Levels Of Collagenase Research Articles

Melatonin: a potential therapeutic agent for alleviating renal tubular epithelial cell interstitial transformation.

Melatonin (Mel) has been documented to modulate epithelial-mesenchymal transition (EMT) in cellular systems. The interstitial transformation of renal tubular epithelial cells constitutes a key pathogenic mechanism underlying renal fibrosis. This study aims to elucidate the role of Mel in the EMT process of renal tubular epithelial cells. A rat model of unilateral ureteral obstruction (UUO) was developed through unilateral ureteral ligation, followed by treatment with Mel (5, 10, and 20 mg/kg). Subsequent analyses included histopathological examination, measurement of creatinine and blood urea nitrogen levels, immunofluorescence analysis of fibronectin (FN), and immunohistochemical analysis of alpha-smooth muscle actin (α-SMA). Transforming growth factor-beta 1 (TGF-β1) initiates a fibrotic response in NRK-52E cells, which is subsequently treated with Mel (0. 1, 1, and 10 μmol/L). Evaluates cell viability, migration, and the expression of EMT related proteins, including FN, α-SMA, collagenase I, and E-cadherin. To validate the regulatory interaction between miR-153-3p and Forkhead transcription factor o subfamily member 3A (FOXO3A), cells are transfected with miR-153-3p mimics or siRNA targeting FOXO3A (si-FOXO3A). In results Mel exhibits a dose-dependent capacity to ameliorate renal injury and rectify glomerular structural abnormalities in UUO rat models. In comparison to UUO model rats, melatonin significantly reduced the expression levels of FN and α-SMA. In vitro studies demonstrated that Mel inhibited the activity and migratory behavior of cells, as well as the protein expression levels of FN, α-SMA, and collagenase I in NRK-52E cells. Relative to the control group, UUO rats and TGF-β1-induced NRK-52E cells exhibited elevated expression of miR-153-3p and reduced FOXO3A. In vitro models further revealed that upregulation of miR-153-3p or downregulation of FOXO3A can negate the protective effects of Melatonin on TGF-β1-induced EMT. Concluding, Mel inhibits miR-153-3p, thereby promoting the transcription of FOXO3A in UUO rats, which alleviates renal injury and attenuates TGF-1-induced EMT in cells.

γ-Mangosteen, an autophagy enhancer, prevents skin-aging via activating KEAP1/NRF2 signaling and downregulating MAPKs/AP-1/NF-κB-mediated MMPs

BackgroundMangosteens, a naturally occurring xanthones, found abundantly in mangosteen fruits. The anti-skin aging potential of γ-mangosteen (GM) remains unexplored; therefore, we investigated the UVB-induced anti-skin aging of GM via activation of autophagy. HypothesisWe hypothesized that GM exerts antioxidant and anti-aging capabilities both in vitro and in vivo through activation of autophagy as well as control of KEAP1/NRF2 signaling and MAPKs/AP-1/NF-κB-mediated MMPs pathways. MethodsThe anti-skin aging effects of GM were studied using HDF cells and a mice model. Various assays, such as DPPH, ABTS, CUPRAC, FRAP, and ROS generation, assessed antioxidant activities. Kits measured antioxidant enzymes, SA-β-gal staining, collagen, MDA content, si-RNA experiments, and promoter assays. Western blotting evaluated protein levels of c-Jun, c-Fos, p-IκBα/β, p-NF-κB, MAPK, MMPs, collagenase, elastin, KEAP1, NRF2, HO-1, and autophagy-related proteins. ResultsGM exhibited strong antioxidant, collagenase and elastase enzyme inhibition activity surpassing α- and β-mangosteen. GM competitively inhibited elastase with a Ki value of 29.04 µM. GM orchestrated the KEAP1-NRF2 pathway, enhancing HO-1 expression, and suppressed UVB-induced ROS in HDF cells. NRF2 knockdown compromised GM's antioxidant efficacy, leading to uncontrolled ROS post-UVB. GM bolstered endogenous antioxidants, curbing lipid peroxidation in UVB-exposed HDF cells and BALB/c mice. GM effectively halted UVB-induced cell senescence, and reduced MMP-1/-9, while elevated TIMP-1 levels, augmented COL1A1, ELN, and HAS-2 expression in vitro and in vivo. Additionally, it suppressed UVB-induced MAPKs, AP-1, NF-κB phosphorylation. Pharmacological inhibitors synergistically enhanced GM's anti-skin aging potential. Moreover, GM inhibited UVB-induced mTOR activation, upregulated LC3-II, Atg5, Beclin 1, and reduced p62 in both UVB induced HDF cells and BALB/c mice, while blocking of autophagy successfully halt the GM effects against the UVB-induced increase of cell senescence, degradation of collagen through upregulation of MMP-1, underscoring GM's substantial anti-skin aging impact via autophagy induction in vitro and in vivo. ConclusionTogether, GM has potent antioxidant and anti-skin aging ingredients that can be used to formulate skin care products for both the nutraceutical and cosmeceutical industries.

Luteolin ameliorates cornea stromal collagen degradation and inflammatory damage in rats with corneal alkali burn

Corneal alkali burn (AB) is a blindness-causing ocular trauma commonly seen in clinics. An excessive inflammatory reaction and stromal collagen degradation contribute to corneal pathological damage. Luteolin (LUT) has been studied for its anti-inflammatory effects. In this study, the effect of LUT on cornea stromal collagen degradation and inflammatory damage in rats with corneal alkali burn was investigated. After corneal alkali burn, rats were randomly assigned to the AB group and AB + LUT group and received an injection of saline and LUT (200 mg/kg) once daily. Subsequently, corneal opacity, epithelial defects, inflammation and neovascularization (NV) were observed and recorded on Days 1, 2, 3, 7 and 14 post-injury. The concentration of LUT in ocular surface tissues and anterior chamber, as well as the levels of collagen degradation, inflammatory cytokines, matrix metalloproteinases (MMPs) and their activity in the cornea were detected. Human corneal fibroblasts (HCFs) were co-cultured with interleukin (IL)-1β and LUT. Cell proliferation and apoptosis were assessed by CCK-8 assay and flow cytometry respectively. Measurement of hydroxyproline (HYP) in culture supernatants was used to quantify the amount of collagen degradation. Plasmin activity was also assessed. ELISA or real-time PCR was used to detect the production of matrix metalloproteinases (MMPs), IL-8, IL-6 and monocyte chemotactic protein (MCP)-1. Furthermore, the immunoblot method was used to assess the phosphorylation of mitogen-activated protein kinases (MAPKs), transforming growth factor-β-activated kinase (TAK)-1, activator protein-1 (AP-1) and inhibitory protein IκB-α. At last, immunofluorescence staining helped to develop nuclear factor (NF)-κB. LUT was detectable in ocular tissues and anterior chamber after intraperitoneal injection. An intraperitoneal injection of LUT ameliorated alkali burn-elicited corneal opacity, corneal epithelial defects, collagen degradation, NV, and the infiltration of inflammatory cells. The mRNA expressions of IL-1β, IL-6, MCP-1, vascular endothelial growth factor (VEGF)-A, and MMPs in corneal tissue were downregulated by LUT intervention. And its administration reduced the protein levels of IL-1β, collagenases, and MMP activity. Furthermore, in vitro study showed that LUT inhibited IL-1β-induced type I collagen degradation and the release of inflammatory cytokines and chemokines by corneal stromal fibroblasts. LUT also inhibited the IL-1β-induced activation of TAK-1, mitogen-activated protein kinase (MAPK), c-Jun, and NF-κB signaling pathways in these cells. Our results demonstrate that LUT inhibited alkali burn-stimulated collagen breakdown and corneal inflammation, most likely by attenuating the IL-1β signaling pathway. LUT may therefore prove to be of clinical value for treating corneal alkali burns.

Biochemical Pilot Study on Effects of Pomegranate Seed Oil Extract and Cosmetic Cream on Neurologically Mediated Skin Inflammation in Animals and Humans: A Comparative Observational Study.

The presence of phenobarbital and formaldehyde in drugs, food, and beverages can lead to various health issues, including inflammation, oncogenesis, and neurological distress. Psychological stress leads to mood fluctuations and the onset of skin inflammation. Skin inflammation has a range of causes, including chemicals, heavy metals, infection, immune-related disorders, genetics, and stress. The various treatments for skin inflammation include medical and cosmetic creams, diet changes, and herbal therapy. In this study, we investigated the effects of Avocom-M and pomegranate seed oil extract (PSOE) against phenobarbital- and formaldehyde-induced skin biochemical changes in rats. We analyzed the constituents of PSOE using gas chromatography-mass spectrometry and inductively coupled plasma-mass spectrometry. We also observed biochemical changes in the skin of human volunteers with and without TROSYD and PSOE as a skin cream. We compared the biochemical changes in human volunteers' skin before treatment and 21 days after the treatment stopped. The outcomes showed an improvement in the rats' biochemical status, due to PSOE and Avocom-M treatment. The human volunteers treated with TROSYD and PSOE showed substantial amelioration of skin inflammation. PSOE, Avocom-M, and TROSYD produced beneficial effects by reducing the levels of cyclooxygenase-2, lipid peroxidation, tyrosinase, hyaluronidase, elastase, collagenase, and nitric oxide in the animals tested on and in human volunteers.

Novel mannich-based derivative of 2-mercaptobenzimidazole (AK7): a new candidate for the treatment of inflammatory arthritis owing to its NF-κB1 inhibitory potential.

This study investigated the anti-arthritic potential of novel mannich-based derivatives of 2-mercaptobenzimidazole (AK7 and AK9) in rats. The compounds were characterized by NMR and FTIR spectroscopies and their acute anti-inflammatory effects were measured by carrageenan (CRG)-induced paw edema model. The most potent doses of AK7 and AK9 were subsequently evaluated in the complete Freund's adjuvant (CFA)-induced inflammatory arthritis model. AK7 and AK9 inhibited CRG-induced inflammation in a dose-dependent fashion and a similar reduction in CFA-induced paw inflammation was observed. Moreover, X-ray and histopathological analyses of AK7-treated animals displayed normal joint structure whereas AK9, despite of its anti-inflammatory effects, failed to protect against cartilage destruction. Interestingly, biochemical analysis revealed a better safety profile for AK7 than for AK9 and methotrexate. Both compounds suppressed mRNA levels of pro-inflammatory mediators (IRAK1, NF-κB1, TNF-α, IL1B) while only AK7 reduced the transcript levels of interstitial collagenase (MMP1). Molecular docking analysis of AK7 and AK9 with TNF-α and MMP1 also supported the experimental data. These findings clearly highlight the beneficial effects of AK7 in the prevention and/or treatment of inflammatory arthritis.

Cellular-scale sex differences in extracellular matrix remodeling by valvular interstitial cells.

Males acquire calcific aortic valve disease (CAVD) twice as often as females, yet stenotic valves from females display significantly higher levels of fibrosis compared to males with similar extent of disease. Fibrosis occurs as an imbalance between theproduction and degradation of the extracellular matrix (ECM), specifically type I collagen. This work characterizes ECM production and remodeling by male and female valvular interstitial cells (VICs) to better understand the fibrocalcific divergence between sexes evident in CAVD. Male and female VICs were assessed for gene and protein expression of myofibroblastic markers, ECM components, matrix metalloproteinases (MMPs), and tissue inhibitors of MMPs (TIMPs) via qRT-PCR and western blot. Overall metabolic activity was also measured. Activity assays for collagenase and gelatinase were performed to examine degradation behavior. Male VICs produced greater levels of myofibroblastic markers while female VICs showed greater metabolic activity and collagen production. In general, females displayed a greater level of MMP expression and production than males, but no sex differences were observed in TIMP production. Male VICs also displayed a greater level of collagenase and gelatinase activity than female VICs. This work displays sex differences in ECM remodeling by VICs that could be related to the sexual dimorphism in ECM structure seen in clinical CAVD.

Electron-Beam Irradiated Recombinant Human Collagen-Phosphorylcholine Corneal Implants Retain Pro-Regeneration Capacity

Sterilization of biodegradable, collagen-based implants is challenging as irradiation sterilization methods can alter their mechanical properties. Electron beam (EB) irradiation is a terminal sterilization method that has been used for biologically-derived implants. Here, recombinant human collagen type III-phosphorylcholine (RHCIII-MPC) hydrogels were irradiated with EB doses of 17, 19, or 21 kGy and their subsequent biocompatibility and ability to promote regeneration in rabbit corneas was evaluated. Unirradiated hydrogels stored in 1% chloroform in phosphate-buffered saline (C-PBS) were the controls. There were no significant differences between irradiated and non-irradiated samples in optical or physical properties (tensile strength, modulus, elasticity), or the ability to support cell growth. However, irradiated implants were more sensitive to high levels of collagenase than unirradiated controls and the C-PBS implants had increased cell growth compared to EB and controls at 72 h. Corneal implants e-beamed at 17 kGy or e-beamed and subsequently frozen (EB-F) to increase shelf-life showed no adverse biological effects of the irradiation. EB, EB-F, and C-PBS implanted corneas all rapidly re-epithelialized but showed mild neovascularization that resolved over 6 months. The regenerated neo-corneas were transparent at 6 months post-operation. In vivo confocal microscopy confirmed normal morphology for the epithelium, stroma, sub-basal nerves and unoperated endothelium. Histology showed that all the regenerated corneas were morphologically similar to the normal. Immunohistochemistry indicated the presence of a differentiated corneal epithelium and functional tear film. In conclusion, the e-beamed corneal implants performed as well as non-irradiated control implants, resulting in fully regenerated neo-corneas with new nerves and without blood vessels or inflammation that may impede vision or corneal function. Therefore, a complete validation study to establish EB irradiation as an effective means for corneal implant sterilization prior to clinical application is necessary as a next step.

Mechanisms that limit regression of myocardial fibrosis following removal of left ventricular pressure overload.

Left ventricular pressure overload (LVPO) can develop from antecedent diseases such as aortic valve stenosis and systemic hypertension and is characterized by accumulation of myocardial extracellular matrix (ECM). Evidence from patient and animal models supports limited reductions in ECM following alleviation of PO, however, mechanisms that control the extent and timing of ECM regression are undefined. LVPO, induced by 4 wk of transverse aortic constriction (TAC) in mice, was alleviated by removal of the band (unTAC). Cardiomyocyte cross-sectional area, collagen volume fraction (CVF), myocardial stiffness, and collagen degradation were assessed for: control, 2-wk TAC, 4-wk TAC, 4-wk TAC + 2-wk unTAC, 4-wk TAC + 4-wk unTAC, and 4-wk TAC + 6-wk unTAC. When compared with 4-wk TAC, 2-wk unTAC resulted in increased reactivity of collagen hybridizing peptide (CHP) (representing initiation of collagen degradation), increased levels of collagenases and gelatinases, decreased levels of collagen cross-linking enzymes, but no change in CVF. When compared with 2-wk unTAC, 4-wk unTAC demonstrated decreased CVF, which did not decline to control values. At 4-wk and 6-wk unTAC, CHP reactivity and mediators of ECM degradation were reduced versus 2-wk unTAC, whereas levels of tissue inhibitor of metalloproteinase (TIMP)-1 increased. ECM homeostasis changed in a time-dependent manner after removal of LVPO and is characterized by early increases in collagen degradation, followed by a later dampening of this process. Tempered ECM degradation with time is predicted to contribute to the finding that normalization of hemodynamic overload alone does not completely regress myocardial fibrosis.NEW & NOTEWORTHY In this study, a murine model demonstrated persistent interstitial fibrosis and myocardial stiffness following alleviation of pressure overload.

Toxicity study in a pig model of intraperitoneal collagenase as an \u201cenzymatic scalpel\u201d directed to break stroma in order to generate a new perspective for peritoneal carcinomatosis approach: an experimental research

BackgroundThis study aimed to measure the toxicity resulting from collagenase administration to the peritoneal cavity in a pig model as a preliminary step to break down the stroma surrounding tumors.MethodsEight pigs were treated with 2 different collagenase concentrations previously tested in rats by our group. Time and temperature were controlled using a peritoneal lavage system (PRS System, Combat Medical Ltd.) identical to that used in human surgeries through hyperthermic intraperitoneal chemotherapy (HIPEC); 2 additional pigs were treated with peritoneal lavage only. Samples of blood and peritoneal fluid were collected pre-treatment, immediately after treatment, and 24 h postoperatively. In addition, histological studies and blood collagenase levels were measured.ResultsNo complications were observed during the surgeries. Intraoperative images evidenced the release of peritoneal tissue during collagenase treatment. After surgery, the animals showed no signs of pain. Diet and mobility were normal at 4 h postoperatively, and there were no significant differences in hematologic or biochemical parameters. Quantification of MMP1 and MMP2 in all samples as measured by absorbance showed no differences in blood collagenase levels between pre-treatment, post-treatment, and 24 h postoperatively.None of the animals treated with collagenase showed peritoneal adhesions during the second surgery. Histologically, peritoneal organs and serous structures did not show any microscopic alterations associated with collagenase treatment in any group.ConclusionLavage of the peritoneal cavity with doses of up to 100,000 collagen digestion units/animal for 30 min is safe and removes connective tissue from the peritoneal cavity.

Curcumin inhibition of bleomycin-induced changes in lung collagen synthesis, deposition and assembly.

Idiopathic pulmonary fibrosis is characterized by progressive lung tissue remodeling and disproportionate deposition of collagenous proteins with limited therapeutic interventions. The purpose of this study was to determine whether curcumin inhibits bleomycin (BLM)-induced increases in synthesis, degradation and cross-linking of lung collagen in rats. Following a single intratracheal instillation of BLM to rats (0.75 U/100g, sacrificed 3, 5, 7, 14 and 28days post-BLM), lung collagen synthesis (determined by incorporation of 3H-proline) and deposition (determined by lung hydroxyproline content) progressively increased at days 7, 14 and 28 post-BLM injection. Lung lavage fluid hydroxyproline and collagenase levels (a measure of collagen turnover) were increased in BLM rats compared with control groups. In addition, BLM instillation resulted in increased concentrations of collagenase and collagenolytic cathepsin in the lungs. Furthermore, increased cross-linking (as determined by aldehyde content of acid soluble collagen), and decreased susceptibility of fibrotic lung insoluble collagen to denaturing agents occurred in BLM-injured lungs. Significant increases in alveolar macrophage (AM) release of transforming growth factor-β1 (TGF-β1) were noted at various time points (days 3, 5, 7, 14 and 28 post-BLM) during the development and progression of lung fibrosis in rats. Curcumin treatment to BLM rats (300mg/kg 10days before and daily thereafter throughout the experimental time period) was associated with marked reductions in lung collagen synthesis and deposition, BALF and lung collagenase activity, BALF hydroxyproline content and lung collagenolytic levels. Additionally, reduced levels of collagen cross-linking and enhanced susceptibility of insoluble lung collagen to denaturing agents were observed in curcumin-treated BLM rats. Finally, curcumin inhibited BLM-induced increases in AM production of TGF-β1. Our data demonstrate for the first time that curcumin prevents fibrotic deposits by modulating collagen turnover, assembly and deposition in BLM-instilled rat lungs, and that curcumin treatment protects against BLM activation of macrophages by suppressing the release of TGF-β1.

Neutrophil to Lymphocyte Ratio Reflects the Proteolytic Activity of the Abdominal Aortic Aneurysm Wall

Background: This study aims to evaluate the local proteolytic activity from the level of Abdominal Aortic Aneurysm (AAA) wall and correlate the obtained values with the preoperative values of NLRs (Neutrophil-Lymphocyte Ratio), evaluating a possible association between the two variables and, implicitly, between the local proteolysis process and the systemic inflammatory response of those patients diagnosed with AAA. Methods: The current study is monocentric, observational, and prospective, taking place at the Department of Cardiovascular Surgery, Cluj-Napoca, Romania. Patients undergoing elective or emergency classical surgery for unruptured AAA or ruptured AAA were included in the study. During classical surgery, samples from the infrarenal aortic aneurysmal wall were collected in a standardized manner, from the central part of the anterior wall from uAAA and rAAA and were analyzed by gel zymography. Results: The concentration of MMP2 was similar in the ruptured/non-ruptured group, without any statistical significance. In the MMP-9 case, we obtained a mean of 821.35 U arb/µg at the level of unruptured aneurysmal wall and 1411.57 U arb/µg at the level of the ruptured aneurysmal wall. According to the ANOVA test, there is a significant difference between the two categories of aneurysms. The same correlation was observed regarding both the zymogen category, pro-MMP-2, as well as pro-MMP-9: they expressed significant higher quantities of inactive enzymes in rAAA. We splitted the study population into two categories: patients who presented preoperative NLR values < 5 and > 5. MMP-2 collagenase levels did not register statistical differences between the two groups, p = 0.3236. High levels of MMP-9 are positively associated with increased values of NLR, the NLR<5 group had an MMP-9 mean of 902.41(473.71) U arb/µg, statistically lower than the MMP-9 mean indicated in the NLR>5 group, 1474(521.21) U arb/µg. Similarly, MMP-2 and MMP-9 zymogens were found in statistically higher quantities (p < 0.05) in the NLR>5 group of patients. Conclusions: This is the first study that analyzes a possible correlation between the local proteolytic activity at the site of the dilated aneurysmal aortic wall and circulating levels of NLR. Following the results obtained, we conclude that the group of patients presenting with NLR>5 preoperatively, as in the rAAA group, significantly greater levels of MMP-9 and inactive proenzymes were identified. Local metalloproteinase MM9 activity is proportional to the systemic inflammatory activity. Concomitantly, we hypothesize that the increased sensitivity of NLR as a prognostic marker in AAA pathology, which is ensured and confirmed by its strong association with local proteolytic activity, directly implied in the evolution of the disease.

Matrix metalloproteinases in children with uncomplicated compression fractures of the spine

The aim of the study was to determine changes in the content of matrix metalloproteinases (MMP) and their tissue inhibitor in children with uncomplicated compression fractures of the spine (UCFS). Materials and methods. Eighty-five children, including 69 patients with UCFS (average age 12.3 ± 2.6 years), were comprehensively examined. The reference group consisted of 16 children (average age 11.8 ± 2.7 years) without spinal pathology. During the diagnostic period for 1-3 days, changes in the MMP content and their tissue inhibitor (TIMP-1) in blood serum were determined by the enzyme immunoassay method in all children after trauma. Results. It was found that in the acute period after spinal injury, the blood levels of gelatinases (MMP-2 and MMP-9), stromelysin (MMP-3), and collagenases (MMP-8) significantly increased compared to their levels in children of the reference group. At the same time, the levels of TIMP-1 and the ratio of MMP/TIMP-1 concentrations in the blood of patients with UCFS significantly decreased compared to the control, which indicates the predominance of the proteolytic effect of MMP. Analysis of changes in the content of MMP in the blood in UCFS boys and girls did not reveal significant differences in the levels of the studied MMP and TIMP-1, except for a significant increase in the concentrations of stromelysin (MMP-3) in the blood serum of boys compared with its level in girls and the control. With different severity of the course of UCFS in children, a significant increase in MMP concentrations associated with an increase in the severity of the injury was revealed, and a substantial decrease in the content of TIMP-1 in the blood of patients compared to its levels in children with 1-2 degrees of severity and control. Conclusion. The established patterns indicate that the determination of the content of MMR and TIMP-1 in the blood in UCFS children allows monitoring the course of the reparative process after injury to the vertebral bodies in children.

Ameliorative Effects of Loganin on Arthritis in Chondrocytes and Destabilization of the Medial Meniscus-Induced Animal Model.

Arthritis is a common inflammatory disease that causes pain, stiffness, and joint swelling. Here, we investigated the ameliorative effects of loganin on arthritis in vitro and in vivo. A single bioactive compound was fractionated and isolated from Cornus officinalis (CO) extract to screen for anti-arthritic effects. A single component, loganin, was identified as a candidate. The CO extract and loganin inhibited the expression of factors associated with cartilage degradation, such as cyclooxygenase-2 (COX-2), matrix metalloproteinase 3 (MMP-3), and matrix metalloproteinase 13 (MMP-13), in interukin-1 beta (IL-1β)-induced chondrocyte inflammation. In addition, prostaglandin and collagenase levels were reduced following treatment of IL-1β-induced chondrocytes with loganin. In the destabilization of the medial meniscus (DMM)-induced mouse model, loganin administration attenuated cartilage degeneration by inhibiting COX-2, MMP-3, and MMP-13. Transverse micro-CT images revealed that loganin reduced DMM-induced osteophyte formation. These results indicate that loganin has protective effects in DMM-induced mice.

Effect of Selenium nanoparticles in wound healing

Background: Wound healing is a complex process necessary to repairing damaged tissues and preventing infection. Selenium nanoparticles were known by their antioxidant and antimicrobial effect that is important in wound healing. Aim: the aim of this study was to investigate the effect of selenium nanoparticles in reducing and accelerating the wound healing time in mice. Methods: Se NPs were synthesized by a simple wet chemical method. Then the experimental animals were divided into the following groups: Negative control group, Positive control group, selenium nanoparticles control group, wounded mice treated with selenium nanoparticles group. At the end of experiment, mice were scarified& skin tissue samples were collected for biological analysis [Vascular Endothelial cell Growth Factor, Collagenase I and Nitric oxide] and histopathological study. Results: The results showed that the percentage of wound area has been significantly reduced in Se NPs treated group compared to the positive control. The level of Vascular Endothelial Growth Factor and Collagenase I in Se NPs groups were significantly increased compared to positive control while nitric oxide was significantly decreased in Se NPs treated group. Skin tissue showed effect of selenium nanoparticles by regenerating and recavering skin layer. Conclusion: Selenium nanoparticles have an important role in accelerating and reducing wound healing time.

Impact of resistance exercises and nano-curcumin on synovial levels of collagenase and nitric oxide in women with knee osteoarthritis

BackgroundOsteoarthritis is the most common joint damage that leads to cartilage destruction, pain, and disability. The aim of this study is to investigate the effects of resistance exercises and Nano curcumin supplementation on synovial levels of collagenase-2 and NO in patients with knee osteoarthritis.MethodsForty women with primary knee osteoarthritis were divided into four groups including control, exercise, supplementation, and supplementation /exercises. The resistance exercise group performed exercises in 16 weeks (three sessions per week). The supplement group consumed one Nano curcumin capsule (1000 mg) per day for 16 weeks. Exercise and supplementation group also performed resistance exercises along with consuming Nano curcumin. Synovial fluid samplings were done in two stages of pre-test (24 h before training and supplementation) and post-test (48 h after the last session of treatment and supplementation). The obtained synovial fluid was used to measure the synovial level of collagenase -II and nitric oxide.ResultsAlthough, there were not any significant correlations between resistance exercise, nanocurcumum supplementation, and synovial levels of collagenase-2 and NO among women with knee osteoarthritis, we observed interesting reduced levels of both markers following the supplementation.ConclusionsIt can be concluded that the Nano curcumin supplementation can be an effective method to reduce the levels of these inflammatory factors in synovial fluids of cases with knee Osteoarthritis. Longer duration of the protocol may have more promoting results.Trial registrationThe project was registered at Iranian Registry of Clinical Trials (Trial registration number: IRCT20161208031300N1).

Amelioration of Rheumatoid Arthritis by Anacardium occidentale via Inhibition of Collagenase and Lysosomal Enzymes.

Anacardium occidentale (cashew) has been used in the traditional system of medicine for curing many inflammatory disorders. The present study investigates the antiarthritic effects of cashew leaves extract using the rat model of FCA-induced rheumatoid arthritis. Arthritic rats were treated with 100 and 200 mg/kg b.w. ethanolic extract of cashew leaves. Animals were sacrificed at day 23, and before sacrificing the animals, gross pathological changes were observed. Histopathology of ankle joint was evaluated with hematoxylin and eosin staining, whereas the serum levels of C-reactive protein (CRP) were evaluated by the agglutination method. Inflammatory cells and other hematological parameters were assessed by employing an automated hemocytometer and chemistry analyzer. Rheumatoid factor (Rf) and lysosomal enzymes levels were determined in blood. Results indicated that A. occidentale significantly decreased the CPR levels, macroscopic arthritic score, and rheumatoid factor as compared to the diseased group. Histopathological evaluation showed significant attenuation in bone erosion, joint inflammation, and pannus formation by plant extract. Treatment with A. occidentale significantly suppressed the levels of acid phosphatase, β-galactosidase, β-glucuronidase, N-acetylglucosaminidase, and collagenase. Moreover, A. occidentale significantly raised the HB levels and RBCs counts which were found depleted in the diseased group. The raised counts of total leukocytes, platelets, neutrophils, lymphocytes, and monocytes were also significantly decreased by treatment with plant extract. Comparative analysis showed that higher dose of A. occidentale demonstrated superior amelioration of rheumatoid arthritis as compared to low dose. In conclusion, A. occidentale possesses significant antiarthritic potential, which may be attributed to the suppression of lysosomal enzymes and collagenase levels.

Rare Case of Collagenous Colitis

With an incidence of 5.2 per 100,000, collagenous colitis has a female preponderance and peaks at the age of 50–70 years and female to male ratio of 9:1. It is associated with HLA B8/HLA DR3 and shows increased expression of HLA1. The underlying pathogenesis is decreased levels of interstitial collagenase which leads to reduced matrix degradation which in turn results in accumulation of matrix proteins and this causes defective sodium and chloride absorption. Diagnosis is based upon microscopic analyses of colonic biopsies. The characteristic histopathological finding is the collage band thickening that mainly occurs in the caecum and transverse colon in over 80% cases and in the rectum is lesser than 30%. Blood and urine cultures are usually normal and endoscopy and radiographic evidences are absent. We hereby report a case who presented with a history of intermittent watery diarrhea associated with occasional low-grade fever that spanned over about five years and was finally diagnosed as a case of collagenous colitis.

Ursonic acid exerts inhibitory effects on matrix metalloproteinases via ERK signaling pathway

Ursonic acid is a pentacyclic triterpenoid compound that can be extracted from Ziziphus jujuba Mill., a traditional medicine. Matrix metalloproteinases (MMPs) are involved in cancer metastasis and skin aging. Regulation of various MMPs is closely associated with mitogen-activated protein kinases (MAPKs), including ERK, p38, and JNK MAPKs. In this study, we investigated the possibility of ursonic acid as an anti-cancer/anti-skin aging agent targeting MMPs. Cytotoxic effects of ursonic acid were analyzed by cell counting kit-8 (CCK-8) assay. Invasive abilities of ursonic acid-treated A549 and H1299 non-small cell lung cancer (NSCLC) cells were tested with Boyden chamber assay. Effects of ursonic acid on MMPs were analyzed by zymography assays and quantitative real time polymerase chain reaction (qRT-PCR). We also conducted flow cytometry and western blot analysis to elucidate the mechanisms of MMP regulation by ursonic acid. Our results revealed that ursonic acid inhibited transcriptional expression of gelatinases (MMP-2 and MMP-9) via inhibition of ERK and CREB signaling pathways in NSCLC cells. Moreover, ursonic acid reduced mRNA levels of collagenase (MMP-1) via suppression of ERK and c-Fos signaling pathways in HaCaT keratinocytes. These results suggest that ursonic acid could be a potential candidate for development of an effective novel anti-cancer and anti-wrinkle agent.

Dynamics of Matrix Metalloproteinases in Local Cold Injury

The relevance of cold injury is due to its high specific weight in the structure of injuries, complexities of complex therapy, and unsatisfactory results of treatment. It is known that in the pathogenesis of local cold injury, the leading role belongs to endothelial dysfunction, which secretes a huge amount of biologically active substances, including matrix metalloproteinases.Aim: to determine the dynamics of the content of matrix metalloproteinases of the second subfamily (MMP-2, MMP-9) in the serum of patients with local cold injury.Materials and methods. The study included 80 patients (60 men, 20 women) with frostbite of the III–IV degree of the foot to the level of the lower third of the leg in the late reactive period and the period of granulation and epithelization.Results. In the late reactive period in patients with cryopreservation, the level of MMP-2 was 3.4 times higher relative to the control (p = 0.011), on day 30 MMP-2 values did not differ from the control parameters (p = 0.103). The level of collagenase B (MMP-9) in patients with local cold injury on day 5 was 14.5 times higher than the control parameters (p = 0.002), on day 30 – 12.5 times higher compared to the control group (p = 0.000094).Conclusion. During the analysis of the data obtained, we can think about the dual nature of collagenases during the processes of inflammation. In a situation of distress, collagenases prevent endothelial dysfunction by eliminating the cellular structures of the endothelium to ensure adequate metabolism in tissues. Their importance as markers of failure of adequate flow of reparative processes is not excluded.

SNEV(Prp19/PSO4) deficiency increases PUVA-induced senescence in mouse skin.

Senescent cells accumulate during ageing in various tissues and contribute to organismal ageing. However, factors that are involved in the induction of senescence in vivo are still not well understood. SNEVP rp19/ PSO 4 is a multifaceted protein, known to be involved in DNA damage repair and senescence, albeit only in vitro. In this study, we used heterozygous SNEV +/− mice (SNEV‐knockout results in early embryonic lethality) and wild‐type littermate controls as a model to elucidate the role of SNEVP rp19/ PSO 4 in DNA damage repair and senescence in vivo. We performed PUVA treatment as model system for potently inducing cellular senescence, consisting of 8‐methoxypsoralen in combination with UVA on mouse skin to induce DNA damage and premature skin ageing. We show that SNEVP rp19/ PSO 4 expression decreases during organismal ageing, while p16, a marker of ageing in vivo, increases. In response to PUVA treatment, we observed in the skin of both SNEVP rp19/ PSO 4 and wild‐type mice an increase in γ‐H2AX levels, a DNA damage marker. In old SNEVP rp19/ PSO 4 mice, this increase is accompanied by reduced epidermis thickening and increase in p16 and collagenase levels. Thus, the DNA damage response occurring in the mouse skin upon PUVA treatment is dependent on SNEVP rp19/ PSO 4 expression and lower levels of SNEVP rp19/ PSO 4, as in old SNEV +/− mice, result in increase in cellular senescence and acceleration of premature skin ageing.