Paw Inflammation Research Articles

EHDA engineering of Piroxicam-PVP components for pharmaceutical dosages

This study aimed to prepare piroxicam loaded polyvinyl pyrollidone (PVP) particulate mat, fabricated mat incorporated tablets and coated polyvinyl alcohol (PVA) films. The mat, engineered by using electrohydrodynamic atomisation (EHDA) technique, was evaluated for morphological features and particle size using scanning electron microscopy. In vitro drug release, ex vivo skin permeation and in vivo anti-inflammatory studies of oral (i.e., tablets) and topical (i.e., films) dosage forms were conducted to assess the efficiency of prepared formulations. The EHDA processed piroxicam-PVP components exhibited spherical shape, uniform surface and particle size of 3.67 ± 0.70 μm. The piroxicam mat incorporated oral and coated topical formulations exhibited release percentage of 73.79 ± 1.9% and 80.11 ± 2.3% respectively over a time period of 180 min. During ex vivo permeation study, treatment of rat skin using 500 μm microneedles led to a higher permeation of piroxicam (from coated films i.e., 75.54 ± 2.2% as compared to unpierced skin i.e., 58.23 ± 2.7% with a gradient of 0.28 and 0.21 respectively within 240 min. During in vivo anti-inflammatory study, a 52.58 ± 3.2% decrease in paw inflammation was observed in mat incorporated tablets administered group within 360 min. In comparison, topical application of coated films led to a 67.28 ± 2.9% decrease in inflammation. Microneedle treatment followed by topical application of coated films resulted in an 81.43 ± 3.1% reduction in paw inflammation over 360 min. Micropiercing of the skin resulted in breaching of uppermost skin barrier layer stratum corneum leading to an increased piroxicam permeation and rapid therapeutic response. It was concluded that EHDA processed mat incorporated tablets and coated topical films can serve as a promising choice for improving the solubility of poorly soluble piroxicam and oral/topical anti-inflammatory therapy.

Modulation of immune cell function, IDO expression and kynurenine production by the quorum sensor 2-heptyl-3-hydroxy-4-quinolone (PQS).

Many invasive micro-organisms produce 'quorum sensor' molecules which regulate colony expansion and may modulate host immune responses. We have examined the ability of Pseudomonas Quorum Sensor (PQS) to influence cytokine expression under conditions of inflammatory stress. The administration of PQS in vivo to mice with collagen-induced arthritis (CIA) increased the severity of disease. Blood and inflamed paws from treated mice had fewer regulatory T cells (Tregs) but normal numbers of Th17 cells. However, PQS (1μM) treatment of antigen-stimulated lymph node cells from collagen-immunised mice in vitro inhibited the differentiation of CD4+IFNγ+ cells, with less effect on CD4+IL-17+ cells and no change in CD4+FoxP3+Tregs. PQS also inhibited T cell activation by anti-CD3/anti-CD28 antibodies. PQS reduced murine macrophage polarisation and inhibited expression of IL1B and IL6 genes in murine macrophages and human THP-1 cells. In human monocyte-derived macrophages, IDO1 gene, protein and enzyme activity were all inhibited by exposure to PQS. TNF gene expression was inhibited in THP-1 cells but not murine macrophages, while LPS-induced TNF protein release was increased by high PQS concentrations. PQS is known to have iron scavenging activity and its suppression of cytokine release was abrogated by iron supplementation. Unexpectedly, PQS decreased the expression of indoleamine-2, 3-dioxygenase genes (IDO1 and IDO2), IDO1 protein expression and enzyme activity in mouse and human macrophages. This is consistent with evidence that IDO1 inhibition or deletion exacerbates arthritis, while kynurenine reduces its severity. It is suggested that the inhibition of IDO1 and cytokine expression may contribute to the quorum sensor and invasive actions of PQS.

Hyaluronic acid-coated proglycosomes for topical delivery of tofacitinib in rheumatoid arthritis condition: Formulation design, in vitro, ex vivo characterization, and in vivo efficacy studies

Tofacitinib (TF) is a selective oral jakanib approved by the USFDA for the treatment of rheumatoid arthritis (RA). To overcome the adverse effects of orally administered TF, topical delivery can be a suitable choice. The therapeutic efficacy of TF can be improved through the high affinity of natural ligands (hyaluronic acid and chondroitin sulphate) to CD44 receptors on the macrophages or other immune cells in the dermal region. Thus, the present research work was inspired by the possibility to develop and evaluate the potential of hyaluronic acid-coated proglycosomes (HA-TF-PG) as the carrier for site-specific dermal delivery. The normal-PG (N-TF-PG) and HA-TF-PG showed particle sizes of <250 nm. The HA-TF-PG demonstrated 3.15-fold higher retention of TF in the viable dermis layers than the conventional formulation. The in vivo pharmacodynamic study, cytokines, and radiographic study on Complete Freud's Adjuvant-induced arthritic rat model revealed that HA-TF-PG exhibited a significant (P < 0.001) reduction in inflammation in arthritic rat's paw compared to the conventional TF. The developed HA-TF-PG treated groups showed significantly lowered CD44 levels compared to FD-gel and N-TF-PG i.e. 2.28 and 1.32-fold respectively (p < 0.001). In conclusion, The HA-TF-PG can be developed as an effective carrier for the site-specific dermal drug delivery system of TF to treat RA.

Topical Calendula officinalis L. inhibits inflammatory pain through antioxidant, anti-inflammatory and peripheral opioid mechanisms

ObjectiveThe present study evaluated the antinociceptive activity of Calendula officinalis L. (Ca) cream on inflammatory hypernociception. MethodsCreams with different Ca concentrations were tested for their ability to ameliorate pain-related behavior and edema in rats using formalin test, carrageenan (Cg)-induced acute inflammation model, bradykinin (BK)-induced acute inflammation model, and complete Freund’s adjuvant (CFA)-induced chronic inflammation model. Using the formalin test, we also examined the implication of peripheral opioid receptors in the antinociceptive mechanisms of Ca cream, by means of Q-naloxone, a peripherally acting nonselective opioid antagonist. Furthermore, the effects of Ca cream compared with diclofenac on BK-induced edema were examined when the kininase II in tissue was preserved or inhibited by captopril. The local production of redox biomarkers in formalin model, tumor necrosis factor-α (TNF-α) in Cg model and histopathological changes in CFA model were also evaluated. ResultsA single application of Ca cream at a dose of 10% or 30% (w/w) decreased the formalin-induced licking/biting behavior during both phases of this test in a Q-naloxone-sensitive manner. This effect was associated with the reduction of oxidative stress in the injured paw and the edema associated with the later phase of formalin-induced pain. A single application of Ca cream (10% or 30%) reduced paw edema and thermal hypernociception in Cg-induced acute inflammation, corresponding with a local decrease in TNF-α. Ca cream also inhibited BK-induced spontaneous nociceptive behavior and paw inflammation in both paradigms studied. Repeated applications of Ca cream also decreased CFA-induced chronic inflammation, mechanical hypernociception and histopathological changes in the paw. ConclusionThese results reveal the topical antinociceptive and antiedematogenic effects of Ca cream. A modulatory action on peripheral opioid receptors associated with its antioxidant mechanism may be involved, at least in part, in its analgesic effects. These findings may have an impact on the clinical management of painful inflammatory diseases.

Alloferon-1 ameliorates acute inflammatory responses in λ-carrageenan-induced paw edema in mice

Alloferon-1 have been proposed as an effective peptide to enhance antitumoral immunity, antiviral defense and anti-inflammatory activity. This work aimed to assess anti-inflammatory effects of alloferon-1 against acute inflammation and histopathological deformations in λ-carrageenan-induced paw edema in mice. Systemic pretreatment with alloferon-1 (22.0 mg/kg) intraperitoneally injected mice showed a significant reduction in paw thickness and vascular permeability. Alloferon-1 prevented λ-carrageenan-evoked exudation and the neutrophil influx to the mouse pleura and the neutrophil migration into carrageenan-stimulated mouse air pouches based on the histopathological changes in the paw tissues. Administration of alloferon-1 also suppressed the expression of the inflammatory cytokines in the inflamed paw tissues such as tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein 1 (MCP1), interleukin-5 (IL-5), etc. detected by Luminex liquid chip. Collectively, the present study provides evidences for the marked anti-inflammatory effects of alloferon-1 which might represent new therapeutic options for the treatment of acute inflammatory diseases.

Geographical variation in the chemical composition, anti-inflammatory activity of the essential oil, micromorphology and histochemistry of Schinus terebinthifolia Raddi

Ethnopharmacological relevanceSchinus terebinthifolia Raddi (Anacardiaceae) is rich in essential oil, distinguished by a predominance of monoterpenes and sesquiterpenes, it being widely used in traditional medicine for the treatment of inflammations. Aim of studyThis study's objective was to investigate the chemical composition of the essential oil of S. terebinthifolia (EOST) collected in six states of Brazil, evaluate its anti-inflammatory effects in mice, and analyze the histochemistry and micromorphology of leaves and stems. Materials and methodsAerial parts of S. terebinthifolia were collected in six states of Brazil, and the essential oil was extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC–MS). The histochemistry and micromorphology of leaves and stems were performed using standard reagents, light and field emission scanning electron microscopy, beyond energy-dispersive X-ray spectroscopy. The EOST were evaluated for anti-inflammatory activity and hyperalgesia using the carrageenan-induced paw edema methodology. ResultsThe EOST showed variation across the six states in its yield (0.40%–0.86%) and chemical composition: hydrocarbon monoterpenes (28.76%–47.73%), sesquiterpenes, (31.43%–41.76%), oxygenated monoterpenes (14.31%–19.57%), and oxygenated sesquiterpenes (4.87%–14.38%). Both α-pinene and limonene were predominant constituents of essential in five regions, except for one state where α-phellandrene and limonene were the dominant components. A comprehensive description of the leaf and stem micromorphology and histochemistry was performed. In the in vivo testing, all EOST samples exerted antiedematogenic and anti-hyperalgesic effects, when tested in a carrageenan-induced paw inflammation (mechanical and thermal hyperalgesia) model with oral doses of 30 mg/kg. ConclusionOur results indicate that the EOST samples collected in six Brazilian states differed in their chemical composition but not their anti-inflammatory and antihyperalgesic effects, which was correlated with the synergistic effect of its components, collaborating the etnhopharmacologycal use of this plant due to its an anti-inflammatory effect. Also, micromorphology and histochemistry of leaves and stems presented in this study provide anatomical and microchemical information, which aids species identification.

The effect of 1.3 bis(p-Hydroxyphenyl)urea compound on IL-6, IL-1β, TNF-α and COX-2 protein expression on λ-Carrageenan-induced rats

Objectives: This study aims to determine the protein expression of IL-6, IL-1β, TNF-α, and COX-2 using an immunohistochemistry method based on the anti-inflammatory activity of intraplantar carrageenan solution in male rat paws. Materials and methods: Animals were provided with 1.3 bis(p-Hydroxyphenyl)urea at 12.5, 25, 50, 100, and 200 mg/kg BW. This was administered approximately one hour before the 1% carrageenan induction. As a control measure, diclofenac sodium and CMC sodium 0.5% was similarly administered. After 360 mins, all of the rats were sacrificed, and the inflammatory tissue in the rat’s paw was acquired. The immunohistochemistry preparations were carried out to observe the expression of IL-6, IL-1β, TNF-α, and COX-2. Results: The result showed a decrease in the expression of IL-6, IL-1β, TNF-α, and COX-2, all groups given 1.3 bis(p-Hydroxyphenyl)urea and diclofenac sodium compared to the control group CMC sodium 0.5% CMC sodium 0.5%. From the results of the mean difference test, it can be seen that the IL-1β expression at doses of 50, 100, and 200 mg/Kg BW was not significantly different from that of diclofenac sodium (p &amp;gt; 0.05). Meanwhile, the expression of COX-2, TNF-α, and IL-6 at doses of 100 and 200 mg/Kg BW was not significantly different from that of diclofenac sodium (p &amp;gt; 0.05). Conclusion: The 1.3 bis(p-Hydroxyphenyl)urea has anti-inflammatory activity by decreasing the percentage of IL-6, IL-1β, TNF-α, and COX-2 protein expression in rat’s paw inflammation tissue.

Thymoquinone counteracts oxidative and inflammatory machinery in carrageenan-induced murine paw edema model.

Thymoquinone (TQ) is an active constituent in Nigella sativa (black cumin) and is extensively reported for its distinguished antioxidant and anti-inflammatory bioactivities. Despite the local protective response of acute inflammation, it contributes to the development of various disease conditions such as cell death, organ damage, or carcinogenesis. Hence, in this study, the effects of orally administered TQ (50mg/kg and 100mg/kg) for 14days against edema development, oxidative stress, and inflammation were investigated in paw edema induced by carrageenan in mice. Indomethacin (10mg/kg) was used as a reference drug. The results revealed that TQ reduced the paw edema volume in a time-dependent manner, attenuated acetic acid-provoked writhing movements, and reduced xylene-triggered ear edema. Hematological findings revealed marked normalization of altered counts of WBCs, and platelets. Furthermore, paw tissue levels of malondialdehyde and nitric oxide showed marked decreases together with increases in nuclear factor erythroid 2-related factor 2, glutathione, superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase after TQ administration. Additionally, TQ decreased pro-inflammatory mediators, such as interleukin-1 beta, tumor necrosis factor-alpha, interleukin-6, monocyte chemoattractant protein-1, C-reactive protein, myeloperoxidase, and nuclear factor kappa-B in the inflamed paw tissue. Moreover, appreciable decreases were recorded in cyclooxygenase-2 and its product prostaglandin E2 and the immune reaction of tumor necrosis factor-alpha in TQ-treated mice. Histopathological findings further validated the potential antiedematous, anti-inflammatory power of TQ in inflamed tissues. Conclusively, the results encourage the potent application of TQ to subside acute inflammatory events because of its striking antioxidant and anti-inflammatory properties in inflamed paw tissue.

Anti-inflammatory activity and toxicity evaluation of 1,3-bis(p-hydroxyphenyl)urea

Background: Inflammation is a normal protective response caused by tissue damage through physical trauma, chemical damage, or invasion of pathogenic microorganisms. 1,3-bis(p-hydroxyphenyl)urea is a modified p-aminophenol compound, which is considered to have strong analgesic activity based on cyclooxygenase-2 inhibition and has fewer hepatotoxic side effects. In-silico test showed 1,3-bis(p-hydroxyphenyl)urea has COX-1 and TNF- binding activity, so it has the potential to be developed as an anti-inflammatory agent. Anti-inflammatory activity was tested using mice. Toxicity test was conducted to test the safety of 1,3-bis(p-hydroxyphenyl)urea. Methods: Anti-inflammatory test was carried out by measuring the percentage of inflammation in rat paws using a plethysmometer after administration of 1,3-bis(p-hydroxyphenyl)urea induced by carrageenan 1%, and then histology was performed to observe the number of neutrophils. A toxicity test using OECD guidelines carried out acute toxicity for 24 hours and was observed for 14 days. The subchronic toxicity test was carried out for 28 days, followed by 42 days in the satellite group. Results: Analysis of rat paw inflammation volume showed 1,3-bis(p-hydroxyphenyl)urea could suppress inflammation after carrageenan-1% induction. The group given {1.3 bis (p-Hydroxyphenyl)urea} and sodium diclofenac 2.25 mg/kg BW had a significant difference in results (p&lt;0.05) with 0.5% Na CMC group. Doses of 50, 100, and 200 mg/kg BW showed no significant difference (p&gt;0.05) with diclofenac sodium. The number of neutrophils also decreased compared to the 0.5% Na CMC group. The acute toxicity test of 1,3-bis(p-hydroxyphenyl)urea did not cause toxic symptoms and death up to a dose of 5000 mg/kg BW. The microscopic results of subchronic toxicity of liver tissue experienced hydropic degeneration at a dose of 1000 mg/kg BW; in the lungs causes congestion, and microscopic renal tissue undergoes Bowman space dilatation and tubular lumen dilatation, but this condition is reversible. Conclusion: This 1,3-bis(p-hydroxyphenyl)urea compound had an anti-inflammatory activity and relatively low toxicity.

Anti-inflammatory potential of berberine-rich extract via modulation of inflammation biomarkers.

Berberine-rich extract (BRE) prepared from Berberis lycium root bark using green extraction approach and its marker compound berberine has a broad spectrum of clinical applications. Berberine's potential pharmacological effects include anticancer, antidiarrheal, antidiabetic, antimicrobial and anti-inflammatory activities. In current work, BRE and berberine were evaluated for their therapeutic prospects in inflammation models. The comparative effect of BRE and berberine against inflammation was determined through in vitro chemiluminescence technique. The in vivo anti-inflammatory evaluation of BRE and berberine (25, 75, and 125 mg/kg) compared to diclofenac (10mg/kg) was performed in carrageenan and formaldehyde-induced inflammation in Wistar rats. Histopathological and biochemical studies were conducted to find the comparative anti-inflammatory potential of BRE and berberine on pathological hallmarks induced by formaldehyde. Moreover, the modulatory effects on inflammatory biomarkers were also investigated through qPCR. ELISA (enzyme-linked immunoassay test assay) was performed to investigate the expression of pathological protein biomarkers like TNF-α and IL-6 and levels of antioxidant enzymes were estimated in liver homogenates. Both BRE and berberine markedly (p < .001) reduced paw diameter and inflammation in carrageenan and formaldehyde-induced inflammation. The levels of antioxidant enzymes were recovered (p < .001) by BRE and berberine treatments, and compared to the formaldehyde-treated inflammation model. Both BRE and berberine remarkably downregulated the mRNA and protein expression of inflammatory biomarkers. BRE similar to berberine mitigated the level of antioxidant enzymes in liver homogenate. The undertaken study suggests that BRE, a natural, green, and therapeutically bioequivalent to berberine could be used as an economical phytomedicine in the treatment of inflammatory disorders. PRACTICAL APPLICATIONS: Anti-inflammatory drugs like NSAIDS are associated with serious adverse effects like gastrointestinal ulcer, worsening of preexisting cardiovascular disorders, and renal failure. Therefore, there is a constant demand to develop novel, inexpensive therapeutic strategies to treat the inflammatory disorder with the least harmful effects. Pure phytochemicals with anti-inflammatory potential are costly and hard to isolate, therefore green microwave-assisted extraction technique is developed to get the rich bioequivalent extract. Berberis lycium a medicinal plant with berberine as a major bioactive constituent, has wide acceptance in traditionally used medicine and as food. Pharmacological studies revealed its hepatoprotective, anticancer, antidiabetic, and antihypertensive activities. BRE was prepared by green microwave-assisted extraction and enrichment by resin column to get a higher yield of berberine. The comparative anti-inflammatory effect of BRE and berberine was determined by in vitro and in vivo studies. Results obtained from this experimental work contribute beneficial guidance that reinforces the use of the BREto treat inflammatory disorders.

Amelioration of injury-induced tissue acidosis by a nonsteroidal analgesic attenuates antinociceptive effects of the pH-dependent opioid agonist NFEPP

Opioid agonists are powerful drugs for managing pain. However, their central side effects are limiting their use and drugs with similar potency, but a lower risk profile are needed. (±)-N-(3-fluoro-1-phenethylpiperidine-4-yl)-N-phenylpropionamide (NFEPP) is a novel opioid agonist that preferentially activates opioid receptors at acidic extracellular pH. NFEPP was designed to activate peripheral opioid receptors in injured tissue, therefore precluding side effects elicited at normal pH in brain or intestinal wall. Considering the common combination of opioids and nonsteroidal anti-inflammatory drugs (NSAIDs) in multimodal analgesia, we investigated the interaction between NFEPP and a widely prescribed prototypical NSAID, diclofenac (DCF), in a rat model of unilateral hindpaw inflammation induced by complete Freund’s adjuvant. We evaluated the effects of systemically applied DCF on the paw tissue pH, on the expression of inflammatory mediators in immune cells from inflamed paws and on the expression of opioid receptors in dorsal root ganglia. Additionally, we investigated the antinociceptive efficacy of NFEPP injected into the inflamed paws after DCF treatment. We found that DCF reduced inflammation-induced nociceptive responses and tissue acidosis, but did not change the mRNA expression of IL-1β, TNF-α, IL-6, IL-4, NGF, or of mu-, delta-, or kappa-opioid receptors. The treatment with DCF moderately reduced the antinociceptive efficacy of NFEPP, suggesting a correlation between an increase in local tissue pH and the decreased antinociceptive effect of this pH-sensitive opioid agonist.

Novel Bile Salt Stabilized Vesicles-Mediated Effective Topical Delivery of Diclofenac Sodium: A New Therapeutic Approach for Pain and Inflammation

The oral delivery of diclofenac sodium (DNa), a non-steroidal analgesic, anti-inflammatory drug, is associated with various gastrointestinal side effects. The aim of the research was to appraise the potential of transdermal delivery of DNa using bilosomes as a vesicular carrier (BSVC) in inflamed paw edema. DNa-BSVCs were elaborated using a thin-film hydration technique and optimized using a 31.22 multilevel categoric design with Design Expert® software 10 software (Stat-Ease, Inc., Minneapolis, MI, USA). The effect of formulation variables on the physicochemical properties of BSVC, as well as the optimal formulation selection, was investigated. The BSVCs were evaluated for various parameters including entrapment efficiency (EE%), vesicle size (VS), zeta potential (ZP) and permeation studies. The optimized BSVC was characterized for in vitro release, Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM) and incorporated into hydrogel base. The optimized DNa-BSVC gel effectiveness was assessed in vivo using carrageenan-induced paw edema animal model via cyclooxygenase 2 (COX-2), interleukin 6 (IL-6), Hemooxygenase 1 (HO-1) and nuclear factor-erythroid factor2-related factor 2 (Nfr-2) that potentiate anti-inflammatory and anti-oxidant activity coupled with histopathological investigation. The resulting vesicles presented VS from 120.4 ± 0.65 to 780.4 ± 0.99 nm, EE% from 61.7 ± 3.44 to 93.2 ± 2.21%, ZP from −23.8 ± 2.65 to −82.1 ± 12.63 mV and permeation from 582.9 ± 32.14 to 1350.2 ± 45.41 µg/cm2. The optimized BSVCs were nano-scaled spherical vesicles with non-overlapped bands of their constituents in the FTIR. Optimized formulation has superior skin permeability ex vivo approximately 2.5 times greater than DNa solution. Furthermore, histological investigation discovered that the formed BSVC had no skin irritating properties. It was found that DNa-BSVC gel suppressed changes in oxidative inflammatory mediators (COX-2), IL-6 and consequently enhanced Nrf2 and HO-1 levels. Moreover, reduction of percent of paw edema by about three-folds confirmed histopathological alterations. The results revealed that the optimized DNa-BSVC could be a promising transdermal drug delivery system to boost anti-inflammatory efficacy of DNa by enhancing the skin permeation of DNa and suppressing the inflammation of rat paw edema.

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

Evaluating the anti-arthritic potential of walnut (Juglans regia L.) in FCA induced Sprague Dawley rats.

Rheumatoid arthritis (RA) is an autoimmune progressive disease, associated with many pathophysiological consequences. Owing to the adverse effects and higher costs of pharmaceuticals, people are now looking for complementary and alternative remedies. In this milieu, the present study was designed to explore the therapeutic potential of walnuts against FCA-induced arthritis in rat models. Purposely, 50 Sprague Dawley rats were housed in a well-ventilated animal room and separated into 5 groups of 10 rats each. The rats were categorized as G0 (negative control), G1 (positive control, i.e., FCA induced untreated arthritic rats), G2 (arthritic rats treated with MTX), G3 (arthritic rats treated with walnut feed), and G4 (arthritic rats treated with walnut extract), with an efficacy trial lasting for 42 days. The physical analysis explicated that paw swelling was significantly improved by 10%-12.8% in treatment groups after the intervention when compared with positive control. Moreover, biochemical analyses revealed significantly lower levels of ESR, CRP, and RF in rats treated with walnut-based interventions when compared to positive control. ESR values were decreased by 62.4% and 69.92% in G3 and G4 , whereas CRP levels were improved by 56.20% and 77.78% in G3 and G4 when compared with G1 . Likewise, RF values decreased in G2 , G3 , and G4 by 64.71%, 55.88%, and 69.24%, respectively when compared to G1 . The histological examination demonstrated the potential role of walnut-based interventions in reducing the severity of disease by decreasing cell infiltration, bone erosion, and paw inflammation. Meanwhile, the gene expression analysis revealed that walnut-based interventions protected the paw joints from damage by downregulating the RANKL-OPG pathway. Conclusively, walnut feed and extract may serve as potent anti-arthritic interventions with no side effects. PRACTICAL APPLICATIONS: Plant-based therapeutics are effective in the prevention and management of various chronic diseases. The current research explored the anti-arthritic potential of walnuts. Walnut feed and extract effectively reduced the serum arthritic biomarkers as well as downregulated the genes involved in bone destruction. Thus, the inclusion of dietary ingredients having therapeutic potential such as walnuts may be synchronized in clinical practices to ameliorate arthritis.

Synergistic antinociceptive effects of concomitant NAAA and peripheral FAAH inhibition

The intracellular lipid amidases, fatty acid amide hydrolase (FAAH) and N-acylethanolamine acid amidase (NAAA), terminate the actions of anandamide and palmitoylethanolamide (PEA), two antinociceptive and anti-inflammatory lipid-derived mediators. Here we show, confirming prior research, that small-molecule inhibitors of peripheral FAAH (compound URB937) and systemic NAAA (compound ARN19702) individually attenuate, in male CD-1 mice, pain-related behaviors and paw inflammation in the formalin and carrageenan tests. More importantly, isobolographic analyses revealed that the combination of URB937 and ARN19702 produced substantial synergistic (greater than additive) antinociceptive effects in both models as well as additive anti-inflammatory effects in the carrageenan test. Together, the findings uncover a functional interplay between FAAH and NAAA substrates in the control of nociception, which might be exploited clinically to develop safe and effective pain management strategies

PPAR-γ alleviates the inflammatory response in TNF-α-induced fibroblast-like synoviocytes by binding to p53 in rheumatoid arthritis.

Rheumatoid arthritis (RA) is characterized by synovial inflammation, synoviocyte expansion and damage to cartilage and bone. We recently reported that peroxisome proliferator-activated receptor (PPAR)-γ inhibited the proliferation and activation of fibroblast-like synoviocytes (FLS), and was downregulated in RA synovial. In this study we investigated the role of PPAR-γ in RA and the underlying mechanisms. Adjuvant-induced arthritis (AIA) was induced in rats; from D15, AIA rats were orally administered pioglitazone (30 mg·kg-1·d-1) or rosiglitazone (4 mg·kg-1·d-1) for 14 days. Collagen-induced arthritis (CIA) was induced in wild-type and Ppar-γ+/- mice. We showed that the expression of PPAR-γ was significantly reduced, whereas that of TNF-α was markedly increased in human RA FLS. In CIA mice, knockdown of PPAR-γ expression (Ppar-γ+/-) aggravated the ankle inflammation. Similarly, T0070907 (a PPAR-γ antagonist) or si-PPAR-γ promoted the activation and inflammation of TNF-α-induced FLS in vitro. On the contrary, administration of PPAR-γ agonist pioglitazone or rosiglitazone, or injection of ad-Ppar-γ into the ankle of AIA rat in vivo induced overexpression of PPAR-γ, reduced the paw swelling and inflammation, and downregulated activation and inflammation of FLS in RA. Interesting, injection of ad-Ppar-γ into the ankle also reversed the ankle inflammation in Ppar-γ+/- CIA mice. We conducted RNA-sequencing and KEGG pathway analysis, and revealed that PPAR-γ overexpression was closely related to p53 signaling pathway in TNF-α-induced FLS. Co-IP study confirmed that p53 protein was bound to PPAR-γ in RA FLS. Taken together, PPAR-γ alleviates the inflammatory response of TNF-α-induced FLS by binding p53 in RA.

Unani Formulation Habb-e-Suranjan: A Treasure of Biological Activities.

Habb-e-Suranjan (HES), an Unani formulation, has been studied for its anti-inflammatory properties in both in vitro and in vivo experiments. HES is recommended for arthritis, gout, and joint pain. The current endeavor is an attempt to put it to the test and verify its efficacy scientifically. It was tested for DPPH, hydroxyl, and nitric oxide scavenging activities. It was shown that HES had the greatest TAC and FRAC values when compared to catechin and ascorbic acid. HES exhibited DPPH and hydroxyl radical scavenging activity that was dose-dependent. Incubation of sodium nitroprusside solutions in PBS at 25°C for 150 min resulted in the production of nitric oxide. Nitric oxide production was effectively decreased by HES. Anti-inflammatory medications boosted the migration of PMN cells toward the chemoattractant FMLP in an agarose experiment of PMN chemotaxis. In carrageenan-induced rat paw edema, in the HES-treated group, paw thickness was 3.021 ± 0.084 at t = 0, but it showed an increase in paw inflammation after one hour, i.e., 3.195 ± 0.082 cm which again showed a decrease in paw thickness up to 4th hour, i.e., 3.018 ± 0.078, 2.98 ± 0.032, and 2.684 ± 0.061 at t = 2, 3, and 4, respectively. It showed again getting back to the normal thickness of paw at t = 24 hrs, i.e., 3.029 ± 0.118 cm. It is concluded that the formulation is potent enough and can be used effectively for the treatment of inflammation and associated health issues. Moreover, there is much scope to evaluate its effectiveness using different in vitro and in vivo models.

Oral administration of botryosphaeran [(1 → 3)(1 → 6)-β-d-glucan] reduces inflammation through modulation of leukocytes and has limited effect on inflammatory nociception.

Several biological activities of the fungal exopolysaccharide (1 → 3)(1 → 6)-β-d-glucan (botryosphaeran) have been described in the literature, but its effects on inflammation have not been evaluated. This study aimed to investigate the action of botryosphaeran on experimental mice models of carrageenan-induced acute pleurisy and acute paw edema, and complete Freund's adjuvant-induced persistent paw edema. All botryosphaeran doses tested (1.0, 2.5, 5.0, and 10.0 mg/kg birth weight [b.w.], orally administered) reduced leukocyte recruitment, nitric oxide (NO) levels, and protein extravasation in the pleural cavity. Botryosphaeran (5 mg/kg b.w.) did not diminish edema and mechanical hyperalgesia in the paw within 4 h; however, cold allodynia was alleviated within the first 2 h. In the persistent paw inflammation model, the effects of daily oral administration of botryosphaeran (5 mg/kg b.w.) were evaluated over 3 and 7 days. The fungal β-glucan significantly reduced the levels of the cytokines, tumor necrosis factor(TNF)-α, interleukin (IL)-6), and IL-10, in the paw homogenates in both protocols, while paw edema and the levels of advanced oxidation protein products (AOPP) only diminished on Day 7. No effect in mechanical hyperalgesia was observed. Oral treatment for 3 or 7 days also decreased the plasma levels of NO, AOPP, TNF-α, and IL-10. On Day 7, the number of leukocytes in the blood was also reduced by this treatment. Importantly, botryosphaeran did not induce inflammation in mice when administered alone over 7 days. This study demonstrated the anti-inflammatory and antinociceptive potential of botryosphaeran in these experimental models, making this fungal β-glucan a new possibility for complementary treating acute and chronic inflammation.

Anti-inflammatory activity of Atavijambira (Atalantia monophylla DC.) root and stem

Abstract Background: Atalantia monophylla DC. family Rutaceae, a promising drug, has multifaceted ethnomedicinal claims to treat rheumatism, arthritis, paralysis, cough, skin diseases, etc., In traditional practice, root and stem are administered to treat rheumatism and arthritis, respectively. Aim: The aim of this study was to evaluate the anti-inflammatory activity of the root and stem of A. monophylla against carrageenan-induced acute paw edema in wistar albino rats. Materials and methods: Shade dried root and stem were grinded separately into a fine powder (120#) by the mechanical grinder. Test drugs were prepared in Phanta (hot infusion) form and used as fresh (A. monophylla root Phanta [AMRP] and A. monophylla stem Phanta [AMSP]). Difference between the groups was statistically determined by one-way ANOVA, followed by Dunnett’s multiple “t” test for unpaired data. Results: AMSP showed a significant decrease in paw inflammation at all-time interval (P &lt; 0.05), whereas AMRP produced nonsignificant decrease in paw inflammation at 1 h (43.53%) and 5 h (25.74%) while significant (P &lt; 0.05) anti-inflammatory activity at 3 h (36.35%) in comparison with the normal control group. Conclusion: The present study concluded that A. monophylla stem is having significant anti-inflammatory activity in comparison to its root when administered in Phanta form against carrageenan-induced acute inflammation in rats.

Liquid Biopsy-Based Biomarkers of Inflammatory Nociception Identified in Male Rats.

Physicians are challenged in treating pain patients due to the lack of quantifiable, objective methods of measuring pain in the clinic; pain sensation is multifaceted and subjective to each individual. There is a critical need for point-of-care quantification of accessible biomarkers to provide objective analyses beyond the subjective pain scales currently employed in clinical care settings. In the present study, we employed an animal model to test the hypothesis that circulating regulators of the inflammatory response directly associate with an objective behavioral response to inflammatory pain. Upon induction of localized paw inflammation, we measured the systemic protein expression of cytokines, and activity levels of matrix metalloproteinases (MMPs) that are known to participate in the inflammatory response at the site of injury and investigated their relationship to the behavioral response across a 24 h period. Intraplantar injection with 1% λ-carrageenan induced a significant increase in paw thickness across this timespan with maximal effects observed at the 8 h timepoint when locomotor activity was also impaired. Expression of the chemokines C-X-C motif chemokine ligand 1 (CXCL1) and C-C motif chemokine ligand 2 (CCL2) positively correlated with paw inflammation and negatively correlated with locomotor activity at 8 h. The ratio of MMP9 to MMP2 activity negatively correlated with paw inflammation at the 8 h timepoint. We postulate that the CXCL1 and CCL2 as well as the ratio of MMP9 to MMP2 activity may serve as predictive biomarkers for the timecourse of inflammation-associated locomotor impairment. These data define opportunities for the future development of a point-of-care device to objectively quantify biomarkers for inflammatory pain states.