Expression Of Anti-inflammatory Cytokines Research Articles

Efficacy of caerulomycin A in modulating macrophage polarization and cytokine response in a murine model of lipopolysaccharide-induced sepsis

BackgroundSepsis is characterized by an excessive immune response. Modulation of the immune response, particularly macrophage polarization, may provide therapeutic benefit. The effects of Caerulomycin A (caeA), a known STAT1 phosphorylation inhibitor, on macrophage polarization and inflammatory markers were explored using a lipopolysaccharide (LPS)-induced sepsis mouse model.MethodsA sepsis model was established in C57BL/6 mice induced by intraperitoneal injection of LPS, and the survival rate of mice was observed after treatment with different doses of caeA to determine the optimal therapeutic dose. For in-vitro assays using the RAW264.7 macrophage cell line, the concentration of caeA that was non-toxic to cell survival was screened using the MTT assay, followed by the analyses by qRT-PCR, ELISA, Western blot and flow cytometry for M1/M2 type macrophage markers (CD86, NOS2, CD206, ARG1) and inflammatory factors (IL-1β, IL-6, TNF-α, IL-4, and IL-10) expression. In addition, the phosphorylation levels of STAT1 and STAT6 in the JAK–STAT signaling pathway were detected.ResultsThe results of in-vivo experiments showed that caeA treatment (20 mg/kg) significantly increased the survival of LPS-induced septic mice and decreased the expression of M1-type macrophage markers (CD86 and NOS2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) while increasing the expression of M2-type markers (CD206 and ARG1) and anti-inflammatory cytokines (IL-4 and IL-10) expression. In in-vitro experiments, 20 μM caeA effectively inhibited LPS-induced polarization of M1-type macrophages without affecting the activity of RAW264.7 cells, and caeA significantly inhibited the phosphorylation of STAT1 yet enhanced the phosphorylation level of STAT6, as detected by Western blot.ConclusionsCaeA effectively modulates macrophage polarization and attenuates the inflammatory response in septic mice, possibly by affecting the JAK–STAT signaling pathway. These findings support further exploration of the potential of caeA as a therapeutic agent for sepsis.

Immunomodulatory effects of calcium phosphate microspheres: influences of particle size on macrophage polarization and secretion patterns.

This study investigated the immunomodulatory effects of calcium phosphate (CaP) microspheres, focusing on how particle size influenced macrophage polarization and cytokine secretion patterns. SEM analysis revealed that HA microspheres predominantly exhibited a spherical shape with distinct sizes and sub-micro-sized pores. The average particle sizes for the S1, S2, and S3 groups were 17.36 μm, 27.59 μm, and 47.14 μm, respectively. In vitro experiments demonstrated that small-sized S1 microspheres were more readily phagocytosed by macrophages, leading to a pro-inflammatory M1 phenotype characterized by increased gene expression of iNos and inflammatory cytokines (IL-1β, IL-6, TNF-α), and a higher proportion of CCR7+ M1 macrophages. In contrast, the larger S2 and S3 microspheres favored an anti-inflammatory M2 phenotype, with higher expression of Arg and anti-inflammatory cytokines (IL-10), and greater proportions of CD206+ M2 macrophages. Additionally, HA microspheres were injected into mouse quadriceps muscles, revealing significant differences in immune cell infiltration and tissue response. The S1 microspheres induced a prolonged and more severe inflammatory response, while the S2 and S3 microspheres were embedded in cell-rich tissue with minimal inflammation or fibrosis. It indicated the potential of larger microspheres (S2 and S3) to create a more favorable immune microenvironment that supported faster and more effective tissue healing. These findings underscore the importance of optimizing microsphere size to achieve desired immunomodulatory effects, thereby enhancing their clinical efficacy.

Designing and evaluating a novel blood-brain barrier in vitro model of teleost for reproducing alterations in brain infecting

A new blood-brain barrier (BBB) in vitro model of tilapia (Oreochromis niloticus) was successfully established by co-culture with brain microvascular endothelial cell line TVEC-01 and brain astrocyte cell line TA-02 of tilapia. Experiments with the expression levels of BBB-related genes, TEER value detection and four-hour water-leaking test have shown that the BBB in vitro model has an excellent barrier effect. In bacterial penetration experiments, the pathogenic strain of Streptococcus agalactiae was able to pass through the BBB in vitro model and initiate a series of immune responses, among which TA-02 contributed to the expression of pro-inflammatory cytokines and TVEC-01 contributed to the expression of anti-inflammatory cytokines, providing an excellent research tool and theoretical basis for further study of meningitis. Moreover, the qRT-PCR and transmission electron microscopy revealed that the pathogenic strain of S. agalactiae effectively penetrated the BBB in vitro model of tilapia during early-stage infection without destroying tight junction integrity. This suggested that, in the initial phases of infection, the pathogenic strain of S. agalactiae may breach the BBB via a transcellular pathway rather than a paracellular pathway. Summarily, a novel BBB in vitro model of tilapia was successfully designed and evaluated for reproducing alterations in brain infecting.

Azithromycin-loaded PLGA microspheres coated with silk fibroin ameliorate inflammation and promote periodontal tissue regeneration.

Periodontitis, a widespread inflammatory disease, is the major cause of tooth loss in adults. While mechanical periodontal therapy benefits the periodontal disease treatment, adjunctive periodontal therapy is also necessary. Topically applied anti-inflammatory agents have gained considerable attention in periodontitis therapy. Although azithromycin (AZM) possesses excellent anti-inflammatory properties, its bioavailability is limited owing to poor water solubility and the absence of sustained release mechanisms. Herein, we synthesized biodegradable microspheres (AZM@PLGA-SF) for sustained AZM release to locally ameliorate periodontal inflammation and facilitate periodontal tissue regeneration. AZM was encapsulated in poly (lactic-co-glycolic acid) (PLGA) microspheres (AZM@PLGA) using single emulsion-solvent evaporation, followed by surface coating with silk fibroin (SF) via electrostatic adsorption, reducing the initial burst release of AZM. In vivo, local treatment with AZM@PLGA-SF microspheres significantly reduced periodontal inflammation and restored periodontal tissue to healthy levels. Mechanically, the formulated microspheres regulated the periodontal inflammatory microenvironment by reducing the levels of pro-inflammatory cytokines (tumor necrosis factor -α, interleukin [IL]-6, interferon-γ, IL-2, and IL-17A) in gingival crevicular fluid and promoted the expression of anti-inflammatory cytokines (IL-4 and IL-10). AZM@PLGA-SF microspheres demonstrated excellent biological safety. Therefore, we introduce an anti-inflammatory therapy for periodontitis with substantial potential for mitigating periodontal inflammation and encouraging the repair and regeneration of periodontal tissues.

The role of probiotics in restoring the Th1 to Th2 ratio in women experiencing recurrent implantation failure; a double-blind randomized clinical trial.

Among the complex causes of infertility, Recurrent Implantation Failure (RIF) stands out as a challenging condition. Immunological aberrations are closely implicated in RIF, particularly the imbalance between T helper 1 (Th1) and T helper 2 (Th2) cytokines. Interest in harnessing probiotics to enhance fertility outcomes in RIF-affected women is growing. This study aimed to investigate the potential of probiotics in restoring the Th1 to Th2 ratio and enhancing implantation outcomes in women experiencing RIF. A double-blinded randomized controlled trial enrolled 50 women with a record of at least three RIFs and 50 women with successful pregnancies as a control group in our study. RIF patients were randomly assigned to either the probiotic intervention group (n=25) or the placebo group (n=25). The probiotic group administered two tablets per day for 6months the probiotic tablets that contained prebiotic. At the same time, the placebo group received an identical-looking placebo. Flow cytometry technique was used to evaluate Th1 and Th2 cell frequencies. Gene expression of IL-4, IL-10, TNF-α and IFN-ɣ was analyzed using QRT-PCR. Serum concentrations of IL-4, IL-10, TNF-α and IFN-ɣ cytokines were evaluated using the ELISA technique. Flow cytometry analysis revealed significantly higher Th1 cell percentages in RIF patients compared to healthy controls, while healthy controls exhibited more significant Th2 cell proportions. Probiotic intervention led to a reduction in Th1 cells and an increase in Th2 cells in RIF patients. ELISA analysis indicated higher proinflammatory and lower anti-inflammatory cytokines compared to controls. After probiotic treatment, TNF-α and IFN-ɣ levels decreased, while IL-10 and IL-4 levels increased in RIF patients. QRT-PCR analysis showed no significant differences in cytokine gene expression between RIF patients before intervention. After probiotic therapy, pro-inflammatory cytokine expression decreased, and anti-inflammatory cytokine expression increased. Placebo treatment resulted in limited changes. Probiotic intervention effectively modulated the Th1 to Th2 ratio in RIF patients, evidenced by reduced Th1 cell percentages, enhanced Th2 cell populations, and cytokine level adjustments. The probiotic intervention improved implantation outcomes in women with RIF, as evidenced by a higher clinical pregnancy rate in the treatment group compared to the placebo group (p=0.037). These findings highlight the potential of probiotics in restoring immune balance and improving implantation outcomes in women with RIF.

Antioxidant, Anti-Inflammatory, and Anticancer Activities of Five Citrus Peel Essential Oils.

Citrus peel essential oil (CPEO) is favored by people for its aromatic scent, while also possessing numerous bioactive compounds that are advantageous to human health. This study evaluated the antioxidant, anti-inflammatory, and anticancer activities of CPEOs through cell experiments. The results showed that CPEOs could increase the activity of the antioxidant enzyme system and nonenzymatic defence system in H2O2-treated RAW 264.7 cells by reducing cellular lipid peroxidation. CPEOs also reduced the nitric oxide production induced by lipopolysaccharide treatment in RAW 264.7 cells while decreasing proinflammatory cytokines expression and increasing anti-inflammatory cytokine expression. Wound healing assays, flow cytometry, and quantitative real-time fluorescent quantitative PCR (qRT-PCR) revealed that CPEOs could induce apoptosis in U87 cells through the mitochondrial apoptotic pathway. These findings indicate that CPEOs possess excellent antioxidant, anti-inflammatory, and anticancer activity potential, making them suitable for use in functional antioxidant and anti-inflammatory foods and nutritional health products.

Imperatorin, a natural furanocoumarin alleviates chronic neuropathic pain by targeting GABAergic tone in an animal model of spinal cord injury.

Chronic neuropathic pain (CNP) caused by spinal cord injury (SCI) has long-term implications, that result in functional deficit. In this study, we explored imperatorin, a natural furanocoumarin isolated from the dried pulp of Aegle marmelos L. Imperatorin (10 mg/kg, i.p.) was administered to injured animals for seven consecutive days. It was discovered that imperatorin reduced the CNP by upregulating the expression of anti-inflammatory chemokines and cytokines like IL-10 and IL-12, and increasing the expression of GABA with p-value < 0.001 as compared to vehicle at DPI-28 and DPI-42. Also, GABA receptor expression was increased with p-value < 0.01 as compared to vehicle at DPI-28. Additionally, it downregulated the expression of various proinflammatory mediators like IL-6 (p-value < 0.01), CCL-2 (p-value < 0.01), IL-1β (p-value < 0.01), and CCL-3 (p-value < 0.001) as compared to vehicle at DPI-1. Furthermore, imperatorin reduced scar formation by decreasing fibrosis and gliosis post-SCI and also lessened pain behaviour, suggesting it helps reduce chronic neuropathic pain (CNP).

BPA Exacerbates Zinc Deficiency-Induced Testicular Tissue Inflammation in Male Mice Through the TNF-α/NF-κB/Caspase8 Signaling Pathway.

Bisphenol A (BPA) is an endocrine-disrupting chemical that is toxic to reproduction. Zinc (Zn) plays an important role in male reproductive health. Zn deficiency (ZD) can co-exist with BPA. In order to investigate the specific mechanism of reproductive damage caused by BPA exposure in ZD male mice, a mouse model of ZD, BPA exposure, and their combined exposure was established in this study. Forty 4-week-old SPF male ICR mice with an average body weight of 31.7 ± 4.2g were divided into four groups including normal Zn diet group 30mg/(kg•d), BPA exposure group 150mg/(kg•d), zinc deficiency diet group 7.5mg/(kg•d), and BPA + ZD combined exposure group (BPA 150mg/(kg•d) + ZD 7.5mg/(kg•d)). The mice were kept for 8weeks. The results showed that the testicular tissue structure was disturbed, and semen quality, serum Zn, testicular tissue Zn, and testicular tissue free Zn ions were decreased in the BPA-exposed and ZD groups. The expression of zinc transporters (ZIP7, ZIP8, ZIP13, and ZIP14) in testicular tissue was changed. The expressions of pro-inflammatory cytokines including TNF-α and IL-1β as well as inflammatory pathway-related proteins (IKB-α, p-IKB-α, NF-κB, p-NF-κB, Caspase8, and Caspase3) were increased, while the expressions of anti-inflammatory cytokines (TGF-β and IL-10) were decreased. The changes in the above indexes in the BPA + ZD group were more obvious. Both BPA exposure and ZD can induce testicular tissue inflammation through the TNF-α/NF-κB/Caspase8 signaling pathway, and BPA further aggravates zinc deficiency-induced testicular tissue inflammation and apoptosis damage.

Improving dexamethasone drug loading and efficacy in treating rheumatoid arthritis via liposome: Focusing on inflammation and molecular mechanisms.

Rheumatoid arthritis (RA) is a chronic autoimmune disease that affects approximately 0.46% of the global population. Conventional therapeutics for RA, including disease-modifying antirheumatic drugs (DMARDs), nonsteroidal anti-inflammatory drugs (NSAIDs), and corticosteroids, frequently result in unintended adverse effects. Dexamethasone (DEX) is a potent glucocorticoid used to treat RA due to its anti-inflammatory and immunosuppressive properties. Liposomal delivery of DEX, particularly when liposomes are surface-modified with targeting ligands like peptides or sialic acid, can improve drug efficacy by enhancing its distribution to inflamed joints and minimizing toxicity. This study investigates the potential of liposomal drug delivery systems to enhance the efficacy and targeting of DEX in the treatment of RA. Results from various studies demonstrate that liposomal DEX significantly inhibits arthritis progression in animal models, reduces joint inflammation and damage, and alleviates cartilage destruction compared to free DEX. The liposomal formulation also shows better hemocompatibility, fewer adverse effects on body weight and immune organ index, and a longer circulation time with higher bioavailability. The anti-inflammatory mechanism is associated with the downregulation of pro-inflammatory cytokines like tumor necrosis factor-α (TNF-α) and B-cell-activating factor (BAFF), which are key players in the pathogenesis of RA. Additionally, liposomal DEX can induce the expression of anti-inflammatory cytokines like interleukin-10 (IL-10), which has significant anti-inflammatory and immunoregulatory properties. The findings suggest that liposomal DEX represents a promising candidate for effective and safe RA therapy, with the potential to improve the management of this debilitating disease by providing targeted delivery and sustained release of the drug.

Characteristics and Biological Activities of a Novel Polysaccharide R1 Isolated from Rubus chingii Hu.

Raspberry (Rubus chingii Hu) is a Chinese herb that is rich in nutrients and has anti-inflammatory, antibacterial, antioxidant, anti-allergic, hypoglycemic, and other effects. A water-soluble polysaccharide was extracted from raspberry by using hot water extraction then purified by DEAE-Sepharose Fast Flow column chromatography. The structural characteristics of the polysaccharide (R1) are as follows: the molar ratio of the monosaccharide composition is Ara:Gal:Xyl:Glc:Man = 31.15:27.64:13.61:13.48:10.60; the molecular weight is 32,580 Da; the methylation results show that 5-Araf is the main chain and there is a presence of 3,6-Galp, 4-Xylp, and 2,3,5-Araf branches, and that terminal Araf (T-Araf) is the major telomeric sugar. It contains α and β glycosidic bonds and is highly branched, with the presence of a helical structure. In the in vitro antioxidant assay, R1 showed the highest scavenging of superoxide anion radicals at 70.38%, followed by the scavenging of DPPH radicals at 52.9% and the scavenging of hydroxyl radicals at 29.28%. In immunomodulation and anti-cancer experiments, R1 did not significantly inhibit or promote RAW264.7 cells but was able to increase the expression of anti-inflammatory cytokines in a concentration-dependent manner. It also significantly inhibited cancer cell survival. R1 enhances immunity by limiting the proliferation of cancer cells primarily through direct inhibition while promoting the secretion of pro-inflammatory cytokines. These findings reveal the potential benefits of raspberry polysaccharides and provide evidence for developing immunologically functional products from raspberry polysaccharides.

Neuroprotective effect of L-DOPA-induced interleukin-13 on striatonigral degeneration in cerebral ischemia

Levodopa (L-DOPA) treatment is a clinically effective strategy for improving motor function in patients with ischemic stroke. However, the mechanisms by which modulating the dopamine system relieves the pathology of the ischemic brain remain unclear. Emerging evidence from an experimental mouse model of ischemic stroke, established by middle cerebral artery occlusion (MCAO), suggested that L-DOPA has the potential to modulate the inflammatory and immune response that occurs during a stroke. Here, we aimed to demonstrate the therapeutic effect of L-DOPA in regulating the systemic immune response and improving functional deficits in mice with ischemia. Transient MCAO led to progressive degeneration of nigrostriatal dopamine neurons and significant rotational behavior in mice. Exogenous L-DOPA treatment attenuated the striatonigral degeneration and reversed motor behavioral impairment. Notably, treatment with L-DOPA significantly increased IL-13 but reduced IFN-γ in infarct lesions. To investigate the role of IL-13 in motor behavior, we stereotaxically injected anti-IL-13 antibodies into the infarct area of the mouse brain one week after MCAO, followed by L-DOPA treatment. The intervention reduced dopamine, IL-13, and IL-10 levels and exacerbated motor function. IL-13 is potentially expressed on CD4 T cells, while IL-10 is mainly expressed on microglia rather than astrocytes. Finally, IL-13 activates the phagocytosis of microglia, which may contribute to neuroprotection by eliminating degenerating neurons. Our study provides evidence that the L-DOPA-activated dopamine system modulates peripheral immune cells, resulting in the expression of anti-inflammatory and neuroprotective cytokines in mice with ischemic stroke.

DDEL-16. MICROMESH FOR REVAMPING THE ‘COLD’ TUMOR MICROENVIRONMENT IN GLIOBLASTOMA IMMUNOTHERAPY

Abstract High-grade gliomas have poor prognoses with limited treatment options, typically maximal safe resection and adjuvant radiochemotherapy. Novel therapies often face challenges due to low brain penetrance and poor bioavailability. To address these issues, a conformable, compartmentalized polymeric implant called microMESH has been developed. microMESH provides localized, sustained release of drugs, small inhibitors, antibodies, and nanomedicines, enabling novel therapeutic combinations. When applied to the tumor margins, microMESH adheres to the tissue, releasing its therapeutic cargo over weeks and reducing systemic toxicity. Its biocompatibility, cytotoxicity, and anti-tumor efficacy were tested on murine cancer cells (CT-2A) and orthotopic glioblastoma syngeneic models. Exposing primary bone marrow-derived monocytes (BMDM) to CT-2A conditioned medium increased the expression of anti-inflammatory cytokines (IL-10 and IL-4) by 2 times without significantly affecting the pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), indicating that CT-2A cells tend to establish an immunosuppressive ‘cold’ tumor microenvironment. Next, the co-incubation of CT-2A cells with BMDM followed by the administration of the immune checkpoint inhibitor aCD47 resulted in a cancer cell viability drop by 50% already at 50 μg/ml aCD47 after 1 day. Additionally, pre-treatment with docetaxel (DTXL) further decreased cell viability, suggesting a synergism between aCD47 and the chemotherapeutic drug. Flow cytometry showed that 50 nM DTXL at 3 days post-incubation was responsible for a strong translocation of the ‘immunogenic dell death’ protein calreticulin on the cell membrane, potentially explaining the synergism. Over 2 months, the intracranially deployed microMESH biodegraded without inducing any toxic effect. In orthotopic CT-2A tumors, aCD47/DTXL–microMESH (0.75 mg/kg, single intracranial application) showed a 40-day survival compared to 22 days for iv DTXL (3 mg/kg qod), 24 days for ip aCD47 (5 mg/kg qod), and 18 days for untreated mice. microMESH helped identify and enabled novel combination therapies directly at the tumor bed, demonstrating superior efficacy over systemic therapies.

TLR2/TLR5 Signaling and Gut Microbiota Mediate Soybean-Meal-Induced Enteritis and Declined Growth and Antioxidant Capabilities in Large Yellow Croaker (Larimichthys crocea)

Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims to elucidate the molecular basis of soybean-meal-induced enteritis and its impact on intestinal microbiota in L. crocea. To this end, four isonitrogenous and isolipidic diets with varying soybean meal levels (0% FM, 15% SBM15, 30% SBM30, and 45% SBM45) were administered to L. crocea for 8 weeks. The results indicated that the SBM30 and SBM45 treatments significantly hindered fish growth, digestive efficiency, and protein utilization. Furthermore, high soybean meal levels (SBM30 and SBM45) activated intestinal Toll-like receptors (TLR2A, TLR2B, TLR5, and TLR22), stimulating C-Rel and mTOR protein expression and elevating ERK phosphorylation. This led to increased pro-inflammatory cytokine production (IL-1β, IL-6, and TNF-α) and decreased anti-inflammatory cytokine expression (IL-4/13A, IL-4/13B, and TGF-β), suggesting a potential signaling pathway for soybean-meal-induced enteritis. Furthermore, enteritis induced by high soybean meal levels led to oxidative damage, evident from increased MDA levels and decreased antioxidant enzyme activities (SOD and CAT). The SBM30 and SBM45 treatments increased Firmicutes and Bacteroidetes abundance in fish gut microbiota, while Proteobacteria abundance decreased. This microbiota shift may enhance soybean meal nutrient utilization, yet high soybean meal concentrations still impair growth. A soybean-meal-rich diet promotes harmful bacteria like Rhodococcus and depletes probiotics like Ralstonia, increasing disease risks. L. crocea has limited tolerance for soybean meal, necessitating advanced processing to mitigate anti-nutritional factors. Ultimately, exploring alternative protein sources beyond soybean meal for fish meal replacement is optimal for L. crocea.

Development and Optimization of a Redox Enzyme-Based Fluorescence Biosensor for the Identification of MsrB1 Inhibitors.

MsrB1 is a thiol-dependent enzyme that reduces protein methionine-R-sulfoxide and regulates inflammatory response in macrophages. Therefore, MsrB1 could be a promising therapeutic target for the control of inflammation. To identify MsrB1 inhibitors, we construct a redox protein-based fluorescence biosensor composed of MsrB1, a circularly permutated fluorescent protein, and the thioredoxin1 in a single polypeptide chain. This protein-based biosensor, named RIYsense, efficiently measures protein methionine sulfoxide reduction by ratiometric fluorescence increase. We used it for high-throughput screening of potential MsrB1 inhibitors among 6868 compounds. A total of 192 compounds were selected based on their ability to reduce relative fluorescence intensity by more than 50% compared to the control. Then, we used molecular docking simulations of the compound on MsrB1, affinity assays, and MsrB1 activity measurement to identify compounds with reliable and strong inhibitory effects. Two compounds were selected as MsrB1 inhibitors: 4-[5-(4-ethylphenyl)-3-(4-hydroxyphenyl)-3,4-dihydropyrazol-2-yl]benzenesulfonamide and 6-chloro-10-(4-ethylphenyl)pyrimido[4,5-b]quinoline-2,4-dione. They are heterocyclic, polyaromatic compounds with a substituted phenyl moiety interacting with the MsrB1 active site, as revealed by docking simulation. These compounds were found to decrease the expression of anti-inflammatory cytokines such as IL-10 and IL-1rn, leading to auricular skin swelling and increased thickness in an ear edema model, effectively mimicking the effects observed in MsrB1 knockout mice. In summary, using a novel redox protein-based fluorescence biosensor, we identified potential MsrB1 inhibitors that can regulate the inflammatory response, particularly by influencing the expression of anti-inflammatory cytokines. These compounds are promising tools for understanding MsrB1's role during inflammation and eventually controlling inflammation in therapeutic approaches.

Effects of Conditioned Media From Human Umbilical Cord-Derived Mesenchymal Stem Cells on Tenocytes From Degenerative Rotator Cuff Tears in an Interleukin 1β-Induced Tendinopathic Condition.

Evidence suggests that mesenchymal stem cells (MSCs) are safe for treating different tendinopathies. Synovial fluid is a pooled environment of biomarkers from the inflammatory and degenerative joint cavity. Understanding the effects of synovial fluid on MSCs is important, as it is the first microenvironment that administered MSCs encounter. Several studies have reported that exposure to osteoarthritic synovial fluid-activated MSCs increased the release of soluble factors; however, the paracrine effects of shoulder synovial fluid-stimulated umbilical cord-derived MSCs (SF-UC-MSCs) on tendinopathy have yet to be investigated. To assess the effects of the conditioned media from SF-UC-MSCs on tenocytes from degenerative rotator cuff tears in an interleukin-1β (IL-1β)-induced tendinopathic condition. Controlled laboratory study. UC-MSCs were isolated and cultured from healthy, full-term deliveries by cesarean section. Tenocytes were isolated and cultured from patients with degenerative rotator cuff tears. Conditioned media were obtained from UC-MSCs stimulated with synovial fluid. To evaluate the gene expression of proinflammatory and anti-inflammatory cytokines, enzymes and their inhibitors, matrix molecules, and growth factors, the tenocytes were cultured with IL-1β and 50% of the conditioned media from the SF-UC-MSCs; quantitative, real-time, reverse transcriptase polymerase chain reaction was also performed. A prostaglandin E2 (PGE2) assay was performed to investigate the PGE2 level secreted by the tenocytes. Western blotting was performed to examine protein synthesis of collagen type I and III. Cell viability, senescence, and apoptosis assays were also performed. The conditioned media from the SF-UC-MSCs interfered with the inflammatory gene expression on tenocytes induced by IL-1β, but it increased the gene expression of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-3. Meanwhile, the conditioned media decreased the PGE2 level on cells induced by IL-1β. It did increase the type I/III ratio of gene expression and protein synthesis, mainly through the induction of type I collagen. Conditioned media of SF-UC-MSCs reversed senescence and apoptosis induced by IL-1β. Study findings indicated that the conditioned media from SF-UC-MSCs had anti-inflammatory effects and cytoprotective effects on IL-1β-treated tenocytes from degenerative rotator cuff tears. UC-MSCs have useful potential for the treatment of tendinopathy in practice.

Shikonin delivery nano system based on calcium alginate for targeted therapy of inflammatory bowel disease by modulating macrophage polarization

Effective delivery of therapeutic drugs to colonic epithelial cells and macrophages is crucial for mitigating inflammation and repairing the intestinal barrier in inflammatory bowel disease (IBD). We developed a pH-triggered colonic targeted delivery nano system utilizing natural polymers, including chitosan (CS), hyaluronic acid (HA), and calcium alginate (ALG), encapsulating the active component shikonin (SK) to structure SK-loaded ALG/HA/CS delivery nano system (SK@LAS). The oral administration of SK@LAS specifically delivered SK in the colon via pH sensitivity and colonic targeting. SK@LAS was internalized by CD44 receptor-mediated endocytosis into macrophages. Furthermore, significant therapeutic effects of SK@LAS were observed in acute and chronic mouse colitis models, including reduced pro-inflammatory cytokines and mediators, increased expression of anti-inflammatory cytokines, and suppression of M1 polarization. Furthermore, SK@LAS upregulated the expression of tight junction proteins, thereby restoring intestinal barrier function. This colonic targeted delivery nano system exhibits promise as an oral drug delivery approach for the treatment of IBD.

New Nanovesicles from Prickly Pear Fruit Juice: A Resource with Antioxidant, Anti-Inflammatory, and Nutrigenomic Properties.

Plant-derived nanovesicles represent a novel approach in the field of plant-derived biomaterials, offering a sustainable and biocompatible option for various biomedical applications. The unique properties of these vesicles, such as their ability to encapsulate bioactive compounds, make them suitable for therapeutic, cosmetic, and nutraceutical purposes. In this study, we have, for the first time, successfully bio-fabricated vesicles derived from Opuntia ficus-indica (FicoVes) using an efficient and cost-effective method. Characterized by a size of approximately of 114 nm and a negative zeta potential of -20.9 mV, FicoVes exhibited excellent biocompatibility and hemocompatibility, showing no reduction in the viability of human and animal cells. Our results showed that FicoVes possess significant antioxidant properties as they reduced ROS generation in TBH-stimulated cells. FicoVes displayed anti-inflammatory properties by reducing the expression of pro-inflammatory cytokines (Il 1β, TNF α) and enhancing the expression of anti-inflammatory cytokines (IL4, IL10) following an inflammatory stimulus. Furthermore, FicoVes accelerated epithelial wound closure in L929 fibroblast monolayers in a dose-dependent manner, highlighting their potential role in tissue repair. This study establishes FicoVes as a promising candidate for nutrigenomic applications, particularly in the context of inflammation-related disorders and wound healing. Further research, including in vivo studies, is essential to validate these findings and fully explore their therapeutic potential.

Influence of titanium nanoparticles on cytotoxicity and inflammatory cytokines expression in gingival fibroblasts - An in vitro study

Background Despite the success of titanium (Ti) implants in dental rehabilitation, emerging evidence implicates the release of titanium oxide nanoparticles (TiO2NPs) from the implant surface as a potential contributor to the initiation of the pathogenic process leading to peri-implantitis and ultimately failure of the implants. However, a comprehensive investigation to elucidate the dose-dependent effects of TiO2NPs on cytotoxicity and inflammatory responses is lacking. The present study was undertaken to evaluate the effect of different concentrations of TiO2NPs on the cytotoxicity and inflammatory cytokine expression in Human Gingival Fibroblasts (HGFs). Methods HGFs were isolated from gingival tissue samples obtained from periodontally and systemically healthy subjects. Ti standard solution for ICP was diluted to create concentrations of (0.001 ppm, 0.01 ppm, 0.1 ppm, 1 ppm, 10 ppm, and 100 ppm) for cell culture media containing titanium. HGFs were then cultured in these varying concentrations for specific time periods (days 1, 3, 5, and 7) to assess cell viability. A cytotoxicity assay was performed to determine the levels and expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and anti-inflammatory cytokines (IL-10, IL-12, TGF-β) using qRT-PCR and ELISA techniques. Results Our findings demonstrate a concentration and duration-dependent decrease in HGF viability upon exposure to titanium nanoparticles. Notably, a 50% reduction in cell viability was observed at the highest concentration (100 ppm). qRT-PCR analysis revealed a significant upregulation of pro-inflammatory cytokines, particularly IL-1β and IL-6, in HGFs exposed to titanium. Interestingly, the expression of anti-inflammatory cytokines (IL-10, IL-12, and TGF-β) remained comparable or even equivalent compared to controls across different titanium concentrations. Conclusions The study revealed a concentration and duration-dependent influence on HGF viability and cytokine profile, suggesting potential cytotoxicity and modulation of the inflammatory response mediated by TiO2NPs. Further research is necessary to elucidate the underlying mechanisms and their implications for dental implant biocompatibility.

Functional Role of Piezo1 in the Human Eosinophil Cell Line AML14.3D10: Implications for the Immune and Sensory Nervous Systems.

Mechanosensitive ion channels, particularly Piezo channels, are widely expressed in various tissues. However, their role in immune cells remains underexplored. Therefore, this study aimed to investigate the functional role of Piezo1 in the human eosinophil cell line AML14.3D10. We detected Piezo1 mRNA expression, but not Piezo2 expression, in these cells, confirming the presence of the Piezo1 protein. Activation of Piezo1 with Yoda1, its specific agonist, resulted in a significant calcium influx, which was inhibited by the Piezo1-specific inhibitor Dooku1, as well as other nonspecific inhibitors (Ruthenium Red, Gd3+, and GsMTx-4). Further analysis revealed that Piezo1 activation modulated the expression and secretion of both pro-inflammatory and anti-inflammatory cytokines in AML14.3D10 cells. Notably, supernatants from Piezo1-activated AML14.3D10 cells enhanced capsaicin and ATP-induced calcium responses in the dorsal root ganglion neurons of mice. These findings elucidate the physiological role of Piezo1 in AML14.3D10 cells and suggest that factors secreted by these cells can modulate the activity of transient receptor potential 1 (TRPV1) and purinergic receptors, which are associated with pain and itch signaling. The results of this study significantly advance our understanding of the function of Piezo1 channels in the immune and sensory nervous systems.

Cultured fecal microbial community and its impact as fecal microbiota transplantation treatment in mice gut inflammation

The fecal microbiome is identical to the gut microbial communities and provides an easy access to the gut microbiome. Therefore, fecal microbial transplantation (FMT) strategies have been used to alter dysbiotic gut microbiomes with healthy fecal microbiota, successfully alleviating various metabolic disorders, such as obesity, type 2 diabetes, and inflammatory bowel disease (IBD). However, the success of FMT treatment is donor-dependent and variations in gut microbes cannot be avoided. This problem may be overcome by using a cultured fecal microbiome. In this study, a human fecal microbiome was cultured using five different media; growth in brain heart infusion (BHI) media resulted in the highest microbial community cell count. The microbiome (16S rRNA) data demonstrated that the cultured microbial communities were similar to that of the original fecal sample. Therefore, the BHI-cultured fecal microbiome was selected for cultured FMT (cFMT). Furthermore, a dextran sodium sulfate (DSS)-induced mice-IBD model was used to confirm the impact of cFMT. Results showed that cFMT effectively alleviated IBD-associated symptoms, including improved gut permeability, restoration of the inflamed gut epithelium, decreased expression of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1, IL-6, IL-12, and IL-17), and increased expression of anti-inflammatory cytokines (IL-4 and IL-10). Thus, study’s findings suggest that cFMT can be a potential alternative to nFMT.Key points• In vitro fecal microbial communities were grown in a batch culture using five different media.• Fecal microbial transplantation was performed on DSS-treated mice using cultured and normal fecal microbes.• Cultured fecal microbes effectively alleviated IBD-associated symptoms.Graphical