ABSTRACT
OBJECTIVE: Although several surgical techniques have been developed for treatment of gingival recession (GR), the underlying wound healing process remains relatively unexplored. This systematic review aimed to investigate the expression of wound healing markers in gingival crevicular fluid (GCF) before and after surgical treatment of GR. DESIGN: Randomized clinical trials (RCTs) reporting changes in the expression of GCF markers following any root coverage surgical procedure were identified from 4 electronic databases and manual searches followed by data extraction and result synthesis. The risk of bias (RoB) was assessed using Cochrane RoB 2.0 tool. Overall certainty of evidence was summarized using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) tool. RESULTS: Four RCTs comprising 100 patients and investigating 15 biomarkers were included. Post-surgery, GCF levels of cytokines and inflammatory proteins were raised during the first 2-10 days of healing. MMP-8 levels increased during the first week followed by a gradual decline. RoB was found to be high for all studies and the overall certainty of evidence was very low. CONCLUSION: A limited number of studies with large methodological variations precluded reliable conclusions. Well-designed studies powered for GCF markers' levels that follow a standardized protocol for GCF sampling and processing are needed to draw conclusive evidence.
Subject(s)
Biomarkers , Gingival Crevicular Fluid , Gingival Recession , Randomized Controlled Trials as Topic , Wound Healing , Gingival Crevicular Fluid/chemistry , Humans , Wound Healing/physiology , Biomarkers/metabolism , Gingival Recession/surgery , Gingival Recession/metabolism , Cytokines/metabolism , Matrix Metalloproteinase 8/metabolismABSTRACT
OBJECTIVES: The purpose of this prospective cohort study is to evaluate the effect of peri-implant phenotype (PPh) on the severity of peri-implant diseases and the results of non-surgical mechanical treatment (NSMT), along with calprotectin (CLP) and MMP-8(matrix metalloproteinase-8) levels. MATERIALS AND METHODS: 77 implants from 39 patients were included. The implants were categorized Group-1(peri-implant mucositis), Group-2(peri-implantitis).Baseline (0. Month-PrT) clinical parameters (PD, GI, PI, BOP, CAL) and radiographic bone loss were documented, and peri-implant crevicular fluid (PICF) samples were collected. Various intruments and methodologies were employed to assess PPh components (mucosa thickness, supracrestal tissue height, keratinized mucosa) and peri-implant attached mucosa (AM). NSMT was applied to diseased implant sites. All clinical parameters were reassessed again by taking PICF samples at the 6th month-after treatment (PT). In PICF samples obtained from both groups, MMP-8 and CLP levels were evaluated using the ELISA test. RESULTS: PrT-PD,PrT-GI,PrT-CAL and PrT-BOP percentage values in Group-2 were significantly higher than Group-1.PrT-PD,PrTPI scores are significantly higher in thin biotype implants. All components of the PPh and AM were significantly lower in thin biotype. Intra-group time-dependent changes of MMP-8 and CLP were significant in both groups (p < 0.05). When the relationship between thin and thick biotype and biochemical parameters was evaluated, the change in PrT-PT didn't show a significant difference (p > 0.05). CONCLUSIONS: PPh plays a role in influencing the severity of peri-implant diseases. However, the impact of phenotype on NSMT outcomes was similar in both groups. CLINICAL RELEVANCE: The PPh should be considered when planning implant surgery.
Subject(s)
Gingival Crevicular Fluid , Leukocyte L1 Antigen Complex , Matrix Metalloproteinase 8 , Peri-Implantitis , Phenotype , Humans , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/analysis , Female , Prospective Studies , Peri-Implantitis/metabolism , Male , Middle Aged , Gingival Crevicular Fluid/chemistry , Leukocyte L1 Antigen Complex/analysis , Dental Implants , Enzyme-Linked Immunosorbent Assay , Biomarkers , Stomatitis/metabolism , Periodontal Index , Adult , AgedABSTRACT
The utilization of platelet-rich plasma (PRP) has exhibited potential as a therapeutic approach for the management of diabetic foot ulcers (DFUs). However, it is currently not well understood how the diabetic environment may influence PRP-derived exosomes (PRP-Exos) and their potential impact on neutrophil extracellular traps (NETs). This study aims to investigate the effects of the diabetic environment on PRP-Exos, their communication with neutrophils, and the subsequent influence on NETs and wound healing. Through bulk-seq and Western blotting, we confirmed the increased expression of MMP-8 in DFUs. Additionally, we discovered that miRNA-26b-5p plays a significant role in the communication between DFUs and PRP-Exos. In our experiments, we found that PRP-Exos miR-26b-5p effectively improved diabetic wound healing by inhibiting NETs. Further tests validated the inhibitory effect of miR-26b-5p on NETs by targeting MMP-8. Both in vitro and in vivo experiments showed that miRNA-26b-5p from PRP-Exos promoted wound healing by reducing neutrophil infiltration through its targeting of MMP-8. This study establishes the importance of miR-26b-5p in the communication between DFUs and PRP-Exos, disrupting NETs formation in diabetic wounds by targeting MMP-8. These findings provide valuable insights for developing novel therapeutic strategies to enhance wound healing in individuals suffering from DFUs.
Subject(s)
Diabetic Foot , Exosomes , Extracellular Traps , Matrix Metalloproteinase 8 , MicroRNAs , Platelet-Rich Plasma , Wound Healing , Animals , Humans , Male , Mice , Diabetes Mellitus, Experimental/metabolism , Diabetic Foot/therapy , Diabetic Foot/metabolism , Diabetic Foot/genetics , Exosomes/metabolism , Extracellular Traps/metabolism , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/genetics , Mice, Inbred C57BL , MicroRNAs/genetics , MicroRNAs/administration & dosage , Neutrophils/metabolismABSTRACT
The aim of this study was to test the molecular expression profile (senescence-associated secretory phenotype; SASP) in gingival crevicular fluid (GCF) prior to surgery in relation to the distribution of clinical success of periodontal regeneration. Forty consecutive patients presenting sites with residual probing pocket depth (PPD) ≥ 6 mm and intrabony defects ≥ 3 mm were treated through a minimally invasive surgical technique. Pre-operatively, GCF was sampled for inflammatory biomarker analysis related to SASP [interleukin (IL)-1ß, IL-6, and IL-12; matrix-metalloproteinases (MMP)-8 and -9]. Better or worse responders were classified depending on the achievement of a composite outcome measure at 1-year [COM; PPD ≤ 4 mm and clinical attachment gain (CAL) gain ≥ 3 mm]. Correlation analyses and logistic regression models were performed. Periodontal regeneration led to significant improvements in mean clinical and radiographic parameters. Teeth achieving COM presented significantly lower amounts of SASP factors compared with non-successful teeth. Higher CAL gain, PPD reduction, and radiographic bone fill were negatively correlated with IL-1ß and MMP-8 and -9 (p < 0.001), while IL-12 showed a direct relationship with CAL gain (p = 0.005) and PPD reduction (p = 0.038). Sites expressing higher SASP expression in the GCF before periodontal regeneration achieved worse clinical and radiographic outcomes.
Subject(s)
Biomarkers , Gingival Crevicular Fluid , Humans , Gingival Crevicular Fluid/metabolism , Male , Female , Middle Aged , Adult , Regeneration , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/genetics , Phenotype , Matrix Metalloproteinase 9/metabolism , Matrix Metalloproteinase 9/genetics , Inflammation/metabolism , Treatment Outcome , Interleukin-1beta/metabolism , AgedABSTRACT
OBJECTIVE: The objective of this study was to investigate the effectiveness of testing for active matrix metalloproteinase-8 (aMMP-8) by a quantitative point-of-care (PoC), chairside lateral flow immunotest and azurocidin, in the peri-implant sulcular fluid (PISF), as biomarkers for the presence or absence of peri-implant diseases. BACKGROUND: Current research indicates that proinflammatory cytokines and extracellular matrix-degrading enzymes may be of value to diagnose and predict peri-implant disease initiation and progression, but more data are needed. METHODS: Eighty patients with implants were recruited. PISF samples were collected and quantitatively analyzed for aMMP-8 (chairside) and azurocidin with ELISA. Radiographic assessments and clinical indices (probing depth, probing attachment level, bleeding on probing, and plaque) were recorded after sampling. Kruskal-Wallis test and pairwise post hoc Dunn-Bonferroni test were used to relate aMMP-8 levels and azurocidin levels to clinical parameters. The diagnostic ability of aMMP-8 (ng/mL) and azurocidin was analyzed by receiver operator curve analysis. Area under the curve (AUC) was calculated and the Spearman's rho, and the coefficient of determination (R2) were used to calculate the correlations between aMMP-8, azurocidin, and periodontal parameters. RESULTS: Statistically significant differences were observed for aMMP-8 levels but not for azurocidin between healthy implants, implants with mucositis, and those with peri-implantitis (13.65 ± 7.18, 32.33 ± 21.20, and 73.07 ± 43.93 ng/mL, respectively), (Kruskall-Wallis test p < .05). The aMMP-8 test with a threshold of 20 ng/mL has a sensitivity of 71.7% and a specificity of 77.8% to identify peri-implantitis and healthy implants, respectively. AUC was found to be 0.814, and the accuracy of the method reaches 73.8%. Above a cutoff value of 33.7 ng/mL of aMMP-8, the accuracy of the test to detect peri-implantitis reaches 77.5% in relation to 62.5% of BoP from the same site. CONCLUSION: Taken collectively, present data indicate that the aMMP-8 PoC lateral flow immunotest can be a beneficial, adjunctive diagnostic quantitative tool for real-time screening for peri-implant diseases.
Subject(s)
Biomarkers , Dental Implants , Gingival Crevicular Fluid , Matrix Metalloproteinase 8 , Peri-Implantitis , Humans , Matrix Metalloproteinase 8/analysis , Matrix Metalloproteinase 8/metabolism , Biomarkers/analysis , Biomarkers/metabolism , Male , Female , Middle Aged , Peri-Implantitis/diagnosis , Peri-Implantitis/metabolism , Aged , Dental Implants/adverse effects , Gingival Crevicular Fluid/chemistry , Gingival Crevicular Fluid/metabolism , Adult , Enzyme-Linked Immunosorbent Assay/methods , Periodontal Index , ROC Curve , Blood Proteins , Antimicrobial Cationic PeptidesABSTRACT
ADGRF5 (GPR116) has been identified as a facilitator of breast cancer cell migration and metastasis, yet the underlying mechanisms remain largely elusive. Our current study reveals that the absence of ADGRF5 in breast cancer cells impairs extracellular matrix (ECM)-associated cell motility and impedes in vivo tumor growth. This correlates with heightened expression of matrix metalloproteinase 8 (MMP8), a well-characterized antitumorigenic MMP, and a shift in the polarization of tumor-associated neutrophils (TANs) towards the antitumor N1 phenotype in the tumor microenvironment (TME). Mechanistically, ADGRF5 inhibits ERK1/2 activity by enhancing RhoA activation, leading to decreased phosphorylation of C/EBPß at Thr235, hindering its nuclear translocation and subsequent activation. Crucially, two C/EBPß binding motifs essential for MMP8 transcription are identified within its promoter region. Consequently, ADGRF5 silencing fosters MMP8 expression and CXCL8 secretion, attracting increased infiltration of TANs; simultaneously, MMP8 plays a role in decorin cleavage, which leads to trapped-inactivation of TGF-ß in the TME, thereby polarizing TANs towards the antitumor N1 neutrophil phenotype and mitigating TGF-ß-enhanced cell motility in breast cancer. Our findings reveal a novel connection between ADGRF5, an adhesion G protein-coupled receptor, and the orchestration of the TME, which dictates malignancy progression. Overall, the data underscore ADGRF5 as a promising therapeutic target for breast cancer intervention.
Subject(s)
Breast Neoplasms , Cell Movement , Matrix Metalloproteinase 8 , Receptors, G-Protein-Coupled , Animals , Female , Humans , Mice , Breast Neoplasms/pathology , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Line, Tumor , Disease Progression , Extracellular Matrix/metabolism , Gene Expression Regulation, Neoplastic , Interleukin-8/metabolism , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/genetics , Mice, Nude , Neutrophils/metabolism , Receptors, G-Protein-Coupled/metabolism , rhoA GTP-Binding Protein/metabolism , Transforming Growth Factor beta/metabolism , Tumor MicroenvironmentABSTRACT
The hypothesis that physiological changes in women can affect periodontal tissues is the subject of this study, and inflammatory markers such as matrix metalloproteinase-8 can measure susceptibility to inflammation. The study aimed to analyze MMP-8 levels in periodontal sites of postpartum women and women without a history of pregnancy, comparing health parameters and periodontal disease. This is a case-control study with 40 participants, 20 cases (women in the postpartum period) and 20 controls (women without any pregnancy), who underwent clinical periodontal examination and the collection of crevicular gingival fluid. The ELISA test was used to detect MMP-8 levels. Postpartum women had worse periodontal parameters, such as bleeding index on probing, number of sites with CAL ≥ 3, and fewer teeth present. In the group of women without a history of pregnancy, a significantly lower MMP-8 level was observed in healthy sites and a higher one was observed in periodontal pockets (p < 0.01). In contrast, in postpartum women, MMP-8 levels were elevated in both healthy sites and periodontal pockets (p > 0.01). The MMP-8 levels in gingival fluid appear to be related to periodontal clinical parameters and may be a possible marker of enzymatic changes involved in periodontal tissue destruction in postpartum women.
Subject(s)
Gingival Crevicular Fluid , Matrix Metalloproteinase 8 , Postpartum Period , Humans , Female , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/analysis , Adult , Case-Control Studies , Gingival Crevicular Fluid/enzymology , Pregnancy , Periodontal Diseases/enzymology , Biomarkers/metabolism , Young AdultABSTRACT
OBJECTIVE: We aimed to determine whether the semi-quantitative metalloproteinase-8 (MMP-8) bedside test is a worthwhile indicator in reflecting the severity of of intra-amniotic inflammation (IAI) and in predicting adverse pregnancy outcomes. STUDY DESIGN: This retrospective cohort study comprised 76 singleton-pregnant women admitted to the Seoul National University Bundang Hospital with a diagnosis of preterm premature rupture of membranes (preterm PROM) between 20 weeks 0 days and 33 weeks 6 days of gestation who underwent trans-abdominal amniocentesis to confirm intra-amniotic infection by positive results for aerobic/anaerobic bacteria, fungi, and genital mycoplasma and evaluate lung maturity. The semi-quantitative MMP-8 rapid test kit employs a colourimetric assay to quantify MMP-8 levels in amniotic fluid (AF), expressing results from 0 to 100 percent. Participants were divided into three groups: group 1, including negative MMP-8 test with colour scale of 0 % (negative, n = 17); group 2, including positive MMP-8 test with colour scale < 51 % (weak positive, n = 21); and group 3, including positive MMP-8 test with colour scale of 51 %-100 % (strong positive, n = 38). RESULTS: Approximately 78 % (59/76) of the participants showed a positive MMP-8 test result; all culture-proven AF samples (33.3 % [25/75]) yielded positive MMP-8 test, categorizing these patients into either group 2 or group 3. A significant trend was observed where the rate of positive culture-proven samples increased with the progression from group 1 (negative) to group 3 (strong positive). Both white blood cell counts in AF and maternal serum C-reactive protein levels were found to escalate with the progression of test results from negative to strong positive. This progression was associated with an increased risk of spontaneous preterm birth within 48 h, 7 days, and 14 days from amniocentesis and within 34 weeks of gestation. CONCLUSION: The more the test results progress from negative to strong positive, the shorter the interval from amniocentesis to delivery becomes, and the higher the risk of intra-amniotic infection, spontaneous preterm delivery, and other perinatal complications. This relationship highlights the critical value of the semi-quantitative MMP-8 rapid test in predicting adverse pregnancy outcomes in patients with preterm PROM.
Subject(s)
Amniotic Fluid , Fetal Membranes, Premature Rupture , Matrix Metalloproteinase 8 , Humans , Female , Pregnancy , Fetal Membranes, Premature Rupture/diagnosis , Matrix Metalloproteinase 8/analysis , Matrix Metalloproteinase 8/metabolism , Retrospective Studies , Adult , Amniotic Fluid/microbiology , Pregnancy Outcome , Chorioamnionitis/diagnosis , Amniocentesis , Predictive Value of Tests , Biomarkers/analysis , Premature Birth/diagnosisABSTRACT
Psychosocial stress has profound effects on the body, including the immune system and the brain1,2. Although a large number of pre-clinical and clinical studies have linked peripheral immune system alterations to stress-related disorders such as major depressive disorder (MDD)3, the underlying mechanisms are not well understood. Here we show that expression of a circulating myeloid cell-specific proteinase, matrix metalloproteinase 8 (MMP8), is increased in the serum of humans with MDD as well as in stress-susceptible mice following chronic social defeat stress (CSDS). In mice, we show that this increase leads to alterations in extracellular space and neurophysiological changes in the nucleus accumbens (NAc), as well as altered social behaviour. Using a combination of mass cytometry and single-cell RNA sequencing, we performed high-dimensional phenotyping of immune cells in circulation and in the brain and demonstrate that peripheral monocytes are strongly affected by stress. In stress-susceptible mice, both circulating monocytes and monocytes that traffic to the brain showed increased Mmp8 expression following chronic social defeat stress. We further demonstrate that circulating MMP8 directly infiltrates the NAc parenchyma and controls the ultrastructure of the extracellular space. Depleting MMP8 prevented stress-induced social avoidance behaviour and alterations in NAc neurophysiology and extracellular space. Collectively, these data establish a mechanism by which peripheral immune factors can affect central nervous system function and behaviour in the context of stress. Targeting specific peripheral immune cell-derived matrix metalloproteinases could constitute novel therapeutic targets for stress-related neuropsychiatric disorders.
Subject(s)
Depressive Disorder, Major , Matrix Metalloproteinase 8 , Monocytes , Stress, Psychological , Animals , Humans , Mice , Depressive Disorder, Major/blood , Depressive Disorder, Major/enzymology , Depressive Disorder, Major/genetics , Depressive Disorder, Major/metabolism , Extracellular Space/metabolism , Matrix Metalloproteinase 8/blood , Matrix Metalloproteinase 8/deficiency , Matrix Metalloproteinase 8/genetics , Matrix Metalloproteinase 8/metabolism , Mice, Inbred C57BL , Monocytes/chemistry , Monocytes/immunology , Monocytes/metabolism , Nucleus Accumbens/metabolism , Nucleus Accumbens/pathology , Parenchymal Tissue/metabolism , Single-Cell Gene Expression Analysis , Social Behavior , Social Isolation , Stress, Psychological/blood , Stress, Psychological/genetics , Stress, Psychological/immunology , Stress, Psychological/metabolismABSTRACT
There is an intrinsic relationship between psychiatric disorders and neuroinflammation, including bipolar disorder. Ouabain, an inhibitor of Na+/K+-ATPase, has been implicated in the mouse model with manic-like behavior. However, the molecular mechanisms linking neuroinflammation and manic-like behavior require further investigation. CCAAT/Enhancer-Binding Protein Delta (CEBPD) is an inflammatory transcription factor that contributes to neurological disease progression. In this study, we demonstrated that the expression of CEBPD in astrocytes was increased in ouabain-treated mice. Furthermore, we observed an increase in the expression and transcript levels of CEBPD in human primary astrocytes following ouabain treatment. Transcriptome analysis revealed high MMP8 expression in human primary astrocytes following CEBPD overexpression and ouabain treatment. We confirmed that MMP8 is a CEBPD-regulated gene that mediates ouabain-induced neuroinflammation. In our animal model, treatment of ouabain-injected mice with M8I (an inhibitor of MMP8) resulted in the inhibition of manic-like behavior compared to ouabain-injected mice that were not treated with M8I. Additionally, the reduction in the activation of astrocytes and microglia was observed, particularly in the hippocampal CA1 region. Excessive reactive oxygen species formation was observed in ouabain-injected mice, and treating these mice with M8I resulted in the reduction of oxidative stress, as indicated by nitrotyrosine staining. These findings suggest that MMP8 inhibitors may serve as therapeutic agents in mitigating manic symptoms in bipolar disorder.
Subject(s)
Neuroinflammatory Diseases , Ouabain , Animals , Humans , Mice , Astrocytes/metabolism , CCAAT-Enhancer-Binding Protein-delta/metabolism , Matrix Metalloproteinase 8/metabolism , Ouabain/toxicityABSTRACT
BACKGROUND: Although the current role of cytokines and neuroinflammation in glaucoma remains obscure, it represents an expanding field in research. The purpose of this study was to analyze cytokines in the aqueous humor (AH) of glaucoma patients and in retinas from an ex vivo glaucoma animal model, to aid in determining the role of neuroinflammation in glaucoma. METHODS: AH samples were collected from 20 patients during cataract surgeries (controls: n = 10, age = 70.3 ± 9.742; glaucoma: n = 10, age: 66.5 ± 8.073) in Shanghai East Hospital, an affiliate of Tongji University, between September 2018 and March 2019 and analyzed in duplicate by Luminex cytokine polystyrene color bead-based multiplex assay. Retinas from female Sprague-Dawley rats (n = 6) were harvested ex vivo and cultured with or without 60 mmHg of hydrostatic pressure for 24 hours. Retinal ganglion cells (RGCs) were quantified using Brn3a staining. Cytokines in the retina and culture medium were analyzed by rat cytokine array (Abcam). RESULTS: At baseline, patients with primary angle closure glaucoma (PACG) have significantly lower levels of IL-6 and IP-10 and a higher level of PDGF-BB in their AH, compared to the controls. Postoperatively, patients with PACG have significantly higher levels of IL-1ra, IL-13, and MIP-1α and a lower level of IL-6. Elevated hydrostatic pressure led to significant RGC loss in the retina, ex vivo, as well as the upregulation of ciliary neurotrophic factor (CNTF), IL-6, IL-10, IL-4, and TIMP-1 alongside the downregulation of PDGF-AA, MMP-8, TNF-α, and IFN-γ. Furthermore, eight cytokines were detected as being downregulated in the culture medium, including PDGF-AA, MMP-8, and IL-4. CONCLUSIONS: Proinflammatory cytokines showed changes in both AH and ex vivo. Further studies are needed on the role of these cytokines and their corresponding signaling pathways in both neurodegeneration and glaucoma.
Subject(s)
Glaucoma, Open-Angle , Glaucoma , Humans , Female , Rats , Animals , Middle Aged , Aged , Aged, 80 and over , Interleukin-6/metabolism , Aqueous Humor/metabolism , Neuroinflammatory Diseases , Interleukin-4/metabolism , Matrix Metalloproteinase 8/metabolism , Glaucoma, Open-Angle/metabolism , Glaucoma, Open-Angle/surgery , Rats, Sprague-Dawley , China , Glaucoma/metabolism , Cytokines/metabolism , Models, Animal , RetinaABSTRACT
This study aims to investigate the intervention effect of the aqueous extract of Epimedium sagittatum Maxim on the mouse model of bleomycin(BLM)-induced pulmonary fibrosis, so as to provide data support for the clinical treatment of pulmonary fibrosis. Ninety male C57BL/6N mice were randomized into normal(n=10), model(BLM, n=20), pirfenidone(PFD, 270 mg·kg~(-1), n=15), and low-, medium-, and high-dose E. sagittatum extract(1.67 g·kg~(-1), n=15; 3.33 g·kg~(-1), n=15; 6.67 g·kg~(-1), n=15) groups. The model of pulmonary fibrosis was established by intratracheal instillation of BLM(5 mg·kg~(-1)) in the other five groups except the normal group, which was treated with an equal amount of normal saline. On the day following the modeling, each group was treated with the corresponding drug by gavage for 21 days. During this period, the survival rate of the mice was counted. After gavage, the lung index was calculated, and the morphology and collagen deposition of the lung tissue were observed by hematoxylin-eosin(HE) and Masson staining, respectively. The levels of reactive oxygen species(ROS) in lung cell suspensions were measured by flow cytometry. The levels of glutathione peroxidase(GSH-Px), total superoxide dismutase(T-SOD), and malondialdehyde(MDA) the in lung tissue were measured. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling(TUNEL) was employed to examine the apoptosis of lung tissue cells. The content of interleukin-6(IL-6), chemokine C-C motif ligand 2(CCL-2), matrix metalloproteinase-8(MMP-8), transforming growth factor-beta 1(TGF-ß1), alpha-smooth muscle actin(α-SMA), E-cadherin, collagen â , and fibronectin in the lung tissue was measured by enzyme-linked immunosorbent assay(ELISA). The expression levels of F4/80, Ly-6G, TGF-ß1, and collagen â in the lung tissue were determined by immunohistochemistry. The mRNA levels of CCL-2, IL-6, and MMP-7 in the lung tissue were determined by qRT-PCR. The content of hydroxyproline(HYP) in the lung tissue was determined by alkaline hydrolysation. The expression of α-SMA and E-cadherin was detected by immunofluorescence, and the protein levels of α-SMA, vimentin, E-cadherin in the lung tissue were determined by Western blot. The results showed the aqueous extract of E. sagittatum increased the survival rate, decreased the lung index, alleviated the pathological injury, collagen deposition, and oxidative stress in the lung tissue, and reduced the apoptotic cells. Furthermore, the aqueous extract of E. sagittatum down-regulated the protein levels of F4/80 and Ly-6G and the mRNA levels of CCL-2, IL-6, and MMP-7 in the lung tissue, reduced the content of IL-6, CCL-2, and MMP-8 in the alveolar lavage fluid. In addition, it lowered the levels of HYP, TGF-ß1, α-SMA, collagen â , fibronectin, and vimentin, and elevated the levels of E-cadherin in the lung tissue. The aqueous extract of E. sagittatum can inhibit collagen deposition, alleviate oxidative stress, and reduce inflammatory response by regulating the expression of the molecules associated with epithelial-mesenchymal transition, thus alleviating the symptoms of bleomycin-induced pulmonary fibrosis in mice.
Subject(s)
Epimedium , Pulmonary Fibrosis , Mice , Male , Animals , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/drug therapy , Pulmonary Fibrosis/metabolism , Transforming Growth Factor beta1/metabolism , Epimedium/metabolism , Fibronectins/metabolism , Matrix Metalloproteinase 7/metabolism , Matrix Metalloproteinase 7/pharmacology , Matrix Metalloproteinase 7/therapeutic use , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 8/pharmacology , Matrix Metalloproteinase 8/therapeutic use , Vimentin/metabolism , Interleukin-6/metabolism , Mice, Inbred C57BL , Lung , Collagen/metabolism , Bleomycin/toxicity , RNA, Messenger/metabolism , Cadherins/metabolismABSTRACT
Renal fibrosis is tightly associated with chronic kidney disease, irrespective of the underlying pathogenesis. We previously demonstrated mild antifibrotic effects of targeting the P2X7 receptor in a pyelonephritis model. Reduced P2X7 R-activation elevated the neutrophil-to-macrophage ratio, resulting in less matrix accumulation without affecting the initial tissue healing. Here, we test if this P2X7 R-dependent modification of matrix accumulation also applies to a noninfectious fibrosis model of unilateral ureteral obstruction (7dUUO) and whether the response is gender-dependent. We found that P2X7 -/- mice show reduced fibrosis compared to wild type after 7dUUO: the effect was most pronounced in females, with a 55% decrease in collagen deposition after 7dUUO (p < 0.0068). P2X7 R deficiency did not affect early fibrosis markers (TGF-ß, α-SMA) or the renal infiltration of neutrophils. However, a UUO-induced increase in macrophages was observed in wildtypes only (p < 0.001), leaving the P2X7 -/- mice with ≈50% fewer CD68+ cells in the renal cortex (p = 0.018). In males, 7dUUO triggered an increase in diffusely interstitial scattering of the profibrotic, macrophage-attracting metalloproteinase MMP8 and showed significantly lower MMP8 tissue expression in both male and female P2X7 -/- mice (p < 0.0008). Thus, the P2X7 R is advocated as a late-stage fibrosis moderator by reducing neutrophil-dependent interstitial MMP8 release, resulting in less macrophage infiltration and reduced matrix accumulation.
Subject(s)
Kidney Diseases , Ureteral Obstruction , Female , Mice , Male , Animals , Ureteral Obstruction/pathology , Matrix Metalloproteinase 8/metabolism , Disease Models, Animal , Kidney/metabolism , Kidney Diseases/pathology , Fibrosis , Macrophages/metabolism , Mice, Inbred C57BLABSTRACT
OBJECTIVE: The objective of the study was to determine the concentration of matrix metalloproteinase 9 (MMP-9) and changes in the presence of periodontopathogens in the gingival crevicular fluid before and after tooth preparation with the subgingival and equigingival finish line position. PATIENTS AND METHODS: The clinical prospective study included 20 subjects with an indication for upper canine preparation, with the subgingival (group 1) and equigingival finish line (group 2). Samples were taken in four observation intervals: 5 minutes before (control samples), as well as 15 minutes, 24 and 72 hours after tooth preparation (experimental samples). Measurement of MMP-9 was done using Enzyme-linked Immunosorbent Assay (ELISA). The presence of bacteria in the gingival fluid was proven by the Polymerase chain reaction (PCR) analysis. RESULTS: The MMP-9 values did not differ statistically significantly between the groups (p=0.524). The MMP-9 values showed a statistically significant difference in the given observation period (p<0.001) with a significant linear increase in values (p<0.001). A significant quadratic trend recorded a decrease in the MMP-9 values 15 minutes after preparation, and an increase 24 hours after preparation, without a significant difference in the interaction between groups (p=0.392). After preparation, a significant difference in the presence of periodontopathogens was confirmed, i.e., a decrease in the presence of Prevotella intermedia (p=0.025) and Tannerella forsythia (p=0.016) in group 1, and an increase in the presence of Aggregatibacter actinomycetemcomitans in both groups (p=0.029, p=0.026). CONCLUSIONS: The study is a good basis for determining the influence of tooth preparation on gingival inflammation, with therapeutic (choice of preparation technique) and preventive significance regarding the protection of the periodontal tissue from possible iatrogenic damage.
Subject(s)
Gingival Crevicular Fluid , Matrix Metalloproteinase 9 , Humans , Gingival Crevicular Fluid/metabolism , Matrix Metalloproteinase 9/analysis , Prospective Studies , Matrix Metalloproteinase 8/analysis , Matrix Metalloproteinase 8/metabolismABSTRACT
Children with single ventricle heart disease (SVHD) experience morbidity due to inadequate pulmonary blood flow. Using proteomic screening, our group previously identified members of the matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinase (TIMP), and fibroblast growth factor (FGF) families as potentially dysregulated in SVHD. No prior study has taken a targeted approach to mapping circulating levels of these protein families or their relationship to pulmonary vascular outcomes in SVHD. We performed a prospective cohort study of 70 SVHD infants pre-Stage 2 palliation and 24 healthy controls. We report targeted serum quantification of 39 proteins in the MMP, TIMP, and FGF families using the SomaScan platform. Clinical variables were extracted from the medical record. Twenty of 39 tested proteins (7/14 MMPs, 2/4 TIMPs, and 11/21 FGFs) differed between cases and controls. On single variable testing, 6 proteins and no clinical covariates were associated with both post-Stage 2 hypoxemia and length of stay. Multiple-protein modeling identified increased circulating MMP 7 and MMP 17, and decreased circulating MMP 8 and FGFR2 as most associated with post-Stage 2 hypoxemia; increased MMP 7 and TIMP 4 and decreased circulating MMP 1 and MMP 8 were most associated with post-operation length of stay. The MMP, TIMP, and FGF families are altered in SVHD. Pre-Stage 2 imbalance of extracellular matrix (ECM) proteins-increased MMP 7 and decreased MMP 8-was associated with multiple adverse post-operation outcomes. Maintenance of the ECM may be an important pathophysiologic driver of Stage 2 readiness in SVHD.
Subject(s)
Heart Diseases , Matrix Metalloproteinase 8 , Child , Humans , Infant , Matrix Metalloproteinase 8/metabolism , Matrix Metalloproteinase 7/metabolism , Tissue Inhibitor of Metalloproteinase-1/metabolism , Prospective Studies , Proteomics , Extracellular Matrix/metabolism , Biomarkers , Extracellular Matrix Proteins/metabolism , Heart Diseases/metabolismABSTRACT
OBJECTIVES: To test the effect of locally delivered doxycycline (DOX) administered 2 weeks prior to minimally invasive periodontal regeneration in terms of presurgical inflammatory status and cytokine expression profile in the gingival crevicular fluid (GCF). Secondary aim was to assess the early wound healing index (EHI) at 2 weeks after surgery. BACKGROUND: It is hypothesized that healing after periodontal regeneration is dependent on preoperative soft tissue condition, and that local antibiotics may improve the site-specific inflammatory status at short time. METHODS: Sites associated with periodontal intrabony defects requiring regenerative surgery and showing bleeding on probing (BoP) were included. At T0, experimental sites were randomly treated with subgingival instrumentation with or without topic DOX application. After 2 weeks (T1), defects were approached by means of minimally invasive surgical technique. GCF was sampled at both T0 and T1 for inflammatory biomarker analysis. Two weeks after surgery, the EHI was evaluated (T2). RESULTS: Forty-four patients were included. At T1, the number of BoP+ sites was statistically significantly less in the test group (27.3% vs. 72.7%; p < .01). The total amount of interleukin (IL)-1ß (p < .001), matrix-metalloproteinases (MMP)-8 (p < .001), and MMP-9 (p = .010) in the GCF significantly decreased in the test group at T1, with relevant differences compared to controls. At T2, the EHI had an average value of 1.45 ± 0.86 in the test group while in the control, it was 2.31 ± 1.43 (p = .027). A statistically significantly positive correlation was observed between the amount of IL-1ß and MMP-9 and EHI scores. CONCLUSIONS: Within the limitations of this study, sites treated with DOX showed improved clinical and molecular inflammatory parameters before surgery, as well as soft tissue healing 2 weeks after surgery.
Subject(s)
Doxycycline , Matrix Metalloproteinase 9 , Humans , Doxycycline/therapeutic use , Anti-Bacterial Agents/therapeutic use , Wound Healing , Matrix Metalloproteinase 8/metabolism , Gingival Crevicular Fluid/metabolismABSTRACT
Objective: To investigate the effects of colony-stimulating factor 1 receptor (CSF-1R) inhibitor pexidartinib (PLX3397) on the senescence of bone marrow-derived macrophages (BMDM) stimulated by lipopolysaccharide (LPS). Methods: BMDM were isolated and cultured from femurs and tibiae of 10 male C57BL/6 mice aged 6-8 weeks (obtained from Laboratory Animal Center of Guizhou Medical University). They were divided into blank control group, LPS group (treated with 1 µg/ml LPS for 24 h) as well as low, medium and high concentration PLX3397 pretreatment groups (treated with 100, 500 and 1 000 nmol/L PLX3397 for 4 h respectively followed by 1 µg/ml LPS for 24 h). The corresponding markers of macrophages were detected by flow cytometry. Cell viability was detected by cell counting kit-8 and cellular senescence was detected by senescence-associated-ß-galactosidase (SA-ß-gal) staining. Meanwhile, protein expressions of cycle-dependent kinase inhibitor p16, p21 and CSF-1R were detected by Western blotting, and the expressions of p16 and p21 were detected by intracellular immunofluorescence. Real-time fluorescence quantitative PCR (RT-qPCR) was used to investigate the mRNA levels of senescence-associated secretory phenotype (SASP) genes including interleukin (IL), IL-1ß, chemokine-1/10 (CXCL-1/10), matrix metalloproteinase-8 (MMP-8), and transforming growth factor-ß (TGF-ß). Results: The rate of SA-ß-gal positive staining in medium and high concentration PLX3397 pretreatment groups [(39.33±4.93)% and (36.33±3.06)% respectively] were significantly downregulated compared with LPS group [(52.00±3.00)%] (P=0.020, P=0.005). The expression of CSF-1R protein in low, medium and high concentration PLX3397 pretreatment groups were (0.74±0.18, 0.61±0.07, 0.54±0.06), all of which were significantly lower than that in LPS group (1.16±0.08) (P=0.013, P=0.002, P<0.001). The expression levels of CSF-1R mRNA in low, medium and high concentration PLX3397 pretreatment groups (1.04±0.06, 0.90±0.05, 1.18±0.08) showed similar trend (2.90±0.25) (P<0.001). The average fluorescence intensity of p16 in all PLX3397 pretreatment groups were 49.76±3.65, 48.21±1.72, 47.99±1.26 respectively, which were significantly lower than that in LPS group (66.88±5.85) (P=0.001, P<0.001, P<0.001). The average fluorescence intensity of p21 in medium and high concentration PLX3397 pretreatment groups were (34.43±3.62, 30.13±0.86), significantly lower than that in LPS group (46.82±5.33) (P=0.043, P=0.007). The expression of p16 protein in low, medium and high concentration PLX3397 pretreatment groups (0.56±0.04, 0.55±0.04, 0.35±0.19) were significantly lower than that in LPS group (0.98±0.10) (P=0.003, P=0.002, P<0.001), as well the expression of p21 protein (0.69±0.20, 0.42±0.08, 0.26±0.14) (P=0.032, P=0.002, P<0.001). According to the results of RT-qPCR, the expressions of IL-6, IL-1ß, CXCL-1, CXCL-10 and MMP-8 in PLX3397 pretreatment groups were significantly lower than those in LPS group (P<0.001), while the expression of TGF-ß increased (P<0.001). Conclusions: LPS could induce the cell senescence, increase the secretion of SASP and aggravate local inflammation by activating the CSF-1R on the cell surface of bone marrow-derived macrophages. CSF-1R inhibitor PLX3397 might attenuate CSF-1R activation associated with LPS and inhibit the senescence of bone marrow-derived macrophages induced by LPS.
Subject(s)
Lipopolysaccharides , Macrophage Colony-Stimulating Factor , Mice , Animals , Male , Lipopolysaccharides/pharmacology , Macrophage Colony-Stimulating Factor/metabolism , Matrix Metalloproteinase 8/metabolism , Mice, Inbred C57BL , Macrophages , Transforming Growth Factor beta/metabolism , RNA, Messenger/metabolismABSTRACT
PROBLEM: To investigate whether altered expression of various inflammation-, angiogenesis-, and extracellular matrix-related mediators in cervicovaginal fluid (CVF) could be independently associated with acute histological chorioamnionitis (HCA), microbial-associated HCA, and funisitis in women with preterm premature rupture of membranes (PPROM). METHOD OF STUDY: Clinical data of 102 consecutive singleton pregnant women with PPROM at 23+0 to 34+0 weeks were retrospectively analyzed. CVF samples were collected upon admission. Levels of APRIL, DKK-3, IGFBP-1/2, IL-6/8, lipocalin-2, M-CSF, MIP-1α, MMP-8/9, S100A8A9, TGFBI, TIMP-1, TNFR2, uPA, and VDBP were determined by ELISA. Placentas were histologically examined after birth. RESULTS: Multivariate logistic regression analyses showed that: (1) elevated CVF levels of IL-8 and TNFR2 were independently associated with acute HCA; (2) elevated CVF levels of IL-6, IL-8, M-CSF, MMP-8, and TNFR2 were independently associated with microbial-associated HCA; and (3) elevated CVF IL-8 and MMP-8 levels were independently associated with funisitis when adjusted for gestational age. Areas under the curves of the aforementioned CVF biomarkers ranged within 0.61-0.77, thereby demonstrating poor to fair diagnostic capacity for these clinical endpoints. HCA risk significantly increased as the CVF levels of each inflammatory mediator increased (P for trend < 0.05). CONCLUSIONS: Herein, we identified several inflammatory biomarkers (IL-6/8, M-CSF, MMP-8, and TNFR2) in the CVF that are independently associated with acute HCA, microbial-associated HCA, and funisitis in women with PPROM. Furthermore, the degree of inflammatory response in the CVF, based on the levels of these proteins, demonstrated a direct relationship with HCA risk (especially risk severity).
Subject(s)
Chorioamnionitis , Fetal Membranes, Premature Rupture , Infant, Newborn , Female , Pregnancy , Humans , Chorioamnionitis/pathology , Macrophage Colony-Stimulating Factor , Receptors, Tumor Necrosis Factor, Type II/metabolism , Retrospective Studies , Interleukin-6/metabolism , Interleukin-8/metabolism , Matrix Metalloproteinase 8/metabolism , Fetal Membranes, Premature Rupture/metabolism , Biomarkers/metabolism , Amniotic Fluid/metabolismABSTRACT
OBJECTIVE: The aim of this study was to evaluate if a similar catabolic and inflammatory gene pattern exists between the synovium, hyaline cartilage, and blood of patients with the knee joint tissues and if one precedes the other. DESIGN: A total of fifty-eight patients (34 females and 24 males) with a mean age of 44.7 years (range, 18-75) underwent elective knee arthroscopy due to previously diagnosed pathology. Full blood samples were collected preoperatively from synovium and cartilage samples intraoperatively. Real time PCR with spectrophotometric analysis was performed. Following genes taking part in ECM (extracellular matrix) remodeling were selected for analysis: MMP-1, MMP-2, MMP-8, MMP-9, MMP-13, MMP-14, ADAMTS-4 (Agg1) and ADAMTS-5 (Agg2) proteases, TIMP-1, and TIMP-2 - their inhibitors - and IL-1 and TNF-α cytokines. RESULTS: Analysis revealed a strong and significant correlation between gene expression in synovial and systemic blood cells (p <0.05 for all studied genes) with ADAMTS-4, ADAMTS-5, IL-1, TNF-α and TIMP-2 expression most positively correlated with an R>0.8 for each. An analysis between chondrocytes and systemic blood gene expression shown no significant correlation for all genes. Bivariate correlation of International Cartilage Repair Society grading and genes expression revealed significant associations with synovial MMP-1, MMP-2, MMP-8, MMP-9, IL-1, TNF-α and TIMP-2. CONCLUSION: We suggest that the synovial tissue is the first responder for knee joint stress factors in correlation with the response of blood cells. The chondrocyte's genetic response must be further investigated to elucidate the genetic program of synovial joints, as an organ, during OA development and progression.
Subject(s)
Cartilage Diseases , Cartilage, Articular , Emergency Responders , Male , Female , Humans , Adult , Matrix Metalloproteinase 1/metabolism , Tissue Inhibitor of Metalloproteinase-2/genetics , Tissue Inhibitor of Metalloproteinase-2/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Tumor Necrosis Factor-alpha/metabolism , Matrix Metalloproteinase 8/metabolism , Cartilage, Articular/pathology , Knee Joint/pathology , Synovial Membrane/metabolism , Cartilage Diseases/pathology , Interleukin-1/metabolismABSTRACT
It is difficult to evaluate the pre-symptomatic state of mental disorders and prevent its onset. Since stress could be a trigger of mental disorders, it may be helpful to identify stress-responsive biomarkers (stress markers) for the evaluation of stress levels. We have so far performed omics analyses of the rat brain and peripheral blood after various kinds of stress and have found numerous factors that respond to stress. In this study, we investigated the effects of relatively moderate stress on these factors in the rat to identify stress marker candidates. Adult male Wistar rats underwent water immersion stress for 12 h, 24 h, or 48 h. Stress caused weight loss and elevated serum corticosterone levels, and alterations regarded as anxiety and/or fear-like behaviors. Reverse-transcription PCR and Western blot analyses revealed significant alterations in the expressions of hippocampal genes and proteins by the stress for no longer than 24 h, such as mitogen-activated protein kinase phosphatase 1 (MKP-1), CCAAT/enhancer-binding protein delta (CEBPD), small ubiquitin-like modifier proteins 1/sentrin-specific peptidase 5 (SENP5), matrix metalloproteinase-8 (MMP-8), kinase suppressor of Ras 1 (KSR1), and MKP-1, MMP-8, nerve growth factor receptor (NGFR). Similar alterations were observed in three genes (MKP-1, CEBPD, MMP-8) in the peripheral blood. The present results strongly suggest that these factors may serve as stress markers. The correlation of these factors in the blood and brain may enable the evaluation of stress-induced changes in the brain by blood analysis, which will contribute to preventing the onset of mental disorders.