogenic effects, suggesting that adalimumab exerts disease-modifying effects on the microenvironment beyond inflammatory cytokine suppression. Sex-dependent differences in vascular regression underscore the importance of considering sex as a biological variable in HS pathogenesis and treatment response. These results highlight the significance of vascular interactions in HS and support adalimumab as a disease-modifying treatment. These exploratory findings require confirmation in longitudinal studies with paired biopsies. Full article

Early prevention of pathological changes underlying gastric cancer (GC) development is a critical strategy, offering the most effective opportunity to limit malignant progression and improve patient outcomes. We have previously demonstrated that Helicobacter pylori (Hp) (cagA⁺vacA⁺) contributes to GC development by activating gastric fibroblasts toward CAF-like phenotype, eliciting aggressive, cancer stem cells (CSCs)-related malignant transformation of LGR5⁺ normal epithelial cells. A key mediator of these processes appears to be the NF-κB/STAT3 axis. Therefore, our aim was to investigate the protective role of hydrogen sulfide (H₂S) as a potential novel strategy for counteracting Hp-induced fibroblast reprogramming. Human fibroblasts were infected with Hp (cagA+vacA+) for 120 h. The fast-releasing H₂S donor NaHS (50, 100, 200 and 400 µM) was added every 24 h. Activation markers, corresponding signaling pathways, H₂S release and activities of H₂S-metabolizing enzymes were determined. NaHS reduced Hp-induced fibroblast activation and their pro-inflammatory, pro-tumorigenic markers, which was associated with the inhibition of NF-κB/STAT3 axis and Twist expression. Additionally, it modulated sulfur metabolism while preserving sulfur-enzyme homeostasis. NaHS limited Hp adhesion (high doses), reduced reinfection-induced activation and increased sensitivity of Hp to metronidazole. These findings suggest that H₂S signaling may represent a modulatory factor of NF-κB/STAT3-driven inflammatory responses during Hp infection and warrant further investigation. Full article

Breast cancer remains a leading cause of cancer-related morbidity and mortality globally, highlighting the urgent need for novel therapeutic strategies. This study investigates the molecular mechanisms underlying the anti-proliferative potential of Cannabis sativa dichloromethane extract (C. sativa DCM) on oxidative stress, apoptosis, and invasion in human breast cancer cells. Key biomarkers, such as antioxidant enzymes (Superoxide Dismutase (SOD) and Glutathione (GSH)), the transcription factor Nrf2, apoptotic proteins (p53, caspase-8 and 9), metalloproteinase (MMP-1 and MMP-9), and Transforming Growth Factor Beta (TGF-β) were examined. Cytotoxicity was assessed using an MTT assay in the MDA-MB-231 and MCF-7 breast cancer cell lines, with comparisons to normal skin fibroblasts (HS27). Oxidative stress biomarkers were quantified using enzymatic assays and ELISA kits, while apoptotic and anti-metastatic factors were determined by Western blotting. Results demonstrated that C. sativa DCM extract induced significant cell death in a concentration-dependent manner, with IC50 values of 75.46 ± 0.132 μg/mL for MDA-MB-231 and 78.68 ± 0.50 μg/mL for MCF-7 cells. The extract decreased SOD and GSH levels while increasing p53 and caspase activity, confirming apoptosis activation. Additionally, C. sativa DCM inhibited migration and invasion by downregulating MMP-1, MMP-9, and TGF-β. The anti-proliferative potential of C. sativa DCM in breast cancer cells is mediated through a continuous biological pathway involving oxidative stress modulation, apoptotic signaling, and anti-invasive effects. Phytochemical analysis revealed terpenoids and steroids, including compounds like cannabidiol and tetrahydrocannabinol acid. These findings suggest that C. sativa DCM extract holds potential as an anti-breast cancer therapeutic and warrants further preclinical and clinical investigations. Full article

Background/Objectives: Hypertrophic scar (HS) is a fibroproliferative disorder characterized by excessive fibroblast activation and collagen deposition. The role of PIWI-interacting RNAs (piRNAs) in HS pathogenesis has not been defined. This study aimed to identify HS-related piRNAs, clarify their molecular mechanisms, and evaluate their therapeutic potential. Methods: High-throughput piRNA sequencing was performed on hypertrophic scar and matched normal tissues, followed by validation in patient-derived samples and dermal fibroblasts using quantitative reverse transcription PCR. Functional assays, including proliferation, apoptosis, migration, and invasion assays, were conducted after transfection with piRNA mimics or inhibitors. RNA sequencing, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes enrichment analyses, as well as dual-luciferase reporter and rescue assays, were used to identify and confirm molecular targets. Results: Sequencing revealed piR-hsa-022095 as one of the most significantly upregulated piRNAs in HS. Its inhibition suppressed fibroblast viability, migration, and invasion while inducing apoptosis and G₀/G₁ arrest. Transcriptomic profiling identified cell-cycle–related genes as major downstream targets, with KLF11 emerging as the principal effector. piR-hsa-022095 targets the 3′ UTR of KLF11, repressing its expression and thereby facilitating fibroblast proliferation. Restoration of KLF11 reversed the pro-fibrotic effects of piR-hsa-022095, confirming its functional role in HS pathogenesis. Conclusions: This study identifies piR-hsa-022095 as a novel regulator implicated in HS formation through repression of KLF11. The piR-hsa-022095–KLF11 axis may represent a previously unrecognized regulatory pathway involved in hypertrophic scar formation, providing new insights into the molecular mechanisms underlying HS pathogenesis. Full article

The article presents the results of structural, corrosion, microbiological, biological, and genotoxicity studies on the effect of graphene oxide deposited on a flat titanium foil surface, intended for use, in general, implantology and other medical applications. The methodology of graphene oxide (GO) deposition involved a surface cleaning process combined with RF plasma activation, followed by the application of a thin layer of dispersed aqueous GO suspension using a spin coater. The graphene oxide layer was uniformly deposited on the surface, which was confirmed by SEM imaging. Corrosion studies were carried out in an electrochemical cell filled with a buffered solution prepared to mimic the composition of physiological intracellular fluids. It was demonstrated that the deposition of graphene oxide on the titanium surface limited the access of electrolyte and oxygen. Surface activation and deposition of the aqueous graphene oxide suspension contributed to improved adhesion, condition, growth, and proliferation of fibroblast cell lines Hs 895.T and Hs 895.Sk. The inhibition zone analysis revealed a bacteriostatic effect against Pseudomonas aeruginosa and Staphylococcus aureus. Moreover, no genotoxicity changes were observed. Full article

This study presents a novel hydrogel formulation combining mupirocin, a broad-spectrum antibiotic, with keratinocyte growth factor (KGF) to enhance wound healing through antibacterial action and tissue regeneration. Mupirocin was encapsulated in hydroxypropyl β-cyclodextrin (HP-β-CD) and stabilized with poly(amidoamine) dendrimers (PAMAM). Molecular docking studies assessed mupirocin’s binding to PAMAM and its interaction with isoleucyl-tRNA synthetase. Physicochemical properties—including zeta potential, particle size, and surface tension—were characterized, and drug release kinetics were evaluated using Franz diffusion cells. In vitro assays on human dermal fibroblasts (HS27) included proliferation, scratch wound healing, and flow cytometry to assess cellular behavior. Antibacterial efficacy was determined via the Kirby–Bauer disk diffusion method. Results showed strong binding of mupirocin to its target enzyme, enhanced by KGF. The hydrogel exhibited favorable properties: surface tension of 24.7 dyne/cm, zeta potential of −24.79 mV, and particle size of ~119 nm, indicating high stability. Franz diffusion revealed sustained drug release compared to commercial mupirocin. Cellular assays demonstrated significant fibroblast migration and proliferation, with flow cytometry confirming increased wound healing markers. The formulation showed potent antimicrobial activity, including against Methicillin-resistant Staphylococcus aureus (MRSA), highlighting its promise for infected wound treatment and advanced clinical wound care. Full article

The Atlantic blue crab (Callinectes sapidus) is an opportunistic invasive species in the Mediterranean that is negatively affecting biodiversity, fisheries, and tourism. In Italy, it is appreciated for its good meat quality, but the processing yield is low (21.87 ± 2.38%), generating a significant amount of by-products (72.45 ± 4.08%), which are underutilized. Valorizing this biomass is in line with circular economy principles and can improve both environmental and economic sustainability. This study aimed to valorize Atlantic blue crab by-products (BCBP), producing protein hydrolysates and assessing their in vitro bioactivities, in order to plan applications in animal food and related sectors. BCBP hydrolysates were obtained by enzymatic hydrolysis using Alcalase and Protamex enzymes. The treatment with Alcalase resulted in a higher degree of hydrolysis (DH = 23% in 205 min) compared to Protamex (DH = 14% in 175 min). Antioxidant activity of the hydrolisates was evaluated through DPPH, ABTS, reducing power and FRAP assays, as well as in vitro test in fibroblasts (HS-68). At 10 mg/mL, hydrolysates from both enzymes exhibited the maximum radical scavenging activity in DPPH and ABTS assays. In HS-68 cells, 0.5 mg/mL hydrolysates protected against H₂O₂-induced oxidative stress, showing a cell viability comparable to cells treated with 0.5 mM N-acetyl cysteine (NAC), as an antioxidant. Statistical analyses were performed using one-way ANOVA followed by Student–Newman–Keuls (SNK) or Games–Howell post hoc tests, with significance set at p < 0.05. Overall, both enzymes efficiently hydrolyzed BCBP proteins, generating hydrolysates with significant antioxidant activity and cytoprotective effects. These results demonstrate the potential to produce high-quality bioactive compounds from BCBPs, suitable for food, nutraceutical, and health applications. Scaling up this valorization process represents a viable strategy to improve sustainability and add economic value to the management of this invasive species, turning a problem in a resource. Full article

Skin repair is essential in the treatment of burns and wounds. After an injury, the concept of tissue engineering emerges to restore skin function and facilitate wound healing. This field often involves the use of biodegradable and biocompatible materials as a primary scaffold for tissue regeneration. In this study, a PHB/Collagen wound dressing mat loaded with the antimicrobial peptide LL37 was developed via electrospinning. The polymer solutions were prepared by dissolving polyhydroxybutyrate (PHB) biopolymer extracted from Cereibacter sphaeroides, commercial PHB, and marine collagen in hexafluoroisopropanol (HFIP). The resulting nanofibers were characterized using Field-Emission Scanning Electron Microscopy (FE-SEM), Thermogravimetric Analysis (TGA), X-Ray Diffractometry (XRD), and an Optical Tensiometer. Antibacterial activity assessments were conducted against Staphylococcus aureus (ATCC 29213) and Escherichia coli (ATCC 25922). Degradability studies were carried out in DMEM medium, cytotoxicity tests were performed on the L929 fibroblast cell line, and the wound healing effect was investigated on the HS2 keratinocyte cell line. To evaluate the properties of the designed material under in vitro conditions, the morphology of cells on the nanofiber was examined using an inverted light microscope. The findings demonstrated that the nanofibers were biocompatible in vitro and exhibited no toxic effects. And, compared to the control groups, the 5.56 nmol LL37-loaded PHB/Collagen nanofibers significantly enhanced wound closure by 15–30% and effectively reduced the viability of S. aureus and E. coli by 20–25% and approximately 80–85%, respectively. These results highlight the therapeutic potential of LL37-loaded PHB/Collagen nanofibers for use in wound healing applications. Full article

Background/Objectives: Idiopathic pulmonary fibrosis (IPF) is a progressive disease characterized by lung scarring, impaired function, and high mortality. Effective therapies to reverse fibrosis are lacking. This study aims to uncover the molecular mechanisms of IPF, explore diagnostic biomarkers, and identify therapeutic targets. Methods: Multi-omics data were integrated to identify biomarkers with causal associations to IPF using Mendelian randomization and transcriptomic analysis. Machine learning was employed to construct a diagnostic model, and single-cell transcriptomic analysis determined gene expression patterns in fibrotic lung tissue. Results: Seven core genes (GREM1, UGT1A6, CDH2, TDO2, HS3ST1, ADGRF5, and MPO) were identified, showing strong diagnostic potential (AUC = 0.987, 95% CI: 0.972–0.987). These genes exhibited distinct distribution patterns in fibroblasts, endothelial cells, epithelial cells, macrophages, and dendritic cells. Conclusions: This study highlights key genes driving IPF, involved in pathways related to metabolism, immunity, and inflammation. However, their utility as fluid-based biomarkers remains unproven and requires protein-level validation in prospective cohorts. By integrating genomic, immunological, and cellular insights, it provides a framework for targeted therapies and advances mechanism-based precision medicine for IPF. Full article