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The increase of coarse particulate matter (PM 10 ) due to industrialization and urban sprawl has been identified as a significant contributor to air pollution and a threat to human skin health and premature aging. The objective was to analyze the antioxidant effect of phenolic-enriched extracts (PHE) obtained from black bean (BB) and pinto bean (PB) varieties (Phaseolus vulgaris L.) and pure phenolic compounds (rutin, catechin, and gallic acid) in two human dermal fibroblasts cell lines exposed to PM 10 . Petunidin-3-O-glucoside was the most abundant anthocyanin, with 57 ± 0.9 mg/g dry extract (DE) in PHE-BB. Gallic acid was the prevalent phenolic acid with 8.2 ± 2.8 mg/g DE in PHE-BB (p < 0.05). Hs27 and Hs68 cell lines were exposed to PM 10 (100 μg/mL) to induce oxidative stress; PHE-BB reduced it by 69% ± 12 and PHE-PB by 80% ± 5 relative to PM 10 treatment (p < 0.05). Delphinidin-3-O-glucoside showed the highest binding affinity in adenosine monophosphate-activated protein kinase (AMPK) with -9.0 kcal/mol and quercetin-3-D-galactoside with -6.9 kcal/mol in sirtuin 1 (Sirt1). Rutin increased the expression of Sirt1 by 30% (p < 0.05) in the Hs27 cell line treated with PM 10 . Common bean extracts can potentially reduce oxidative stress induced by PM 10 in human dermal fibroblasts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Natural dissipative assembly (DSA) often exhibit energy-driven shifts in natural functions. However, creating man-made DSA that can mimic such biological activities transformation remains relatively rare. Herein, we introduce a cytomembrane-like dissipative assembly system based on chiral supramolecules. This system employs benzoyl cysteine in an out of equilibrium manner, enabling the shifts in biofunctions while minimizing material use. Specifically, aroyl-cystine derivatives primarily assemble into stable M-helix nanofibers under equilibrium conditions. These nanofibers enhance fibroblast adhesion and proliferation through stereospecific interactions with chiral cellular membranes. Upon the addition of chemical fuels, these functional nanofibers temporarily transform into non-equilibrium nanospheres, facilitating efficient drug delivery. Subsequently, these nanospheres revert to their original nanofiber state, effectively recycling the drug. The programmable function-shifting ability of this DSA establishes it as a novel, fuel-driven drug delivery vehicle. And the bioactive DSA not only addresses a gap in synthetic DSAs within biological applications but also sets the stage for innovative designs of 'living' materials., (© 2024 Wiley-VCH GmbH.)

Background: Idiopathic pulmonary fibrosis (IPF) is a chronic and highly fatal disease characterized by excessive accumulation of extracellular matrix (ECM), foci of myofibroblasts, and a usual pattern of interstitial pneumonia. As suggested by international guidelines, the treatment for this disease involves supportive therapies, as there is currently no effective treatment. Plant-derived nanovesicles have emerged as a new treatment for various diseases and have been tested in cellular and murine models., Methods and Results: This research aimed to test the use of Allium sativum nanovesicles (AS-NV) in a murine model of IPF induced by bleomycin. AS-NV reduced the amount of collagen and restored lung architecture in the mouse model. AS-NV was tested on human lung fibroblasts, which do not affect the viability of healthy cells. AS-NV treatment decreases the mRNA levels of genes related to fibrosis, inflammation, and ECM deposition (Mmp2,Timp-2,Vegf,Pcna,Col1a1,Tgf-β,α-Sma,IL-1β,and Hif1a) in bleomycin-induced idiopathic pulmonary fibrosis., Conclusions: This research highlights the anti-inflammatory and antifibrotic activity of AS-NV, which contributes to plant nanovesicle mechanisms in IPF; however, more AS-NV studies are needed to identify alternative treatments for idiopathic pulmonary fibrosis., Competing Interests: Declarations Ethical approval All the experimental animal studies were performed with the approval of the Ethics Committee of the Faculty of Medicine and Surgery – Autonomous University “Benito Juarez” of Oaxaca with registration number 0047-CEI-2022. Informed consent Not applicable. Competing interests The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Wound healing is a complex and dynamic process supported by several cellular events. Around 13 million individuals globally suffer from chronic wounds yearly, for which dressings with excellent antimicrobial activity and cell viability (>70%, as per ISO 10993) are needed. Excessive use of silver can cause cytotoxicity and has been linked to increasing antimicrobial resistance. In this study, HDI Ag foam was synthesized using a safer hexamethylene diisocyanate-based prepolymer (HDI prepolymer) instead of commonly used diisocyanates like TDI and MDI and substantially lower Ag content than that incorporated in other Ag foams. In vitro characteristics of the HDI Ag foam were evaluated in comparison with leading clinically used foam-based dressings. All dressings underwent a detailed characterization in accordance with industrially accepted BS EN 13726 standards. The HDI Ag foam exhibited highest antimicrobial efficiency against S. aureus and P. aeruginosa (static condition), with the lowest amount of Ag (0.2 wt%) on the wound contact surface. The extracts from HDI Ag foam showed superior cell viability (>70%), when tested on the L929 mouse fibroblast cell line. Measurements of moisture vapor transmission, fluid handling, physico-chemical and mechanical properties ensured that the HDI foam was clinically acceptable for chronic wound patients., (© 2024 IOP Publishing Ltd. All rights, including for text and data mining, AI training, and similar technologies, are reserved.)

While myofibroblasts are the key cause of abnormal extracellular matrix accumulation, the origin of which has not yet been fully elucidated. Recently, it has been found that macrophage-myofibroblast transformation (MMT) defined by the expression of both macrophage markers (F4/80 or CD68) and myofibroblast markers (α-SMA) is one of its important sources. In the process of MMT, it is unclear whether epor is involved. In this study, when BMDM was induced by tgf-β1, the number of F4/80 + α-SMA + cells increased, the cells polarized toward M2, and the expression of tgf-β1 increased. After the activation of epor, the number of F4/80 +α-SMA + cells and the polarization level of M2 were further increased. At the same time, we found that the conditioned medium from MMT cells could induce the activation of 3T3 cells with increased the expression of α-SMA and col-1. In contrast, the number of F4/80+α-SMA + cells, the polarization of M2, and the expression of Tgf-β1 decreased after epor was inhibited by siRNA. Our results demonstrate that the activation of epor in BMDMs could promote the transformation of macrophage-myofibroblast induced by TGF-β1., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Context: Proptosis in thyroid eye disease (TED) can result in facial disfigurement and visual dysfunction. Treatment with insulin-like growth factor I receptor (IGF-IR) inhibitors has been shown to be effective in reducing proptosis but with side effects., Objective: To test the hypothesis that inhibition of IGF-IR indirectly and more selectively with PAPP-A inhibitors attenuates IGF-IR signaling in TED. Informed consent was obtained from patients with TED undergoing surgery, and retro-orbital tissue was collected for fibroblast isolation and culture. Operations were performed in Mayo Clinic operating suites. Cell culture was performed in a sterile tissue culture facility. Retro-orbital tissue was collected from 19 patients with TED., Methods: Treatment of TED fibroblasts with proinflammatory cytokines. Flow separation of CD34- and CD34+ orbital fibroblasts, the latter representing infiltrating fibrocytes into the orbit in TED. PAPP-A expression and proteolytic activity, IGF-I stimulation of phosphatidylinositol 3 kinase/Akt pathway, and inhibition by immuno-neutralizing antibodies against PAPP-A, CD34+ status, and associated PAPP-A and IGF-IR expression were measured., Results: Proinflammatory cytokines markedly increased PAPP-A expression in TED fibroblasts. IGF-IR expression was not affected by cytokine treatment. Inhibition of PAPP-A's proteolytic activity suppressed IGF-IR activation in orbital fibroblasts from patients with TED. TED fibroblasts that were CD34+ represented ∼80% of the cells in culture and accounted for ∼70% of PAPP-A and IGF-IR-expressing cells., Conclusion: These results support a role for PAPP-A in TED pathogenesis and indicate the potential for novel therapeutic targeting of the IGF axis., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Endocrine Society.)

Background: Bacterial keratitis is a common cause of blindness. Antibiotic treatment leads to the rapid release of lipopolysaccharide (LPS), which can activate corneal fibroblasts and cause persistent and excessive inflammatory responses. The anti-inflammatory drugs currently used to treat keratitis have serious side effects. Therefore, the ability of sodium butyrate (NaB), which can suppress the production of proinflammatory cytokines and promote the production of anti-inflammatory cytokines, to ameliorate keratitis was assessed in the present study., Methods: The effect of NaB on the viability of primary human corneal fibroblasts was assayed with a CCK-8 kit. Cell migration was assessed by an in vitro scratch assay. Cell phenotypes were assessed by Western blotting and immunofluorescence staining. An antibody array was used to measure the production of proinflammatory cytokines and chemokines., Results: At 0-1 mM, NaB had no significant effect on cell viability, promoted the expression of the keratocyte marker keratocan and inhibited the fibroblast marker vimentin. Inhibition of cell migration was observed in the wound healing assay. By targeting the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signalling pathway, NaB decreased the levels of inflammation-related cytokines and chemokines whose expression was induced by LPS., Conclusions: NaB maintained the keratocyte phenotype, inhibited cell migration, and relieved LPS-induced inflammatory responses through the JAK/STAT signalling pathway., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Irreversible pulmonary fibrosis induced by paraquat is the most prevalent cause of death in patients with paraquat poisoning. Pulmonary fibrosis is characterized by abnormal deposition of extracellular matrix (ECM). Currently, the role of fibrotic ECM microenvironment in paraquat-induced pulmonary fibrosis has not been established., Methods: Rat pulmonary fibrosis model was induced by paraquat, ATN-161 (an integrin-β 1 antagonist) was given to investigate their effect on Rat survival and pulmonary fibrosis. Lungs were decellularized to generate normal and fibrotic acellular ECM scaffolds using Triton and SDS. Fibroblasts were cocultured with ECM scaffolds to established 3D culture systems to investigate the relationship between fibrotic ECM and the differentiation of fibroblasts. Then we explored the effect of fibrotic ECM microenvironment systematically promoting on integrin-β1/FAK/ERK1/2 pathway and established 3D culture systems to investigate the relationship between fibrotic ECM and the differentiation of fibroblasts., Results: Antagonism of integrin-β 1 could alleviate paraquat-induced pulmonary fibrosis and ameliorate survival status of rats. Compared to normal ECM, fibrotic extracellular microenvironment promoted the differentiation of fibroblasts to myofibroblasts. Antagonism of integrin-β 1 could also ameliorate the promotion of fibrotic extracellular microenvironment on differentiation of fibroblasts to myofibroblasts. Fibrotic ECM microenvironment promotes fibroblasts transforming into myofibroblasts through integrin-β 1 /FAK/ERK1/2 signaling pathway. Moreover, this phenomenon holds independent on exogenous integrin-β 1 ., Conclusions: Activation of integrin-β 1 /FAK/ERK1/2 pathway aggravates paraquat-induced pulmonary fibrosis depend on fibrotic ECM and integrin-β 1 may be a prospective therapeutic target for paraquat-induced pulmonary fibrosis in the future., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Rapid regeneration of the injured tissue or organs is necessary to achieve the usual functionalities of the damaged parts. However, bacterial infections delay the regeneration process, a severe challenge in the personalized healthcare sector. To overcome these challenges, 3D-printable multifunctional hydrogels of Zn/tannic acid-reinforced glycol functionalized chitosan for rapid wound healing were developed. Polyphenol strengthened intermolecular connections, while glutaraldehyde stabilized 3D-printed structures. The hydrogel exhibited enhanced viscoelasticity (G'; 1.96 × 10 4  Pa) and adhesiveness (210 kPa). The dual-crosslinked scaffolds showed remarkable antibacterial activity against Bacillus subtilis (∼81 %) and Escherichia coli (92.75 %). The hydrogels showed no adverse effects on human dermal fibroblasts (HDFs) and macrophages (RAW 264.7), indicating their superior biocompatibility. The Zn/TA-reinforced hydrogels accelerate M2 polarization of macrophages through the activation of anti-inflammatory transcription factors (Arg-1, VEGF, CD163, and IL-10), suggesting better immunomodulatory effects, which is favorable for rapid wound regeneration. Higher collagen deposition and rapid re-epithelialization occurred in scaffold-treated rat groups vis-à-vis controls, demonstrating superior wound healing. Taken together, the developed multifunctional hydrogels have great potential for rapidly regenerating bacteria-infected wounds in the personalized healthcare sector., Competing Interests: Declaration of competing interest The authors declare that no conflicts of financial interests or personal relationships have influenced the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Ethnopharmacological Relevance: Blumea balsamifera (L.) DC. (BB), the source of Blumea balsamifera oil (BBO), is an aromatic medicinal plant, renowned for its pharmacological properties and its traditional use in Southeast Asian countries such as China, Thailand, Vietnam, Malaysia, and the Philippines for centuries. Traditionally, BB has been used as a raw herbal medicine for treating various skin conditions like eczema, dermatitis, athlete's foot, and wound healing for skin injuries., Aim of the Study: This research aimed to explore the inhibitory effects of BBO on skin aging using two models: in vitro analysis with human dermal fibroblasts (HDF) under UVB-induced stress, and in vivo studies on UVA-induced dorsal skin aging in mice. The study sought to uncover the mechanisms behind BBO's anti-aging effects, specifically, its impact on cellular and tissue responses to UV-induced skin aging., Materials and Methods: We applied doses of 10-20 μL/mL of BBO to HDF cells that had been exposed to UVB radiation to simulate skin aging. We measured cell viability, and levels of reactive oxygen species (ROS), SA-β-gal, pro-inflammatory cytokines, and matrix metalloproteinases (MMPs). In addition, we investigated the involvement of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling pathways in mediating the anti-aging effects of BBO. Histopathological and biochemical analyses were conducted in a mouse model to examine the effects of BBO on UV-induced photoaging., Results: UV exposure accelerated aging, and caused cellular damage and inflammatory responses through ROS-mediated pathways. In HDF cells, BBO treatment countered the UVB-induced senescence, and the recovery of cell viability was correlated to notable reductions in SA-β-gal, ROS, pro-inflammatory cytokines, and MMPs. Mechanistically, the anti-aging effect of BBO was associated with the downregulation of the JNK/NF-κB signaling pathways. In the in vivo mouse model, BBO exhibited protective capabilities against UV-induced photoaging, which were manifested by the enhanced antioxidant enzyme activities and tissue remodeling., Conclusions: BBO effectively protects fibroblasts from UV-induced photoaging through the JNK/NF-κB pathway. Recovery from photoaging involves an increase in dermal fibroblasts, alleviation of inflammation, accelerated synthesis of antioxidant enzymes, and slowed degradation of ECM proteins. Overall, BBO enhances the skin's defensive capabilities against oxidative stress, underscoring its potential as a therapeutic agent for oxidative stress-related skin aging., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Recent research found artesunate could inhibit ocular fibrosis; however, the underlying mechanisms are not fully known. Since the ocular fibroblast is the main effector cell in fibrosis, we hypothesized that artesunate may exert its protective effects by inhibiting the fibroblasts proliferation. TGF-β1-induced ocular fibroblasts and glaucoma filtration surgery (GFS)-treated rabbits were used as ocular fibrotic models. Firstly, we analyzed fibrosis levels by assessing the expression of fibrotic marker proteins, and used Ki67 immunofluorescence, EdU staining, flow cytometry to determine cell cycle status, and SA-β-gal staining to assess cellular senescence levels. Then to predict target genes and pathways of artesunate, we analyzed the differentially expressed genes and enriched pathways through RNA-seq. Western blot and immunohistochemistry were used to detect the pathway-related proteins. Additionally, we validated the dependence of artesunate's effects on HO-1 expression through HO-1 siRNA. Moreover, DCFDA and MitoSOX fluorescence staining were used to examine ROS level. We found artesunate significantly inhibits the expression of fibrosis-related proteins, induces cell cycle arrest and cellular senescence. Knocking down HO-1 in fibroblasts with siRNA reverses these regulatory effects of artesunate. Mechanistic studies show that artesunate significantly inhibits the activation of the Cyclin D1/CDK4-pRB pathway, induces an increase in cellular and mitochondrial ROS levels and activates the Nrf2/HO-1 pathway. In conclusion, the present study identifies that artesunate induces HO-1 expression through ROS to activate the antioxidant Nrf2/HO-1 pathway, subsequently inhibits the cell cycle regulation pathway Cyclin D1/CDK4-pRB in an HO-1-dependent way, induces cell cycle arrest and senescence, and thereby resists periorbital fibrosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Salvia miltiorrhiza Bunge (S. miltiorrhiza) is an important Traditional Chinese herbal Medicine (TCM) used to treat cardio-cerebrovascular diseases. Based on the pharmacodynamic substance of S. miltiorrhiza, the aim of present study was to investigate the underlying mechanism of S. miltiorrhiza against cardiac fibrosis (CF) through a systematic network pharmacology approach, molecular docking and dynamics simulation as well as experimental investigation in vitro., Materials and Methods: A systematic pharmacological analysis was conducted using the Traditional Chinese Medicine Pharmacology (TCMSP) database to screen the effective chemical components of S. miltiorrhiza, then the corresponding potential target genes of the compounds were obtained by the Swiss Target Prediction and TCMSP databases. Meanwhile, GeneCards, DisGeNET, OMIM, and TTD disease databases were used to screen CF targets, and a protein-protein interaction (PPI) network of drug-disease targets was constructed on S. miltiorrhiza/CF targets by Search Tool for the Retrieval of Interacting Genes/Proteins (STING) database. After that, the component-disease-target network was constructed by software Cytoscape 3.7. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed for the intersection targets between drug and disease. The relationship between active ingredient of S. miltiorrhiza and disease targets of CF was assessed via molecular docking and molecular dynamics simulation. Subsequently, the underlying mechanism of the hub compound on CF was experimentally investigated in vitro., Results: 206 corresponding targets to effective chemical components from S. miltiorrhiza were determined, and among them, there were 82 targets that overlapped with targets of CF. Further, through PPI analysis, AKT1 and GSK3β were the hub targets, and which were both enriched in the PI3K/AKT signaling pathway, it was the sub-pathways of the lipid and atherosclerosis pathway. Subsequently, compound-disease-genes-pathways diagram is constructed, apigenin (APi) was a top ingredients and AKT1 (51) and GSK3β (22) were the hub genes according to the degree value. The results of molecular docking and dynamics simulation showed that APi has strong affinities with AKT and GSK3β. The results of cell experiments showed that APi inhibited cells viability, proliferation, proteins expression of α-SMA and collagen I/III, phosphorylation of AKT1 and GSK3β in MCFs induced by TGFβ1., Conclusion: Through a systematic network pharmacology approach, molecular docking and dynamics simulation, and confirmed by in vitro cell experiments, these results indicated that APi interacts with AKT and GSK3β to disrupt the phosphorylation of AKT and GSK3β, thereby inhibiting the proliferation and differentiation of MCFs induced by TGFβ1, which providing new insights into the pharmacological mechanism of S. miltiorrhiza in the treatment of CF., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction. Ecological concerns and the depletion of petroleum resources have driven the exploration of biodegradable 3D-printing materials derived from bio-renewable sources, such as polylactic acid (PLA) and polyhydroxyalkanoates (PHA). This study aimed to compare the potential cytotoxic effects of a biodegradable PLA/PHA blend filament, a conventional photopolymer (MED610), and a combination of MED610 with a support material (SUP705) before and after steam sterilization in vitro, with a focus on their application in the production of surgical guides. Materials and Methods. PLA/PHA, MED610, and SUP705 (both in their pure and steam-sterilized forms; n = 6 per group) were assessed for their cytotoxic effects on human fibroblasts using the neutral red uptake assay. Positive controls included zinc diethyldithiocarbamate and zinc dibutyldithiocarbamate, while high-density polyethylene served as a negative control. A stock solution of the extraction medium was used as the vehicle control (VC). Results. Significant differences in cell viability were observed between pure PLA/PHA (1.2 ± 0.24) and MED610 (0.94 ± 0.08) ( p = 0.005). However, both materials exhibited non-cytotoxicity, with cell viability exceeding 70% compared to VCs. SUP705 (0.58 ± 0.42) demonstrated significantly reduced cell viability compared to PLA/PHA ( p = 0.001) and MED610 ( p = 0.007). After steam sterilization, no significant difference in cell viability was noted between MED610 (1.0 ± 0.08) and PLA/PHA (1.2 ± 0.25) ( p = 0.111). While both materials remained non-cytotoxic after sterilization, SUP705 (0.60 ± 0.45) exhibited cytotoxic effects compared to MED610 ( p = 0.006) and PLA/PHA ( p < 0.001). Steam sterilization did not induce significant cytotoxic effects in the investigated materials ( p = 0.123). Conclusion. Pure and steam-sterilized PLA/PHA and MED610 were not cytotoxic, supporting their potential use in the production of surgical guides. However, the observed cytotoxicity of SUP705 suggests caution in scenarios requiring sterile conditions, as the removal of support material from complex printed parts may be challenging. The consideration of PLA/PHA is recommended in such settings to ensure biocompatibility., (Creative Commons Attribution license.)

Pathological conditions such as oxidative stress or inflammation may alter the homeostasis of adventitia triggering vascular wall remodeling and abnormal angiogenesis, what can lead to development of atherosclerosis. Growth differentiation factor-15 (GDF-15) is a stress-responsive cytokine and metabolic regulator, but its role in angiogenesis is not yet fully defined. Here we utilized an organ-on-a-chip technology to analyze endothelial sprouting in an adventitia-resembling microenvironment. We analyzed angiogenic responses to growth factor gradient across the extracellular matrix-resembling fibrin gel and in cell co-culture in response to GDF-15-treated adventitial fibroblasts. We observed that GDF-15 enhanced the pro-angiogenic effect of vascular endothelial growth factor. On the other hand, GDF-15-treated adventitial fibroblasts decreased endothelial sprouting. GDF-15 seems to indirectly affect endothelial cells and, depending on the microenvironment, its effect can be either pro- or anti-angiogenic., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Fibroblast activation plays an important role in the occurrence and development of idiopathic pulmonary fibrosis (IPF), which is a progressive, incurable, and fibrotic lung disease. However, the underlying mechanism of fibroblast activation in IPF remains elusive. Here, we showed that the expression levels of STX11 and SNAP25 were downregulated in the lung tissues from patients with IPF and mice with bleomycin (BLM)-induced lung fibrosis as well as in the activated fibroblasts. Upregulation of STX11 or SNAP25 suppressed TGF-β1-induced activation of human lung fibroblasts (HLFs) via promoting autophagy. However, they failed to suppress fibroblast actviation when autophagy was blocked with the use of chloroquine (CQ). In addition, STX11 or SNAP25 could inhibit TGF-β1-induced fibroblast proliferation and migration. In vivo, overexpression of STX11 exerted its protective role in the mice with BLM-induced lung fibrosis. STX11 and SNAP25 mutually promoted expression of each other. Co-IP assay indicated that STX11 has an interaction with SNAP25. Mechanistically, STX11-SNAP25 interaction activated fibroblast autophagy and further inhibited fibroblast activation via blocking the PI3K/AKT/mTOR pathway. Overall, the results suggested that STX11-SNAP25 interaction significantly inhibited lung fibrosis by promoting fibroblast autophagy and suppressing fibroblast activation via blocking the PI3K/ATK/mTOR signaling pathway. Therefore, STX11 serves as a promising therapeutic target in IPF., Competing Interests: Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

The present work consist of the synthesis of photo-cross-linkable materials, based on unsaturated polyesters (UPs), synthesized from biobased monomers from renewable sources such as itaconic acid and 1,4-butanediol. The UPs were characterized to assess the influence of polycondensation reaction temperature and cross-linking time on their final properties. For this purpose, different UV irradiation exposure periods were tested. Homogeneous, uniform, and transparent films were obtained after 1, 3, and 5 min of UV exposure. These cross-linked films were then characterized. All materials presented high gel content, which was dependent on the reaction's temperature. The thermal behaviors of the UPs were shown to be similar. In vitro hydrolytic degradation tests showed that the materials can undergo degradation in phosphate-buffered saline (PBS) at pH 7.4 and 37 °C, ensuring their biodegradability over time. Finally, to assess the applicability of the polyesters as biomaterials, their cytocompatibility was determined by using human dermal fibroblasts.

This study aimed to investigate the cytotoxicity and apoptotic activity of different endodontic sealers: Sealapex, Apexit Plus, AH Plus, MTA-Fillapex and TotalFill BC Sealer in the culture of human periodontal ligament fibroblast (HPDLF) cells. The sealers were mixed, set for 24 h, and then covered with culture medium to obtain extracts, which were diluted to 1:0, 1:1, 1:2, 1:4, and 1:8. Simultaneously, HPDLF cells (1 × 10 4 ) were seeded in 96-well plates and incubated for 24 h at 37 °C 5% CO 2 conditions. The cells were then exposed to 100 µL of diluted extract medium. Cytotoxicity was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to check cell viability, while apoptosis was assessed by TUNEL assay. Statistical analyses were performed using Kruskal-Wallis and Dunn post hoc tests with Mann Whitney U. In MTT and TUNEL assay cells were treated with sealers both 24 and 72 h. All materials showed higher toxicity at 72 h compared to 24 h. AH Plus exhibited the highest cytotoxicity, followed by MTA-Fillapex, Apexit Plus, Sealapex, while TotalFill BC Sealer had the lowest cytotoxicity. Consuquently, it was considered that TotalFill BC Sealer had the lowest cytotoxic potency when compared to other sealers, so it can be considered biocompatible., (© 2024. The Author(s).)

Research on keloid is limited by the lack of proper in vitro and animal model reflecting in vivo status. Based on heterogeneity of keloid and important role of endothelial cells in its pathogenesis, a novel 3D in vitro keloid spheroid prepared with keloid fibroblasts and endothelial cells was evaluated in this study. Commercial cell lines of keloid fibroblasts and endothelial cells were used at various cellular ratios to generate keloid spheroids to determine the optimal condition. Keloid spheroids from three keloid patients were also made and their usefulness as in vitro models, including their responses to drugs, were assessed. Spheroids with higher endothelial cell proportions exhibited increased viability and propagation ability. Patient-derived keloid spheroids showed heterogeneity which might reflect individual clinical conditions. The optimal ratio of fibroblasts to endothelial cells was determined to be 4:1 for keloid spheroids based on gene expression and viability analyses. Patient-derived keloid spheroid showed better keloidal changes in genetic expressions than 2D monolayer culture. Spheroids exhibited varied responses and resistance to each drug used for keloids, depending on the cell type used. 3D keloid spheroids might provide an effective in vitro model for investigating disease pathogenesis and appropriate treatment modalities for future precision medicine., (© 2024. The Author(s).)

Nitric oxide (NO), a potential therapeutic antiaging molecule, modulates various physiological and cellular processes. However, alterations in endogenous NO levels brought on by aging impact multiple organ systems and heighten susceptibility to age-related skin diseases. This correlation underscores the importance of investigating NO-based antiaging interventions. Nonthermal plasma-generated NO is a promising avenue for cosmetic and regenerative medicine due to its capacity to stimulate cellular growth. Herein, we examine the potential of plasma-generated nitric oxide water (NOW) as a bioactive agent in human dermal fibroblasts, emphasizing gene expression patterns linked to extracellular matrix (ECM) breakdown and cellular senescence. The findings of our study indicate that administering NOW at lower dosages enhances cell migration and proliferation. Moreover, the genetic signatures associated with ECM synthesis, antioxidant defense, and antisenescence pathways have been analyzed in NOW-exposed cells. Notably, the downregulation of ECM-degrading enzyme transcripts─collagenase, elastase, and hyaluronidase─suggests NOW's potential in mitigating the intrinsic skin aging phenomena, emphasizing the promise of NO-based interventions in advancing antiaging strategies within regenerative medicine.

The burgeoning incidence of thyroid cancer globally necessitates a deeper understanding of its etiological factors. Emerging research suggests a link to environmental contaminants, notably perfluoroalkyl carboxylates (PFACs). This study introduces a novel biomaterial-based approach for modeling thyroid cancer and assesses PFAC exposure-related health risks. This biomaterial-centric methodology enabled a realistic simulation of long-term, low-dose PFAC exposure, yielding critical insights into their carcinogenic potential. Initially, the no observed adverse effect level concentration of 10 μM for four different PFACs, determined using cytotoxicity tests in 2D cell cultures, was employed with thyroid cancer organoids. Specifically, these organoids were exposed to 10 μM of PFACs, refreshed every 3 days over a period of 21 days. The impact of these PFACs on the organoids was assessed using western blotting and immunofluorescence, complemented by high-content screening imaging. This evaluation focused on thyroid-specific biomarkers, epithelial-mesenchymal transition markers, and the proliferation marker Ki-67. Findings indicated significant alterations in these markers, particularly with long-chain PFACs, suggesting an increased risk of thyroid cancer progression and metastasis upon prolonged exposure. This research advances our understanding of thyroid cancer pathology within the context of environmental health risks by investigating the effects of low-dose, long-term exposure to PFACs on human thyroid cancer organoids. The findings reveal the potential carcinogenic risk associated with these substances, emphasizing the urgent need for stricter regulatory controls., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Purpose: In thyroid eye disease (TED), activation and proliferation of orbital fibroblasts (OFs) promotes remodeling and causes an increase in the volume of orbital tissue. Platelet-derived growth factors (PDGFs) are elevated in TED and promote OF activation. The aryl hydrocarbon receptor (AHR), a ligand activated nuclear receptor, is important in regulating OF activation. AHR ligands have been evaluated as therapeutic agents for inflammatory diseases. Here, we hypothesize that AHR ligands will block PDGF-induced signaling in TED OFs., Methods: OFs from both non-TED and TED patients were treated with PDGFβ, with or without the AHR ligands 6-Formylindolo[3,2-b]carbazole (FICZ) or tapinarof. Cell viability was measured by the Alamar Blue assay. Cell proliferation was quantified using the BrdU assay. Cell lysates were collected and analyzed by Western blotting and real-time quantitative PCR (RT-qPCR) to measure PDGF and AHR signaling. Scratch assays were used to measure OF migration., Results: PDGFβ induced proliferation in TED OFs significantly more than in non-TED OFs. Additionally, PDGFβ increased phosphorylation of AKT and expression of thymidylate synthase (TYMS). PDGFβ dependent proliferation and downstream signaling were attenuated by FICZ or tapinarof. TYMS and other PDGF target genes were upregulated by PDGFβ and reduced by AHR activation. PDGFβ induced TED OF migration while both FICZ and tapinarof diminished this effect., Conclusions: PDGF signaling led to increased proliferation and activation of TED OFs. Treatment of TED OFs with the AHR ligands, FICZ and tapinarof, mitigated PDGF induced effects. These studies support the concept that AHR and PDGF signaling could form the basis for new TED therapeutics.

Objective: The objectives of this study were to develop a tooth-binding graphene quantum dots silver nanocomposites (ALN-GQDs-Ag) and evaluate their antibacterial, mineralising, and discolouring properties for the prevention of dental caries., Methods: In this study, ALN-GQDs-Ag were developed by synthesising nano silver (Ag) with graphene quantum dots (GQDs) and functionalised GQDs with alendronate (ALN). ALN-GQDs-Ag were characterised by transmission electron microscopy (TEM), zeta potential analysis, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and Raman spectroscopy. The cytotoxicity of ALN-GQDs-Ag against human gingival fibroblasts (HGF-1) and stem cells from human exfoliated deciduous teeth (SHED) was examined using a colorimetric assay with reference to silver nitrate solution. The affinity of ALN-GQDs-Ag for hydroxyapatite particles was investigated using inductively coupled plasma spectroscopy (ICP). The antibacterial properties of ALN-GQDs-Ag against Streptococcus mutans were evaluated by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and colony-forming units counting (CFUs). The mineralisation properties of ALN-GQDs-Ag on human dentine were assessed using micro-computed tomography (micro-CT), scanning electron microscopy (SEM), and Fourier transform infrared in a biochemical cycling model. The discolouring properties of ALN-GQDs-Ag on artificial dentine caries were determined using spectrophotometry., Results: TEM, Zeta potential, XPS, FTIR, and Raman spectroscopy confirmed the synthesis of stable spherical ALN-GQD-Ag nanocomposites with a 10.3 ± 5.5 nm diameter. The colorimetric assay demonstrated that ALN-GQDs-Ag were less cytotoxic than silver nitrate to HGF-1 and SHED (p<0.001). ICP showed that ALN-GQDs-Ag were bound to hydroxyapatite. SEM, CLSM, and CFUs showed that ALN-GQDs-Ag was bactericidal and inhibited biofilm growth of Streptococcus mutans . Micro-CT, SEM, and FTIR showed that ALN-GQDs-Ag repressed dentine demineralisation under a cariogenic challenge. Spectrophotometry revealed no significant discolouration of dentine caries in the ALN-GQDs-Ag., Conclusion: This study developed a biocompatible and tooth-binding ALN-GQDs-Ag with promising antibacterial, mineralising, and non-discolouring properties. ALN-GQDs-Ag could be a novel anti-caries agent for preventing dentine caries if translated for clinical use., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could influence the work reported in this study., (© 2024 Yin et al.)

Background: When inflammation occurs in periodontal tissues, a dynamic cellular crosstalk interacts between gingival fibroblasts and bone marrow mesenchymal stem cells (BMSCs), which plays a crucial role in the biological behaviour and differentiation of the cells. Recently, flavonoids are increasingly recognized for their therapeutic potential in modulating inflammation and osteogenic differentiation. Owing to their varied molecular structures and mechanisms, there are more needs that flavonoid compounds should be identified by extensive screening. However, current drug research mostly relies on static, single-type cell cultures. In this study, an innovative bionic microfluidic chip system tailored for both soft and hard tissues was developed to screen for flavonoids suitable for treating periodontitis., Methods: This study developed a microfluidic system that bionically simulates the soft and hard structures of periodontal tissues. Live/dead staining, reactive oxygen species (ROS) staining, and RT-qPCR analysis were employed. These techniques evaluated the effects of flavonoid compounds on the levels of inflammatory factors and ROS contents in HGF and HBMSC under LPS stimulation. Additionally, the impact of these compounds on osteogenic induction in HBMSC and the exploration of the underlying mechanisms were assessed., Results: The microfluidic chip used in this study features dual chambers separated by a porous membrane, allowing cellular signal communication via bioactive factors secreted by cells in both layers under perfusion. The inflammatory response within the chip under LPS stimulation was lower compared to individual static cultures of HGF and HBMSC. The selected flavonoids-myricetin, catechin, and quercetin-significantly reduced cellular inflammation, decreased ROS levels, and enhanced osteogenic differentiation of BMSCs. Additionally, fisetin, silybin, and icariside II also demonstrated favorable outcomes in reducing inflammation, lowering ROS levels, and promoting osteogenic differentiation through the Wnt/β-catenin pathway., Conclusions: The bionic microfluidic chip system provides enhanced capabilities for drug screening and evaluation, delivering a more precise assessment of drug efficacy and safety compared to traditional in vitro methods. This study demonstrates the efficacy of flavonoids in influencing osteogenic processes in BMSCs primarily through the Wnt/β-catenin pathway. These results uncover the potential of flavonoids as therapeutic medicine for treating periodontitis, meriting further research and development., (© 2024. The Author(s).)

Nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) has emerged as a promising target for developing drugs to tackle renal fibrosis. In this study, a series of chromone derivatives were designed and synthesized. Additionally, we established a NOX4 overexpression model using the NRK-49F rat renal fibroblasts cell line and identified compound 14m as highly active through the assessment of intracellular reactive oxygen species (ROS) levels in this model. The drug affinity responsive target stability (DARTS) assay illuminated the robust binding stability of 14m with NOX4. Mechanistic studies further substantiated its efficacy in ameliorating fibrosis and inflammation. This investigation positions 14m as a noteworthy NOX4 inhibitor, shedding light on its regulatory role in renal fibroblasts. Importantly, it diversifies the structural landscape of NOX4 inhibitors, offering novel lead compounds for future development., (© 2024 John Wiley & Sons Ltd.)

Aims: Although the genetic locus of X-linked dystonia parkinsonism (XDP), a neurodegenerative disease endemic in the Philippines, is well-characterized, the exact mechanisms leading to neuronal loss are not yet fully understood. Recently, we demonstrated an increase in myeloperoxidase (MPO) levels in XDP postmortem prefrontal cortex (PFC), suggesting a role for inflammation in XDP pathogenesis. Therefore, we hypothesized that inhibiting MPO could provide a therapeutic strategy for XDP., Methods: MPO activity was measured by using an MPO-activatable fluorescent agent (MAFA) in human postmortem PFC. Reactive oxygen species (ROS) and MPO activity were measured in XDP-derived fibroblasts and SH-SY5Y cells following MPO inhibition., Results: MPO activity was significantly increased in XDP PFC. Additionally, treatment of cell lines with postmortem XDP PFC resulted in a significant increase in ROS levels. To determine whether increases in MPO activity caused increases in ROS, MPO content was immunodepleted from XDP PFC, which resulted in a significant decrease in ROS in SH-SY5Y cells. Consistently, the treatment with verdiperstat, a potent and selective MPO inhibitor, significantly decreased ROS in both XDP-derived fibroblasts and XDP PFC-treated SH-SY5Y cells., Conclusions: Collectively, our results suggest that MPO inhibition mitigates oxidative stress and may provide a novel therapeutic strategy for XDP treatment., (© 2024 The Author(s). CNS Neuroscience & Therapeutics published by John Wiley & Sons Ltd.)

Osteoarthritis (OA) is the most common form of arthritis and a leading cause of pain and disability in adults. A central feature is progressive cartilage degradation and matrix fragment formation driven by the excessive production of matrix metalloproteinases (MMPs), such as MMP-13, by articular chondrocytes. Inflammatory factors, including interleukin 6 (IL-6), are secreted into the joint by synovial fibroblasts, and can contribute to pain and inflammation. No therapeutic exists that addresses the underlying loss of joint tissue in OA. To address this, we developed and utilized a cell-based high-throughput OA drug discovery platform using normal human chondrocytes treated with a recombinant fragment of the matrix protein fibronectin (FN-f) as a catabolic stimulus relevant to OA pathogenesis and a readout using a fluorescent MMP-13 responsive probe. The goal was to test this screening platform by identifying compounds that inhibited FN-f-induced MMP-13 production and determine if these compounds also inhibited catabolic signaling in OA chondrocytes and synovial fibroblasts. Two pilot screens of 1344 small molecules revealed five "hits" that strongly inhibited FN-f induced MMP-13 production with low cytotoxicity. These included RO-3306 (CDK1 inhibitor (i)), staurosporine (PKCi), trametinib (MEK1 and MEK2i), GSK-626616 (DYRK3i), and edicotinib (CSF-1Ri). Secondary testing using immunoblots and cells derived from OA joint tissues confirmed the ability of selected compounds to inhibit chondrocyte MMP-13 production and FN-f stimulated IL-6 production by synovial fibroblasts. These findings support the use of this high throughput screening assay for discovery of disease-modifying osteoarthritis drugs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Coryell et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The present study aimed to evaluate the anti-staphylococcal, antibiofilm, cytotoxicity and trypanocidal activity, mechanisms of parasite death and immunomodulatory effect of CrataBL encapsulated into liposomes (CrataBL-Lipo). CrataBL-Lipo were prepared by the freeze-thaw technique and characterized. Anti-staphylococcal and antibiofilm activities of CrataBL and CrataBL-Lipo were evaluated against standard and clinical strains of Staphylococcus aureus susceptible and resistant. Thus, broth microdilution method was performed to determine the Minimum Inhibitory Concentration (MIC). Antibiofilm activity at subinhibitory concentrations was evaluated using the crystal violet staining method. Cytotoxicity of CrataBL-Lipo was verified in L929 fibroblasts and J774A.1 macrophages by determining the inhibitory concentration necessary to kill 50 % of cells (IC 50 ). Trypanocidal activities of CrataBL-Lipo was evaluated in Trypanosoma cruzi and the efficacy was expressed as the concentration necessary to kill 50 % of parasites (EC 50 ). The mechanisms of parasite death and immunomodulatory effect of CrataBL-Lipo were evaluated using flow cytometry analysis. CrataBL-Lipo presented Ø of 101.9 ± 1.3 nm (PDI = 0.245), ζ of +33.8 ± 1.3 mV and %EE = 80 ± 0.84 %. CrataBL-Lipo presented anti-staphylococcal activity (MIC = 0.56 mg/mL to 0.72 mg/mL). CrataBL-Lipo inhibited 45.4 %-75.6 % of biofilm formation. No cytotoxicity of CrataBL-Lipo was found (IC 50  > 100 mg/L). CrataBL-Lipo presented EC 50 of 1.1 mg/L, presenting autophagy, apoptosis and necrosis as death profile. In addition, CrataBL-Lipo reduced the production of IL-10 and TNF-α levels, causing an immunomodulatory effect. CrataBL-Lipo has a therapeutic potential for the treatment of staphylococcal infections and Chagas disease exhibiting a high degree of selectivity for the microorganism, and immunomodulatory properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Some skin wounds often have many exudate. Ordinary single layer electrospunning nanofiber wound dressings often don't have enough capacity to absorb them. Therefore, a separable double layer electrospunning nanofiber dressing was developed in this work. The dressing had a separable feature that allowed the upper layer to be separated and removed after it had absorbed a significant amount of wound exudate. This dressing consisted of an upper layer of super hydrophilic sodium polyacrylate nanofibers and a bottom layer of 3D-structure coaxial nanofibers with encapsulated Astragaloside (AS). The results showed that nanofibers had better morphology. The water absorption rate, water vapor transmission rate and free radical scavenging rate of the double-layer dressings were 1461.71 ± 39.72 %, 1193.63 ± 134 g·m -2 ·day -1 , and 63.35 ± 3.65 %, respectively. The double-layer nanofiber dressing achieved 65.69 ± 2.62 % and 75.10 ± 6.26 % inhibition against Staphylococcus aureus and Escherichia coli, respectively. The double-layer dressing had proliferative, migratory, and adhesive effects on L929 fibroblasts. And the double-layer dressing resulted in a 96.78 ± 1.0 % wound healing rate in rats after giving a 14 days treatment. Therefore, the 3D-structure separable double-layer wound dressing designed and prepared in this study was effective in promoting wound healing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Natural substances and bioactive agents possess great potential in wound care based on their ability to promote healing and prevent infection. This study focused on the fabrication of antibacterial wound dressings by combining gelatin (G), tragacanth gum (TG), and galbanum essential oil (GEO) as a loaded drug. TG addition resulted in more elastic and flexible films besides enabling encapsulation of the hydrophobic GEO into the biopolymeric matrix. GEO was utilized as an antibacterial and a wound-healing enhancer for open wounds such as incisions. Field emission scanning electron microscopy (FE-SEM) analysis revealed a porous film structure after GEO incorporation. Higher GEO concentration caused reduced swelling and slower degradation. Water vapor transfer rate varied from 596 to 894 g/m 2 .day, making the films suitable for wound dressings. GEO release exhibited a two-phase profile with prolonged diffusion-controlled release for a higher content of GEO. The films demonstrated dose-dependent antimicrobial activity against S. aureus and E. coli strains. Effectiveness and noteworthy application of this research were approved by scratch assay on human dermal fibroblast cells, and films with 3 % GEO showed 79.42 % wound closure, which is significantly higher than the control sample (55.15 %), indicating promoted cell migration and promising wound healing properties., Competing Interests: Declaration of competing interest The authors declare no competing interests. The authors declare that no funds, grants, or other financial support were received during the research and preparation of this manuscript., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Acyl-CoA Oxidase-1 (ACOX1) deficiency (MIM 264470) is an autosomal recessive disease characterized by impairments in the desaturation of acyl-CoAs to 2-trans-enoyl-CoAs, which is the first step in the catalysis of the β-oxidative breakdown of very long chain fatty acids (VLCFA) occuring in peroxisomes. The deleterious accumulation of VLCFA in several organs, including the brain, is a key biochemical feature of this disease which has devastating neurological consequences. ACOX1 deficiency is ultra-rare; as such, few studies have been conducted to determine the leading causes of symptoms or uncover new therapeutics. When confronted with one such case, we decided to bring drug discovery tools to the patient's bedside in an attempt to identify a cure. A skin biopsy was performed on a young patient with ACOX1 deficiency, following which screening technologies and mass spectrometry analysis techniques were applied to design a cellular assay that enabled the direct measurement of the effect of small molecules on the patient's primary fibroblasts. This approach is particularly well adapted to inherited metabolic disorders such as ACOX1 deficiency. Through the evaluation of a proprietary library of repurposable drugs, we found that the anthelmintic drug niclosamide led to a significant reduction in VLCFA in vitro. This drug was subsequently administered to the patient for more than six years. This study outlines the screening and drug selection processes. Additionally, we present our comprehensive clinical and biochemical findings that aided in understanding the patient's natural history and analysis of the progression of the patient's symptoms throughout the treatment period. Although the patient's overall lifespan was extended compared to the average age at death in severe early onset cases of ACOX1 deficiency, we did not observe any definitive evidence of clinical or biochemical improvement during niclosamide treatment. Nonetheless, our study shows a good safety profile of long-term niclosamide administration in a child with a rare neurodegenerative disease, and illustrates the potential of individualized therapeutic strategies in the management of inherited metabolic disorders, which could benefit both patients and the broader scientific and medical communities., Competing Interests: Declaration of competing interest Terence Beghyn is Apteeus co-founder and chief executive officer. Camille Moreau and Adrien Paquot were members of the Apteeus operational team. Apteeus is a French biotechnology company. Part of the project described in this article was funded by Apteeus. Apteeus, Terence Beghyn, Camille Moreau and Adrien Paquot declare that they have no link with Bayer, which discovered niclosamide, or with several companies that currently market the drug. Apteeus, Terence Beghyn, Camille Moreau and Adrien Paquot received no funding from these companies., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objectives: Lithium disilicate (LS) ceramic emerges as a compelling option for customized implant abutments. However, ensuring its safety and reliability requires clarification on key aspects, notably its impact on inflammation and potential for cell adhesion. This study delves into these considerations, examining the influence of LS ceramic on cytokine release and the transcriptional profile of human gingival fibroblasts (hGFs) in direct contact with various LS surfaces., Methods: hGFs were cultured on LS disks featuring three distinct surfaces (unpolished, polished, and polished glaze), while titanium disks served as reference material and cells cultured directly on plates as controls. The surface of the disks was analyzed using a scanning electron microscope. The cell metabolism was analyzed by MTT test, cytokine release by MAGPIX and the expression of genes related to cell adhesion was evaluated by qPCR., Results: The disks exhibited similar topography with smooth surfaces, except for the unpolished LS disks, which had an irregular surface. Contact with LS surfaces did not substantially reduce cell metabolism. Moreover, it generally decreased cytokine release compared to controls, particularly pro-inflammatory mediators like IL-1β, IL-6, and TNF-α. Significantly increased expression of genes related to cell adhesion to LS was observed, comparable to titanium, the gold standard material for implant abutments., Significance: This study unveils that LS ceramic not only fails to trigger pro-inflammatory cytokine release, but also significantly enhances gene expression associated with cell adhesion. These mechanisms are closely linked to gene pathways such as PTK2, SRC, MAPK1, and transcription factors ELK-1 and MYC. In summary, the findings underscore LS ceramic's potential as a biocompatible material for implant abutments, shedding light on its favorable inflammatory response and enhanced cell adhesion properties., Competing Interests: Declaration of Competing Interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Engineered matrices with aligned microarchitectures are pivotal in regulating the fibroblast-to-myofibroblast transition, a critical process for wound healing and scar reduction. However, developing a three-dimensional (3D) aligned matrix capable of effectively controlling this transition remains challenging. Herein, we developed a cell-adaptive hydrogel with highly oriented microporous structures, fabricated through bioprinting of thermo/ion/photo-crosslinked gelatin methacrylate/sodium alginate (GelMA/SA) incorporating shear-oriented polyethylene oxide (PEO) filler. The synergistic interactions among GelMA, PEO, and SA yield a homogeneous mixture conducive to the printing of biomimetic 3D constructs with anisotropic micropores. These anisotropic micropores, along with the biochemical cues provided by the GelMA/PEO/SA scaffolds, enhance the oriented spreading and organization of fibroblasts. The resultant spread and aligned cellular morphologies promote the transition of fibroblasts into myofibroblasts. By co-culturing human keratinocytes on the engineered dermal layer, we successfully create a bilayer skin construct, wherein the keratinocytes establish tight junctions accompanied by elevated expression of cytokeratin-14, while the fibroblasts display a highly spread morphology with increased fibronectin expression. The printed hydrogels accelerate full-thickness wound closure by establishing a bioactive microenvironment that mitigate inflammation and stimulate angiogenesis, myofibroblast transition, and extracellular matrix remodeling. This anisotropic hydrogel demonstrates substantial promise for applications in skin tissue engineering., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

In this work, chitosan/collagen-based membranes loaded with 2,3-dihydrobenzofuran (2,3-DHB) were developed through a simple solvent-casting procedure for use in the treatment of cutaneous Leishmaniasis. The obtained membranes were characterized by elemental analysis, FTIR, TG, DSC, and XRD. Porosity, swelling, mechanical properties, hydrophilicity, and antioxidant activity were analyzed. In addition, assessment to the biocompatibility, through fibroblasts/keratinocytes and in vitro wound healing essays were performed. The obtained results show that the new 2,3-DHB loaded chitosan/collagen membrane presented high porosity and swelling capacity as well as maximum strength, hydrophilicity, and antioxidant activity higher in relation to the control. The tests of antileishmanial activity and the AFM images demonstrate great efficacy of inhibition growth of the parasite, superior to those from the standard therapeutic agent that is currently used: Amphotericin B. The new membranes are biocompatible and stimulated the proliferation of keratinocytes. SEM images clearly demonstrate that fibroblasts were able to adhere, maintained their characteristic morphology. The healing test evidenced that the membranes have adequate environment for promoting cell proliferation and growth. As the conventional treatments often use drugs with high toxicity, the as-developed new membranes proved to be excellent candidate to treat cutaneous Leishmaniasis and can be clearly indicated for further advanced studies in vivo., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Edvani Curti Muniz reports financial support was provided by National Council for Scientific and Technological Development. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Pulmonary fibrosis (PF) is a common, severe, chronic, and progressive pulmonary interstitial disease characterized by rapid disease progression and high mortality. Despite the Food and Drug Administration (FDA)'s approval of two antifibrotic drugs, nintedanib and pirfenidone, effectively halting the progression of pulmonary fibrosis remains challenging. Histone deacetylase (HDAC) inhibitors have indeed emerged as an important class of antitumour drugs. However, their application in the treatment of fibrotic diseases is still relatively limited. Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) has the potential to inhibit fibrotic processes by inducing fibroblast apoptosis. In this study, we designed and synthesized a series of histone deacetylase 6 (HDAC6) inhibitors that activate TRAIL, among which compound 7e exhibited potent inhibitory activity against HDAC6, with an IC 50 of 42.90 ± 4.96 nM and superior antiproliferative effects on fibroblasts. Therefore, we further investigated its anti-pulmonary fibrosis effect in mouse models of both idiopathic pulmonary fibrosis (IPF) and silicosis. Our results suggest that compound 7e is a promising candidate for the treatment of pulmonary fibrosis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction/objectives: JAK/STAT signaling inhibition exerts therapeutic effects on angiogenesis in rheumatoid arthritis (RA). However, whether the inhibitory effect differs among JAK inhibitors because of differing selectivity is unknown. Therefore, we compared the inhibitory effects of tofacitinib, baricitinib, peficitinib, upadacitinib, and filgotinib on angiogenesis., Method: RA-derived fibroblast-like synoviocytes (RA-FLS) were seeded on type I collagen gel, and human umbilical vein endothelial cells (HUVECs) were directly added. The control and aforementioned JAK inhibitors were added to the medium, followed by stimulation with interleukin (IL)-6 and soluble IL-6 receptor (sIL-6R). Each JAK inhibitor's concentration was determined based on estimated blood concentrations. The vascular endothelial growth factor (VEGF) concentration was evaluated with an enzyme-linked immunosorbent assay using the medium from the first exchange. A migration assay was performed, and HUVEC migration was evaluated using CD31 fluorescence immunostaining., Results: Hematoxylin-eosin staining showed that compared with the non-JAKi treatment group, the JAKi treatment group markedly degenerated in the sub-lining and deep lining, with decreased lymphocyte infiltration and neovascularization [Rooney's score subscale, non-JAKi vs JAKi (median, 6.5 vs 2.5, p = 0.005)]. In vitro, IL-6 and sIL-6R administration increased VEGF production from RA-FLS and promoted neovascularization in HUVECs, and JAK-inhibitor administration, which decreased VEGF production from RA-FLS and suppressed HUVEC migration, inhibited neovascularization in RA-FLS and HUVEC co-cultures., Conclusions: The JAK inhibitors suppressed IL-6-induced angiogenesis via decreased VEGF production and HUVEC migration in RA-FLS and HUVEC co-cultures. No significant differences were observed among the JAK inhibitors, whose anti-angiogenic effect may be an important mechanism for RA treatment. Key Points • JAK inhibitors inhibit angiogenesis in RA by reducing VEGF production from RA-derived fibroblast-like synoviocytes. • Our study provides new insights into RA treatment by elucidating the anti-angiogenic effect of JAK inhibitors., (© 2024. The Author(s), under exclusive licence to International League of Associations for Rheumatology (ILAR).)

Yangtze finless porpoises (YFP) accumulate high levels of per- and polyfluoroalkyl substances (PFASs). However, the health impacts of PFASs to YFP are still unknown because it is technically and ethically unfeasible to use the critically endangered YFP in toxicological exposures. To uncover the potential toxicities of PFASs to YFP, this study exposed a YFP umbilical cord fibroblast cell line to perfluorobutane sulfonate (PFBS), an emerging PFASs pollutant in the aquatic environments. After exposure, the cytotoxicity and mechanisms of PFBS were explored. Our preliminary experiments found that PFBS compromised the cell viability in a concentration and duration dependent manner. In an exposure of 48-h duration, the maximum no observed effect concentration (NOEC) of PFBS was determined to be 400 µM. High-throughput proteomics were then conducted to identify the differentially expressed proteins in YFP cells exposed to 400 µM PFBS for 48 h. The results found that PFBS exposure significantly perturbed the proteome fingerprints of YFP umbilical cord fibroblast cells. Functional annotation of differential proteins showed that PFBS had the potential to impair a variety of biological processes associated with the immunity, oxidative stress, metabolism, and proteolysis. Consistently, the intracellular levels of reactive oxygen species (ROS) and proinflammatory cytokine IL-1β were significantly increased by PFBS in YFP umbilical cord fibroblast cells. Overall, this study highlights the toxic effects of emerging PFASs on YFP and provides reference data to evaluate the health risks of aquatic pollution under the context of national YFP protection. To our knowledge, this is the first omics study using YFP umbilical cord fibroblast cells in ecotoxicology of PFASs, which is applicable to various cetacean species and pollutants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Prolonged exposure of human dermal fibroblasts (HDF) to ultraviolet (UV) radiation triggers the production of reactive oxygen species by upregulating the expression of matrix metalloproteinases (MMPs), causing type-I collagen degradation and photoaging. A sulfated (1 → 3)/(1 → 4) mannogalactan exopolysaccharide (BVP-2) characterized as [→3)-α-Galp-{(1 → 4)-α-6-O-SO 3 -Manp}-(1 → 3)-α-6-O-SO 3 -Galp-(1→] was isolated from seaweed-associated heterotrophic bacterium Bacillus velezensis MTCC13097. Whole genome analysis of B. velezensis MTCC13097 (Accession number JAKYLL000000000) revealed saccharine biosynthetic gene clusters for exopolysaccharide production. BVP-2 administered cells showed noteworthy reduction in mitochondrial superoxide (∼85 %, p < 0.05) and ROS production (62 %) than those exhibited by UV-A irradiated HDF cells. Oxidative imbalance in HDF cells (after UV-A exposure) was recovered with BVP-2 treatment by significantly downregulating nitric oxide (NO) production (98.6 μM/mL, 1.9-fold) and DNA damage (⁓67 %) in comparison with UV-A induced cells (191.8 μM/mL and 98.7 %, respectively). UV-irradiated HDF cells showed a ∼30-50 % downregulation in the expression of MMPs (1, 2, and 9) following treatment with BVP-2. Considerable amount of sulfation (18 %) along with (1 → 3)/(1 → 4) glycosidic linkages in BVP-2 could be pivotal factors for down-regulation of the intracellular MMP-1, which was further supported by molecular docking and structure-activity studies. The (1 → 3)/(1 → 4)-linked bacterial exopolysaccharide (BVP-2) might be used as prospective natural lead to attenuate and mitigate UV-A-induced photoaging., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Stress concentration surrounding wounds drives fibroblasts into a state of high mechanical tension, leading to the delay of wound healing, exacerbating pathological fibrosis, and even causing tissue dysfunction. Here, an innovative skin stress-shielding hydrogel wound dressing is reported that makes the wound sites shrink as a response to body temperature and then remolds the stress micro-environment of wound sites to reduce the formation of skin scars. Composed of a modified natural temperature-sensitive polymer cross-linked with polyacrylic acid networks, this hydrogel wound dressing has demonstrated a substantial decrease in scar area for full-thickness wounds in rat models. The physical forces exerted by the wound dressing are instrumental in attenuating the activation and transduction of fibroblasts within the wound sites, thereby mitigating the excessive deposition of the extracellular matrix (ECM). Notably, the wound dressing significantly down-regulates the expression of transforming growth factor-β1(TGF-β1) and collagen I, while concurrently exerting a dramatic inhibitory effect on the integrin-focal adhesion kinase (FAK)/phosphorylated-FAK (p-FAK) signaling pathway. Collectively, the fabrication of functional hydrogels with a stress-shielding profile is a new route for achieving scar-less wound healing, thus offering immense potential for improving clinical outcomes and restoring tissue integrity., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Background: Silica particles can cause silicosis, a disease characterized by diffuse fibrosis of the lungs. Various signaling pathways composed of different types of cells and cytokines are involved in the development of silicosis. Exosomes have become a research hotspot recently. However, the role of exosomal microRNA (miRNA) in silicosis remains unclear., Methods: In this study, we generated exosomal miRNA sequences from exosomes isolated from bronchoalveolar lavage fluid (BALF) of silicosis patients and the control group by high-throughput sequencing. Functional annotation and analysis of miRNA identified key target miRNAs. Levels of target miRNAs were analyzed in patient and animal samples and cells. Effects of increased miRNA were assessed through protein levels in target signaling pathways in cells treated with silica, miRNA mimics, and inhibitors., Results: Our study identified 40 up-regulated and 70 down-regulated miRNAs, with miR-552-3p and its putative target gene Caveolin 1 (CAV1) as targets for further research. We found that the levels of exosomal miR-552-3p increased in silicosis patients' BALF samples, silicosis model mice, and A549 cells exposed to silica. Inhibition of miR-552-3p suppressed the expression of fibrosis markers. The increased miR-552-3p leads to the up-regulation of fibronectin and α-smooth muscle actin (α-SMA) and the suppression of caveolin 1 in fibroblast cells. Mitogen-activated protein kinase (MAPK) signaling pathways are activated in cells treated with silica and miR-552-3p mimics., Conclusions: These results help to understand exosomal miRNA-mediated intercellular communication and its key role in fibroblast activation and silicosis., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jie Meng reports financial support was provided by National Natural Science Foundation of China., (Copyright © 2024. Published by Elsevier B.V.)

Curcumin (Cur) and resveratrol (Rsv) have already been proposed for both anti-tumor and wound healing applications and contrasting results have been published regarding their anti- or pro-angiogenic activity; depending on the final application, an anti- or a pro-angiogenic activity is required. In the present study, a comparison of Cur and Rsv loaded electrospun fibers based on collagen and polycaprolactone (PCL) mixture was performed in order to make a contribution to understanding whether the two polyphenols have anti or pro-angiogenic activity. Despite their hydrophobic character, the two polyphenols affected morphology and wettability of the fibers, and Rsv-loaded fibers resulted larger and more quickly wettable. After hydration, collagen/PCL fibers loaded with both Cur and Rsv exhibited higher elongation and better deformation with respect to the unloaded fibers. Fourier transformed infrared spectroscopy and thermal analysis showed interactions between the polyphenols and collagen. Both fiber formulations resulted biocompatible with an increase of fibroblast number during 7 days of culture; confocal microscopy analyses demonstrated that Cur released by the fibers was internalized by the cells which remained vital and adherent. Chick embryo chorioallantoic membrane assay showed that both fibers had anti-angiogenic behavior, suggesting that an anti-cancer application more than a wound healing one could be envisaged., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Collagen and hyaluronic acid are essential components of the dermis that collaborate to maintain skin elasticity and hydration due to their unique biochemical properties and interactions within the extracellular matrix. Prolonged exposure to glucocorticoids can induce skin aging, which manifests as diminished collagen content and hyaluronic acid levels in the dermis. Nerol, a monoterpene alcohol found in essential oils, was examined in this study for its potential to counteract glucocorticoid-induced skin aging and the underlying mechanism behind its effects. Our findings reveal that non-toxic concentrations of nerol treatment can reinstate collagen content and hyaluronic acid levels in human dermal fibroblasts treated with dexamethasone. Mechanistically, nerol mitigates dexamethasone-induced oxidative stress by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. The protective effects of nerol were significantly abrogated when the Nrf2 pathway was inhibited using the specific inhibitor ML385. In conclusion, nerol protects human dermal fibroblasts against glucocorticoid-induced skin aging by ameliorating oxidative stress via activation of the Nrf2 pathway, thereby highlighting its potential as a therapeutic agent for preventing and treating glucocorticoid-induced skin aging., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Metabolic dysregulation is a key driver of cellular senescence, contributing to the progression of systemic aging. The heterogeneity of senescent cells and their metabolic shifts are complex and unexplored. A microfluidic SlipChip integrated with surface-enhanced Raman spectroscopy (SERS), termed SlipChip-SERS, is developed for single-cell metabolism analysis. This SlipChip-SERS enables compartmentalization of single cells, parallel delivery of saponin and nanoparticles to release intracellular metabolites and to realize SERS detection with simple slipping operations. Analysis of different cancer cell lines using SlipChip-SERS demonstrated its capability for sensitive and multiplexed metabolic profiling of individual cells. When applied to human primary fibroblasts of different ages, it identified 12 differential metabolites, with spermine validated as a potent inducer of cellular senescence. Prolonged exposure to spermine can induce a classic senescence phenotype, such as increased senescence-associated β-glactosidase activity, elevated expression of senescence-related genes and reduced LMNB1 levels. Additionally, the senescence-inducing capacity of spermine in HUVECs and WRL-68 cells is confirmed, and exogenous spermine treatment increased the accumulation and release of H 2 O 2 . Overall, a novel SlipChip-SERS system is developed for single-cell metabolic analysis, revealing spermine as a potential inducer of senescence across multiple cell types, which may offer new strategies for addressing ageing and ageing-related diseases., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)

Bioprinted hydrogels are extensively studied to provide an artificial matrix for 3D cell culture. The success of bioprinting hydrogels relies on fine-tuning their rheology and composition to achieve shear-thinning behavior. However, a challenge arises from the limited viscoelastic and stiffness range accessible from a single hydrogel formulation. Nevertheless, hydrogel mechanical properties are recognized as essential cues influencing cell phenotype, migration, and differentiation. Thus, it is crucial to develop a system to easily modulate bioprinted hydrogels' mechanical behaviors. In this work, we modulated the viscoelastic properties and stiffness of bioprinted hydrogels composed of fibrinogen, alginate, and gelatin by tuning the crosslinking bath solution. Various concentrations of calcium ionically crosslinked alginate, while transglutaminase crosslinked gelatin. Subsequently, we characterized the mechanical behavior of our bioprinted hydrogels from the nanoscale to the macroscale. This approach enabled the production of diverse bioprinted constructs, either with similar elastic behavior but different elastic moduli or with similar elastic moduli but different viscoelastic behavior from the same hydrogel formulation. Culturing fibroblasts in the hydrogels for 33 days revealed a preference for cell growth and matrix secretion in the viscoelastic hydrogels. This work demonstrates the suitability of the method to decouple the effects of material mechanical from biochemical composition cues on 3D cultured cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Calprotectin, a calcium-binding protein, plays a crucial role in inflammation and has been associated with various inflammatory diseases, including asthma. However, its regulation and impact on steroid hyporesponsiveness, especially in severe asthma, remain poorly understood., Methods: This study investigated the regulation of calprotectin proteins (S100A8 and S100A9) by IL-17 and its role in steroid hyporesponsiveness using in vitro and in vivo models. Calprotectin expression was assessed in primary bronchial fibroblasts from healthy controls and severe asthmatic patients, as well as in mouse models of steroid hyporesponsive lung inflammation induced by house dust mite (HDM) allergen and cyclic-di-GMP (cdiGMP) adjuvant. The effects of IL-17A stimulation on calprotectin expression and steroid response markers in bronchial epithelial and fibroblast cells were examined. Additionally, the therapeutic potential of paquinimod, a calprotectin inhibitor, in mitigating airway inflammation and restoring steroid response signatures in the mouse model was evaluated., Results: The results demonstrated upregulation of calprotectin expression in asthmatic bronchial fibroblasts compared to healthy controls, as well as in refractory asthma samples compared to non-refractory asthma. IL-17 stimulation induced calprotectin expression and dysregulated glucocorticoid response signatures in lung epithelial and fibroblast cells. Treatment with paquinimod reversed IL-17-induced dysregulation of steroid signatures, indicating the involvement of calprotectin in this process. In the HDM/cdiGMP mouse model, paquinimod significantly attenuated airway inflammation and hyperresponsiveness, and restored steroid response signatures, whereas dexamethasone showed limited efficacy. Mechanistically, paquinimod inhibited MAPK/ERK and NF-κB pathways downstream of calprotectin, leading to reduced lung inflammation., Conclusion: These findings highlight calprotectin as a potential therapeutic target regulated by IL-17 in steroid hyporesponsive asthma. Targeting calprotectin may offer a promising approach to alleviate airway inflammation and restore steroid responsiveness in severe asthma. Further investigations are warranted to explore its therapeutic potential in clinical settings and elucidate its broader implications in steroid mechanisms of action., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Objective: To develop a novel and biocompatible copper tetraamine fluoride (CTF) with antibacterial and nondiscolouring properties., Method: This study used copper fluoride and ammonia solution to develop CTF solution. The CTF was characterized by X-ray photoelectron spectroscopy (XPS). Cytotoxicity was evaluated by stem cells from human exfoliated deciduous teeth (SHED) and human gingival fibroblasts (HGF-1). The fluoride concentration was determined using ion-selective electrode. The alkalinity was measured by a pH electrode. The human dentine blocks were treated with CTF and then incubated with Streptococcus mutans to evaluate the antimicrobial and discolouring effects. The silver diamine fluoride (SDF) was employed as the positive control, and water was the negative control. The colony-forming units (CFUs) and confocal laser scanning microscopy (CLSM) were used to examine the kinetics and viability of the biofilm. The discolouring property on dentine was assessed by spectrophotometry. One-way analysis of variance with the Bonferroni post hoc test was performed to assess and compare the data., Results: XPS confirmed synthesis of CTF solution. The half-maximal inhibitory concentration of CTF on SHED and HGF-1 was 195±16 ppm and 137±11 ppm. The fluoride concentration was 121,000±5,000 ppm. The pH value was 9. Log 10 CFU of the CTF, SDF and water group were 5.0 ± 0.2, 4.9 ± 0.1 and 7.4 ± 0.1 (p < 0.001, CTF, SDFWater). Spectrophotometry showed that the ΔE of the CTF, SDF and water group were 5 ± 2, 6 ± 3 and 45±2 (p < 0.001, CTF, Water

An important objective of researchers is to develop a perfect wound dressing that can effectively treat different kinds of wounds. Natural substances with beneficial qualities, such as plant extracts and biopolymers are an ideal aid for wound care. Hydrogels based on biopolymers offer a lot of promising applications for topical use and are biocompatible, hydrophilic and non-toxic. When employed alone or in conjunction with other active agents, herbal extracts have a great deal of use in the healing of wounds. This study comprises Ruellia tuberosa extract loaded with carboxymethyl cellulose and guar gum hydrogels that have potential anti-bacterial, antioxidant, anti-inflammatory and hemocompatibility. Using mouse fibroblast cells (L929), the MTT (3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) test was conducted to assess the biocompatibility. Furthermore, the scratch wound assay using the L929 fibroblast cell line of mouse was employed to assess the in vitro wound healing potential of the synthesised composite hydrogels., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: To determine the biological effects of arecoline on oral submucous fibrosis (OSF)., Design: The differential genes between OSF tissue and normal oral tissue were collected form GSE64216 dataset, analyzed by Gene Expression Omnibus (GEO) database. Real-time PCR and immunohistochemistry were used to analyze the expression of IL-4 gene and protein in oral tissue. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of exocrine IL-4 protein in human oral fibroblasts (HOF) pre-treated by arecoline. Cell Counting Kit-8 (CCK-8) and transwell assays were used to analyze the proliferation and migration of HOF cells, respectively. After IL-4 was knocked down by short hairpin (sh) plasmid, the proliferation and migration of HOF cells were detected. Flow cytometry was used to analyze the proportion of M2-macrophages. Real-time PCR and immunohistochemistry were used to verify the expression of biomarker proteins of macrophages in OSF tissues., Results: The expression of IL-4 gene and protein were both up-regulated in OSF tissue. Arecoline could enhance the expression of IL-4 gene and exocrine protein in HOF cells, and promote the proliferation and migration of HOF cells. While knockdown of IL-4 could inhibit arecoline-induced proliferation and migration in HOF cells. The results of flow cytometry showed that recombinant human IL-4 (rhIL-4) protein could increase the proportion of M2-macrophages. Similarly, the results of real-time PCR and immunohistochemistry showed the expression of ARG1 (Biomarker proteins of M2-macrophage) was up-regulated in OSF tissues., Conclusion: Arecoline promotes activation of fibroblasts and polarization of M2-macrophages by up-regulating the expression of IL-4., Competing Interests: Declaration of Competing Interest All authors no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objective: To assess the corrosion products and cytotoxicity of generic and proprietary fixed lingual retainers (FLRs)., Materials and Methods: Seven FLRs were investigated. Wires were submersed in solution for 34 days, at 37°C, under constant agitation. A proportion of this solution was analyzed to determine the concentration of metallic ions leaching off the wires. The remainder was diluted to 5%, 10% and 20% followed by exposure to human gingival fibroblasts and analysis of cytotoxicity of the wires., Results: Three wires (Dentaflex, Universal, and AZDent) released excessive concentrations of lead, two wires (MeshMark and Orthoflex) released excessive concentrations of nickel, and one wire (Universal) released excessive concentrations of molybdenum into solution. No statistically significant difference was found between the wires analyzed (P = .24). Slight cytotoxicity was noted in only one wire (Dentaflex) at a 20% dilution of eluent. This was also the wire which released the highest concentration of lead into solution. All other wires, at all concentrations, were deemed noncytotoxic, but five samples overall were deemed statistically significant (P < .0024). A statistically significant difference existed between wires (P = .013) and concentrations analyzed (P < .001)., Conclusions: Metals were released in differing quantities from all wires, with some elemental concentrations measuring more than that deemed acceptable in drinking water in Australia. A trend toward increased cell viability across samples was found with only one demonstrating cytotoxicity. There was no indication that generic FLRs were more or less biocompatible than their proprietary counterparts., (© 2024 by The EH Angle Education and Research Foundation, Inc.)

Objectives: The aim of these studies was to characterise the molecular effects of a tool JAK1 inhibitor on cultured primary fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA) through both total and individual cell analysis., Methods: RA-FLS cultures from 6 (Bulk RNA-seq) or 4 (ScRNA-seq) donors were pre-treated with various concentrations (100 nM and 1μM) of ABT-317 with/without exposure to 25% SEB-conditioned PBMC medium to mimic the RA inflammatory milieu. Cells were subjected to both bulk RNA-seq (36 libraries) and single cell RNA-seq (scRNA-seq; 24 libraries) to identify biological processes impacted by CM and ABT-317 treatments., Results: In our bulk RNA-seq analysis, a total of 2,605 differentially expressed genes (DEGs) were identified between CM-stimulation and unstimulated groups, while 1,122 DEGs were found between ABT-317 1μM and DMSO in CM-stimulated groups using thresholds of log2 (fold change) ≥ |0.58| and FDR ≤ 10%. Both bulk and single cell mRNA analysis of RA-FLS treated with a combination of CM and ABT-317 demonstrated the expected changes in inflammatory pathways such as interferon and IL-6 signalling. However, other non-inflammation associated pathways were also altered by ABT-317. In addition, the single cell analysis highlighted that FLS segregate into distinctive clusters upon combination CM and ABT-317 treatment, suggesting JAK inhibition can drive RA-FLS into multiple heterogenous cell populations. Interestingly, one of the unique RA-FLS clusters that emerged from the CM and ABT-317 treatment showed matrix metalloproteinase-3 (MMP3)high expression as well as several gene signatures that are not found in any other ABT-317 derived clusters., Conclusions: JAK inhibition with ABT-317 is effective at globally inhibiting CM-induced pro- and non-inflammatory pathways in FLS cultures, but also results in several distinct fibroblast populations with unique gene-associated pathways. This study advances the molecular understanding of JAK1 inhibitor effects on fibroblasts that may contribute to clinical efficacy.

Methacrylated biopolymers are unique and attractive in preparing photocrosslinkable hydrogels in biomedical applications. Here we report a novel chitosan (CS) derivative-based injectable hydrogel with anti-inflammatory capacity via methacrylation modification. First, ibuprofen (IBU) was conjugated to the backbone of CS by carbodiimide chemistry to obtain IBU-CS conjugate, which converts water-insoluble unmodified CS into water-soluble IBU-CS conjugate. The IBU-CS conjugate did not precipitate at the pH of 7, which was beneficial to subsequent chemical modification with methacrylic anhydride to prepare IBU-CS methacrylate (IBU-CS-MA) with significantly higher methacrylation substitution. Photocrosslinkable in situ gel formation of injectable IBU-CS-MA hydrogel was verified using lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) initiator under visible light. The IBU-CS-MA hydrogel showed good cytocompatibility as revealed by encapsulating and in vitro culturing murine fibroblasts within hydrogels. It promoted macrophage polarization toward M2 phenotype, as well as downregulated pro-inflammatory gene expression and upregulated anti-inflammatory gene expression of macrophages. The hydrogel also significantly reduced the reactive oxygen specifies (ROS) and nitrogen oxide (NO) produced by lipopolysaccharides (LPS)-stimulated macrophages. Upon subcutaneous implantation in a rat model, it significantly mitigated inflammatory responses as shown by significantly lower inflammatory cell density, less cell infiltration, and much thinner fibrous capsule compared with CS methacrylate (CS-MA) hydrogel. This study suggests that IBU-CS conjugate represents a feasible strategy for preparing CS-based methacrylate hydrogels for biomedical applications., (© 2024 Wiley Periodicals LLC.)