Matrix Metalloproteinase-9 Protein Research Articles

4-Year Pulp Survival in a Randomized Trial on Direct Pulp Capping

IntroductionThis study aimed to assess pulp survival in a randomized trial on pulp lavage in adult nonpainful posterior teeth with carious pulp exposure. The treatment included complete caries excavation, direct pulp capping with mineral trioxide aggregate, and immediate restoration with composite resin. MethodsFluid was collected from the pulp wound to assess matrix metalloproteinase-9 (MMP-9) and total protein values. Before pulp capping, cavities were randomly (block randomization, n = 48) washed with a physiological saline or a sodium hypochlorite solution (2.5% NaOCl). Treatment outcome was assessed clinically (cold test) and radiographically after at least 1 year and again after at least 3 years. Painful failures were differentiated from nonpainful failures. Pulp survival was estimated using the Kaplan-Meier method including 95% confidence intervals (CIs) up to 1500 days. ResultsFrom the 96 patients originally enrolled, 73 individuals could be followed continuously. The clinical observations indicated a beneficial and sustained effect of pulp lavage with 2.5% NaOCl over a control treatment with physiological saline solution on estimated pulp survival 1500 days postintervention, with 7% (95% CI, 1%–40%) in the saline group versus 55% (95% CI, 30%–100%) in the NaOCl group. High MMP-9/total protein values in pulpal fluid collected from the exposed site indicated early and painful treatment failures yet were not associated with failures that occurred more than 250 days after intervention. ConclusionsThe low 4-year success rates reported here challenge the concept of direct pulp capping in the cases that were included. NaOCl lavage did not only increase the survival of affected pulps substantially but also particularly diminished painful failures (33% in the NaOCl group vs 62% in the saline group). The lack of the predictive value of MMP-9 assessments beyond early treatment failures points to inflammatory states of the pulp tissue under deep caries, which are not related to neutrophil infiltration.

Evidence Accumulates: Patients with Ascending Aneurysms Are Strongly Protected from Atherosclerotic Disease.

Ascending thoracic aortic aneurysms may be fatal upon rupture or dissection and remain a leading cause of death in the developed world. Understanding the pathophysiology of the development of ascending thoracic aortic aneurysms may help reduce the morbidity and mortality of this disease. In this review, we will discuss our current understanding of the protective relationship between ascending thoracic aortic aneurysms and the development of atherosclerosis, including decreased carotid intima-media thickness, low-density lipoprotein levels, coronary and aortic calcification, and incidence of myocardial infarction. We also propose several possible mechanisms driving this relationship, including matrix metalloproteinase proteins and transforming growth factor-β.

Assessment of the Antioxidant Properties of Horned Turban (Turbo cornutus) Viscera, Marine By-Products with Potential Application in Nutraceuticals

Horned turban (Turbo cornutus) is an edible gastropod that occurs along the intertidal zone and basalt coastline, and is an important marine resource in Jeju, Korea. However, T. cornutus viscera are mostly discarded following processing. In this study, the antioxidant activity of viscera and muscle extracts was compared. In addition, the protective effect of T. cornutus viscera ethanol extract (TVEE) against H2O2-induced oxidative stress in human dermal fibroblasts (HDFs) was investigated. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and hydrogen peroxide scavenging activity of the viscera (IC50 = 0.64 ± 0.64 and 0.76 ± 0.01 mg/mL, respectively) showed higher activity than that in muscle. TVEE increased the reactive oxygen species (ROS) scavenging effect and cell viability in H2O2-induced HDFs without cytotoxicity. Furthermore, the fluorescence intensity of HDFs was high in those from H2O2-induced intracellular ROS production, but TVEE treatment decreased ROS production. H2O2 activated the mitogen-activated protein kinase (MAPK) signaling and matrix metalloproteinase 2 (MMP-2) in HDFs. However, MMP2 expression was confirmed to be reduced via MAPK (ERK, JNK, and p38) signaling phosphorylation. In conclusion, various antioxidant effects were confirmed in T. cornutus viscera, which instead of being discarded as marine by-products, can be applied as nutraceuticals in various industries.

Hypoxia ischemia results in blood brain barrier damage via AKT/GSK-3β/CREB pathway in neonatal rats

Previous studies have showed that the permeability of blood brain barrier (BBB) increased after hypoxia ischemia (HI). The current research uncovered the mechanism of altered BBB permeability after hypoxic-ischemic brain damage (HIBD) through AKT/GSK-3β/CREB signaling pathway in neonatal rats. Firstly, Magnetic resonance imaging (MRI) combined with hematoxylin-eosin (H&E) staining was used to assess brain injury. Initial findings showed abnormal signals in T2-weighted imaging (T2WI) and diffusion weighted imaging (DWI). Changes also happened in the morphology of nerve cells. Subsequently, we found that BBB damage is manifested as leakage of immunoglobulin G (IgG) and destruction of BBB-related proteins and ultrastructure. Meanwhile, the levels of matrix metalloproteinase-9 (MMP-9) significantly increased at 24 h after HIBD compared to a series of time points. Additionally, immunohistochemical (IHC) staining combined with Western blot (WB) was used to verify the function of the AKT/GSK-3β/CREB signaling pathway in BBB damage after HI in neonatal rats. Results showed that less Claudin-5, ZO-1, p-AKT, p-GSK-3β and p-CREB, along with more MMP-9 protein expression were visible on the damaged side of the cerebral cortex in the HIBD group in contrast to the sham and HIBD + SC79 groups. Together, our findings demonstrated that HI in neonatal rats might upregulate the levels of MMP-9 protein and downregulate the levels of Claudin-5 and ZO-1 by inhibiting the AKT/GSK-3β/CREB pathway, thus disrupting the BBB, which in turn aggravates brain damage after HI in neonatal rats.

HMGB1 inhibition reduces TDI-induced occupational asthma through ROS/AMPK/autophagy pathway

Exposure to toluene diisocyanate (TDI) can cause pulmonary diseases such as asthma. Inhibition of high mobility group box 1 protein (HMGB1) has been found to be protective against the toxic effects of TDI on human bronchial epithelial (HBE) cells. Here, we evaluated the in vivo positive roles of HMGB1 in the TDI-caused asthma mice and explored its underlying mechanisms in HBE cells. We found that suppression of HMGB1 obviously alleviated airway inflammation, airway hyperresponsiveness, and airway remodeling in the lung tissue of the asthma mice. The in vitro results showed that inhibition of HMGB1 ameliorated TDI-induced reactive oxygen species (ROS) release, inflammatory response, and activation of autophagy in HBE cells. At the molecular level, inhibition of HMGB1 decreased the expressions of HMGB1, Toll-like receptor 4, Vimentin and matrix metalloproteinase-9 proteins, activated NF-κB and NOD-like receptor protein 3 (NLRP3) inflammasome, and increased E-cadherin expression. Importantly, activation of autophagy could lead to the overactivation of NLRP3 inflammasome in TDI-induced asthma. These results suggest that inhibition of HMGB1 can alleviate TDI-induced asthma through ROS/AMPK/autophagy pathways, which may provide valuable evidence for the pathogenesis and therapeutic targets of TDI-induced asthma.

Serum matrix metalloproteinase-9 (MMP9) and amyloid-beta protein precursor (APP) as potential biomarkers in children with Fragile-X syndrome: A cross sectional study

IntroductionFragile-X syndrome(FXS) is a neurological disease caused by abnormal repeats in the 5′untranslated region of the FMR1 gene leading to a defective fragile-X-messenger-ribonucleoprotein-1 (FMRP). Although relatively common in children, it is usually under-diagnosed especially in developing countries where genetic screening is not routinely practiced. So far, FXS lacks a laboratory biomarker that can be used for screening, severity scoring or therapeutic monitoring of potential new treatments. Methods110 subjects were recruited; 80 male children with suspected FXS and 30 matched healthy children. We evaluated the clinical utility of serum matrix metalloproteinase-9(MMP9) and amyloid-beta protein precursor(APP) as potential biomarkers for FXS. ResultsOut of 80 suspected children, 14 had full mutation, 8 had the premutation and 58 children had normal genotypes. No statistically-significant difference was detected between children with different genotypes concerning age of onset(P = 0.658), main clinical presentation(P = 0.388), clinical severity-score(P = 0.799), patient’s disease-course(P = 0.719) and intellectual disability(P = 0.351). Both MMP9 and APP showed a statistically significant difference when comparing different genotype subgroups(P = 0.019 and < 0.001, respectively). Clinically, MMP9 levels were highest in children presenting with language defects, while APP was highest in children with neurodevelopmental delay. In receiver operating curve analysis, comparing full and premutation with the normal genotype group, MMP9 has an area-under-the-curve of 0.701(95 % CI 0.557–0.845), while APP was marginally better at 0.763(95 % CI 0.620–0.906). When combined together, elevated MMP9 or APP had excellent sensitivity > 95 % for picking-up FXS cases in the clinical setting. ConclusionsScreening for circulating biomarkers in the absence of FXS genetic diagnosis is justified. Our study is the first to evaluate both MMP9 and APP in FXS suspected children in a clinical setting and to assess their correlation with disease presentation and severity.

Matrix Metalloproteinase-9, Neuron-specific Enolase, S100 B and Tau Protein Levels in the Patients with Carbon monoxide Poisoning

Background: S100B, NSE, MMP-9, and Tau protein levels increase in cases causing hypoxic cell damage. The diagnosis of the severity of carbon monoxide (CO) poisoning in the early period of these parameters was studied. Material and Method: COHb level measurement was made using a signal capture CO-pulse oximeter (Masimo's SET Rainbow, Masimo's Co, USA) at the first admission of the patients. Then, COHb levels were confirmed by arterial blood gas(ABG) analysis. The patients were divided into two groups as mild and moderate-severe, according to their Glasgow coma scores (GCS) [Mild (14–15); Moderate (9–13) or Severe (3–8)]. The control group was composed of 16 healthy and non-smoking volunteers. Results: The serum S100B protein and MMP-9 values at 0 hr of admission in the hospital and 3hr of treatment were not significantly different in the patient group as compared to the control group. Tau protein levels were significantly higher in the patient group at 0 and 3 hours (p&gt; 0.05) as compared to healthy person. Conclusion: There was no relationship between CO poisoning and MMP-9 and S100B protein levels. NSE and Tau protein were significantly higher in the patient group than the control group. Tau protein may be more useful marker as compared to neuron-specific enolase.

Electric-Double-Layer (EDL)-Gated FET Aptasensors for Highly Sensitive MMP7 Detection

ABSTRACT Biliary Atresia (BA) is a severe liver disease that affects newborns. This disease may cause cholestasis and progressive hepatic failure or even death if not treated immediately. ELISA blood tests for metabolic screening are the current method of diagnosing BA. However, newborns need rapid BA detection for treatment in the future. The increased matrix metalloproteinase-7 (MMP7) cause BA-related liver fibrosis and newborns diagnosed with BA show higher MMP7 levels than those diagnosed with cholestasis. Several platforms have been developed for the fast detection of Biliary Atresia (BA). Here, we developed an electric-double-layer (EDL)-gated field-effect transistor (FET) platform for Biliary Atresia (BA) detection. The core idea of this sensor array is to use a uniquely designed DNA probe to capture MMP7 proteins and monitor the change in drain current caused by the target capturing. Different concentrations of MMP7 spiked and tested over a functionalized sensor. The simple and cost-effective fabrication and ease of use make this sensor a promising tool as a point care detection device.Keywords: EDL, FET, Biliary atresia, MMP7, Biosensors INTRODUCTION Due to the symptomatic similarity to cholestatic liver diseases, it is challenging to diagnose a precise BA for newborns. False diagnosis may delay timely treatment (e.g., surgery) and worsens the prognosis. However, BA's underlying causes are still unclear. A level increase of intrahepatic matrix metalloproteinase-7 (MMP7) signifies BA-related liver fibrosis. BA cases found in newborns revealed higher MMP7 levels than cholestasis. MMP7 is a protease that contributes to tissue remodeling and breaks down extracellular matrix through a signal pathway. Based on the clinical results, MMP7 has emerged as a novel biomarker for BA diagnosis. ELISA was employed as the current method for detecting Biliary Atresia (BA). However, the procedure of ELISA has shortcomings, such as being time-consuming, high cost, and complicated operation. Hence, rapid and precise diagnostic methods need to be developed. In this work, we propose to detect MMP7 proteins using an electric-double-layer (EDL)-gated field-effect transistor (FET). As shown in Figure 1, The aptamer of MMP7 was immobilized on sensor electrodes, allowing a binding with the protein. As the number of the captured analytes increases; the sensor signals, amplified by a FET, will respond to it. As such, we look forward to detecting MMP7 in serum samples collected from newborns and helping the clinical diagnostics by this EDL-gated FET biosensor. MATERIALS AND METHODS An extended gate chip was employed in the sensor array. The electrode surface was cleaned using oxygen plasma, following which, the probe was immobilized on it. The MMP7 aptamer is mixed with TCEP for 30 minutes at room temperature, TCEP was used as a reducing agent which helps in the formation of dithiol bonds (SS), making the attachment of the probe easier. The mixture solution is then dropped and cast over the sensor surface and immobilized at 24 ℃ for 24 hr. The sensor is washed with 0.01× PBS to remove unbound aptamers and was verified with fluorescent imaging. ELECTRICAL MEASUREMENT After the aptamer was immobilized over the sensor surface, a baseline measurement was carried out. The drain current was measured at different voltage biases (Vg=-2V and 3V, at Vd=3.5V). different concentration of MMP7 protein was spiked in 0.01x PBS and tested. A signal was read out every 1min, and 20 measurements were taken for each concentration of the target for 20 minutes. RESULTS AND CONCLUSIONS The platform established is to detect MMP7 protein precisely and further identify BA. Owning a characteristic of electrical amplification, the EDL-gated FET will be utilized to test MMP7 proteins with concentrations ranging from 10 ng/mL to 100 ng/mL, as shown in Figure 2. When testing proteins level at 100 ng/mL, the signal can increase by around 3.5 mA. The tests using EDL-FET were successfully validated for MMP7 detection in serum and 0.01x PBS and the results indicate that the proposed platform had a high sensitivity. The calibration curve (signal vs. analyte's concentration) will be established to quantify the unknown sample. Conclusively, the sensor developed in this study has the potential to be developed as a point-of-care testing device and is suitable for clinical diagnosis. Figure 1

Synergistic effect of Hsp90 inhibitor ginkgolic acids C15꞉1 combined with paclitaxel on nasopharyngeal carcinoma.

Nasopharyngeal cracinoma is a kind of head and neck malignant tumor with high incidence and high mortality. Due to the characteristics of local recurrence, distant metastasis, and drug resistance, the survival rate of patients after treatment is not high. Paclitaxel (PTX) is used as a chemotherapy drug in treating nasopharyngeal carcinoma, but nasopharyngeal carcinoma cells are easy to develop resistance to PTX. Inhibition of heat shock protein 90 (Hsp90) can overcome common signal redundancy and resistance in many cancers. This study aims to investigate the anti-tumor effect of ginkgolic acids C15꞉1 (C15:1) combined with PTX on nasopharyngeal carcinoma CNE-2Z cells and the mechanisms. This experiment was divided into a control group (without drug), a C15:1 group (10, 30, 50, 70 μmol/L), a PTX group (5, 10, 20, 40 nmol/L), and a combination group. CNE-2Z cells were treated with the corresponding drugs in each group. The proliferation of CNE-2Z cells was evaluated by methyl thiazolyl tetrazolium (MTT). Wound-healing assay and transwell chamber assay were used to determine the migration of CNE-2Z cells. Transwell chamber was applied to the impact of CNE-2Z cell invasion. Annexin V-FITC/PI staining was used to observe the effect on apoptosis of CNE-2Z cells. The changes of proteins involved in cell invasion, migration, and apoptosis after the combination of C15꞉1 and PTX treatment were analyzed by Western blotting. Compared with the control group, the C15꞉1 group and the PTX group could inhibit the proliferation of CNE-2Z cells (all P<0.05). The cell survival rates of the C15꞉1 50 μmol/L combined with 5, 10, 20, or 40 nmol/L PTX group were lower than those of the single PTX group (all P<0.05), the combination index (CI) value was less than 1, suggesting that the combined treatment group had a synergistic effect. Compared with the 50 μmol/L C15꞉1 group and the 10 nmol/L PTX group, the combination group significantly inhibited the invasion and migration of CNE-2Z cells (all P<0.05). The results of Western blotting demonstrated that the combination group could significantly down-regulate Hsp90 client protein matrix metalloproteinase (MMP)-2 and MMP-9. The results of double staining showed that compared with the 50 μmol/L C15꞉1 group and the 10 nmol/L PTX group, the apoptosis ratio of CNE-2Z cells in the combination group was higher (both P<0.05). The results of Western blotting suggested that the combination group could decrease the Hsp90 client proteins [Akt and B-cell lymphoma-2 (Bcl-2)] and increase the Bcl-2-associated X protein (Bax). The combination of C15꞉1 and PTX has a synergistic effect which can inhibit cell proliferation, invasion, and migration, and induce cell apoptosis. This effect may be related to the inhibition of Hsp90 activity by C15꞉1.

AdhMMP8 Vector Administration in Muscle: An Alternate Strategy to Regress Hepatic Fibrosis.

The development of several vaccines against the SARS-CoV2 virus and their application in millions of people have shown efficacy and safety in the transfer of genes to muscle turning this tissue into a protein-producing factory. Established advanced liver fibrosis, is characterized by replacement of hepatic parenchyma by tissue scar, mostly collagen type I, with increased profibrogenic and proinflammatory molecules gene expression. Matrix metalloproteinase 8 (MMP-8) is an interstitial collagen-degrading proenzyme acting preferentially on collagen type I when activated. This study was carried out to elucidate the effect of an intramuscularly delivered adenoviral vector containing proMMP-8 gene cDNA (AdhMMP8) in male Wistar rats with experimental advanced liver fibrosis induced by thioacetamide. Therapeutic effects were monitored after 1, 2, or 3 weeks of a single dose (3 × 1011 vp/kg) of AdhMMP8. Circulating and liver concentration of MMP-8 protein remained constant; hepatic fibrosis decreased up to 48%; proinflammatory and profibrogenic genes expression diminished: TNF-α 2.28-fold, IL-1 1.95-fold, Col 1A1 4-fold, TGF-β1 3-fold and CTGF 2-fold; and antifibrogenic genes expression raised, MMP-9 2.8-fold and MMP-1 10-fold. Our data proposes that the administration of AdhMMP8 in muscle is safe and effective in achieving liver fibrosis regression at a comparable extent as when the adenoviral vector is delivered systemically to reach the liver, using a minimally invasive procedure.

A reverse docking approach to explore the anticancer potency of natural compounds by interfering metastasis and angiogenesis

Angiogenesis, which results in the formation of new blood and lymph vessels, is required to serve metastatic cancer progression. Cancer medications may target these two interconnected pathways. Phytocompounds have emerged as promising options for treating cancer. In this study, we used a reverse docking strategy to find new candidate molecules for cancer treatment that target both pathways. Following a literature study, the important cancer-causing proteins vascular endothelial growth factor D (VEGF-D) and basic fibroblast growth factor (bFGF) for angiogenesis and matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) for the metastatic pathway were targeted. Protein Data Bank was used to retrieve the structures of chosen proteins. 22 significant plant metabolites were identified as having anticancer activity. To determine the important protein binding residues, active site prediction was used. Using Lenvatinib and Withaferin A as reference ligands, the binding affinity of certain proteins for plant metabolites was determined by docking analysis. Homoharringtonine and viniferin, both have higher binding affinities when compared to reference ligands, with docking scores of −180.96 and −180.36 against the protein MMP-9, respectively. Moreover, Viniferin showed the highest binding affinity with both MMP-9 and MMP-2 proteins, which were then subjected to a 100-ns molecular dynamic simulation. where they were found to be significantly stable. In pharmacoinformatics investigations, the majority of our compounds were found to be non-toxic for the host. In this study, we suggested natural substances as cutting-edge anticancer treatments that target both angiogenesis and metastasis, which may aid in accelerating drug development and identifying viable therapeutic candidates. Communicated by Ramaswamy H. Sarma

Histological findings in resected leiomyomas following MR-HIFU treatment, single-institution data from seven patients with unfavorable focal therapy

Purpose Magnetic resonance – high-intensity focused ultrasound (MR-HIFU) is a noninvasive treatment option for symptomatic uterine leiomyomas. Currently, pretreatment MRI is used to assess tissue characteristics and predict the most likely therapeutic response for individual patients. However, these predictions still entail significant uncertainties. The impact of tissue properties on therapeutic outcomes remains poorly understood and detailed knowledge of the histological effects of ultrasound ablation is lacking. Investigating these aspects could aid in optimizing patient selection, enhancing treatment effects and improving treatment outcomes. Methods and materials We present seven patients who underwent MR-HIFU treatment for leiomyoma followed by second-line surgical treatment. Tissue samples obtained during the surgery were stained with hematoxylin and eosin, Masson’s trichrome and Herovici to evaluate general morphology, fibrosis and collagen deposition of leiomyomas. Immunohistochemical CD31, Ki-67 and MMP-2 stainings were performed to study vascularization, proliferation and matrix metalloproteinase-2 protein expression in leiomyomas, respectively. Results The clinical characteristics and radiological findings of the leiomyomas prior to treatment as well as qualitative histological findings after the treatment are presented and discussed in the context of current literature. A tentative model for volume reduction is presented. Conclusion These findings provide insights into potential factors contributing to suboptimal therapeutic outcomes and the variability in histological changes following treatment.

Identification the key factor of pulmonary fibrosis following silica nanoparticles exposure based on bioinformatics analysis

Objective: To investigate the main mechanisms of pulmonary fibrosis following silica nanoparticles (SiNPs) exposure through constructing the macrophage-fibroblast model in vitro, which simulated the process of pulmonary fibrosis. Methods: In January 2021, human mononuclear leukemia cells (THP-1) were treated with 0, 25, 50, 100 μg/ml SiNPs for 24 h. The supernatant of THP-1 cells was collected and applied to human embryonic lung fibroblast cells (MRC-5) which divided into control and low, medium and high dose groups at the logarithmic growth stage for 24 h. MRC-5 cell viability was detected by CCK8. The hydroxyproline (Hyp), interleukin 6 (IL-6), interleukin 1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) expression were detected in the supernatants of MRC-5. The changed proteins were detected by liquid-phase mass spectrometry in high dose group. GeneCard database were applied to identity the differential pulmonary fibrosis proteins in high dose group. Gene Ontology (GO) was performed to identity the key biological process in differential pulmonary fibrosis proteins of high dose group. The String database was used to construct the protein-protein interactions (PPI) network of differential pulmonary fibrosis proteins. The APP of CytoHubba was applied to calculate the key protein of differential pulmonary fibrosis proteins in PPI network. Correlation coefficients between key differential pulmonary fibrosis proteins were calculated using Pearson correlation analysis. Western blotting was applied to detect the expression of key proteins of differential pulmonary fibrosis proteins in different groups. Results: CCK8 results showed that MRC-5 cell viability was increasing in low, medium and high dose groups compared with control group (P<0.05). The expression levels of Hyp and IL-1β in different group were increased compared with control group, the expression levels of IL-6 and TNF-α were increased in high dose group compared with control group (P<0.05). GeneCard database identified 26 differential pulmonary fibrosis proteins, which were mainly involved in extracellular matrix hydrolysis, cell inflammatory response, tissue repair, cell proliferation, inflammation response by GO analysis. The APP of CytoHubba was calculated that matrix metalloproteinase 9 (MMP9) and tissue inhibitor metalloproteinase 1 (TIMP1) played an important role in PPI network. The results of correlation analysis showed that MMP9 was correlated with the expression of matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 3 (MMP3), TIMP1 and epidermal growth factor receptor (EGFR) (r=0.97, 0.98, 0.94, 0.93, P<0.05). Western blotting results showed that TIMP1 protein expression was increased in low, medium and high dose groups, while MMP9 protein expression was increased only in high dose group (P<0.05) . Conclusion: Differential expression proteins related with pulmonary fibrosis in MRC-5 cells mainly regulate biological processes of extracellular matrix hydrolysis, tissue repair, and cellular inflammation response following SiNPs exposure. MMP9 and TIMP1 may be the key proteins, which affected the fibrosis process in vitro pulmonary fibrosis model.

2D vanadium disulfide nanosheets assisted ultrasensitive, rapid, and label-free electrochemical quantification of cancer biomarker (MMP-2)

Cancer is one of the most tormenting global health burdens reporting high mortality and morbidity worldwide. Matrix metalloproteinase 2 (MMP-2) protein has elevated expression for most types of cancers, including prostate and breast cancer. Therefore, accurate and specific detection of MMP-2 biomarker is crucial for screening, treatment, and prognosis of related cancer. In this work, we have proposed a label-free electrochemical biosensor for the detection of MMP-2 protein. This biosensor was fabricated using hydrothermally synthesized vanadium disulfide (VS2) nanosheets with monoclonal anti-MMP2 antibodies biofunctionalized using a suitable linker. The VS2 nanomaterials were synthesized hydrothermally at different reaction temperatures (140 °C, 160 °C, 180 °C and 200 °C) generating different morphologies from a 3D bulk cubic structure at 140 °C to 2D nanosheets at 200 °C. Owing to the advantages of 2D VS2 nanosheets with high surface-to-volume ratio, excellent electrochemical response and high antibody loading possibility, it was selected for fabricating an MMP-2 specific biosensor. The antibody–antigen binding event is analyzed by recording electrochemical impedance spectroscopy signals for different target MMP-2 protein concentrations. The sensitivity and lower limit of detection were 7.272 (ΔR/R)(ng ml)−1 cm−2 and 0.138 fg ml−1, respectively in 10 mM phosphate buffer saline for this proposed sensor. Further, interference studies were also performed which demonstrates the sensor to be highly selective against non-specific target proteins. This 2D VS2 nanosheet-based electrochemical biosensor is a sensitive, cost-effective, accurate, and selective solution for cancer diagnosis.

Mesenchymal stem cells facilitate gastric cancer cell growth and oxaliplatin resista nce via upregulation of SMARCA5

Purpose: To evaluate the role of mesenchymal stem cells (MSCs) in gastric cancer (GC), and reveal its underlying mechanisms.&#x0D; Methods: Expression of SMARCA5 was quantified in GES-1, AGS and MKN45 cell lines. In MSCs-GC co-cultured cells, cell viability was assessed using cell counting kit-8 (CCK-8) assay, and colony formation were performed to determine cell proliferation. The invasion of the MSCs-GC cells was evaluated by Transwell assay. The levels of vimentin, snail and slug, as well as Wingless-Int (Wnt)/β-catenin related proteins β-catenin, Axin, c-myc, and matrix metalloproteinase (MMP)-7 were determined using immunoblotting. Oxaliplatin was added to GC cells, and the effect of siSMARCA5 on cell sensitivity to oxaliplatin was investigated after transfection with SMARCA5 siRNA into MSCs-GC cells.&#x0D; Results: SMARCA5 was highly expressed after co-culture with MSCs (p &lt; 0.001). The MSCs facilitated the proliferation of GC cells, and enhanced cell invasion as well as migration (p &lt; 0.001) compared with untreated cells. MSCs also promoted epithelial–mesenchymal transition (EMT) by increasing production of vimentin, snail, and slug (p &lt; 0.001). The MSCs decreased the sensitivity of GC cells to oxaliplatin, while the knockdown of SMARCA5 reversed the effect (p &lt; 0.001). Furthermore, MSCs up-regulated β-catenin, c-myc, and MMP-7 levels, but downregulated Axin expression compared with untreated cells (p &lt; 0.001). However, siSMARCA5 blocked these processes compared with si-NC group (p &lt; 0.001).&#x0D; Conclusion: MSCs facilitate GC cell growth and oxaliplatin resistance by upregulating SMARCA5 and activating Wnt/β-catenin signaling. This may provide an alternative treatment target for patients with GC.

Curcumin inhibits vascular mimicry to prevent thyroid cancer progression through regulation of EPH receptor A2/phosphoinositide 3-kinase

This study explored the impact of curcumin on vascular mimicry in thyroid cancer (TC) cells and underlying mechanism. TC cells (TPC-1) were exposed to curcumin at 15 and 30 μmol/L (low-dose and high-dose group) concentrations, respectively, with control group exposed to culture medium. After treatment, cell migration, invasion, proliferation and apoptosis were detected along with analysis of EphA2 and PI3K expressions. The amount of migrated and invaded cells, EphA2 and PI3K protein expressions, Matrix metalloproteinase 2(MMP2), MMP9, CyclinD1, EphA2 and PI3K of TPC-1 cells in curcumin groups were decreased (P &lt; 0.05). Apoptosis, as well as Cleaved-caspase-3, Bax and p21 expressions increased (P &lt; 0.05). Curcumin dose-dependently suppressed angiogenesis of TPC-1 cells (P &lt; 0.05) and lumen formation. In addition, TPC-1 cells showed cell rearrangement and scattered tubules after conditioned medium of neurotrophic factor (NTF-CM) treatment, while the CM of cancer-associated fibroblasts (CAF-CM) (50%) and CAF-CM (100%) both significantly promoted vasculogenic mimicry in TPC-1 cells, with elevated amount of tubules and junctions (P &lt; 0.05). Moreover, treatment with ALW-II-41-27, which is an EphA2 inhibitor, alleviated the CAF-CM’s effect on vasculogenic mimicry (P &lt; 0.05), and EphA2 knockdown decreased MMP2, MMP9, EphA2 and PI3K protein expressions when decelerating tube formation in NTF-CM and CAF-CM groups (P &lt; 0.05). Collectively, Curcumin suppresses malignant behaviors and hinders vascular mimicry of TC cells by inhibiting the EPHA2/PI3K pathway, thereby preventing malignant progression of TC.

Gold Nanoparticles Inhibit PMA-Induced MMP-9 Expression via microRNA-204-5p Upregulation and Deactivation of NF-κBp65 in Breast Cancer Cells.

Breast cancer (BC) is the most common malignancy in females globally. Matrix metalloproteinase-9 (MMP-9) is crucial to the invasion, progression and spread of BC. Gold nanoparticles (AuNPs) have an anti-tumorigenic role, but their therapeutic role in microRNAs (miRNAs) regulation has not been explored. This study determined the potential of AuNPs against MMP-9 overexpression/production and miRNA-204-5p regulation in BC cells. AuNPs were newly engineered, and their stability was analyzed using the zeta potential, polydispersity index, surface-plasmon-resonance peak and transmission electron microscopy. A bioinformatics algorithm was used to predict the pairing of miRNA in the 3'untranslated-region (3'UTR) of MMP-9 mRNA. TaqMan assays were carried out to quantify miRNA and mRNA, whereas MMP-9-specific immunoassays and gelatin zymography were used to determine protein secretion and activity. The binding of miRNA in MMP-9 mRNA 3'UTR was verified by luciferase reporter clone assays and transfection with anti-miRNAs. In addition, NF-κBp65 activity was determined and confirmed with parthenolide treatment. Engineered AuNPs were highly stable and spherical in shape, with a mean size of 28.3 nm. Tested in MCF-7 BC cells, microRNA-204-5p directly regulates MMP-9. AuNPs inhibit PMA-induced MMP-9 mRNA and protein via hsa-miR-204-5p upregulation. Anti-miR-204 transfected MCF-7 cells demonstrated enhanced MMP-9 expression (p < 0.001), while AuNPs treatment attenuated MMP-9 expression in a dose-dependent manner (p < 0.05). Moreover, AuNPs also inhibit PMA-induced NF-κBp65 activation in anti-hsa-miR-204 transfected MCF-7 cells. Engineered AuNPs were stable and non-toxic to BC cells. AuNPs inhibit PMA-induced MMP-9 expression, production and activation via NF-κBp65 deactivation and hsa-miR-204-5p upregulation. These novel therapeutic potentials of AuNPs on stimulated BC cells provide novel suggestions that AuNPs inhibit carcinogenic activity via inverse regulation of microRNAs.

Prenatal Fumonisin Exposure Impairs Bone Development via Disturbances in the OC/Leptin and RANKL/RANK/OPG Systems in Weaned Rat Offspring.

The goal of the current study was to examine the effects of prenatal exposure to fumonisins (FBs) on bone properties and metabolism in weaned rat offspring divided into groups intoxicated with FBs at either 0 (the 0 FB group), 60 (the 60 FB group), or 90 mg/kg b.w. 0 (the 90 FB group). Female and male offspring exposed to FBs at a dose of 60 mg/kg b.w. had heavier femora. Mechanical bone parameters changed in a sex and FBs dose-dependent manner. Growth hormone and osteoprotegerin decreased in both sexes, regardless of FBs dose. In males osteocalcin decreased, while receptor activator for nuclear factor kappa-Β ligand increased regardless of FBs dose; while in females changes were dose dependent. Leptin decreased in both male FBs-intoxicated groups, bone alkaline phosphatase decreased only in the 60 FB group. Matrix metalloproteinase-8 protein expression increased in both female FBs-intoxicated groups and decreased in male 90 FB group. Osteoprotegerin and tissue inhibitor of metalloproteinases 2 protein expression decreased in males, regardless of FBs dose, while nuclear factor kappa-Β ligand expression increased only in the 90 FB group. The disturbances in bone metabolic processes seemed to result from imbalances in the RANKL/RANK/OPG and the OC/leptin systems.

MMP-13, VEGF, and Disease Activity in a Cohort of Rheumatoid Arthritis Patients

Identifying certain serum biomarkers associated with the degree of rheumatoid arthritis (RA) activity can provide us with a more accurate view of the evolution, prognosis, and future quality of life for these patients. Our aim was to analyze the presence and clinical use of matrix metalloproteinase-13 (MMP-13), along with vascular endothelial growth factor (VEGF) and well-known cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6) for patients with RA. We also wanted to identify the possible correlations between MMP-13 and these serological markers, as well as their relationship with disease activity indices, quality of life, and ultrasonographic evaluation. For this purpose, we analyzed serum samples of 34 RA patients and 12 controls. In order to assess serum concentrations for MMP-13, VEGF, TNF-α, and IL-6, we used the enzyme-linked immunosorbent assay (ELISA) technique. Our results concluded that higher levels of MMP-13, VEGF, TNF-α, and IL-6 were present in the serum of RA patients compared to controls, with statistical significance. We furthermore identified moderately positive correlations between VEGF, MMP-13, and disease activity indices, as well as with the ultrasound findings. We also observed that VEGF had the best accuracy (97.80%), for differentiating patients with moderate disease activity. According to the data obtained in our study, that although MMP-13, TNF-α and C-reactive protein (CRP) have the same sensitivity (55.56%), MMP-13 has a better specificity (86.67%) in the diagnosis of patients with DAS28(4v) CRP values corresponding to moderate disease activity. Thus, MMP-13 can be used as a biomarker that can differentiate patients with moderate or low disease activity. VEGF and MMP-13 can be used as additional parameters, along with TNF-α and IL-6, that can provide the clinician a better picture of the inflammatory process, disease activity, and structural damage in patients with RA. Our data can certainly constitute a start point for future research and extended studies with multicenter involvement, to support the selection of individualized and accurate therapeutic management strategies for our patients.

Knockdown of mechanosensitive adaptor Hic-5 ameliorates post-traumatic osteoarthritis in rats through repression of MMP-13

Osteoarthritis (OA) is the most common joint disease associated with articular cartilage destruction. Matrix metalloproteinase-13 (MMP-13) has an essential role in OA pathogenesis by degradation of collagen II, a major component of articular cartilage. Hydrogen peroxide-inducible clone-5 (Hic-5; TGFB1I1), a transforming growth factor-β-inducible mechanosensor, has previously been reported to promote OA pathogenesis by upregulating MMP-13 expression in mouse osteoarthritic lesions. In our current study, immunohistochemical analysis showed that Hic-5 protein expression was increased in human OA cartilage compared with normal cartilage. Functional experiments demonstrated that Hic-5 and MMP-13 expression was increased by mechanical stress, and mechanical stress-induced MMP-13 expression was suppressed by Hic-5 siRNA in human chondrocytes. Moreover, intracellular localization of Hic-5 shifted to the nucleus from focal adhesions in human chondrocytes subjected to mechanical stress, and nuclear Hic-5 increased MMP-13 gene expression. In vivo, intra-articular injection of Hic-5 siRNA decreased the Osteoarthritis Research Society International score and MMP-13 protein expression in articular cartilage of OA rats. Our findings suggest that Hic-5 regulates transcription of MMP-13 in human chondrocytes, and Hic-5 may be a novel therapeutic target for OA because OA progression was suppressed by intra-articular injection of Hic-5 siRNA in rats.