Search Results (137)

The endometrial immune response in postpartum cows is key to maintaining uterine health and preventing inflammatory diseases such as metritis and endometritis. Appropriate management strategies and diets that enhance the immune response are crucial during the transition period; therefore, diets rich in omega-3 fatty acids have been proposed for their potential beneficial effects on cows. Docosahexaenoic acid (DHA) is an omega-3 fatty acid with anti-inflammatory effects in immune cells; however, its effects on bovine endometrial immunity are not fully known. This study aimed to determine the effect of DHA on the inflammatory response in bovine endometrial (BEND) cells. BEND cells were incubated with DHA without or with lipopolysaccharide (LPS), and the mRNA expressions of prostaglandin-endoperoxide synthase 2 (PTGS2), interleukin (IL)-6, and CXCL8 were analyzed using RT-qPCR. The protein amount of IL-6, or CXCL8, and Resolvin D1 (RvD1) in the cell culture medium were analyzed using ELISA. DHA significantly reduced the expression of LPS-induced IL-6 and PTGS2 but increased LPS-induced CXCL8 expression. In addition, DHA reduced LPS-induced ERK1/2 and Akt phosphorylation, as assessed by immunoblotting. DHA increased the production of RvD1, a metabolite of DHA, at 8 and 24 h. In addition, RvD1 reduced LPS-induced CXCL8 production and increased the phosphorylation of ERK1/2 and Akt. Finally, changes in metabolite levels, such as an increase in 2-hydroxypyridine in DHA-treated cells, were obtained using a metabolomic assay. In conclusion, DHA reduced IL-6 and PTGS2 mRNA expression and IL-6 protein release and increased RvD1 levels in bovine endometrial cells, which suggest that DHA could have beneficial effects on endometrial immunity. The increase in CXCL8 mRNA expression and protein release induced by DHA remains to be studied; however, it could play a role in the innate defensive mechanisms of phagocytes. Full article

Chlorpyrifos (CPF), an organophosphate pesticide, is known to induce pulmonary toxicity through oxidative stress, mitochondrial dysfunction, and inflammation. Astaxanthin (ASX), a xanthophyll carotenoid derived primarily from marine microalgae (Haematococcus pluvialis), possesses strong antioxidant properties and has demonstrated cellular protective effects in numerous oxidative stress studies. However, its efficacy against CPF-induced lung cell damage remains uncharacterized. This study revealed the protective role of ASX, as a pretreatment and co-treatment, against CPF-induced cytotoxicity in human A549 lung adenocarcinoma cells by assessing cell viability, intracellular reactive oxygen species (IROS), total oxidative status (TOS), total antioxidant capacity (TAC), mitochondrial membrane potential (MMP), intracellular calcium ions (Ca²⁺), lactate dehydrogenase (LDH) release, malondialdehyde (MDA) levels, glutathione peroxidase (GPx) activity, superoxide dismutase (SOD) activity, DNA fragmentation, and apoptosis/inflammation-associated gene expression. CPF treatment significantly decreased cell viability and TAC, while elevating IROS, TOS, MMP, intracellular Ca²⁺, and LDH release. CPF also increased MDA levels and suppressed GPx and SOD activities. DNA fragmentation and quantitative polymerase chain reaction (qPCR) analysis revealed upregulation of pro-apoptotic and inflammatory markers such as BCL2-associated X protein (BAX), caspase-3 (CASP3), tumor protein p53 (TP53), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), nuclear factor kappa B (NFκB), and voltage-dependent anion-selective channel protein 1 (VDAC1) and suppression of anti-apoptotic B-cell lymphoma 2 (BCL2) and antioxidant defense genes nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). ASX treatment, particularly when administered as a pretreatment, significantly reversed CPF-induced oxidative and inflammatory responses by restoring SOD, GPx, and TAC levels, reducing IROS, TOS, MDA, and LDH release, and downregulating apoptotic and inflammatory gene expressions. ASX pretreatment notably decreased MMP and intracellular Ca²⁺ levels, indicating protection against mitochondrial dysfunction and calcium dysregulation. ASX upregulated Nrf2 and HO-1 expression and restored the BCL2/BAX balance, suggesting inhibition of mitochondrial-mediated apoptosis. Additionally, ASX significantly attenuated CPF-induced anti-angiogenic effects in the in ovo Hen’s Egg Test Chorioallantoic Membrane (HET-CAM) assay. These findings demonstrate, for the first time, that ASX exerts a broad spectrum of protective effects against CPF-induced cytotoxicity in lung cells, mainly through the stabilization of mitochondrial redox status and modulation of apoptosis- and inflammation-related gene pathways, highlighting ASX as a promising candidate for further therapeutic development. Furthermore, the pronounced efficacy observed in the pretreatment regimen suggests that ASX can be evaluated as a potential nutritional preventive strategy in high-risk populations with occupational or environmental CPF exposure. Full article

Background: Cerebral aneurysm (CA) is a focal or diffuse pathological dilation of the cerebral arterial wall that arises due to various etiological factors. It represents a serious vascular condition, particularly affecting the elderly, and carries a high risk of rupture and neurological morbidity. Clonidine (CL), an α2-adrenergic receptor agonist, has been reported to suppress aneurysm progression; however, its underlying molecular mechanisms, especially in relation to cerebral endothelial dysfunction, remain unclear. This study aimed to investigate the potential of CL to mitigate CA development by modulating apoptosis, inflammation, and oxidative stress in an Angiotensin II (Ang II)-induced endothelial injury model. Methods: Human brain microvascular endothelial cells (HBMECs) were used to establish an in vitro model of endothelial dysfunction by treating cells with 1 µM Ang II for 48 h. CL was administered 2 h prior to Ang II exposure at concentrations of 0.1, 1, and 10 µM. Cell viability was assessed using the MTT assay. Oxidative stress markers, including reactive oxygen species (ROS) and Nitric Oxide (NO), were measured using 2′,7′–dichlorofluorescin diacetate (DCFDA). Gene expression levels of vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP-2 and MMP-9), high mobility group box 1 (HMGB1), and nuclear factor kappa B (NF-κB) were quantified using RT-qPCR. Levels of proinflammatory cytokines; tumor necrosis factor-alpha (TNF-α), Interleukin-6 (IL-6), and interferon-gamma (IFN-γ); were measured using commercial ELISA kits. Results: Ang II significantly increased ROS production and reduced NO levels, accompanied by heightened proinflammatory cytokine release and endothelial dysfunction. MTT assay revealed a marked decrease in cell viability following Ang II treatment (34.18%), whereas CL preserved cell viability in a concentration-dependent manner: 44.24% at 0.1 µM, 66.56% at 1 µM, and 81.74% at 10 µM. CL treatment also significantly attenuated ROS generation and inflammatory cytokine levels (p < 0.05). Furthermore, the expression of VEGF, HMGB1, NF-κB, MMP-2, and MMP-9 was significantly downregulated in response to CL. Conclusions: CL exerts a protective effect on endothelial cells by reducing oxidative stress and suppressing proinflammatory signaling pathways in Ang II-induced injury. These results support the potential of CL to mitigate endothelial injury in vitro, though further in vivo studies are required to confirm its translational relevance. Full article

Objectives: This study investigated the properties of a neutral exopolysaccharide (EPS-LP1) with an average molecular weight of 55,637 Da, isolated from Lactiplantibacillus plantarum DMDL 9010 (LP9010). Results: The composition of EPS-LP1 includes galactose (Gal), glucose (Glu) and mannose (Man) in a molar ratio of 5.35:86.25:8.40. Notably, EPS-LP1 exhibits a smooth and rod-like surface along with thermal stability. Methylation combined with nuclear magnetic resonance analysis revealed that EPS-LP1 structured as t-Galp(1→, →6)-Glcp(1→, 4)-Glcp(1→ and →4,6)-Galp(1→), with relative molar ratio of 1.016:9.874:4.355:78.693:6.062, respectively. In the concentration range of 50 to 400 mg/mL, we observed the absence of cytotoxic effects from EPS-LP1 on RAW264.7 cells. Furthermore, EPS-LP1 demonstrated protective effects on RAW264.7 cells against oxidative damage by reducing the production of reactive oxygen species (ROS), malondialdehyde (MDA), and lactate dehydrogenase (LDH) release. Conversely, an increase in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and concentrations of glutathione (GSH) was observed. Immunoreactivity assays indicated that EPS-LP1 can effectively reduce the production of nitric oxide (NO) and inhibit the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Additionally, it inhibited the activation of the mitogen-activated protein kinase (MAPK)/nuclear factor-kappa B gene binding (NF-kB) signaling pathway. Conclusions: This research provides a foundation basis for further investigations into the neutral exopolysaccharide derived from LP9010. Full article

Psoriasis is a chronic inflammatory skin disease characterized by erythema, infiltration, and scaling, which is mainly caused by interleukin (IL)-17. The use of molecular targeted drugs in specific therapies offers high efficacy; however, high medical costs and a significant risk of side effects highlight the need for novel therapeutic agents. We previously observed that Morus alba extract (MAE) suppressed IL-17-induced CCL20 mRNA expression in normal human epidermal keratinocytes (NHEKs). In this study, we focused on the IL-17 signaling pathway and investigated the effects of pentacyclic triterpenoids, betulinic acid (BA), and ursolic acid (UA), which are present in MAE, on NHEK cells. Real-time reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) revealed that both BA and UA suppressed CCL20 expression, while only UA alone inhibited CCL20 release. ELISA using specific inhibitors demonstrated that both the p38 and extracellular-signal-regulated kinase 1/2 (ERK1/2) pathways were crucial for IL-17-induced CCL20 release in NHEK. UA effectively suppressed ERK1/2 nuclear localization and moderately affected p38 phosphorylation. These results indicated that UA is a potential seed compound for psoriasis treatment through its targeting of the IL-17 pathway. Full article

Background/Objectives: Degenerative arthritis is a chronic inflammatory disease marked by tissue degradation and vascular fibrosis. Macrophages play a central role in the inflammatory response by releasing mediators such as nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), and prostaglandin E2 (PGE2). This study aimed to investigate the anti-inflammatory potential of BTEX-K, a formulation of dried red ginseng combined with alpha-galactosidase, in lipopolysaccharide (LPS)-stimulated cells. Methods: LPS-treated immune cells were used to assess the anti-inflammatory effects of BTEX-K. The levels of NO, IL-6, TNF-α, and PGE2 were measured following BTEX-K treatment. The protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was evaluated. Cytotoxicity assays were conducted to determine whether the observed effects were due to cell viability loss. The involvement of MAPK signaling and NF-κB pathway modulation was examined by analyzing JNK phosphorylation, IκB degradation, and PPAR-γ expression. Results: BTEX-K significantly reduced the production of NO, IL-6, TNF-α, and PGE2 in LPS-treated cells without inducing cytotoxicity. The protein expression levels of iNOS and COX-2 were also suppressed. Furthermore, BTEX-K inhibited the LPS-induced phosphorylation of JNK in the MAPK pathway. It restored IκB levels and suppressed NF-κB activation by preventing the downregulation of PPAR-γ. Conclusions: BTEX-K demonstrates notable anti-inflammatory effects by inhibiting key inflammatory mediators and signaling pathways in immune cells. These findings support its therapeutic potential in mitigating inflammation-related symptoms, including pain, swelling, and redness, commonly seen in degenerative arthritis. Full article

Background/Objectives: Pestalotic acid A (PAA), a polyketide derived from Pestalotiopsis vismiae, an endophyte of the Japanese holly (Ilex crenata), is known to exhibit known antimicrobial activity, but its anti-inflammatory properties remain uncharacterized. This study aimed to investigate the anti-inflammatory effects of PAA in lipopolysaccharide (LPS)-stimulated murine macrophages, RAW264.7 cells. Methods: PAA was isolated from P. vismiae endophytes of Ilex crenata, and its structure was confirmed. RAW264.7 macrophages were treated with 0–50 μM of PAA in the presence of 100 ng/mL LPS. Cell viability was assessed by MTS assay; nitric oxide (NO) production was measured via Griess reagent; interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF) were quantified by enzyme-linked immunosorbent assay. Protein expression of inducible NO synthase (iNOS), nuclear factor (NF)-κB p65 phosphorylation, and related signaling proteins was evaluated by Western blot analysis and immunofluorescence staining. Results: PAA significantly increased macrophage viability and dose-dependently inhibited the release of NO by alleviating the protein expression of iNOS in LPS-treated RAW264.7 cells. Furthermore, PAA suppressed the release of IL-6, IL-1β, and TNF induced by LPS. Western blot and immunofluorescence results also indicated that PAA blocked the p65 subunit phosphorylation of NF-κB, which is one of the underlying mechanisms of the anti-inflammatory action of pestalotic acid A. Conclusions: PAA exerts potent anti-inflammatory effects in LPS-stimulated macrophages via inhibition of the NF-κB pathway, highlighting its potential as a natural therapeutic agent for inflammatory diseases. Full article

This study developed a novel water-soluble Cellulose Acetoacetate (CAA)-chitosan (CS) composite hydrogel drug delivery system. In this system, CAA and CS molecules are cross-linked via dynamic enamine bonds, forming a three-dimensional network structure suitable for drug encapsulation and controlled release. The primary objective was to address the challenges associated with the short half-life and significant fluctuations in therapeutic concentration of cytokine drugs, such as interleukin-2 (IL-2). A hydrogel system with a three-dimensional spatial network structure was successfully constructed via dynamic enamine bonds cross-linking between the acetoacetate groups in CAA molecules and the amino groups in CS. This system exhibits the following characteristics: (1) Dynamic covalent bonds impart adjustable mechanical properties to the hydrogel, enabling precise control over gelation time and mechanical performance; (2) A hierarchical pore structure (average pore size of 100–200 μm) provides a three-dimensional confined space for efficient drug encapsulation, achieving an IL-2 encapsulation efficiency of 83.3 ± 3.1%; (3) In vitro release studies demonstrated that the cumulative release of IL-2 within 72 h ranged from 18.4% to 34.7%, indicating sustained-release behavior. Cell viability assays confirmed that the hydrogel maintained the survival rate of L929 cells above 85% (as determined by the CCK-8 method), and live/dead staining revealed no apparent cytotoxicity. Overall, this three-dimensional network hydrogel based on dynamic covalent bonds represents a promising strategy for low-dose, long-lasting cytokine delivery. Full article

Background: The identification of parasite- and stage-specific antigens is crucial for the development of new diagnostic tests for cystic echinococcosis (CE). We previously analysed the interleukin (IL)-4 response to T-specific peptides corresponding to the immunogenic regions of the five antigen B (AgB) subunits, demonstrating that AgB1 is the most immunogenic protein and that the response to all AgB peptides is associated with viable cysts. However, the response in patients with CE3a (WHO-IWGE) cystic stage was not evaluated and no other immunological factors besides IL-4 were included in the analysis. Methods: Four study groups were defined: “CE3a group” (transitional cysts), “CE3b group” (active cysts), “CE4/CE5 group” (inactive cysts), and “NO CE-group” encompassing patients with non-CE cysts (controls). Whole blood was stimulated in vitro with the five different T-specific peptide pools corresponding to the five AgB subunits and with a pool containing all five peptides’ pools (total pool). IL-4 and other immunological markers were evaluated by ELISA and a multiplex assay, respectively. Results: Twenty-four patients with CE (CE3a-group n = 3; CE3b-group n = 6; CE4/CE5-group n = 15) and 14 subjects with non-CE cysts were enrolled. IL-4 levels in response to AgB1 and AgB3 pools were significantly increased in CE compared to NO CE groups (p = 0.0201, p = 0.0041). Within the CE patients, the highest IL-4 median level was observed in response to the AgB total pool, the AgB3 and AgB4 pools, followed by the AgB1 pool. Moreover, the IL-4 levels in response to the AgB1 pool were found to be significantly higher in the CE3b group compared to the CE4/CE5 group (p = 0.0070), while no differences were found for the CE3a group. As for other cytokines, we found higher IL-7 levels in response to the AgB4 pool in the CE4/CE5 group compared to the CE3b group (p = 0.0012), higher IL-2 levels in response to the AgB1 pool and AgB total pool in CE3b patients compared to controls (p = 0.0016), and higher IL-13 levels in response to the AgB total pool in patients with CE3b and CE4/CE5 cysts compared to NO CE (p = 0.0016; p = 0.0009). Conclusions: These results contribute to a better knowledge of the immune interplay in the presence of CE and may be useful for further exploring the use of recombinant proteins/peptides in cytokine release assays for the diagnosis and follow-up of CE. Full article

Background: The link between periodontal pathogens, inflammation, and neurodegenerative processes, including Alzheimer’s disease (AD), is evident. Porphyromonas gingivalis and Treponema denticola release lipopolysaccharide (LPS), constituting a virulence factor that takes part in the brain inflammatory process. Human gingival fibroblasts (HGF-1) are a source of pro-inflammatory cytokines released during periodontal diseases. Propolis is a rich source of quercetin and apigenin, which exhibit anti-inflammatory and immunomodulatory activities, influencing the concentration of pro-inflammatory cytokines. Considering this aspect, models with stimulated HGF-1, followed by LPS and/or interferon-α (IFN-α), were used. Aim: This study was designed to evaluate the concentrations of selected cytokines produced by HGF-1, which may influence brain inflammation. The immunomodulatory effects of apigenin and quercetin were investigated by measuring the concentration of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-15 (IL-15), and tumour necrosis factor (TNF-α). This study’s novelty is based on insights into the immunomodulatory effects of selected flavonoids by correlating the secretion of pro-inflammatory cytokines by gingival fibroblasts during periodontal disease with inflammatory processes in the brain. The cytotoxicity of apigenin and quercetin was estimated using the MTT assay. Fibroblasts were stimulated with LPS at 200 ng/mL and/or IFN-α at 100 U/mL concentration, followed by incubation with apigenin (25–50 µg/mL) and quercetin (25–50 µg/mL). Cytokine concentrations were measured using the xMAP technology. Results: The most pronounced and statistically significant reduction in cytokine levels, particularly IL-6 and IL-15, was observed for quercetin in both concentrations (25 µg/mL and 50 µg/mL), especially following LPS stimulation. Apigenin in both analysed concentrations also significantly decreased the level of IL-6. These results suggest that quercetin and apigenin may indirectly act as potential immunomodulators in preventing brain inflammation by inhibiting the inflammatory process in periodontitis; however, this should be confirmed in further studies. Full article

To further explore the anti-inflammatory components of the seeds of Dolichos lablab L., a comprehensive phytochemical investigation was conducted using diverse chromatographic and spectrometric technologies, as well as chemical reactions. As a result, ten previously unreported terpenoid glycosides, namely dolilabterpenosides A, B, C₁–C₃, D, E, and F₁–F₃ (1–10), along with four known analogues (11–14), initially identified from Dolichos genus, were obtained. In addition, the lipopolysaccharide (LPS)-induced RAW264.7 cell model was employed to detect the expression levels of nitric oxide (NO), inflammatory cytokines tumour necrosis factor (TNF)-α and interleukin (IL)-1β to assess the anti-inflammatory activities of the obtained compounds. The results of bioactive assay showed that compounds 1, 4–7, and 10–12 showed significant inhibitory activity on NO release in RAW264.7 cells in a dose-dependent manner, and all of them were demonstrated to inhibit the increase in TNF-α and IL-Iβ levels in the supernatant of RAW264.7 cells stimulated by LPS. Full article

The purpose of this study was to examine whether supplementation of ultra- and nanofiltered colostrum-based products, combined with egg yolk extract, nicotinamide mononucleotide (NMN), quercetin, alpha-ketoglutarate, white button mushroom, and celery seed extracts (the formula was patented by 4Life Research Company, USA and named as AgePro), modulate the functional activity of natural killer (NK) cells in vivo. We found that this supplement, taken orally in two capsules twice a day for 30 days, significantly enhanced the cytotoxic activity of NK cells. This was evidenced by the increased NK cell-mediated killing of carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled K562 human myeloid leukemia cells. As expected, this effect was dependent on the ratio between the effector (E) (e.g., peripheral blood mononuclear cells (PBMCs)) and target (T) (e.g., K562) cells, illustrating maximal killing of K562 cells at a 50:1 E/T ratio. Of note, increased NK-mediated killing of K562 cells after taking AgePro correlated with increased perforin release, evidenced by the CD107a degranulation assay. In concordance with these findings, taking of AgePro for 1 month increased production of several cytokines and chemokines, including IL-1β, IL-1Rα, IL-6, IL-8, IL-10, IFN-γ, TNF-α, G-CSF, PDGF-AA, PDGF-AB/BB, GRO, MCP-1, MCP-3, and MIP-1α, in PBMCs co-cultured with K562 cells. Of note, increased production of the cytokines correlated with the activation state of PBMCs, as evidenced by increased expression of the surface activation markers (e.g., the interleukin-2 receptor alpha chain—CD25). A strong correlation was found between NK-based cytotoxic activity and the production of IL-1β, IL-6, TNF-α, and MIP-1α. Importantly, no increase in the aforementioned soluble factors and activation markers was detected in PBMCs cultured alone, thereby illustrating the potent immunoregulatory activity of AgePro only in the presence of the harmful target cells. Hematological parameters also remained unchanged over the entire study period. Collectively, we show herein the significant enhancement of the cytotoxic activity of NK cells against target tumor cells after taking AgePro for 1 month. Notably, this effect was observed for all age groups, including young, adult, and elderly participants. Moreover, a significant improvement in NK cytotoxic activity was also detected for participants with low basal (e.g., before taking AgePro) numbers of NK-mediated killing. The enhancement of NK-based cytotoxicity was associated with an increased release of several cytokines and chemokines involved in regulating a broad spectrum of mechanisms outside the cell-mediated cytotoxicity and killing of target cells. Of note, spontaneous activation of PBMCs, particularly NK cells, was not detected after taking AgePro. Given that spontaneous activation of autoreactive lymphocytes is a feature associated with autoimmunity and taking into account our data illustrating the AgePro-induced activation of NK cells detected only in the presence of the potentially harmful cells, we conclude that our innovative product exhibits potent immunoregulatory activity and high safety profile. Full article

Background/Objectives: Pneumocystis jirovecii pneumonia (PJP) is the most frequently diagnosed AIDS-defining illness in Europe, with especially high mortality in HIV-negative patients caused by delayed diagnosis and low awareness. This study aims to evaluate cytokine release assays (CRA) to facilitate a less invasive and resource-efficient PJP specific diagnostic test. We focus on the P. jirovecii antigens Kexin 1 (KEX1), MSG1, and MSG2, which were identified in prior studies as immunologically relevant. Methods: Whole blood samples from 50 participants—22 healthy individuals and 28 immunocompromised individuals, including 8 with proven PJP—were stimulated in vitro with full-length and partial KEX1, MSG1, MSG2, and a combination of all three antigens (PJ-MIX). Following 24 h incubation at 37 °C, cytokine levels of IL-2, IFN-γ, IL-17A, and IL-17F were measured. Results: Stimulation with full-length KEX1, MSG1, MSG2, and PJ-MIX antigens induced higher IL-2 concentrations in the healthy control group compared to the groups IL-2 baseline levels and to the group of proven PJP cases. Similarly, stimulation with full-length KEX1, MSG1, and PJ-MIX elevated IFN-γ levels in the healthy control group compared to baseline IFN-γ levels. Conclusions: Our findings highlight the potential of IL-2 and IFN-γ release following stimulation with PJ antigens, with PJ-MIX eliciting the strongest and most significant responses, suggesting a cumulative antigen effect. This pilot study establishes a foundation for a PJP-specific CRA, deepening our knowledge of T-cell immunity against PJP. Clinically, such a test could, among other applications, evaluate at-risk patients who should receive prophylaxis and may consequently reduce PJP-related morbidity and mortality. Full article

Today, most anti-acne treatments employ topical and systemic antibiotics such as erythromycin and clindamycin, which induce cutaneous dysbiosis with adverse side effects to the skin’s normal microbiota, consequently leading to the emergence of antimicrobial resistance. In our quest to discover natural anti-acne bioactives as alternatives, we undertook a research program with the aim to identify a new blend of active ingredients based on the monoterpene phenol moiety. Within this program, we evaluated the in vitro anti-acne efficacy of thymol, Curcuma turmerones and their patented combination “Acnocure” in a cosmetic formulation. The minimum inhibitory concentration (MIC) of Acnocure against C. acnes (ATCC 6919), S. aureus (ATCC 6538), S. epidermidis (ATCC 12228) and C. freneyi (CIP 52.16) was determined to be 0.32, 0.26, 0.47 and 0.11 mg/mL, respectively. In the time-kill curve study against C. acnes, Acnocure, containing thymol 0.25% and 0.1% Curcuma turmerone as well as thymol 0.1% and 0.1% Curcuma turmerone in a cosmetic simplex formulation, demonstrated rapid bactericidal activity with a 4.7 log reduction at pH 5.5, occurring within just two hours of the study and lasting for over 24 h. The killing efficacy was similar to our cosmetic reference benchmark, Effaclar DUO serum, used in the same study. Additionally, thymol, Curcuma turmerones and Acnocure were evaluated in an anti-inflammatory efficacy assay in lipopolysaccharide (LPS)-primed U937 macrophages model and demonstrated moderate inhibition of interleukin-1β (IL-1β) at 100 µg/mL and significant inhibition of prostaglandin E-2 (PGE-2) at 1 µg/mL, respectively. Further evidence gathered on thymol and Curcuma turmerones in an IL-1α-stimulated dermal fibroblast model showed >90% inhibition of PGE-2 release between 2 µg/mL and 30 µg/mL concentrations. These promising results position Acnocure as a natural alternative for the replacement of synthetic corticosteroids and antibiotics with potent anti-acne skincare properties. Full article

Inflammation plays a critical role in the pathogenesis of osteoarthritis (OA). The objective of this study was to investigate the anti-inflammatory and chondroprotective properties of Artemisia annua L. water extract (AWE) following the induction of inflammation in cartilage cells (SW1353 cell) through the administration of interleukin-1 beta (IL-1β). We demonstrated significant antioxidant activity, as evidenced by elevated total phenolic and flavonoid content, in addition to robust free radical scavenging capacity, as assessed through DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays. Its cytotoxic effects were assessed at a concentration of 200 μg/mL, where no cytotoxic signs were observed in SW1353 cells treated with IL-1β; the levels of reactive oxygen species (ROS) were notably reduced in a dose-dependent manner. The principal inflammatory markers, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), were significantly diminished by AWE treatment. AWE administration led to a dose-dependent reduction in the expression of key proteins involved in the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) signaling pathways, ultimately resulting in a decrease in the release of matrix metalloproteinases (MMPs), specifically MMP-1 and MMP-13, which are known to contribute to cartilage degradation. Additionally, the levels of degraded collagen type II in the cartilage cells were restored. These findings suggest that reducing oxidative stress and inflammation, along with inhibiting activated MAPK and NF-κB signaling pathways, may ameliorate the progression of IL-1β-induced OA. Furthermore, a molecular docking analysis revealed a strong binding affinity of MMP-13, a critical mediator in the pathogenesis of OA. Six compounds were identified in AWE, corroborating its potential antioxidant and anti-inflammatory effects. Therefore, AWE may serve as a potentially useful therapeutic agent against OA by modulating inflammation-related mechanisms. Full article