Reduction Of Inflammatory Infiltration Research Articles

Sesquiterpene lactones from Cichorium intybus exhibit potent anti-inflammatory and hepatoprotective effects by repression of NF-κB and enhancement of NRF2.

Cichorium intybus is a traditional medicinal herb for hepatitis treatment in China and Europe. Sesquiterpene lactones are the main active ingredients in C. intybus. However, their structure-activity relationship (SAR) and molecular mechanisms of anti-inflammatory and hepatoprotective effects requires further elucidation. To identify new sesquiterpene lactones from C. intybus, and further evaluate their anti-inflammatory effects, SAR, and mechanisms of anti-inflammatory and hepatoprotective properties. Identification of sesquiterpene lactones from C. intybus using chromatographic fractionation, NMR, and mass spectrometry. The repression of inflammation was evaluated in RAW264.7 macrophages incubated with LPS. Western blotting was employed to investigate the anti-inflammatory mechanisms. The hepatoprotective effect was measured in LPS/D-galactosamine (D-GalN)-induced acute hepatitis. We identified 3 new sesquiterpene lactones and 15 known analogues from C. intybus. SAR analysis showed that the α-methylene-γ-lactone moiety was essential for their anti-inflammatory properties. Furthermore, 8-deoxylactucin was identified as the most potent anti-inflammatory component in LPS-induced RAW264.7 macrophages by reduction of nitric oxide production via inhibiting iNOS expression, and suppression of IL-1β, IL-6, and TNF-α expression. Mechanistically, 8-deoxylactucin not only blocked LPS-induced IKKα/β phosphorylation, IκBα phosphorylation and degradation, and NF-κB nuclear accumulation, but also enhanced NRF2 expression and nuclear translocation, HO-1 and NQO1 expression, and reduced ROS generation in vitro. In vivo, 8-deoxylactucin mitigated LPS/D-GalN-induced acute hepatitis, which manifested as reduction in inflammatory infiltration, live injury, serum levels of AST and ALT, and production of pro-inflammatory cytokines and 4-hydroxynonenal. 8-Deoxylactucin, the sesquiterpene lactone of C. intybus, exerted anti-inflammatory and hepatoprotective effects by blocking NF-κB activation and enhancing NRF2 activation.

A combination of gold nanoparticles and laser photobiomodulation to boost antioxidant defenses in the recovery of muscle injuries caused by Bothrops jararaca venom.

108 Swiss male mice were used, divided into nine groups (n = 12) with different combinations of treatments. Animals were inoculated with 250µg of B. jararaca venom at the right gastrocnemius muscle. The treatment started 12h after the venom injection and occurred daily for 7days. AS was administered as recommended by the Ministry of Health. Low-power aluminum gallium indium phosphide (AlGaInP) with continuous emission (wavelength 660nm; average power 30 mW ± 20%; beam size of 0.06310 cm2; and dose of 2J for ± 1min per point). Irradiation was applied to five distinct areas surrounding the wound, at a perpendicular angle to the skin. The groups treated with GNPs received treatment through the application of 1mL of 20nm GNPs (30mg/L). Muscle, heart, and kidneys were removed for histological and biochemical analysis. GNPs can improve the results achieved by PBM and the standard treatment, presenting significant benefits in histological parameters, such as the increase of new blood vessels, fibroblasts, collagen, and a reduction of inflammatory infiltrate. GNPs can be considered an option to accelerate the recovery of Bothrops lesions.

Epidermal keratinocytes-specific PD-L1 knockout causes delayed healing of diabetic wounds

BackgroundDiabetic ulcers (DUs) is a common complication of diabetes, for which the morbidity and mortality increasing annually worldwide. The deficiency of PD-L1 in keratinocytes (KCs) may be linked to the diabetic wound healing impediments. ObjectiveOur study utilized transgenic mice to assess the functions of epidermal KCs-specific PD-L1 in DUs treatment. Methods and resultsEpidermal KCs-specific PD-L1 knockouted mice demonstrate deteriorated healing rates, concomitant with exacerbated inflammatory infiltration and excessive angiogenesis. The streptozotocin-induced murine diabetes model was used to imitate DUs in-vivo context, and the delayed healing was found under diabetic conditions. We then generated transgenic mice overexpressing PD-L1 in the epidermis. PD-L1 overexpression accelerate the DUs healing process accompanied by a reduction in inflammatory infiltration and a corresponding decreasion of angiogenesis. Therefore, overexpression of PD-L1 accelerates the healing process of DUs. ConclusionIn sum, epidermal KCs-specific PD-L1 plays vital roles in epidermis regeneration, inflammatory infiltration, and angiogenesis during DUs restoration, and would not be easily to format fibrous scar. Our study elucidated a new therapeutic idea for slow-healing wound care.

The impact of omega-3 fatty acids in a combined experimental model of periodontitis and metabolic syndrome

Relevance. Periodontitis and metabolic syndrome are interrelated conditions that often aggravate each other through shared pathogenic mechanisms, highlighting the need for integrated immunomodulatory therapeutic approaches. Despite this, existing literature presents inconsistent data regarding the interplay between periodontitis and metabolic syndrome, as well as limited insights into the potential benefits of omega-3 polyunsaturated fatty acids (ω-3 PUFAs) as an adjuvant therapy. Specifically, the role of ω-3 PUFAs in modulating the inflammatory response and mitigating bone resorption remains inadequately explored. Therefore, this study aims to evaluate the inflammatory response and osteoresorption in periodontal tissues under the combined conditions of periodontitis and metabolic syndrome, while also assessing the therapeutic effects of ω-3 PUFAs.Materials and methods. This experimental study was conducted using 30 male Wistar rats, which were randomly assigned to one of five groups: a control group, a periodontitis group, a combined periodontitis and metabolic syndrome group, a periodontitis group treated with omega-3 fatty acids, and a combined periodontitis and metabolic syndrome group treated with omega-3 fatty acids. The study employed several research methodologies. Biochemical analyses were performed to confirm the presence of metabolic syndrome. Histopathological and morphometric assessments were conducted to evaluate the inflammatory response, the extent of osteoresorption, and the number of osteoclasts in the periodontal tissues. Additionally, molecular genetic analysis was used to measure the relative mRNA expression levels of key inflammatory and osteoclastogenic markers, including TNFα, IL-1β, RANK, and OPG.Results. The data demonstrated that metabolic syndrome in laboratory animals significantly exacerbates both the pro-inflammatory response and the extent of lacunar osteoclastic bone resorption. Administration of ω-3 PUFAs at a dose of 40 mg/kg over 30 days led to a statistically significant reduction in inflammatory infiltration within the periodontal tissues (3.17 ± 0.21 in the control group vs. 1.83 ± 0.21 in the treated group, р = 0.001) and a decrease in the number of osteoclasts (3,75 ± 0,45 in the control group vs. 1,75 ± 0,35 in the treated group, р = 0.003). Additionally, there was a notable reduction in the expression levels of TNFα, IL-1β, and RANK mRNA. Similar but less pronounced effects were observed in the group with combined metabolic syndrome following ω-3 PUFA administration.Conclusion. These findings enhance the current understanding of the interplay between periodontitis and metabolic syndrome, highlighting the potential of ω-3 PUFAs as a preventive or therapeutic intervention. ω-3 PUFAs may be effective in mitigating inflammatory responses and bone resorption, both in cases of isolated periodontitis and when accompanied by metabolic syndrome.

Amentoflavone alleviated cartilage injury and inflammatory response of knee osteoarthritis through PTGS2.

The role of amentoflavone on cartilage injury in knee osteoarthritis (KOA) rats and the underlying mechanismwere explored. KOA rat and IL-1β-stimulated chondrocyte models were constructed. MTT, colony formation, and ELISA were performed to determine the cytotoxicity, cell proliferation, and inflammatory factors. The role of PTGS2 in IL-1β-stimulated chondrocytes was also confirmed through transfecting PTGS2 overexpression and silencing plasmids. Further, we analyzed how amentoflavone regulated PTGS2 to improve IL-1β-stimulated chondrocytes in vitro. Additionally, we analyzed the expression of PTGS2 after amentoflavone treatment. In vivo, HE and Safranin-O staining were carried out, and the inflammatory response was detected by ELISA and HE staining. In addition, we also analyzed the regulatory effect of amentoflavone on PTGS2 and explored the mechanism effect of PTGS2 in vitro and in vivo. The results indicated that PTGS2 was the downstream molecule of amentoflavone, which was highly expressed in IL-1β-stimulated chondrocytes and KOA rats, and amentoflavone decreased PTGS2 expression. We also confirmed the potential role of amentoflavone on KOA, which was also characterized by the repair of cartilage injury, reduction of inflammatory infiltration, and improvement of functional disability. Consistent with in vivo results, in vitro experiments gave the same conclusions. Amentoflavone reduced PTGS2 expression in IL-1β-stimulated chondrocytes and inhibited inflammation of chondrocytes via PTGS2. Collectively, the results confirmed that this drug was the potential targeted drug for KOA, whose repair effect on cartilage injury was partly related to PTGS2.

(PM-17) THE ROLE OF PHOSPHODIESTERASE-5 INHIBITORS IN THE TREATMENT OF COMPLEX PENILE WOUNDS: A CASE REPORT AND LITERATURE REVIEW

Abstract Introduction/Objective Phosphodiesterase-5 inhibitors (PDE5i) are traditionally employed for the management of erectile dysfunction and lower urinary tract symptoms. However, their role in the healing of complex penile wounds remains scarcely discussed. This study aims to describe the case of a 27-year-old man who suffered from penile strangulation, treated with sildenafil. Additionally, a literature review concerning the use of PDE5i for the treatment of complex penile injuries will be conducted. Methods A 27-year-old man presented to the emergency room after a self-induced penile strangulation using a ring placed at the base of the penis. Extensive necrosis of the corpus cavernosum was identified through magnetic resonance imaging. Considering the psychosocial implications of penile amputation, organ preservation was chosen, and conservative treatment with sildenafil and daily local dressings was initiated. Thirty days after admission, Doppler ultrasound revealed symmetrical corpus cavernosum with normal echogenicity and usual blood flow patterns. Subsequently, the patient underwent superficial necrotic tissue debridement followed by partial skin grafting in a second stage, leading to hospital discharge after 55 days of hospitalization. Results PDE5 inhibitors enhance nitric oxide bioavailability, promoting vasodilation and improved tissue oxygenation, thereby facilitating various phases of the healing process. Evidence indicates their benefit in microvascular perfusion in skin and soft tissues. Reports suggest satisfactory outcomes of topical PDE5i application in radiation-induced lesions, implying their role in epidermal cell expansion, reduction of inflammatory infiltrates, enhanced vascularization, and decreased collagen deposition. Conclusion This study presents a successful case of sildenafil use in treatment of a complex penile wound, shedding light on an unconventional application of PDE5i. Financing No conflict.

Action of melatonin and physical exercise on the liver of cirrhotic rats: Study of oxidative stress and the inflammatory process.

Cirrhosis is characterized by structural and functional alterations of the liver. Melatonin (MLT) has antioxidant properties. Physical exercise (EX) can reverse muscle loss in cirrhotic patients. The objective was to evaluate the action of MLT and EX on the liver of rats subjected to the experimental model of bile duct ligation (BLD). 48 male Wistar rats were used, divided into groups: Control (CO), CO+MLT, CO+EX, CO+MLT+EX, BDL, BDL+MLT, BDL+EX, and BDL+MLT+EX. The treatments occurred from the 15th to the 28th day. The dose of MLT was 20 mg/kg via I.p (1x/day), and the EX was performed 10 min/day. Blood and liver were collected for analysis. The liver integrity enzymes AST, ALT, and ALP showed a significant reduction in the groups treated with MLT and EX. Histological analyses showed reorganization of the liver parenchyma, reduction of inflammatory infiltrate, and fibrotic nodules. Lipoperoxidation (LPO), the activity of antioxidant enzymes, and nitric oxide metabolites showed a significant reduction in the groups treated with MLT and EX. The expression of TNF-α and NF-kB decreased in the treated groups. Melatonin and physical exercise seem to be effective in restoring the parameters evaluated in this model of experimental cirrhosis.

Protective effect of bamboo root dietary Fibre on hyperlipidaemia mice induced by high-fat diet

ABSTRACT Bamboo root as by-product of bamboo shoot processing is rich in dietary fibre. The protective effects of bamboo root dietary fibre (BRDF) with different intake dose on the hyperlipidaemia mice were investigated. The mice weight in the high dose (HD) group was 31.18 g, lower than that of high fat diet (HFD) group (33.71 g, p < .05), after consuming BRDF. The contents of TC, TG and LDL-C in HD group mice were significantly reduced (p < .05). The mice liver index significantly decreased in the BRDF group compared to the HFD group. TC and TG contents in the liver of HD mice were 0.85 mmol·g−1 and 0.83 mmol·g−1, respectively, significantly lower than 2.25 mmol·g−1 and 1.46 mmol·g−1 in the normal control group (p < .05). H&E staining showed a reduction in inflammatory infiltration. These results suggested that BRDF in the daily diet could reduce blood lipids in vivo and prevent hyperlipidaemia.

Modulation of the tissue composition of regenerative neuroma by dexamethasone and granulocyte colony-stimulating factor

Aim. To evaluate the change of the content of mesenchymal (vimentin-positive) and reparative Schwann (GFAP-positive) cells in the regenerative neuroma of the rat sciatic nerve, taking into account the activity of local homeostasis regulation (CD73 expression) under the influence of dexamethasone (Dex) and granulocyte colony-stimulating factor (GCSF). Materials and methods. Vimentin-, GFAP-, and CD73-positive cells were detected immunohistochemically in the regenerative neuroma of 168 male Wistar rats, and their presence was quantified using statistical methods. Results. The regenerative neuroma of the sciatic nerve is characterized by a stereotypical cellular composition kinetics. Dexamethasone, during the initial stage of neuroma formation (1–7 days), led to a reduction in inflammatory infiltration and accumulation of vimentin+ mesenchymal cells. It significantly accelerated the accumulation of neural GFAP+ reparative Schwann cells and suppressed the expression of CD73. The granulocyte colony-stimulating factor (1–3 days) in the regenerative neuroma resulted in a slight reduction in inflammatory infiltration and an increase in the number of blood vessels penetrating into it. Under these conditions, the speed and quantity of vimentin+ cell accumulation and CD73 expression noticeably increased, as well as the speed of GFAP+ cell accumulation. The deviations in the expression of vimentin, GFAP, and CD73 in the neuroma under the administration of the mentioned substances gradually decreased and, after 8 weeks of the experiment, did not differ significantly from the control. When combined, dexamethasone and granulocyte colony-stimulating factor potentiated each other’s effects and, in addition, led to long-term maintenance of high CD73 expression. At the same time, the most pronounced approximation of the structure of the newly formed part of the nerve to the structure of the intact nerve trunk was observed. Conclusions. The simultaneous action of Dex and GCSF during the initial stages of regenerative neuroma formation, in contrast to their individual actions, not only alters the dynamics of mesenchymal and neural cell accumulation but also modifies the newly formed part of the nerve, bringing its structure closer to the intact state. This phenomenon is accompanied by increased reactivity of neurolemocytes during the axial cylinder germination stage of neuroma regeneration.

Annexin A12–26 hydrogel improves healing properties in an experimental skin lesion after induction of type 1 diabetes

Diabetes mellitus (DM) is characterized by metabolic alterations that involve defects in the secretion and/or action of insulin, being responsible for several complications, such as impaired healing. Studies from our research group have shown that annexin A1 protein (AnxA1) is involved in the regulation of inflammation and cell proliferation. In light of these findings, we have developed a new technology and evaluated its effect on a wound healing in vivo model using type 1 diabetes (T1DM)-induced mice. We formulated a hydrogel containing AnxA12–26 using defined parameters such as organoleptic characteristics, pH, UV–vis spectroscopy and cytotoxicity assay. UV–vis spectroscopy confirmed the presence of the associated AnxA12–26 peptide in the three-dimensional hydrogel matrix, while the in vitro cytotoxicity assay showed excellent biocompatibility. Mice showed increased blood glucose levels, confirming the efficacy of streptozotocin (STZ) to induce T1DM. Treatment with AnxA12–26 hydrogel showed to improve diabetic wound healing, defined as complete re-epithelialization and tissue remodeling, with reduction of inflammatory infiltrate in diabetic animals. We envisage that the AnxA12–26 hydrogel, with its innovative composition and formulation be efficient on improving diabetic healing and contributing on the expansion of the therapeutic arsenal to treat diabetic wounds, at a viable cost.

The protective roles of liraglutide on Kawasaki disease via AMPK/mTOR/NF-κB pathway.

Kawasaki disease (KD) is an acute febrile rash illness among children of unknown etiology, with coronary artery injury. The main purpose of this study was to investigate the protective effects of liraglutide on KD, and elucidate the underlying mechanisms. The candida albicans water-soluble fraction (CAWS)-induced coronary arteritis of mouse KD model in vivo and tumor necrosis factor α (TNF-α) induced endothelial cell injury of human umbilical vein endothelial cell (HUVEC) model in vitro were used to explore the anti-inflammation and anti-apoptosis effects of liraglutide on KD. In vivo results showed that liraglutide could significantly alleviate the coronary artery injury of KD mice, as evidenced by the reduction of inflammatory infiltration around the coronary arteries, downregulation of inflammatory cytokines and chemokines expressions, and decrease of TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) positive cell rates. The results in vitro also displayed that liraglutide could markedly relieve the inflammatory of TNF-α induced HUVECs through downregulating the expressions of inflammatory and chemokine indicators as well as inhibit TNF-α induced HUVEC apoptosis by the less ratio of apoptotic cells, the more loss of mitochondrial membrane potential (△Ψm), the lower level of intracellular reactive oxygen species (ROS), and the more ratio of BCL-2/BAX. Further in vivo and in vitro studies demonstrated that liraglutide could rescue endothelial cell injury through AMPK/mTOR/NF-κB pathway. In conclusion, liraglutide could play protective roles on KD through inhibiting endothelial cell inflammation and apoptosis via the activation of AMPK/mTOR/NF-κB pathway.

Probiotic Lactobacillus rhamnosus EM1107 prevents hyperglycemia, alveolar bone loss, and inflammation in a rat model of diabetes and periodontitis.

This study evaluated the antihyperglycemic, anti-bone-resorptive, and anti-inflammatory efficacy of the probiotic Lactobacillus rhamnosus EM1107 in an experimental model of ligature-induced periodontitis in diabetic rats treated with metformin (Met). A total of 114 male Wistar rats was randomly divided into six groups: (1) control, (2) experimental periodontitis (EP), (3) EP + diabetes mellitus (DM), (4) EP + probiotic (Prob), (5) EP + DM + Prob, and (6) EP + DM + Prob + Met. The animals received probiotic gavage during the 30 days of the experiment. DM was induced on the 14th day of the experiment with a single injection of streptozotocin into the penile vein, followed by ligature for EP induction and Met gavage on the 19th day and euthanasia on the 30th day. Heart blood, gingival and periodontal tissue, and hemimaxillae were collected. Biomolecular analysis, immunoenzymatic assays, histomorphology, and microtomographic analysis were performed. Data were statistically analyzed (p<0.05). There was a significant reduction in interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the Prob groups (p<0.05) as well as in blood glucose levels in the Prob and Met groups (p<0.001). In addition, histomorphological analysis revealed that the Prob groups had a reduction in inflammatory infiltrate. Tartrate-resistant acid phosphatase (TRAP) and microtomographic analyses showed that the EP/DM/Prob/Met group had significantly lower linear and volumetric bone loss than those who had no treatment (p<0.01). SOD and GPx immunostaining decreased in all groups receiving probiotics. The findings suggest the immunoinflammatory efficacy of the probiotic L. rhamnosus EM1107 administered either alone or in association with Met in type 1 DM associated with periodontitis.

Campsiandra laurifolia Reduces Oxidative Stress and Inflammation in Rats with Ulcerative Colitis

AbstractUlcerative colitis (UC) affects the mucosa and submucosa of the large intestine. One of the mechanisms involved in its etiology is oxidative stress (OS), directly involved in the inflammatory process characteristic of UC. The Campsiandra laurifolia, known as acapurana, was described as possessing antioxidant properties. We used 24 male Wistar rats, divided into control (CO), control + acapurana (CO + A), colitis (CL), and colitis + acapurana (CL + A) groups. This study performed histological analysis, measuring anal sphincter pressure (ASP) and lipoperoxidation (LPO). The activity of the antioxidant enzyme superoxide dismutase (SOD) and glutathione (GSH) levels were evaluated. The expression of the nuclear factor kappa B (NFκB) and inducible nitric oxide synthase (iNOS) was analyzed by immunohistochemistry. The statistical analysis used was the one-way analysis of variance (ANOVA), followed by the Student-Newman-Keuls test; values were expressed as mean ± standard error, and the significance level was p &lt; 0.05. In the animals of the CL group, we observed the destruction of the crypts and the presence of mucosal ulcers, edema, and submucosal inflammatory infiltrate, as well as increased damage to the intestinal mucosa, reduced ASP, increased LPO and SOD activity, reduced GSH levels, and increased expression of NFκB and iNOS. The administration of C. laurifolia in the CL + A group was shown to cause regeneration of crypts, reduction of inflammatory infiltrate, reduction of damage to the intestinal mucosa, increase in ASP, and reduction in LPO with the restoration of SOD activity and GSH levels. The immunohistochemistry of NFκB and iNOS was significantly reduced. Therefore, the C. laurifolia aqueous extract appears to exert an antioxidant and anti-inflammatory effect in rats with AA-induced colitis.

Photobiomodulation associated with lipid nanoparticles and hyaluronic acid accelerate the healing of excisional wounds.

Objectives: This article aimed to investigate the effects of the association between photobiomodulation and hyaluronic acid incorporated in lipid nanoparticles in an epithelial lesion model in inflammatory parameters and oxidative stress. Methods: Eighty Wistar rats were randomly assigned to the following groups: epithelial lesion group (EL); EL+PBM; EL+HA; EL+SLNs; EL+SLNs-HA; EL+PBM+HA; EL+PBM+SLNs; EL+PBM+SLNs-HA. The animals were anesthetized with 4% isofluorane after shaving and induced to an epithelial lesion. Topical treatment with a gel containing HA (0.9%) and/or SLNs (10mg/mL) and with laser irradiation occurred daily for 1week. Results: The results showed an increase in wound contraction on the seventh day in the LE + LBM + AH-NPL group, a reduction in pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), an increase in anti-inflammatory cytokines (IL- 4 and IL-10) and TGF-β. The levels of pro-inflammatory cytokine IL-4 and TGF-β also showed an increase in the LE + NPL-AH, LE + FBM + AH, LE + FBM + NPL and LE + FBM + NPL-AH groups. Regarding oxidative stress parameters, the levels of DCF and nitrite decreased in the combined therapy group when compared to the control group, as well as oxidative damage (carbonyl and sulfhydryl). In the antioxidant defense, there was an increase in GSH and SOD in the combination therapy group. Histological analysis showed a reduction in inflammatory infiltrate in the combination therapy group. The number of fibroblasts and the compaction of collagen fibers did not obtain significant responses. Conclusions: Results analyzed together showed that the combined therapy favored the repair process, and that studies can be carried out to enhance the histological analysis therapy favored the tissue repair process and that studies can be carried out to enhance the histological analysis.

IDF21-0365 Treatment with AnxA12-26 gel improves healing properties in an experimental skin lesion after induction of T1DM

Background: Diabetes mellitus (DM) is characterized by metabolic alterations that involve defects in the secretion and/or action of insulin, being responsible for several complications, such as poor healing. In diabetic people, healing is impaired by changes in the inflammatory response and regulatory mechanisms. Recent studies by our research group have shown that the annexin A1 protein (AnxA1) is involved in the regulation of inflammation, apoptosis, cell growth and differentiation. Aim: To develop an economically viable gel of carboxymethylcellulose (CMC) containing the peptide AnxA12-26 and evaluate its effect on skin lesions in type 1 diabetes mellitus (T1DM). Method: The analysis of the gel was performed using organoleptic characteristics, pH and UV-Vi spectroscopy to assess the absorption and fluorescence emission profile of the free and gel-associated peptide. Skin lesions were performed in C57BL/6 control (non-diabetic) and diabetic mice in periods of 3, 7 and 14 days. The following aspects were evaluated: weight and blood glucose; macroscopic analysis; histopathology of scar and adjacent tissue; the expression of ED-1 positive macrophages in the healing process; expression of matrix metalloproteinases (MMP-9) and analysis of the anti-inflammatory protein AnxA1. Results: The gel/AnxA12-26 kept at 4°C showed better results in the parameters of pH and organoleptic characteristics. The UV-Vi spectroscopy analyzes confirmed the presence of the peptide in three-dimensional hydrogel matrix. Diabetic animals, treated or not, showed weight loss and increased blood glucose compared to controls, confirming the efficacy of DM1 induction. Treatment with gel/AnxA12-26 showed evolution of the healing process, reduction of inflammatory infiltrate, re-epithelialization and beginning of remodeling in control and diabetic animals. The effectiveness of the gel/AnxA12-26 was also observed in immunohistochemical analyzes through macrophage modulation, modulation of MMP-9 expression and reduction of endogenous annexin A1 in the control and diabetic groups. Conclusion: Together, our results were innovative in the formulation of the hydrogel with the peptide AnxA12-26, showing efficiency in the cutaneous healing process and indicating its possibility to increase the therapeutic arsenal, at a viable cost, for the treatment of diabetic wounds.

Histopathological and ultrastructural assessment of atovaquone-proguanil hydrochloride combination in chronic murine toxoplasmosis

ABSTRACT Over one billion people worldwide are expected to have Toxoplasma gondii infection with anonymous health problems. Available therapies are ineffective for persistent chronic toxoplasmosis. So, there is an imperative need for effective therapies to eliminate chronic tissue stage. In this study, we aimed to assess the effect of a drug combination of atovaquone and proguanil hydrochloride in the treatment of experimental chronic toxoplasmosis. Fifty Swiss Webster mice were used in the study. Forty mice were infected with Me49 type II cystogenic Toxoplasma gondii strain and allocated into four groups: infected untreated (vehicle-administered), infected and treated with cotrimoxazole (CTX) 370 mg/kg/day, infected and treated with atovaquone (ATV) 100 mg/kg/day, and infected and treated with atovaquone/proguanil (ATV/PROG) 50 mg/kg/day. An additional group of uninfected mice was used as an uninfected control group. Drug treatment was initiated 8 weeks post-infection and continued for two weeks. All mice were sacrificed 12 weeks post-infection. Parasitological and histopathological parameters were assessed. Toxoplasma gondii cysts recovered from brain tissue homogenates of both infected untreated and ATV/PROG-treated groups were examined by scanning electron microscopy. Combined ATV/PROG treatment demonstrated a significant reduction of Toxoplasma gondii cyst count in brain tissue (a reduction rate of 84.87%) compared to untreated group (P < .001). Brain tissues obtained from ATV/PROG treated group showed reduction of inflammatory infiltrate and marked attenuation and deformation of recovered Toxoplasma gondii cysts. We conclude that ATV/PROG drug combination could offer a potential drug therapy for Toxoplasma gondii chronic cystic stage.

Kaempferol protects rats with severe acute pancreatitis through regulating NF-κB and Keap1–Nrf2 signaling pathway

Kaempferol (KF) is an important natural anti-inflammatory flavonol. Acute pancreatitis (AP) is an inflammatory disorder, which in about 20% cases may develop into severe acute pancreatitis (SAP) with a high mortality rate. This research was to study the effects and mechanism of kaempferol on SAP. SAP was induced by sodium taurocholate. The level of cytokines was analyzed by enzyme-linked-immunosorbent serologic assay. The expression of nuclear factor kappa B (NF-κB) and Kelch-like ECH-associated protein 1–nuclear factor erythroid 2-related factor 2 (Keap1–Nrf2) proteins was analyzed by Western blot assay. Pathological changes in the pancreas were evaluated by hematoxylin and eosin staining. Kaempferol attenuated pancreatic injury in SAP rats, including reduction in inflammatory infiltration and necrosis. The level of serum amylase and lipase was also decreased in kaempferol-treated SAP rats. Kaempferol inhibited the expression of inflammatory mediators (nuclear factor-α, Interlukin-1β, and Interlukin-6), and alleviated the oxidative stress characterized by the decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD) levels. Kaempferol decreased the expression of cleaved caspase 3 and anti-apoptotic protein Bcl-2, which indicated that kaempferol could inhibit apoptosis of pancreatic cells in SAP rats. Kaempferol treatment could decrease the expression of p-p65 and the amount of nuclear Nrf2 (Nu-Nrf2), which demonstrated that kaempferol inhibited the NF-κB activation and enhanced the Keap1–Nrf2 pathway. Our research indicated that kaempferol could attenuate the pancreatic injury of SAP by regulating NF-κB and Keap1–Nrf2 signaling pathway. Kaempferol could serve as a natural candidate for treating SAP.

Suppression of Experimental Autoimmune Encephalomyelitis by ILT3.Fc.

Multiple sclerosis (MS) is a chronic autoimmune disease of the CNS that is characterized by demyelination, axonal loss, gliosis, and inflammation. The murine model of MS is the experimental autoimmune encephalopathy (EAE) induced by immunization of mice with myelin oligodendrocyte glycoprotein (MOG)35-55 Ig-like transcript 3 (ILT3) is an inhibitory cell surface receptor expressed by tolerogenic human dendritic cells. In this study, we show that the recombinant human ILT3.Fc protein binds to murine immune cells and inhibits the release of proinflammatory cytokines that cause the neuroinflammatory process that result in paralysis. Administration of ILT3.Fc prevents the rapid evolution of the disease in C57BL/6 mice and is associated with a profound reduction of proliferation of MOG35-55-specific Th1 and Th17 cells. Inhibition of IFN-γ and IL-17A in mice treated with ILT3.Fc is associated with delayed time of onset of the disease and its evolution to a peak clinical score. Neuropathological analysis shows a reduction in inflammatory infiltrates and demyelinated areas in the brains and spinal cords of treated mice. These results indicate that inhibition of Th1 and Th17 development provides effective suppression of EAE and suggests the feasibility of a clinical approach based on the use of ILT3.Fc for treatment of MS. Furthermore, our results open the way to further studies on the effect of the human ILT3.Fc protein in murine experimental models of autoimmunity and cancer.

Effect of natalizumab treatment on metalloproteinases and their inhibitors in a mouse model of multiple sclerosis.

Matrix metalloproteinases (MMPs) regulated by their tissue inhibitors (TIMPs) play a significant role in the pathogenesis of multiple sclerosis (MS) and its mouse model, experimental autoimmune encephalomyelitis (EAE), as they degrade extracellular matrix including vascular basal laminae and by damaging blood-brain barrier (BBB) facilitate transmigration of immune cells into the central nervous system. MMPs are also involved in destruction of myelin sheaths, leading to axonal and neuronal loss. The aim of the present study was to assess whether natalizumab, a transmigration-inhibiting monoclonal antibody against α4β1 integrin, influences expression of MMPs and TIMPs in the central nervous system of mice with EAE. MMP-2 and MMP-9, their respective inhibitors TIMP-2 and TIMP-1 and laminin were assessed by quantitative immunohistochemistry in the spinal cord cryosections of C57BL/6 mice with EAE in the successive phases of the disease (onset, peak and chronic). The percentage of immunopositive areas were calculated in sections encompassing the whole spinal cord cross-sectional area occupied by the gray and white matter. Results obtained in animals administered with 5 mg/kg natalizumab were compared with those collected from control mice receiving 5 mg/kg IgG. Both studied MMPs and both TIMPs were upregulated in control EAE mice. Natalizumab treatment significantly reduced expression of MMPs and increased expression of TIMPs in the peak and chronic phases of the disease. This effect was accompanied by inhibition of laminin degradation in the vascular basal laminae and reduction of inflammatory infiltration. Results of this study demonstrate that in addition to its well known anti-integrin activity counteracting transmigration of immune cells into the central nervous system, natalizumab strengthens this effect by its probably indirect influence on MMPs and TIMPs leading to protection of blood-brain barrier integrity.

Cutaneous wound healing: canine allogeneic ASC therapy

BackgroundWound healing is a complex biological process comprised of a series of sequential events aiming to repair injured tissue. Adult mesenchymal stem cells (MSCs) have been used in cellular therapy in preclinical animal studies; a promising source of MSCs is adipose tissue (AT). In this paper, we evaluated the clinical value and safety of the application of cultured allogenic MSCs from AT for acute and chronic skin wound healing in a canine model.MethodsTwenty-four dogs of different breeds between 1 and 10 years of age with acute and chronic wounds were studied. Morphology of the wounded skin was monitored for changes over time via serial photographs and histopathological studies.ResultsThe percentage of the wounds that exhibited contraction and re-epithelialization were significantly different between wounds treated with adipose mesenchymal stem cells (ASCs) and control wounds; this effect was observed in both acute and chronic conditions. At 90 days, re-epithelization of acute and chronic wounds reached more than 97%. Histopathological study revealed a reduction in inflammatory infiltrate and the presence of multiple hair follicles on day 7 after treatment with ASCs, promoting epidermal and dermal regeneration. To guarantee the safety of our treatment, we determined the serum levels of cytokine markers in our patients. ASC treatment upregulated granulocyte-macrophage colony stimulating factor (GM-CSF) at the gene level, which may contribute to the recruitment of cells that participate in skin repair to the site of injury.ConclusionsThe development of an allogenic ASC therapy to improve wound healing in a canine model could have a clinical impact in human treatment.