Angelica sinensis (dong quai) is a widely used Traditional Chinese Medicine herb whose constituents are associated with antioxidant and anti-inflammatory activity, making it a plausible candidate for skin-care applications. Here, we evaluated whether an Angelica sinensis extract containing 1% ligustilide exerts antioxidant, anti-senescence, antimicrobial, and anti-inflammatory effects in human skin cell models and explored extracellular matrix-related readouts. Human keratinocytes (HaCaT) and dermal fibroblasts (BJ-5ta) were treated with the extract (10, 30, 50 μg/mL); anti-inflammatory activity was assessed by lipopolysaccharide-induced nuclear factor κB (NF-κB) p65 nuclear translocation, antioxidant effects by reactive oxygen species detection with and without hydrogen peroxide challenge, senescence by senescence-associated β-galactosidase after UVB irradiation, and matrix support by collagen type I alpha 1 (COL1A1) immunofluorescence in fibroblasts; scratch-wound assays evaluated migration, and broth microdilution tested activity against Staphylococcus aureus, Staphylococcus epidermidis, Cutibacterium acnes, and Malassezia globosa. The extract reduced lipopolysaccharide-induced NF-κB nuclear localization in keratinocytes, lowered basal and hydrogen peroxide-induced reactive oxygen species at 3 and 24 h in both cell types, and attenuated ultraviolet B (UVB)-induced senescence in keratinocyte and fibroblast models; in fibroblasts, it increased COL1A1 signal, while migration was unchanged. The extract showed strong inhibition of Staphylococcus species and partial, concentration-dependent inhibition of C. acnes and M. globosa after 24 h, with acceptable viability across the tested range. Overall, these results indicate that A. sinensis extract combines anti-inflammatory and antioxidant activity with reduced UVB-associated senescence, fibroblast matrix-supportive signaling, and antimicrobial effects against skin-relevant microbes, supporting further evaluation in more advanced skin models and finished-product formulations before inferring topical performance.

Morus alba L., a member of the Moraceae family, is known for its positive effects on human health, linked to the presence of different classes of secondary metabolites, including flavonoids, stilbenoids, and alkaloids, found in different parts of the plant. Stilbenoids, in particular, are mainly present at the root cortex level and, owing to their valuable activities, have attracted scientific interest in recent years. Since roots are a non-renewable source, in this study, M. alba in vitro callus cultures were established. The biomass with the appropriate growth and texture was selected for juice extraction, and the total phenol, flavonoid, and proanthocyanidin contents, along with the antioxidant activity, were estimated in the juices. The analyses throughout the callus growth cycle revealed the juice of 14-day-old calli to be the richest, resulting in the most active. In this juice, the LC-MS/MS-DAD analysis unveiled the presence of seventeen stilbenoids. Together with the data obtained by the nutritional analysis, the results showed that M. alba cell cultures have the potential to be utilised for producing innovative healthy food materials, bridging the gap between the ever-increasing natural-based-product demand and the need for more environmental, social, and economic development.

Viral infections with gastrointestinal involvement remain a significant global health burden with limited therapeutic options. While probiotics show antiviral potential, their impact on primary human intestinal epithelial defenses is poorly defined. This study utilized human intestinal organoid-derived monolayers (ODMs), generated from the non-inflamed mucosa of patients with inflammatory bowel disease, to examine how Bifidobacterium animalis ssp. lactis BB-12 (BB-12) and Lacticaseibacillus rhamnosus GG (LGG) modulate mucosal antiviral pathways. Unlike conventional Caco-2 cells, ODMs preserved physiological cellular diversity and intact innate signaling. Expression of viral receptors and interferon (IFN)-stimulated genes (ISGs) was quantified by RT-qPCR, while the effector 2'-5'-oligoadenylate synthetase 1 (OAS1) was also assessed by immunofluorescence and flow cytometry. Both probiotic strains modulated IFN-associated pathways; however, BB-12 induced a markedly stronger antiviral transcriptional response than LGG. Notably, OAS1 exhibited cell type-specific regulation; while goblet cells showed high basal levels, both probiotics enhanced OAS1 expression selectively in ileal enterocytes. Despite this shared effect, only BB-12 pretreatment significantly restricted Influenza A (H1N1) replication in ileal ODMs, whereas LGG did not significantly affect viral replication. These findings establish human ODMs as a superior platform for probiotic immunology, suggesting that BB-12 more effectively shapes epithelial antiviral "set-points" and highlighting OAS1 as a sensitive component of a broader antiviral program.

Chitosan is a natural polysaccharide produced by the deacetylation of chitin. It has been employed for several biomedical applications since it is biocompatible and biodegradable. In this work, two synthetic peptides were covalently grafted to chitosan to enhance cellular response. The first sequence, the nonapeptide (351–359) of human vitronectin (HVP), was demonstrated to enhance osteoblast adhesion and migration. The second peptide, GBMP1α, a 22 mer sequence reproducing the fragment (48–69) of BMP-2, increased the osteogenic differentiation of mesenchymal stem cells. The anchoring chemistry involves the formation of a Schiff base between the amino groups of chitosan and aldehyde groups specifically introduced into the peptide backbone, followed by a subsequent reduction step. One of the advantages of this method is the specific anchoring of a bioactive sequence in a single-step reaction carried out in an aqueous solution and mild conditions. Six different 3D-porous scaffolds were designed and obtained by various combinations of both functionalized chitosan and chitosan simultaneously functionalized with both peptides. Scaffolds were characterized through X-ray photoelectron spectroscopy (XPS), nuclear magnetic resonance (NMR), Fourier-transform infrared spectroscopy (FT-IR), and mechanical measurements. Biological assays revealed that functionalized scaffolds exhibited significantly improved adhesion, proliferation, and mineralization in human jaw-derived osteoblasts compared to pristine chitosan. Results from mechanical measurements highlighted the ability to modulate compressive modulus and maximum stress. Notably, scaffolds containing both peptides demonstrated significant synergistic effects in vivo, enhancing material cellular colonization and reducing fibrotic capsule thickness. This study highlights the potential of peptide-functionalized chitosan scaffolds in advancing bone tissue engineering and addressing current limitations in bone repair strategies.

Capparis spinosa L. is a Mediterranean species traditionally used for the treatment of inflammatory and skin-related disorders and increasingly explored as a source of multifunctional ingredients for dermocosmetic applications. In this study, three commercially available C. spinosa fruit extracts obtained through different extraction strategies (two aqueous extracts, CAP1 and CAP2, and one hydroalcoholic extract, CAP3) were comparatively investigated using an integrated phytochemical and biological approach. Phytochemical profiling by LC-DAD-ESI-MS/MS revealed a complex composition dominated by flavonol glycosides (mainly quercetin- and kaempferol-based derivatives), phenylpropanoid esters, and glucosinolates, which define the core chemical signature of caper fruits. While the extracts shared a largely overlapping qualitative profile, marked differences in relative metabolite abundance were related to the extraction solvent, the drug-to-extract ratio, and formulation parameters. The biological relevance of the extracts was evaluated using skin-related cellular models and antimicrobial assays. All extracts showed good cytocompatibility in human keratinocytes and preserved epithelial barrier integrity. Distinct, concentration-dependent effects were observed on intracellular ROS modulation, UVB-induced cellular senescence, collagen type I production in dermal fibroblasts, and antimicrobial activity. CAP1 exhibited the most pronounced antioxidant and antimicrobial effects, CAP2 showed intermediate and balanced activity, whereas CAP3 displayed a more complex redox behaviour combined with marked anti-senescent activity. Overall, these findings demonstrate that extraction strategy critically influences the biological performance of C. spinosa fruit extracts and support their potential as multifunctional dermocosmetic ingredients targeting skin homeostasis, photoaging, and the balance of the skin microbiome (including bacterial and fungal components).

The anion exchanger 1 (AE1), traditionally known for its role in erythrocyte anion transport and acid–base homeostasis, has recently been identified in non-erythroid tissues, suggesting broader physiological functions. In the present study, we investigated for the first time the expression and potential role of AE1 in human ovarian granulosa cells (GCs) obtained from women with endometriosis (ENDO-GCs) or male factor infertility controls (MF-GCs). Cells were cultured in the presence of follicular fluid derived from control (FF-MF) or endometriosis patients (FF-ENDO), and DIDS-sensitive anion exchange activity was pharmacologically inhibited using 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS). AE1 expression was evaluated at both protein and transcript levels together with markers of proliferation, inflammation, and steroidogenesis. The results demonstrate that AE1 is constitutively expressed in GCs and may contribute to granulosa cell homeostasis. Inhibition of DIDS-sensitive anion exchange activity inhibited cell proliferation, shifted cell morphology toward a fibroblast-like phenotype, and reduced estradiol and progesterone secretion and inflammatory (IL-6) gene transcription. Notably, MF-GCs cultured in FF-MF exhibited compensatory upregulation of AE1, whereas ENDO-GCs and cells exposed to FF-ENDO showed impaired adaptive responses and generalized transcriptional suppression. These findings provide preliminary evidence supporting a role for DIDS-sensitive anion exchange activity in granulosa cell physiology and warrant further studies to clarify the specific contribution of AE1 to follicular dysfunction associated with endometriosis. However, the specific mechanistic contribution of AE1 could not be established and will require further functional and genetic investigations.

Botulism is a life-threatening disease caused by botulinum neurotoxins (BoNTs) released by Clostridium bacteria. BoNTs induce a flaccid paralysis by blocking neurotransmitter release from peripheral cholinergic neurons via soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) cleavage. The enteric nervous system (ENS) controls antimicrobial defense reactions within the intestine through neuron-released mediators. Here, we show that subclinical doses of BoNT/A and BoNT/B, which are not sufficient to cause botulism when released in the intestinal lumen, paralyze ENS neurons favoring infection by prototypic bacterial pathogens such as Salmonella enterica and Shigella flexneri. The BoNTs reduce the release of mucin, decreasing the protective barrier against bacterial infection. These findings disclose previously unrecognized functions of BoNTs and roles in intestinal infections. Moreover, they indicate that BoNTs are valuable tools for investigating the ENS neuron involvement in the intestinal defense reactions.

Oral diseases like periodontitis and tooth loss affect billions worldwide, causing alveolar bone resorption and complicating implant placement and bone regeneration. Guided bone regeneration addresses these defects using barrier membranes that block soft tissue infiltration and promote bone growth. CollaTape®, a type I bovine collagen membrane, is widely used for its biocompatibility and resorbability, though its bioactivity and antibacterial properties could be improved. This study compares two functionalization methods for enhancing CollaTape® membranes: adsorption of GBMP1α peptide (a BMP-2 biomimetic) and covalent anchoring of its analogue Aoa-GBMP1α. Both functionalizations were performed at concentrations of 0.25, 0.5, 1, and 1.5 mg ml-1. Optimal conditions were selected basing on osteoblast mineralization assays and resulted to be 0.25 mg ml-1for adsorption and 1.5 mg ml-1for covalent binding. Peptide surface density analysis revealed values of 0.040 μmol cm-2for adsorption and 0.278 μmol cm-2for covalent anchoring. Biological assays assessed mineralization, proliferation, and gene expression (SPP1, RUNX2) in human osteoblasts, and antibacterial activity againstStaphylococcus aureusandEscherichia coli. All functionalized membranes improved osteoblast activity, with adsorption showing superior results. Antibacterial tests showed slight but significant reductions in bacterial colonies, especially for adsorption. Additional mechanical tests via unconfined compression were performed to evaluate the effect of functionalization on the membranes' mechanical properties. These tests confirmed that neither functionalization method compromised the stiffness of the membrane, a critical parameter in clinical applications. Overall, peptide adsorption is a simple and clinically adaptable strategy to enhance CollaTape®'s bioactivity and antibacterial properties while maintaining their original mechanical properties.

Endometriosis (ENDO) and poor ovarian response (POR) represent challenging conditions in assisted reproduction. Both, associated with altered follicular fluid (FF) composition, specifically impact on granulosa cell (GC) function in an incompletely understood way. GCs from male factor (MF, n = 30), ENDO (n = 38), and POR (n = 27) patients were cultured in media supplemented with FF from each group (FF-MF, FF-ENDO, FF-POR). Proliferation, morphology, and secretory activity (cortisol, estradiol, progesterone, IL-6) were assessed. GC proliferation depended primarily on FF origin, being highest with FF-ENDO, intermediate with FF-POR, and lowest with FF-MF. Morphological analysis revealed enrichment of muscle-like and fibroblast-like morphologies under FF-ENDO and FF-POR, suggestive of dysregulated luteinization and extracellular matrix remodeling. Secretory activity reflected a complex interplay between GC origin and FF type: IL-6 was strongly induced by FF-MF and FF-POR but consistently suppressed by FF-ENDO; cortisol and estradiol were generally consumed, while progesterone synthesis was largely confined to MF-GCs, with only variable induction in ENDO-GCs exposed to FF-POR. These findings indicate that pathological FF milieus reprogram GC behavior in distinct ways, with potential consequences for luteal function and oocyte competence. Identifying the molecular mediators of these alterations may guide tailored strategies to improve ART outcomes in ENDO and POR patients.

Background: Osteoarthritis (OA) is a chronic degenerative whole joint disease characterized by cartilage breakdown and inflammation. Galectin-3 (Gal-3), a β-galactoside-binding lectin secreted into the extracellular space, binds to glycosylated components of the extracellular matrix (ECM), modulating cell–matrix interactions and inflammation. This study aims to evaluate the anti-inflammatory effects of Hylach®, a hyaluronic acid (HA) derivative conjugated with lactose-based residues that bind Gal-3, on in vitro inflamed primary human chondrocytes. Methods: Chondrocyte viability, after both Hylach® and HA treatments at different concentrations was assessed using the MTT assay. Two-dimensional and 3D cell cultures exposed to the conditioned medium (CM) of activated U937 monocytes and subsequently treated with Hylach or HA, were analyzed for the expression of IL-1β, IL-6, TNF-α, and Gal-3 at different time points (4, 10, and 24 h). Results: HA and Hylach® did not affect cell viability at any of the tested concentrations. Both molecules reduced the overexpression of Gal-3 and pro-inflammatory molecules in 2D inflamed cell cultures, at both gene and protein levels. Notably, IL-1β, IL-6 and Gal-3 showed a more pronounced inhibitory effect at 4 h, with Hylach demonstrating a stronger reduction compared to native HA. Moreover, in inflamed 3D chondrocyte cultures, Hylach® but not HA, significantly reduced IL-1β, TNF-α and Gal-3 gene expression. Conclusions: Hylach® exerts an early and more potent anti-inflammatory effect in inflamed 2D and 3D chondrocyte cultures when compared to HA. These findings suggest that targeting Gal-3 through selective HA derivatives may represent a promising strategy for modulating both inflammation and matrix remodelling in OA.

Featured Application These findings support the use of Vitis labrusca var. Isabella cell-culture juices as natural products with beneficial potential in gut inflammatory disorders, offering a renewable and scalable support to conventional anti-inflammatory agents.Abstract Inflammatory bowel disease is characterised by chronic mucosal inflammation, oxidative stress, and impaired epithelial barrier function. Current therapies primarily suppress inflammation but do not effectively restore epithelial integrity. In this study, we established in vitro cell cultures of Vitis labrusca var. Isabella to obtain juices that were chemically characterised and assessed for antioxidant and anti-inflammatory activities in human intestinal epithelial cell lines (i.e., Caco-2). Chemical analysis revealed variable levels of stilbenoids, including trans-resveratrol and resveratrol diglucosides depending on culture conditions. The suspension-derived juice grown in darkness (SVMD) significantly reduced lipopolysaccharide-induced IL-1 beta and TNF-alpha release and mitigated oxidative stress in Caco-2 cells by lowering levels of intracellular reactive oxygen species. In Caco-2 monolayers infected with Salmonella enterica, SVMD preserved transepithelial electrical resistance, indicating protection of epithelial barrier integrity, without exerting direct antibacterial effects. These findings demonstrate that V. labrusca cell-culture juices exert potent antioxidant and anti-inflammatory actions and promote epithelial protection through modulation of redox balance. Overall, this study highlights the potential of sustainable cell-culture-derived materials as promising natural products for supporting intestinal homeostasis and managing gut inflammatory disorders.

The protease–antiprotease balance is involved in many biological processes, including blood coagulation, tissue remodeling, inflammation and immune responses. The aim of this study is to determine the balance between SERPINs and some related proteases in the lungs of stable COPD patients. In this cross-sectional study, the expression and localization of human SERPINs (anti-proteases) and some related proteases were measured in the lung parenchyma of mild-moderate COPD (MCOPD, n = 13) patients, control smokers (CS, n = 14) and control nonsmokers (CNS, n = 12) using transcriptome analysis, immunohistochemistry, and ELISA tests. Peripheral lung transcriptomic data showed increased mRNA levels of tissue plasminogen activator (tPA), cathepsin-L and caspase-1 as well as increased SERPINs A6, B3, B5, B11, B13 in the COPD group compared to the CNS group. At the protein level, IHC analysis showed that tPA and cathepsin-L increased in the bronchiolar epithelium and alveolar septa of the CS and COPD groups compared to the CNS group, as well as SERPINB5 and B13 in the alveolar macrophages and alveolar septa of the CS and COPD groups compared to the CNS group. SERPINA6 was shown to be decreased in the bronchiolar epithelium, bronchiolar lamina propria, and alveolar septa of the CS and COPD groups compared to the CNS group and was positively correlated with lung function. SERPINB3 was decreased in the alveolar septa of the CS group compared to the CNS group. The ELISA tests showed that in the total lung extracts, decreased levels of SERPINA6 and increased caspase-1 were shown in the COPD group compared to the CNS or both control groups, respectively. These data show an imbalance, at the protein level, of SERPINs and some related proteases in the lungs of the CS and stable COPD groups. These alterations may play a role in damaging the lung parenchyma of susceptible COPD patients.

Background/Objectives: The rise in multidrug-resistant pathogens such as Pseudomonas aeruginosa (PA), coupled with declining antibiotic development, underscores the need for innovative therapeutic strategies. Repurposing approved drugs provides advantages of safety and rapid development. Since quorum sensing (QS) controls key virulence traits in PA, targeting this pathway represents a promising antivirulence approach. This study aimed to identify and repurpose existing drugs as QS inhibitors. Methods: An in silico docking screen of 3000 FDA-approved or clinically tested compounds was performed against the C4-HSL receptor RhlR. Seventeen candidates were tested in the laboratory strain PAO1 for lactone-dependent signaling inhibition. The most active compound, MK-8245, was further evaluated for effects on growth, cytotoxicity, lactone release, biofilm formation, pyocyanin, elastase, rhamnolipids, and swarming motility. Its activity was also assessed in 20 clinical PA isolates. Results: MK-8245 (40 µM) reduced QS-regulated gene expression by ~60% without affecting viability. In PAO1, it inhibited rhamnolipids (60%), pyocyanin (40%), elastase (25%), biofilm formation, and swarming motility (25%). MK-8245 also enhanced the efficacy of imipenem against biofilms. In clinical isolates, it consistently decreased lactone release (~60%), pyocyanin (~50%), rhamnolipids (~40%), biofilm formation (~30%), and swarming motility (~25%). Conclusions: MK-8245 emerges as a promising antivirulence candidate against P. aeruginosa. By disrupting QS signaling and impairing multiple virulence factors, it attenuates pathogenicity without bactericidal pressure. Its synergy with standard antibiotics and consistent activity in clinical isolates highlight its translational potential and warrant further preclinical evaluation.

Isabella, an ancient hybrid grape originating from Vitis labrusca and Vitis vinifera genotypes, is widely cultivated for various food products and is considered a superfood due to its nutritional profile and high polyphenol content. To overcome the unsustainability of intensive agriculture and establish a new route towards more sustainable and socially fair superfood production, this work validated the establishment of undifferentiated in vitro cultures of V. labrusca var. Isabella. Two callus cell lines have been obtained on two different solid media, exhibiting distinct morpho-chemical characteristics. The total phenolic content and antioxidant activity of the callus juices were statistically different in the two cell lines. The subsequent qualitative–quantitative LC-MS analysis revealed the presence of seven stilbenoid derivatives in one cell line and three in the other; likewise, the total stilbenoid content was statistically different between the two cell lines (5.76 and 23.24 µg/mL of juice in the two cell lines on the 28th day of growth). The Isabella cell cultures possess nutritionally valuable profiles. These results suggest that plant cell culture technology can be a sustainable and viable option for the production of complementary, added-value food materials.

Osteochondral defects are a challenge in orthopaedic surgery due to the complexity and function of cartilage. Within this scenario, this study aimed to develop/characterize bioactive porous supports based on oxidized polyvinyl alcohol (OxPVA), with/without human cartilage-derived decellularized ECM (dECM), as platforms for HM1-SV40 cell adhesion and proliferation. OxPVA scaffolds were fabricated using a particle-leaching technique (gelatin concentrations: 10%, 15% and 25% w/w); Scanning Electron Microscopy (SEM) was used to examine the ultrastructure, and a morphometric study assessed pores number, size and porosity percentage. Fluorescence Recovery after Photobleaching (FRAP) was used to evaluate the interconnectivity of the scaffold pores. To enhance the bioactivity of OxPVA, dECM (25% w/w) was incorporated into the scaffolds; thus, the expression of genes related to collagen synthesis and cartilage differentiation/remodelling in seeded HM1-SV40 cells was analyzed by quantitative PCR; relative protein expression levels of SOX9, ACAN and COMP were also assessed. Composite scaffolds biocompatibility was proved by subcutaneous implantation in Sprague-Dawley. As for bone, 3D-printed polylactic acid (PLA)-based scaffolds with varying geometries (67%, 53% and 40% porosity; 600-1400 µm pores size) were fabricated and tested in vitro. Lower gelatin concentrations led to numerous superficial pores, whereas higher concentrations produced larger, coalescing ones. HM1-SV40 cells showed better adhesion to scaffolds prepared with 25% gelatin. The OxPVA+dECM scaffolds exhibited a homogeneous matrix distribution, further promoting cell interaction, with a reduction in mean pore size versus matrix-free scaffolds. Moreover, OxPVA supports prepared with 25% gelatin + dECM provided a favorable environment supporting chondrogenic differentiation and cartilage matrix deposition. No inflammatory response to the implants was observed in vivo. All PLA supports showed good cell viability; SEM higlighted full-thickness HM1-SV40 cell distribution on and within PLA scaffolds, indicating complete colonization. Further studies are needed to evaluate stem cell differentiation, but bioactive OxPVA and 3D-printed PLA scaffolds show potential for osteochondral regeneration.

A novel hydrogel scaffold for bone regeneration based on chitosan, selected for its biocompatibility, biodegradability, and antimicrobial properties, was covalently functionalized with a bioactive peptide from bone morphogenetic protein-2 (BMP-2) to guide osteoblast growth and proliferation. This study evaluates the impact of incorporating different concentrations (8, 16, or 24% wt/wt) of plant-based micro-fibrillated cellulose or tunicate nanocellulose to improve the mechanical and biological properties of peptide-grafted chitosan hydrogel matrices. While the mechanical properties of the matrices increase with increasing cellulose content, regardless of its source, the behavior of human osteoblasts used in biological tests discriminates between the two types of cellulose and shows better results (proliferation at 2 and 7 days, and mineralization) for the enrichment with tunicate cellulose.

Vernal keratoconjunctivitis (VKC) is a serious eye allergy characterized by poorly understood pathogenic mechanisms and a lack of effective treatments. Autophagy, a process involved in both triggering and suppressing immune and inflammatory responses, plays a role in VKC's pathophysiology. Understanding autophagy's involvement in VKC could lead to new treatment possibilities, such as utilizing specific topical substances to induce or inhibit autophagy and prevent severe complications of this eye condition. In our current protocol, we present a robust methodology established in our laboratory for studying autophagy in primary conjunctival fibroblasts. We assess autophagy through techniques like immunocytochemistry, immunoblotting, and qPCR.

Reviewing the literature published up to October 2024.Sesterterpenoids are one of the most chemically diverse and biologically promising subgroup of terpenoids, the largest family of secondary metabolites. The present review article summarizes more than seven decades of studies on isolation and characterization of more than 1600 structurally novel sesterterpenoids, supplemented by biological, pharmacological, ecological, and geographic distribution data. All the information have been implemented in eight tables available on the web and a relational database https://sesterterpenoids.unige.net/. The interface has two sections, one open to the public for reading only and the other, protected by an authentication mechanism, for timely updating of published results.