MMP-9 Gene Research Articles

Single-Nucleotide Polymorphisms in MMP3, TIMP1, and MTR Genes are Associated With Delayed Deciduous Tooth Eruption.

To analyze whether the single-nucleotide polymorphisms (SNPs) in Matrix metalloproteinases 2, 3, and 9 (MMP2, MMP3, and MMP9), Tissue Inhibitor of Metalloproteinases 1 and 2 (TIMP1 and TIMP2), methionine synthase (MTR) and methionine synthase reductase (MTRR) influence delayed deciduous tooth eruption (DDTE). This cross-sectional study included 1060 biologic unrelated children (aged between 6 and 36months) of both sexes, selected from 25 public schools in Nova Friburgo, Rio de Janeiro, Brazil. Oral examination was conducted and DDTE was defined by the absence of gingival eruption according to a chronology based on the Brazilian population. Genotyping of selected SNPs (rs243847, rs52261, rs17576, rs4898, rs7501477, rs1805087, and rs1801394) was performed using TaqMan real-time PCR with genomic DNA extracted from buccal cells. The association between genotypes and DDTE was evaluated using univariate and multivariate analyses (p < 0.05). A total of 224 children and caregivers were included after the eligibility criteria. The heterozygous genotype for the SNPs MTR (rs11805087) was associated with DDTE in both the univariate (p = 0.004) and multivariate (p < 0.001) codominant models, as well as in the univariate (p = 0.010) and multivariate (p = 0.001) recessive models. TIMP1 (rs4898) and MMP3 (rs522616) were associated with DDTE only in the univariate model (p < 0.05). The SNPs in MTR (rs11805087), MMP3 (rs522616) and TIMP (rs4898) genes are associated with DDTE. The factors affecting the chronology of deciduous tooth eruption has been insufficiently studied. This article brings novel knowledge regarding the role of genetics polymorphisms on timing variation of deciduous tooth eruption. Understanding the factors that impact tooth eruption is crucial for the fields of pediatric dentistry and orthodontics.

Livogrit Mitigates ANIT-Induced Cholestasis-Like Symptoms in an In Vivo model by Curbing Hepatic Inflammation and Regulating BAX, TGF-β, MMP-9 and α-SMA Gene Expression

Livogrit Mitigates ANIT-Induced Cholestasis-Like Symptoms in an In Vivo model by Curbing Hepatic Inflammation and Regulating BAX, TGF-β, MMP-9 and α-SMA Gene Expression

ALLELIC VARIANTS OF MATRIX METALLOPROTEINASE TIPES 1 AND 3 GENES IN PRESCHOOL CHILDREN SUFFERING FROM RECURRENT RESPIRATORY

Matrix metalloproteinases (MMPs) have numerous physiological and pathological effects. In particular, they are involved in extracellular matrix degradation and remodeling, cell differentiation and apoptosis, immune and inflammatory mechanisms, embryogenesis, etc. Aim. To determine the state of interdependence between the allelic variants of the genes MMP-1-1607 (1G/2G) and MMP-3-1171 (5A/6A), on the one hand, and the different frequency of acute respiratory infections (ARI) in preschool children, on the other hand. Materials and methods. A total of 33 children (15 boys and 18 girls), aged 1-6 years, undergoing inpatient treatment for ARI were included in the clinical trial. The number of ARI episodes during the previous year of life was regarded. The distribution of allelic variants was also studied, specifically: 1) 1G/2G of the MMP-1 gene at position 1607 (rs 1799750) and 2) 5A/6A of the MMP-3 gene at position 1171 (rs 35068180). Statistical processing of the obtained digital data was performed using IBM SPSS Statistics 28 licensed software (No. 3922/01/2021). Results. The association between preschool children belonging to the subgroup with more frequent ARI episodes and the 2G/2G genotype for the rs 1799750 polymorphism locus of the MMP-1-1607 gene was demonstrated (OR=7.778; p1=0.008; p2=0.025; CI: 1.561 - 38.756). On the contrary, the 1G/2G genotype was significantly more frequent in patients forming a subgroup with fewer ARI episodes (OR=0.127; p1=0.014; p2=0.039; CI: 0.022 - 0.739). Furthermore, the 1G allele was more frequent in children with fewer ARI episodes, and the 2G allele was more frequent in those with a greater number of such episodes in their history (OR=2.778; p1=0.044; p2=0.080; CI: 1.016 - 7.639). At the same time, there was no association between alleles 5A and 6A and their corresponding genotypes, determined for the rs 35068180 polymorphism locus of the MMP-3 gene at position 1171, and subgroups of children with less frequent and more frequent ARI episodes. Conclusions. The results obtained suggest a significant dependence of the frequency of ARI episodes in preschool children on the studied genotypes (2G/2G and 1G/2G) of the MMP-1 gene (rs 1799750). They should be considered in predicting whether such children are at risk for recurrent course of ARI. The study was conducted within the framework of the initiative research work of the Department of Pediatrics with Childhood Infections of Luhansk State Medical University (Rubizhne, Ukraine) - "Current aspects of the influence of perinatal factors on the development of somatic pathology in children aged 1-14 years" (2017-2021, state registration number 0117U003041). No external funding was received for this study. The study was conducted in accordance with the tenets of the Declaration of Helsinki. The study protocol was approved by the local ethics committees of Bogomolets National Medical University and Luhansk State Medical University. Informed parental consent was obtained for the study in children. No conflict of interests was declared by the authors.

Aloe arborescens Standardized Glycosidic Fraction Suppresses Hepatocarcinoma by Modulating TIMP1, MMP9 Genes Expression, and Inflammation/Ki67/TGFβ1 Pathway.

(1) Background and aim: Aloe arborescens Mill. (A. arborescens) is one of the most widely distributed species in the genus Aloe and has garnered widespread recognition for its anticancer properties. However, the molecular mechanisms underlying these activities have not yet been fully elucidated. This study aimed to explore the effects of the plant polar glycosidic fraction (AAG) on hepatocellular carcinoma (HCC) in an invivo model induced by diethylnitrosamine (DEN). (2) Experimental procedure: The fraction was standardized using HPLC-PDA-MS/MS fingerprinting, and two distinct intragastric AAG dose regimens were examined (10 and 20 mg/kg) in combination with DEN 200 mg/kg. Serum alpha-fetoprotein (AFP), gamma-glutamyl transferase (γ-GGT), glutathione S-transferase placental (GST-P), mRNA expression of metabolic cytochrome enzymes (CYP1A3 and CYP2B2), inflammatory genes (nuclear factor kappa-B p65 subunit; NF-κB p65), metalloproteases 9 (MMP9), tissue inhibitors of metalloproteases (TIMP1), transforming growth factor beta 1 (TGFβ1), and histological features were assessed. (3) Key results and conclusions and implications: AAG was characterized by five major secondary metabolites: saponins, chromones, anthraquinone, and triterpenes. The fraction reduced hepatic malignancy characteristics by diminishing the size and number of altered foci and lowering hepatic cancer biomarkers, such as γ-GGT, AFP, and GST-positive foci. It also reduced the mRNA levels of CYP1A3 and CYP2B2, NF-κB p65, and MMP9, hepatic Ki-67, and TGFβ1 while upregulating TIMP1 levels. This study revealed that AAG exhibited a marked suppressive effect on HCC cell proliferation, displaying a range of mechanistic actions, including decreasing the metabolic activation of cytochrome enzymes, which consequently reduced the production of reactive oxygen species and other genes implicated in cancer development. AAG could be a significant therapeutic candidate for patients diagnosed with hepatocarcinoma.

Extracellular vesicles from platelet-poor plasma possess anti-inflammatory and anti-catabolic effects in chondrocytes stimulated with IL-1β or synovial membrane-conditioned media

BackgroundAlthough osteoarthritis (OA) is the most prevalent form of arthritis, there is still no effective treatment capable of combining immunomodulatory effects with cartilage repair. Extracellular vesicles (EVs) represent a promising new generation of cell-free therapies for OA. Blood-derived products, including plasma, are an easily available and abundant source of EVs with anti-inflammatory and regenerative properties. In this study, our objective was to analyze the effect of platelet poor plasma-derived extracellular vesicles (PPP-EVs) on stimulated OA chondrocytes in vitro. We hypothesize that PPP from healthy donors could be a suitable source of EVs that can modulate the inflammatory environment of OA chondrocytes.MethodsCartilage and synovial membrane (SM) were obtained from patients with OA and whole blood from healthy donors. Synovial membrane-conditioned media (CM / SM) was analyzed using multiplex immunoassays. EVs were isolated from PPP using size exclusion chromatography (SEC) and characterized by nanoparticle tracking analysis (NTA), Western blot, and flow cytometry (FC). The phenotype of the chondrocytes was analyzed using fluorescence microscopy and RT-qPCR. Chondrocytes were stimulated with IL-1β or CM/SM for 24 h. The impact of PPP-EVs on stimulated chondrocyte gene expression was evaluated using RT-qPCR.ResultsThe PPP-EVs isolated by SEC were positive for the tetraspanins CD9, CD63, and CD81. The chondrocyte phenotype was confirmed by positive expression of Collagen II and Aggrecane. CM/SM and IL-1β caused inflammatory changes in chondrocytes, which was observed by increased expression of the genes MMP-1, MMP-3 and MMP-13, RANTES, TSG-6, and YKL-40 compared to the control. PPP-EVs added to stimulated chondrocytes for 24 h significantly decreased the expression of the chondrocyte gene YKL-40, TSG-6 and MMP-1.ConclusionsIn this study, we confirmed that PPP is a suitable source of EVs, which can be efficiently isolated by SEC. We found that PPP-EVs were capable of decreasing the expression of inflammatory genes in OA chondrocytes stimulated with IL-1β or CM/SM. This study provides preliminary results on PPP-EVs as an affordable and promising option to modulate the catabolic microenvironment of OA chondrocytes in vitro.

Leptin/LPS-treated dendritic cells reduce the expression of genes involved in tumor tissue metastasis and angiogenesis in an animal model of breast cancer.

Leptin, an immune-regulating protein, enhances the maturation of dendritic cells (DCs). We previously demonstrated that leptin and lipopolysaccharide (LPS) promote the expression of co-stimulatory molecules on the surface of DCs. Leptin/LPS-treated DCs increased T cell responses against 4T1 breast cancer in mice. Therefore, in the present study, we investigate the effects of a DC vaccine treated with leptin and LPS on the genes involved in tumor metastasis, angiogenesis, and related cytokines in a mouse model of breast cancer. Tumor induction was achieved through subcutaneous injection of 4T1 cells into syngeneic mice. On days 12 and 19, the mouse groups received the DC vaccine treated with leptin and a combination of leptin and LPS. After sacrificing the mice on day 26, the levels of IL-6 and IL-33 in the serum were assayed using the ELISA technique, and the expression levels of the VEGF, CCL2, MMP9, and CCL5 genes in the tumors were measured by Real-Time PCR. Compared to untreated tumor-bearing mice, the leptin-treated mature DC (mDC) group exhibited a significant reduction in the expression of MMP9 (0.33-fold, p = 0.01) and CCL5 (0.81-fold, p = 0.02). The leptin-LPS-treated mDC group showed decreased expression of genes involved in metastasis and tumor growth, including VEGF (0.72-fold, p = 0.03), MMP9 (0.26-fold, p = 0.001), and CCL5 (0.3-fold, p = 0.006), indicating more efficient prevention of metastasis. The CCL2 gene expression levels in both treatment groups showed a slight decreasing trend, but these changes were not statistically significant. The leptin-treated mDC group reduced IL-6 production by approximately 16% (p = 0.02), while treatment with the leptin-LPS-treated mDC significantly decreased IL-6 production by approximately 22% (p = 0.01) and increased IL-33 production by approximately 42% (p = 0.03). The findings of the present study indicate that the leptin-LPS-treated mDC vaccine group reduced the expression of genes and cytokines involved in metastasis and angiogenesis, demonstrating greater efficacy compared to the leptin-treated mDC vaccine group.

Bioinformatics analysis of colorectal cancer transcriptomic data reveals novel prognostic signature and potential biomarker genes

Objective Colorectal cancer (CRC) is a type of digestive system cancer. At the molecular level, some factors, including genetic and epigenetic factors, as well as various signaling pathways such as oxidative stress and inflammation, play an active role in the onset of CRC. Genetic and epigenetic mutations, particularly in oncogenes and tumor suppressor genes, occur during colorectal adenocarcinoma development as a result of a change in gastrointestinal epithelial cell proliferation and self-renewal rates. This study aimed to determine the genes and molecular mechanisms that play a role in the emergence of this disease by analyzing the CRC data. Material and methods Microarray data selected for bioinformatics analysis is Gene Expression data stored with the code GSE110224 in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database. Gene expression analysis, functional clustering analysis, enrichment analysis, and pathway analysis were performed using this data set. Results Analysis of raw transcriptomic data revealed 1770 common DEGs in CRC. While the expression level of 769 of these genes increased, the expression level of 1001 genes decreased. A Protein–protein interaction (PPI) network was created from the first 25 genes with increased expression levels and 11 signature genes were identified. Increased expression of REG1A, MMP3, FOXQ1 and CEMIP genes and decreased expression of AQP8, CA1, CLDN8, PYY, CA4, CEACAM7 and SLC30A10 genes were observed. Conclusions This approach revealed a CRC-specific molecular profile and may provide some guidance for further investigation of potential biomarkers for diagnosis and prognosis prediction of CRC patients.

Systematic pharmacology-based strategy to investigate the mechanism of beta-sitosterol for the treatment of rheumarthritis.

Objective: β-Sitosterol, which is derived from Vladimiriae Radix (VR), is used for the treatment of rheumatoid arthritis (RA), but the pharmacological mechanisms through which β-sitosterol affects RA have not been fully elucidated. Methods: Through the Traditional Chinese Medicine Systems Pharmacology and Analysis (TCMSP), PubChem, SwissTargetPrediction, GeneCards, DisGeNET, and OMIM databases, "β-sitosterol-RA"-related genes were obtained, and a target protein interaction network (protein-protein interaction [PPI]) was constructed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out for the intersecting genes. Discovery Studio 2019 software was used to perform molecular docking on MMP9, CASP3, HSP90AA1, SRC, EGFR, and ALB genes. β-Sitosterol was co-cultured with MH7A cells in three experimental groups: control group (DMSO), positive drug group (methotrexate, 80μmol/L), and drug intervention group (10, 20, 40, 80, and 160μmol/L β-sitosterol). The CCK8 method was used to investigate the inhibitory effect of β-sitosterol on the proliferation of MH7A cells. RT-PCR was used to analyze the mRNA expression of the abovementioned core targets. Results: A total of 41 genes associated with β-sitosterol and RA were obtained, mainly involving the FoxO signaling pathway and PI3K/AKT signaling pathway. The molecular docking results suggested that β-sitosterol could bind effectively to six core targets. The experimental results showed that β-sitosterol could significantly inhibit the excessive proliferation of MH7A cells (p< 0.05). The RT-PCR results showed that the expression of MMP9, HSP90AA1, SRC, EGFR, and ALB core genes in the control group was significantly upregulated, while the CASP3 gene was downregulated. Compared to the control group, the mRNA expression of MMP9, HSP90AA1, SRC, EGFR, and ALB decreased (p< 0.01), while the apoptosis-related gene CASP3 increased in both the drug intervention (80μmol/L β-sitosterol) and positive drug groups (80μmol/L methotrexate). Conclusion: Hence, β-sitosterol could contribute to the inhibition of RA by modulating cell proliferation and regulating the aforementioned six core proteins, potentially through the regulation of the FoxO and PI3K/AKT signaling pathways.

Comparison of biocompatibility of epoxy resin and bioceramic-based root canal sealers.

The purpose of this study is to compare the cytotoxicity of 2 bioceramic-based (Bioserra and AH Plus Bioceramic (Bio)) and 2 epoxy resin-based (Sealart and AH Plus) root canal sealers on the Saos-2 cell line. The cytotoxicity of the root canal sealers was determined using the MTT metabolic activity assay. The mRNA expression levels of Bcl-2, Beclin1, Cas-3, IL-6, LC3, MMP-9 and TNF-α genes were assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and the expression of Beclin1 and LC3 proteins were assessed using western blot (WB) method. The canal sealers' total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were measured using photometric kits. AH Plus Bio significantly increased metabolic activity compared to the 2 resin-containing sealers, but there was no difference with Bioserra. Sealart was clearly the most cytotoxic group. QRT-PCR analysis revealed that the mRNA expressions of Bcl-2, Beclin1, Cas-3, IL-6, LC3, and MMP-9 in the Bioserra group, Bcl-2, Cas-3, and IL-6 in the Sealart group, Beclin1, Cas-3, IL-6, and MMP-9 in the AH Plus group, and Bcl2 in the AH Plus Bio group were stimulated. The OSI results showed no significant difference between the root canal sealer groups. In the study, the best biocompatibility results in MTT, qRT-PCR, WB and OSI were in the AH Plus Bio group. Although Bioserra, the study's other bioceramic paste, was found to be biocompatible according to MTT and OSI measurements, it increased the expression of all genes in qRT-PCR except for TNF-α, and also increased LC3 protein levels in WB. Since periradicular tissues can be directly affected by the materials used in root canal treatment, the sealers with high biocompatibility are being developed. In vitro studies examining the biocompatibility of newly produced root canal sealers are guiding for clinical success.

The Association of Matrix Metalloproteinase-1, -2, -3, -7, and -13 Gene Polymorphisms With Peri-Implantitis in an Iranian Population: A Case-Control Study.

Peri-implantitis (PI) is the most common biological issue surrounding dental implants. According to current knowledge, the aforementioned complication is not equally distributed across different populations, and gene polymorphisms might be one contributing factor. The current study aimed to examine the association between gene polymorphisms of matrix metalloproteinase- (MMP-) 1, -2, -3, -7, and -13 with PI in an Iranian demographic. The study's sample included 50 subjects suffering from PI and 89 healthy controls. From each participant, a venous blood sample of 5 cc was obtained, and DNA was extracted. Gene polymorphisms were investigated using restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) combined with electrophoresis. Statistical analyses were done using the Pearson chi-square test, odds ratio, and t-test via SPSS version 28. The MMP-3 (-1171 5A/6A) and MMP-7 (-181 A/G) gene polymorphisms were significantly different between the patients with PI and healthy controls (PV < 0.001 and =0.025, respectively). MMP-1 (-1607 1G/2G), MMP-2 (-1306 C/T), and MMP-13 (-77 A/G) gene polymorphisms did not, however, differ in terms of prevalence between the two groups (PV > 0.05). Moreover, the presence of the 6 A allele in the MMP-3 (-1171 5A/6A) genotype resulted in a significant decrease in PI risk (PV < 0.001). Gene polymorphisms in the genotypes of MMP-3 (-1171 5A/6A) and MMP-7 (-181 A/G) were differential when comparing PI patients and healthy controls of the studied population.

The Anti-Metastatic Potential of Aronia Leaf Extracts on Colon Cancer Cells.

Numerous studies have demonstrated the health benefits of polyphenols found in aronia fruits; however, little is known about how aronia leaf polyphenols impact colorectal cancer (CRC). This study aimed to evaluate the in vitro anti-metastatic and anti-invasive activity of crude aronia leaf extract (ACE) and purified phenolic-rich aronia leaf extract (APE) against two CRC cell lines (SW-480 and HT-29). Migration and invasion potential of ACE and APE were evaluated. Moreover, ELISA and gelatin zymography were performed to detect translational and activity changes in CRC cells after aronia extracts treatment. We found that a 100 µg/mL concentration of ACE and APE almost entirely downregulated the migration and invasion of SW-480 cells, showing greater effectiveness than HT-29 cells. The observed inhibition was concentration-dependent and statistically significant. Additionally, extracts reduced the product of MMP-2 and MMP-9 gene expression at the protein level and simultaneously inhibited the activity of both MMPs. An APE at 300 µg/mL for SW-480 and 600 µg/mL for HT-29 resulted in a notable reduction in MMP-2 protein synthesis by 72% and 50%, respectively. In contrast, MMP-9 protein synthesis decreased by 48% and 59% in HT-29 cells treated with 300 µg/mL and 600 µg/mL of ACE, respectively. The levels of gelatinase activity were similar for both CRC lines, and the APE tested at a concentration of 300 µg/mL reached almost the IC50 value after 48 h of incubation. Based on the presented results, we provided an experimental foundation for future in vitro and in vivo studies on the potential effects and activities of aronia leaves.

Withaferin-A induced vimentin S56 phosphorylation dissociates NEDD9 signaling loop to regress progressive metastatic melanoma into lung adenocarcinoma

Metastasis is complex and insidious type of disease involves multiple signaling nexus, which have implications in understanding disease pathogenesis. Treatment failure for metastatic cancer is frequently high due to aggressive adaptation of cancerous cells to invade to neighboring organs. Cytoskeleton intermediate filamentous protein Vimentin and scaffolding protein Neural precursor cell expressed Developmentally Down-regulated protein 9 (NEDD9) play a key role in metastatic events by regulating multiple metastatic events. Interaction between these proteins is necessary to promote metastatic progression. Withaferin A (WFA), a natural pharamacophore, known to target Vimentin to induce antitumor potential. However exact molecular mechanism still yet to be elucidated. We hypothesize, Vimentin-NEDD9 signaling nexus is necessary for metastatic progression and targeting this interwoven signaling loop with effective pharamacophore WFA halts metastatic progression of melanoma into lung. To elucidate the same, we carried out gene expression measurement through quantitative Reverses Transcription Polymerase Chain Reaction (qRT-PCR), Immunoblot and Immunohistochemistry. Assessment of interactive signaling by Co-immunoprecipitation, Immunofluorescence, Co-localization and Proximity ligation assay. Phosphorylation studies through transfection of phospho specific mutant constructs generated through site directed mutagenesis. WFA induced cellular behavioral changes by migration, invasion assays and Immunoblot analysis. The B16F10 induced mouse metastatic melanoma model to asses NEDD9-Vimentin expression and anti-metastasis induced by WFA. The results postulates, elevated levels and interaction between NEDD9-Vimentin proteins, have positive correlation in metastatic progression of melanoma into lung in both in-vitro and in-vivo condition, establishing it as therapeutic target. Pharmacologically, WFA targets this complex by extending its activity by not only inducing specific Serine 56 phosphorylation of Vimentin, also dissociates NEDD9 signaling loop to halt Epithelial-mesenchymal transition (EMT) and subsequent metastatic events. Eventually, modulation of the relevant metastatic genes E-Cadherin, N-Cadherin, SNAIL, MMP-2 & MMP-9 resulted in regression of metastatic melanoma progression to lung. The study validates WFA induced S56 phosphorylation is necessary to abrupt the NEDD9-Vimentin metastatic signaling complex to regress aggressive metastatic melanoma. The investigation emphasized more mechanistic approach of WFA. Understanding and targeting such integrative mechanical input in the tumor microenvironment will be a better therapeutic strategy to combat metastasis.

A Versatile Skin-Derived Extracellular Matrix Hydrogel-Based Platform to Investigate the Function of a Mechanically Isolated Adipose Tissue Stromal Vascular Fraction.

Introduction: To accelerate cutaneous wound healing and prevent scarring, regenerative approaches such as injecting a mechanically derived tissue stromal vascular fraction (tSVF) are currently under clinical and laboratory investigations. The aim of our study was to investigate a platform to assess the interaction between skin-derived extracellular matrix (ECM) hydrogels and tSVF and their effects on their microenvironment in the first ten days of culture. Material and Methods: A tSVF mixed with ECM hydrogel was cultured for ten days. After 0, 3, 5, and 10 days of culture viability, histology, immunohistochemistry, gene expression, and collagen alignment and organization were assessed. Results: The viability analysis showed that tSVF remained viable during 10 days of culture and seemed to remain within their constitutive ECM. The fiber analysis demonstrated that collagen alignment and organization were not altered. No outgrowth of capillaries was observed in (immuno)histochemical staining. The gene expression analysis revealed that paracrine factors TGFB1 and VEGFA did not change and yet were constitutively expressed. Pro-inflammatory factors IL1B and IL6 were downregulated. Matrix remodeling gene MMP1 was upregulated from day three on, while MMP14 was upregulated at day three and ten. Interestingly, MMP14 was downregulated at day five compared to day three while MMP2 was downregulated after day zero. Conclusions: Skin-derived ECM hydrogels appear to be a versatile platform for investigating the function of a mechanically isolated adipose tissue stromal vascular fraction. In vitro tSVF remained viable for 10 days and sustained the expression of pro-regenerative factors, but is in need of additional triggers to induce vascularization or show signs of remodeling of the surrounding ECM. In the future, ECM-encapsulated tSVF may show promise for clinical administration to improve wound healing.

Horse Innate Immunity in the Control of Equine Infectious Anemia Virus Infection: A Preliminary Study.

The mechanisms of the innate immunity control of equine infectious anemia virus in horses are not yet widely described. Equine monocytes isolated from the peripheral blood of three Equine infectious anemia (EIA) seronegative horses were differentiated in vitro into macrophages that gave rise to mixed cell populations morphologically referable to M1 and M2 phenotypes. The addition of two equine recombinant cytokines and two EIA virus reference strains, Miami and Wyoming, induced a more specific cell differentiation, and as for other species, IFNγ and IL4 stimulation polarized horse macrophages respectively towards the M1 and the M2 phenotypes. Infection with EIAV reference strains resulted in a morphological transformation of macrophages compatible with the M1 differentiation pattern. All samples were also analyzed by molecular analyses for equine herpesviruses that could have acted as an interference and were found to be negative. The mRNA expression level of the pro-inflammatory genes MMP13 and IL6 in treated equine monocyte-derived macrophages (eMDMs) was evaluated by a SYBR® Green real-time PCR. In this study, MMP13 represented a reliable target gene to evaluate pro-inflammatory status of macrophages in horses because IFNγ and EIAV infection considerably increased its expression. A more in-depth study of the expression genes of both cytokine-induced and virus-induced markers of eMDM polarization may help us to understand whether these markers in horses are the same as those found in other animal species with similar pathways of innate immunity activation. The identified markers of each macrophage population would allow analysis of the differentiation profiles that could provide information on virus infectivity control in equid populations, envisioning their use in therapeutic strategies.

Pre-metastatic niche in oral squamous cell carcinoma: Insights from a transcriptomic meta-analysis

Primary tumors can precondition a pre-metastatic niche, promoting the colonization of circulating neoplastic cells and influencing the secondary tumor microenvironment. Nevertheless, the mechanisms underlying the formation of this niche, as well as perineural invasion in oral squamous cell carcinoma (OSCC), are not well-elucidated. The study aims to identify differentially expressed genes (DEGs) and related pathways associated with pre-metastatic niche and perineural invasion in OSCC. We evaluated metastatic and non-metastatic primary tumor samples, healthy oral tissues, OSCC samples, metastatic lymph nodes from patients with OSCC, and normal lymph node samples. The GEO2R tool was applied to identify mRNAs differentially expressed between tissues exhibiting features of a pre-metastatic niche and normal tissue samples, including selected non-metastatic and metastatic OSCC samples. We also performed an analysis of perineural invasion-negative and perineural invasion-positive tumor samples. Our data revealed that SERPINE1, SPP1, CALCA, and MMP13 genes were upregulated. These upregulated genes are associated with several cancer-related pathways, while downregulated genes are mainly associated with immune responses, axon guidance, and the neurotrophin signaling pathway. Given the upregulation of the circadian rhythm pathway in metastatic lymph nodes, we also performed a correlation analysis that allows users to compute function-specific parameters, with resulting figures dynamically displayed to conveniently access the tumor&amp;rsquo;s immunological, clinical, and genomic features. Downregulation of the circadian rhythm gene PER3 and upregulation of Bhlhe40 were associated with poor survival outcomes. In conclusion, we postulate that during lymph node invasion, OSCCs activate axonal guidance genes, such as SERPENE1, L1AM, CXCR4, and SPP1. As neoplastic cells establish themselves, circadian rhythm genes are upregulated, contributing to immune evasion and promoting tumor growth.

The effects of extracellular matrix-degrading enzymes polymorphisms on intervertebral disc degeneration.

The objective of the current study was to investigate the correlation between polymorphisms in extracellular matrix-degrading enzymes and the risk of intervertebral disc degeneration (IDD) diseases. The databases PubMed, Embase, and Cochrane Database were systematically queried from the inception until March 2023 to ascertain studies that meet the eligibility criteria. Utilizing a standardized data collection form to extract data from individual studies. The data were quantified using odds ratio (OR) along with its corresponding 95% confidence interval (95% CI), following an allelic model of inheritance. The study included a total of nine studies and indicated that the presence of rs17576 in the MMP9 gene was significantly associated with an increased risk of IDD diseases (GG: 1.30, 95% CI [1.09-1.55], p = 0.004). The presence of other polymorphisms in extracellular matrix-degrading enzymes did not exhibit a significant association with the susceptibility to IDD. The current study demonstrated a noteworthy correlation between the GG genotype of MMP-9 rs17576 and susceptibility to IDD. The available evidence is insufficient to substantiate the correlation between other extracellular matrix-degrading enzymes and susceptibility to IDD. The constraints of this analysis necessitate further research involving larger sample sizes across diverse ethnicities to provide a comprehensive understanding of the true impact of these polymorphisms on susceptibility to IDD.

Regulation of Apoptosis, Autophagy, and Metastasis by Luteolin in Human Bladder Cancer EJ138 Cells: An Experimental Study

Background: Chemotherapy remains a primary approach to cancer treatment, widely applied in bladder cancer (BC). However, the various side effects and resistance associated with chemotherapeutic drugs pose significant challenges in BC therapy, prompting interest in natural compounds like luteolin. Studies focus on its effects on key biological processes involved in BC, including metastasis, apoptosis, and autophagy. Objectives: This study investigated the regulation of mRNA expression of genes associated with apoptosis (BCL2, P53), autophagy (ULK1, ATG12), and metastasis (MMP2, MMP9) in malignant BC cells treated with luteolin. Methods: This was an in vitro experimental study. EJ138 BC cells were treated with various concentrations of luteolin, and its impact on cell viability, proliferation, and gene expression was assessed. The cytotoxic effect of luteolin on EJ138 BC cells was evaluated using the MTT assay after 24- and 48-hour treatments with different luteolin concentrations. Flow cytometry was performed to examine luteolin’s anti-proliferative effect, and RT-PCR was used to analyze mRNA expression of BCL2, P53, ULK1, ATG12, MMP2, and MMP9 genes. Results: MTT assay results confirmed that luteolin reduced the proliferation rate of BC cells. Flow cytometry indicated increased cell death in EJ138 BC cells following luteolin treatment. RT-PCR findings demonstrated that luteolin upregulated P53, ULK1, and ATG12 expression while downregulating BCL2 mRNA expression. However, luteolin treatment in EJ138 cells did not significantly alter MMP2 and MMP9 expression levels. Conclusions: These findings indicate that luteolin exerts cytotoxic effects on EJ138 BC cells by dysregulating mRNA expression of genes involved in apoptosis and autophagy. Therefore, luteolin shows potential as an effective anti-cancer agent for BC therapy.

Unveiling the therapeutic potential of anthocyanin/cisplatin-loaded chitosan nanoparticles against breast and liver cancers

BackgroundLiver and breast cancers are among the leading causes of cancer-related deaths worldwide, prompting researchers to seek natural anticancer agents and reduce chemotherapy side effects. Red beetroot (Beta vulgaris Linnaeus), rich in polyphenols and powerful antioxidants, has shown potential in cancer prevention. This study aimed to evaluate the impact of red beetroot-derived anthocyanin (Ant), Ant-loaded chitosan nanoparticles (Ant NPs), cisplatin (Cis), Cis-loaded chitosan (Cis NPs), and Cis + Ant-loaded chitosan NPs on human hepatoma HepG2 and breast adenocarcinoma MCF7 cell lines.MethodsNPs preparation was evaluated by zeta potential, FTIR, and SEM. The cytotoxic, apoptotic, antioxidant, anti-inflammatory, anti-metastatic, and anti-angiogenic effects were assessed by MTT assay, qPCR, AO/EB staining, and flow cytometry.ResultsTreatment with Ant, Ant NPs, Cis, Cis NPs, and Cis + Ant NPs caused cytotoxicity in HepG2 and MCF7 with best effect in Cis-treated cells. The anticancer effects were attributed to mitochondrial-dependent apoptosis (with high Bax and low Bcl2 expression), chromatin disintegration, and cell cycle arrest in G2/M and S phases. All treatments inhibited migration by downregulating the migration-related gene MMP9 and upregulating the anti-migratory gene TIMP1 and decreased the angiogenesis-related gene VEGF and the inflammatory gene TNFα with best results in Cis NPs-treated cells. Interestingly, Ant, Ant NPs, and Cis + Ant NPs increased the antioxidant status (high GSH and upregulated expression of Nrf2 and OH-1) and decreased drug resistance-related MAPK1 and MDR1 genes compared to Cis and Cis NPs-treated cells.ConclusionsAnthocyanin and cisplatin-loaded chitosan nanoparticles effectively combat breast and liver cancers by inducing cancer cell apoptosis, enhancing antioxidant defenses, and reducing inflammation. They also inhibit tumor spread and blood vessel formation through downregulation of MMP9 and VEGF, highlighting their therapeutic potential.

Lipid metabolism-related gene signature predicts prognosis and unveils novel anti-tumor drugs in specific type of diffuse large B cell lymphoma

BackgroundDiffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma which possess highly aggressive and heterogeneous. Despite advances in understanding heterogeneity and development of novel targeted agents, the prognosis of DLBCL patients remains unsatisfied. Lipids are crucial components of biological membranes and signal transduction while accumulating evidence has supported the vital roles of abnormal lipid metabolism in tumorigenesis. Furthermore, some related pathways could serve as prognostic biomarkers and potential therapeutic targets. However, the clinical significance of abnormal lipid metabolism reprogramming in DLBCL has not been investigated. In the current study, we developed a prognostic risk model for DLBCL based on the abnormal expressed lipid metabolism genes and moreover based on our risk model we classified patients with DLBCL into novel subtypes and identified potential drugs for DLBCL patients with certain lipid metabolism profiles.MethodsWe utilized univariate Cox regression analysis to identify the prognosis-related lipid metabolism genes, and then performed LASSO Cox regression to identify prognostic related lipid metabolism related genes. Multivariate cox regression was used to establish the prognostic model. Patients were divided in to high and low risk groups based on the median risk score. Immune cell infiltration and GSEA were used to identify the pathways between high and low risk groups. Oncopredict algorithm was utilized to identify potential drug for high-risk patients. In vitro cell apoptosis and viability analysis were employed to verify the specific tumor inhibition effects of AZD5153.ResultsNineteen survival related lipid metabolism genes TMEM176B, LAYN, RAB6B, MMP9, ATAD3B, SLC2A11, CD3E, SLIT2, SLC2A13, SLC43A3, CD6, SIRPG, NEK6, LCP2, CTTN, CXCL2, SNX22, BCL6 and FABP4 were identified and subjected to build the prognostic model which was further verified in four external microarray cohorts and one RNA seq cohorts. Tumor immune microenvironment analysis and GSEA results showed that the activation of MYC targets genes rather than immunosuppression contribute to the poor survival outcome of patients in the high-risk group. AZD5153, a novel bivalent BET bromodomain inhibitor which could inhibit the transcription of MYC and E2F exhibited specific antitumor function for cells with high-risk score.ConclusionsOur results provide the first lipid metabolism-based gene signature for predicting the survival of patients with DLBCL. Furthermore, by determining novel subtypes with our lipid metabolism prognostic model we illustrated that drugs that compromising MYC target genes rather than immune checkpoint inhibitors may be beneficial to DLBCL patients with certain lipid metabolism profiles.

Lung allograft dysbiosis associates with immune response and primary graft dysfunction

RationaleLower airway enrichment with oral commensals has been previously associated with grade 3 severe primary graft dysfunction (PGD) after lung transplantation (LT). We aimed to determine whether this dysbiotic signature is present across all PGD severity grades, including milder forms, and whether it is associated with a distinct host inflammatory endotype. MethodsLower airway samples from 96 LT recipients with varying degrees of PGD were used to evaluate the lung allograft microbiota via 16S rRNA gene sequencing. Bronchoalveolar lavage (BAL) cytokine concentrations and cell differential percentages were compared across PGD grades. In a subset of samples, we evaluated the lower airway host transcriptome using RNA sequencing methods. ResultsDifferential analyses demonstrated lower airway enrichment with supraglottic-predominant taxa (SPT) in both moderate and severe PGD. Dirichlet Multinomial Mixtures (DMM) modeling identified two distinct microbial clusters. A greater percentage of subjects with moderate-severe PGD were identified within the dysbiotic cluster (C-SPT) than within the no PGD group (48 and 29%, respectively) though this difference did not reach statistical significance (p=0.06). PGD severity associated with increased BAL neutrophil concentration (p=0.03) and correlated with BAL concentrations of MCP-1/CCL2, IP-10/CXCL10, IL-10, and TNF-α (p<0.05). Furthermore, microbial signatures of dysbiosis correlated with neutrophils, MCP-1/CCL-2, IL-10, and TNF-α (p<0.05). C-SPT exhibited differential expression of TNF, SERPINE1 (PAI-1), MPO, and MMP1 genes and upregulation of MAPK pathways, suggesting that dysbiosis regulates host signaling to promote neutrophilic inflammation. ConclusionsLower airway dysbiosis within the lung allograft is associated with a neutrophilic inflammatory endotype, an immune profile commonly recognized as the hallmark for PGD pathogenesis. This data highlights a putative role for lower airway microbial dysbiosis in the pathogenesis of this syndrome.