Matrix Metalloproteinase-2 Staining Research Articles

Histological Effect of Emodin on Acute Vocal Fold Injury in a Rat Model

Emodin has beneficial effects on wound healing and reduces excessive fibrosis during tissue regeneration. Its positive effects on the wound-healing process were demonstrated on human fibroblasts. The aim of the present study was to evaluate the effectiveness of emodin application on acute vocal fold injury. Twenty-four Wistar albino rats were divided into three groups: control, sham, and emodin group. The glottis was examined using a 30°-2.7mm diameter telescope, and vocal folds was unilaterally wounded by a microscissor through the entire layer of the lamina propria down to the vocalis muscle. While no procedure(no acute injury of the vocal fold or an injection of saline/emodin) was applied to the control group, 0.5 cc of saline was injected into the sham group and 0.5 cc of emodin in the emodin group, just lateral to the vocal folds, with 27 gauge injectors. Animals were sacrificed on the 21st day after the procedure. After excised larynx experiments, serial sections were prepared from the vocal fold. Hematoxylin eosin and immunohistochemical staining were performed and fibroblast density, lamina propria thickness, and vessel formation were graded from 0 (none) to 3 (severe reaction). Transforming growth factor-beta 1 (TGF-β1) and matrix metalloproteinase-9 (MMP-9) staining was used for immunohistochemical examinations. Four-point scoring scale for intensity being scored as 0 (no staining) to 3 (severeley stained) to quantify immonuhistochemical reaction. This scoring system was applied to vocal fold epithelium, lamina propria, vessel wall, and vocalis muscle tissues. The groups were compared with the Kruskal Wallis and Dunn tests. Histologically, there was no significant difference (P>0.05) between the sham group and the emodin group in terms of fibroblast density, vessel formation, and lamina propria thickness. These parameters were higher (P<0.05) in both groups compared to the control group. In the lamina propria and vessel wall, MMP-9 staining was more intense in the emodin group than in the sham group. TGF-β1 staining of lamina propria, epithelial tissue, and vocalis muscle was significantly more intense in the emodin group than in the other groups. Emodin induced MMP-9 and TGF-1 staining in the vocalis muscle and epithelium, as well as TGF-1 staining in the lamina propria. In terms of fibroblast density, new vascular creation, and LP thickness in acute vocal fold damage, there was no difference between saline administration and emodin injection. It may increase fibroblast activation in the acute phase of wound healing, but its long-term effects should be further investigated.

Extracellular Vesicles Derived from Adipose-Derived Stem Cells Accelerate Diabetic Wound Healing by Suppressing the Expression of Matrix Metalloproteinase-9.

The healing of diabetic wounds is poor due to a collagen deposition disorder. Matrix metalloproteinase-9 (MMP-9) is closely related to collagen deposition in the process of tissue repair. Many studies have demonstrated that extracellular vesicles derived from adipose-derived stem cells (ADSC-EVs) promote diabetic wound healing by enhancing collagen deposition. In this study, we explored whether ADSC-EVs could downregulate the expression of MMP-9 in diabetic wounds and promote wound healing by improving collagen deposition. The potential effects of ADSC-EVs on MMP-9 and diabetic wound healing were tested both in vitro and in vivo. We first evaluated the effect of ADSC-EVs on the proliferation and MMP-9 secretion of HaCaT cells treated with advanced glycation end product-bovine serum albumin (AGE-BSA) using CCK-8, western blot and MMP-9 enzyme-linked immunosorbent assay(ELISA). Next, the effects of ADSC-EVs on healing, re-epithelialisation, collagen deposition, and MMP-9 concentration in diabetic wound fluids were evaluated in an immunodeficient mouse model via MMP-9 ELISA and haematoxylin and eosin, Masson's trichrome, and immunofluorescence staining for MMP-9. In vitro, ADSC-EVs promoted the proliferation and MMP-9 secretion of HaCaT cells. In vivo, ADSC-EVs accelerated diabetic wound healing by improving re-epithelialisation and collagen deposition and by inhibiting the expression of MMP-9. ADSC-EVs possess the potential of healing of diabetic wounds in a mouse model by inhibiting downregulating MMP-9 and improving collagen deposition. Thus, ADSC-EVs are a promising candidate for the treatment of diabetic wounds.

Expression and Prognostic Value of Small Mothers Against Decapentaplegic 7, Matrix Metalloproteinase 2, and Matrix Metalloproteinase 9 in Resectable Pancreatic Ductal Adenocarcinoma.

Thus far, expression, clinicopathologic, and prognostic implication of small mothers against decapentaplegic 7 (Smad7), matrix metalloproteinase 2 (MMP2), and matrix metalloproteinase 9 (MMP9) in pancreatic ductal adenocarcinoma (PDAC) were rarely investigated or controversial. Expression of Smad7, MMP2, and MMP9 was detected using immunohistochemistry in tissue microarrays based on 322 patients with curatively resected PDAC. Their expression pattern, clinicopathologic, and prognostic relevance were further evaluated. Smad7 expression was found to be lower in tumor than in adjacent nontumor tissues, whereas tumoral MMP2 and MMP9 staining scores were much higher than in adjacent nontumor ones. Furthermore, Smad7 was negatively associated with serum carbohydrate antigen 19-9 level. Univariate survival analyses showed that patients with high Smad7 tumors had significantly better disease-specific survival (P = 0.0007), whereas MMP2 and MMP9 predicted poor disease-specific survival (P = 0.0211 and 0.0404). In multivariate Cox regression test, Smad7 was an independent prognostic indicator (P = 0.021). In addition, these 3 proteins were also prognostic in many subgroups. Smad7 and MMP2/9 significantly predict good or poor prognosis in resectable PDAC, respectively. Therefore, the genes might serve as a tool or targets for molecular therapy in PDAC.

The elevated expression of TLR4 and MMP9 in human abdominal aortic aneurysm tissues and its implication

BackgroundToll-like receptor 4 (TLR4) and matrix metalloproteinase 9 (MMP9) have been investigated to play significant roles in the formation of abdominal aortic aneurysm (AAA). But the reports on the expression pattern of TLR4 and MMP9 in human AAA specimens were relatively scant. The aim of this study was to make a detailed analysis of TLR4 and MMP9 expression in situ and their association with clinical parameters involved in human AAA.Methods40 AAA specimens were obtained from full-thickness aneurysmal tissues at the maximal dilation area during the open surgical repair, and 8 non-aneurysmal abdominal aortas from transplant donors served as controls. Expression of TLR4 and MMP9 protein was determined by immunohistochemistry.ResultsThere were increased levels of TLR4 and MMP9 expression in human AAA tissues. Compared with macrophages or SMCs, lymphocytes showed a higher positive rate of TLR4 and MMP9 staining, and an elevated ratio of high MMP9 expression (all P < 0.05). There existed a significant association between TLR4 and MMP9 expression (r = 0.767, P < 0.001), and both TLR4 and MMP9 levels were statistically related to circulating CRP. Moreover, TLR4 expression in situ indicated a positive correlation with its serum level (r = 0.654, P = 0.006). Multiple analysis revealed that high TLR4 expression in situ was associated with the risk of large AAA (OR = 6.211, 95%CI = 1.226–31.480, P = 0.027), while high MMP9 expression was correlated to the presence of thrombus within AAA (OR = 5.494, 95%CI = 1.181–25.562, P = 0.030), separately compared with their low expression.ConclusionsThis study confirmed the overexpression of TLR4 and MMP9 in human AAA tissues, and their close relationship implying in the pathogenesis of AAA. We further provided evidence that TLR4 had a potential effect on AAA size and MMP9 could influence the occurrence of thrombus within AAA.

Dexamethasone-loaded thermo-sensitive hydrogel attenuates osteoarthritis by protecting cartilage and providing effective pain relief.

BackgroundWe utilized the destabilization of medial meniscus (DMM)-induced mice to illustrate the osteoarthritis (OA) suppressing and pain-relieving effects of a novel prolonged-release intra-articular (IA)-dexamethasone-loaded thermo-sensitive hydrogel (DLTH).MethodsThe effects of temperature and pH on DLTH formation and in vitro DLTH release profile were assessed. C57BL/6J mice were randomly divided into three groups: Ctrl group, Model group and DLTH group. The DLTH group received joint injections of 10 µL DLTH (1 mg/kg) into the right knee once a week from week 2 to week 11. We performed micro-computed tomography (Micro-CT) and histological analyses of safranin O-fast green, hematoxylin and eosin, and tartrate-resistant acid phosphatase in knee joints. We also carried out immunohistochemical (IHC) staining for matrix metalloproteinase-9 (MMP-9), MMP-13, and a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) in cartilage and Ki-67 in synovia. Pain behavioral testing was carried out in all mice. The serum content of prostaglandin E2 (PGE2) and real-time polymerase chain reaction (PCR) of inflammatory cytokines and pain-related factors in dorsal root ganglia (DRGs) were evaluated.ResultsIt took 20 minutes to form DLTH at pH 7.0 and 37 °C. The cumulative release profiles of dexamethasone (Dex) from DLTH at 37 °C revealed a rapid release in the first 24 h and a sustained slow release for 7 days. In vivo study illustrated that DLTH attenuated OA bone destruction and ameliorated synovitis and progression of OA in DMM-induced mice. The chondroprotective effects of DLTH were mediated by decreased expressions of MMP-9, MMP-13, and ADAMTS-5. The results showed that IA-DLTH exerted pain-relieving effects in OA mice. Upregulation of nociceptive response time (NRT) and downregulations of serum PGE2, inflammatory factors, and pain-related mediators in DRGs of mice in the DLTH group were recorded.ConclusionsData presented in this study elucidated that DLTH exhibited a long and lasting Dex release and it is a potential sustainable drug delivery system (DDS) to treat OA locally. IA-DLTH injection exerted chondroprotective and pain-relieving effects in DMM-induced arthritis. The involvement of MMP-9, MMP-13, ADAMTS-5, and inflammatory and pain-related factors, may account for the suppression of OA progression and pain.

Conjunctival Matrix Metalloproteinase-9 Clinical Assessment in Early Ocular Graft versus Host Disease.

Purpose Graft versus Host Disease (GVHD) typically affects the ocular surface, with a presentation resembling Dry Eye Disease (DED). Although the etiopathology is not completely known, the conjunctiva might be a key site of T-cell activation. The differential diagnosis might be tricky at early stages, because of the lack of dedicated clinical and laboratory tests. To meet these needs, we evaluated the suitability of ocular surface matrix metalloproteinase-9 (MMP-9) clinical test. Methods Consecutive GVHD patients, referred to IRCCS San Raffaele Scientific Institute, were recruited. DED patients served as controls. MMP-9 was tested through InflammaDry immunoassay kit in both groups; Ocular Surface Disease Index (OSDI) questionnaire, tear osmolarity, fluorescein Tear Break-up Time (TBUT), corneal and conjunctival staining, and Schirmer test I were also collected. Parametric and nonparametric statistical tests were used to analyze the intergroup differences; Receiver Operating Characteristics (ROC) curve analysis was carried out to perform sensitivity and specificity evaluations. Results Forty-five GVHD and 40 DED patients were included. MMP-9 expression was significantly higher in GVHD group than in DED (84.4% vs 33%, p ≤ 0.001). Corneal and conjunctival staining scores resulted worse in GVHD than in DED (0.95 ± 1.16 vs 0.40 ± 0.63, p=0.02; 0.77 ± 0.42 vs 0.40 ± 0.49, p=0.0005, respectively). No significant differences regarded the other collected parameters. GVHD group was characterized by positive correlations between MMP-9 and conjunctival staining (rho = 0.55, p=0.0002) and between MMP-9 and OSDI (rho = 0.3, p=0.01); a faint inverse correlation was found between MMP-9 and Schirmer test (rho = −0.25, p=0.04). Conclusion MMP-9 has a role in physiologic cellular remodeling; when a proinflammatory stimulus occurs, MMP-9 molecules are overreleased in the extracellular matrix. The positive expression of MMP-9 in GVHD may be interpreted as the consequence of a T-cell aggression against self-antigens and may be considered a reliable biomarker to detect ocular surface inflammation in GVHD, even in early stages of the disease.

Expression of Matrix Metalloproteinase-9 in Different Histopathological Variants of Ameloblastoma: A Cross-sectional Study

Introduction: Ameloblastomas are benign and the most common odontogenic neoplasms with many histopathological subtypes depending on the predominant pattern. They are known for their aggressive behaviour. As ameloblastomas have a high rate of recurrence, it is necessary to understand the biological behaviour of these neoplasms. Matrix Metalloproteinase-9 (MMP-9) is an enzyme that belongs to Metalloproteinases family and is known to degrade the Extracellular Matrix (ECM) and facilitate tumour progression. Evaluation of the expression of MMP-9 in ameloblastomas could contribute in understanding its biological behaviour. Aim: To analyse the expression of MMP-9 in different histopathological variants of ameloblastoma. Materials and Methods: A cross-sectional observational study was done in the Department of Oral Pathology and Microbiology, Bharati Vidyapeeth Deemed to be University, Dental College and Hospital, Pune, India. A total of 30 cases were selected for this study. The study was carried out for a duration of two years from April 2018 to April 2020. The MMP-9 expression was studied by immunohistochemical staining. The statistical comparison was done using Chi-square test between groups, p-value &lt;0.05 was considered significant. The data was statistically analysed using Statistical Package for Social Sciences (SPSS) version 22.0, IBM Corporation, USA for MS Windows. Results: A total of 30 paraffin embedded archival tissue blocks were selected for this study. Among them nine cases were of Plexiform Ameloblastoma, eight of Unicystic Ameloblastoma, three of Acanthomatous Ameloblastoma, four of Desmoplastic Ameloblastoma and six were of Follicular Ameloblastomas. The MMP-9 showed variable expression in different histopathological subtypes of ameloblastoma. This difference was statistically significant between Plexiform and Acanthomatous as well as Plexiform and Follicular variants (p&lt;0.05). A 66.7% (6 out of 9 samples) of Plexiform Ameloblastoma showed intense staining for MMP-9. Conclusion: Expression of MMP-9 varies in different histopathological variants of ameloblastoma and may not have an association with biological behaviour and aggressiveness.

Matrix Metalloproteinase-2 Expression as a Prognostic Marker in Breast Carcinomas

Introduction: Matrix Metalloproteinase-2 (MMP-2) is over expressed in a variety of malignant tumours and their expression and activity are often associated with tumour aggressiveness and a poor prognosis. It serves as a prognostic marker in breast carcinoma regardless of patient age, disease stage, malignancy grade, or hormone receptor status and modulation of MMP-2 expression and activation provides a new mechanism for breast cancer treatment. Aim: To evaluate the expression of MMP-2 in breast carcinoma tumour cells and peritumoural stroma and also to analyse the findings with the existing other prognostic markers of mammary carcinoma. Materials and Methods: The present study was a retrospective study conducted on paraffin blocks of 90 cases of invasive breast carcinoma specimens received in the Department of Pathology, Sri Raamachandra Institute of Higher Education and Research a tertiary care centre in Chennai, Tamil Nadu, India, from January 2012 to June 2017. Immunohistochemical staining for MMP-2, Oestrogen Receptor (ER), Progesterone Receptor (PR) and Human Epidermal Growth Factor Receptor 2 (HER2) was done. Statistical analysis was done on the data collected by using the software GNU-PSPP version 0.10.1. Pearson Chi-square test was used to determine significant clinicopathological differences between MMP-2 expression in positive and negative tumours. Differences were considered statistically significant when p-value was &lt;0.05. Results: The study included 90 cases of histological proven invasive breast carcinoma. The study parameters include age, clinical staging, histopathological grade, lymphnode status, molecular subtype and MMP-2 expression in invasive breast carcinoma. Out of 90 cases, 62 cases were positive for MMP-2 and 18 cases were positive for peritumoural stroma, 30 cases were negative for MMP-2 in tumoural cells and 72 cases were negative surrounding the peritumoural stroma. Conclusion:Present study showed high MMP-2 immunohistochemical expression in breast carcinomas. There was a statistically significant association of increased MMP-2 expression with tumour stage. There was also a statistically significant association of MMP-2 in the tumour and stroma with High Grade Ductal Carcinoma In Situ (HGDCIS). There was an increased expression of MMP-2 in Luminal A subtype. Low expression was seen in the other molecular subtypes. In view of these findings and association with other studies in the literature, the present study demonstrates that the expression of MMP-2 in tumour and stromal cells could serve as a parameter of poor prognosis in breast cancer.

Immunohistochemical and Zymographic Analyses of Matrix metalloproteinase-2 (MMP-2) and Matrix Metalloproteinase-9 (MMP-9) in Bovine Papillomatous Digital Dermatitis

Bovine digital dermatitis (BDD) is an important health issue of dairy cattle. Painful papillomatous tissue proliferations are observed in hind feet of affected animals. The pathogenesis of this papillomatous lesion in BDD is still poorly understood. Matrix metalloproteinases are a group of enzymes for tissue remodeling in both health and disease. In the submitted study, immune expressions of MMP-2 and MMP-9 enzymes and their zymographic activities were evaluated. Higher immune staining of MMP-2 and MMP-9 were found in BDD samples when compared with healthy skin samples. The differences observed in MMP-2 and MMP-9 staining intensity and distributions were statistically significant (p&amp;lt; 0.05, p&amp;lt;0.05 respectively). Zymographic analyses revealed skin samples from both healthy and affected animals had pro-active and active forms of MMP-2 and MMP-9 enzymes. Immunohistochemistry results indicate that significant increase in MMP-2 and MMP-9 enzymes in BDD samples. These results suggest MMP-2 and MMP-9 might have roles in the pathogenesis of BDD.

Immunohistochemical analysis of matrix metalloproteinase-9 predicts papillary thyroid carcinoma prognosis.

The present study aimed to explore the association between immunohistochemical matrix metalloproteinase-9 (MMP-9) expression and the clinicopathological characteristics of patients with papillary thyroid carcinoma (PTC), and to determine whether it may be used as a diagnostic or prognostic tool for PTC. Immunohistochemical staining of MMP-9 was performed in thyroid tissues obtained from 112 patients with PTC and 42 subjects with benign thyroid nodules (BTNs). The receiver operating characteristic curve was used to evaluate the legitimacy of MMP-9 as a diagnostic tool for PTC, and a predictor for structurally persistent/recurrent disease (SPRD) and disease status. Cox regression was applied to identify the risk factors of disease status and SPRD. The present study revealed that MMP-9 was overexpressed in PTC tissues, compared with in BTN tissues. Furthermore, MMP-9 scores yielded an area under the curve (AUC) of 0.842 (95% CI, 0.776–0.908) for differentially diagnosing PTC from BTN. In addition, the MMP-9 score was greater if patients previously had central lymph node metastasis, lateral lymph node metastasis or an advanced tumor-node-metastasis stage (III+IV). When MMP-9 was employed to predict disease status and SPRD, an AUC of 0.811 (95% CI, 0.706–0.917) and 0.806 (95% CI, 0.620–0.992) was obtained, respectively. A tumor size of >2 cm and an MMP-9 staining score of ≥6 were independent risk factors for predicting disease status, whereas vascular invasion and an MMP-9 staining score of ≥8 were risk factors for predicting SPRD. Furthermore, an MMP-9 staining score of ≥6 and ≥8 indicated shortened disease-free survival and survival without SPRD, respectively. In conclusion, the assessment of MMP-9 expression in thyroid carcinoma samples may represent a potential and supplementary tool for the diagnosis and prognostic prediction of PTC.

Eosinophil-derived neurotoxin enhances airway remodeling in eosinophilic chronic rhinosinusitis and correlates with disease severity.

Eosinophilic chronic rhinosinusitis (ECRS) is a subtype of chronic rhinosinusitis (CRS) that is characterized by intractable nasal polyp formation. Eosinophil-derived neurotoxin (EDN) is an eosinophil granule protein that is closely related to allergic inflammation, but the pathological implications of EDN in ECRS remain unknown. In this study, we evaluated the function of EDN in ECRS pathogenesis and assessed its potential as a disease activity marker. Serum EDN levels were significantly higher in patients with ECRS than in those with other nasal and paranasal diseases, and were positively correlated with clinical disease activity. Production of EDN from isolated human eosinophils was induced by stimulation with IL-5 in vitro. Human nasal epithelial cells were stimulated with EDN, and the resultant changes in gene expression were detected by RNA sequencing. Pathway analysis revealed that the major canonical pathway affected by EDN stimulation was 'regulation of the epithelial-mesenchymal transition pathway'; the only gene in this pathway to be up-regulated was matrix metalloproteinase 9 (MMP-9). Consistent with this, immunostaining analysis revealed intense staining of both EDN and MMP-9 in nasal polyps from patients with ECRS. In conclusion, our data demonstrate that serum EDN level is a useful marker for the evaluation of ECRS severity. Furthermore, EDN induces production of MMP-9 from the nasal epithelium, which may be involved in the pathogenesis of ECRS.

Iron Overload Exacerbates the Risk of Hemorrhagic Transformation After tPA (Tissue-Type Plasminogen Activator) Administration in Thromboembolic Stroke Mice.

Background and Purpose- Recanalization with tPA (tissue-type plasminogen activator) is the only pharmacological therapy available for patients with ischemic stroke. However, the percentage of patients who may receive this therapy is limited by the risk of hemorrhagic transformation (HT)-the main complication of ischemic stroke. Our aim is to establish whether iron overload affects HT risk, to identify mechanisms that could help to select patients and to prevent this devastating complication. Methods- Mice fed with control or high-iron diet were subjected to thromboembolic stroke, with or without tPA therapy at different times after occlusion. Blood samples were collected for determination of malondialdehyde, matrix metalloproteinases, and fibronectin. Brain samples were collected 24 hours after occlusion to determine brain infarct and edema size, hemorrhage extension, IgG extravasation, and inflammatory and oxidative markers (neutrophil infiltration, 4-hydroxynonenal, and matrix metalloproteinase-9 staining). Results- Despite an increased rate of recanalization, iron-overload mice showed less neuroprotection after tPA administration. Importantly, iron overload exacerbated the risk of HT after early tPA administration, accelerated ischemia-induced serum matrix metalloproteinase-9 increase, and enhanced basal serum lipid peroxidation. High iron increased brain lipid peroxidation at most times and neutrophil infiltration at the latest time studied. Conclusions- Our data showing that iron overload increases the death of the compromised tissues, accelerates the time of tPA-induced reperfusion, and exacerbates the risk of HT may have relevant clinical implications for a safer thrombolysis. Patients with stroke with iron overload might be at high risk of HT after fibrinolysis, and, therefore, clinical studies must be performed to confirm our results.

Early changes in the apparent diffusion coefficient and MMP-9 expression of a cervical carcinoma U14 allograft model following irradiation.

A cervical carcinoma allograft model was designed to assess the correlation between early changes in the apparent diffusion coefficient (ADC) values on diffusion-weighted magnetic resonance imaging (DW-MRI) and the expression of matrix metalloproteinase-9 (MMP-9) in tumors. BALB/c mice with U14 tumor allografts on the right rear flank were irradiated with a single 20 Gy dose. All tumor-bearing mice were subjected to DW-MRI, followed by calculation of the ADC values and characterization of the T1 and T2 relaxation time constants. Pre- and post-irradiation ADC values were compared with the tumor volume, and the immunohistochemical staining of MMP-9 and hematoxylin-eosin (HE) staining of tumor allografts. However, no correlations between the pre-treatment ADC values and changes in tumor volumes following irradiation were observed. Notably, the mean ADC value was significantly higher in the irradiated tumors (0.756±0.102×10-3 mm2/sec) as compared with those in the untreated tumors (0.501±0.052×10-3 mm2/sec; P=0.002; r=0.682). Additionally, immunohistochemical staining demonstrated that MMP-9 expression in the irradiated tumors was significantly increased. The mean ADC value was significantly higher in the irradiated tumors with high MMP-9 expression levels (0.815±0.112×10-3 mm2/sec), as compared with in the untreated tumors with low MMP-9 expression levels (0.631±0.068×10-3 mm2/sec). Quantitative analysis determined that the ADC values were correlated with MMP-9 expression (r=0.752; P=0.003). Combined, these results suggest that radiation-induced increases in MMP-9 expression levels may be responsible for early changes in the mean ADC value and the response to irradiation in cervical carcinoma.

High Grade T1 Papillary Urothelial Bladder Cancer Shows Prominent Peritumoral Retraction Clefting.

Differentiation of noninvasive from invasive papillary urothelial carcinoma can be challenging due to inability of proper orientation and thermal damage of transurethrally obtained material. The aim of this study was to analyze the presence and extent of peritumoral retractions in pT1 compared to pTa papillary urothelial carcinoma. Since peritumoral retractions may result from altered expression profiles of extracellular matrix proteins, we additionally analyzed the expression of matrix metalloproteinase 2 (MMP-2) and interleukin 8 (IL-8) in these tumors. The study comprised 50 noninvasive (pTa) and 50 invasive (pT1) cases of transurethrally obtained primary papillary urothelial carcinomas. The invasive nature of nests showing peritumoral retractions was confirmed immunohistochemically using antibody against collagen IV. Staining for MMP-2 and IL-8 was evaluated semiquantitatively using immunohistochemical staining index, calculated by multiplying the percentage of positive cells and staining intensity. Peritumoral retractions were found in 32% of pT1 carcinomas but in none of the pTa carcinomas. All tumors showing peritumoral retraction were high grade tumors. There was no statistically significant correlation between the expression of MMP-2 or IL-8 and the presence of peritumoral retractions or stage of the tumor (pTa vs. pT1). A statistically significant but weak correlation was found between MMP-2 and IL-8 expression (χ2-test, p=0,015). There was no statistically significant correlation between the presence of peritumoral retractions or MMP-2 expression and tumor recurrence and progression. Our study shows that, in doubtful cases, when differentiating between pTa and pT1 stages of papillary urothelial carcinoma, the presence of peritumoral retractions could favor the diagnosis of invasive neoplasm.

Correlation of matrix metalloproteinase-2 and -9 expression with recurrences in primary spontaneous pneumothorax patients

Primary spontaneous pneumothorax (PSP) is a common benign disorder. However, unpredictable recurrence is a major concern for most patients. The aim of the present study was to assess the role of matrix metalloproteinase-2 (MMP-2) and MMP-9 in alveolar macrophages of patients with PSP and its relationship with recurrence. Ninety-two patients who received needlescopic video-assisted thoracoscopic surgery (NVATS) wedge resection of lung with identifiable blebs for PSP were enrolled for the study. Immunohistochemistry was performed to evaluate the expression of MMP-2 and MMP-9 in lung tissues of patients with PSP. The result was correlated with clinicopathological variables and recurrence rates by the chi-square test. The value of MMP-2 and MMP-9 for overall recurrence was evaluated by univariate and multivariable Cox regression analyses. The MMP-2 and MMP-9 staining was predominantly observed in alveolar macrophages of patients with PSP. We found that MMP-2 (recurrence: P<0.001; smoking status: P=0.029) and MMP-9 (recurrence: P=0.001; smoking status: P=0.045) expression in PSP, especially male patients, was significantly correlated with recurrence and smoking status. In the multivariate analyses, MMP-2 [hazard ratio (HR) =2.83; 95% confidence interval (CI): 1.37-5.85, P=0.005) and MMP-9 (HR =2.25; 95% CI: 1.19-4.24, P=0.013) were statistically significant risk factors for overall recurrence in PSP patients. High expression levels of MMP-2 and MMP-9 showed a positive correlation with recurrence in PSP patients. Further studies are required to test whether inhibition of MMP-2 and MMP-9 expression renders a promising approach for reducing the risk of PSP recurrence in the future.

Association of MMP-2 and MMP-9 expression with recurrences in primary spontaneous pneumothorax

Primary spontaneous pneumothorax (PSP) is a common benign problem. However, PSP recurrence is still a troublesome complication for most patients. This study intended to determine the role of matrix metalloproteinase-2 (MMP-2) and MMP-9 in type II pneumocytes of patients with PSP and its relation with recurrence. Ninety-one patients who had undergone needlescopic video-assisted thoracoscopic surgery wedge resection of lung with identifiable blebs for PSP were included in this study. Immunohistochemical (IHC) staining was used to measure the expression of MMP-2 and MMP-9 in lung tissues of PSP patients. The results were further correlated with clinicopathological parameters and recurrence rates using chi-square or Fisher's exact test. The value of MMP-2 and MMP-9 for overall recurrence was analyzed by univariate and multivariable Cox regression model. IHC data revealed that MMP-2 and MMP-9 staining was predominantly observed in type II pneumocytes of patients with PSP. We found that MMP-2 and MMP-9 expression in PSP, especially male PSP patients, was significantly correlated with recurrence. In the univariate and multivariate analyses, MMP-2 and MMP-9 were statistically significant risk factors for overall recurrence in PSP patients. Therefore, high expression levels of MMP-2 and MMP-9 in type II pneumocytes show a positive correlation with PSP recurrence risk. Further studies are needed to validate whether reduction of MMP-2 and MMP-9 expression may be a promising way for decreasing the risk of PSP recurrence in the future.

Coexpressed High Levels of VEGF-C and Active MMP-9 Are Associated With Lymphatic Spreading and Local Invasiveness of Papillary Thyroid Carcinoma.

Papillary thyroid carcinoma (PTC) usually has a good prognosis, but some patients develop an aggressive course of the disease, leading to a poor outcome. Vascular endothelial growth factor C (VEGF-C) and matrix metalloproteinase 9 (MMP-9) have been shown to play roles in tumor progression in various human malignancies. We analyzed VEGF-C and active MMP-9 expression profiles in PTC samples using immunohistochemistry and Western blotting. Immunohistochemistry showed positive staining for VEGF-C and active MMP-9 in 83% and 57% of PTCs, respectively (n = 60), with a positive correlation between their expression levels (Spearman, P < .001). Concomitant high expression of VEGF-C and active MMP-9 correlated with the presence of lymph node metastasis (P = .005), pT status (P = .004), pTNM tumor stage (P = .005), and particularly the degree of tumor infiltration (P < .001, Fisher exact test). Densitometric analysis of Western blot bands confirmed correlation between VEGF-C and active MMP-9 expression (Wilcoxon and Spearman tests) and significant association with the clinicopathologic parameters (Mann-Whitney and Kruskal-Wallis tests). Association of coexpressed high levels of VEGF-C and active MMP-9 with lymphatic spreading and local invasiveness of PTC suggests their potential usefulness as predictive biomarkers of aggressive PTC behavior.

Worse prognosis in breast cancer patients can be predicted by immunohistochemical analysis of positive MMP-2 and negative estrogen and progesterone receptors.

Breast cancer is the most cause of death, and approximately 90% of these deaths are due to metastases. Matrix metalloproteinase-2 (MMP-2) gelatinase activity is able to degrade a major constituent of the tumor microenvironment, type IV collagen. Two well-established proteins used as markers in clinical practice for breast cancer are the receptors for estrogen (ER) and progesterone (PR). Although the presence of these receptors has been associated with a better prognosis, loss of these proteins can occur during tumor progression, with subsequent resistance to hormone therapy. To study the correlation among MMP-2, ER, and PR, as well as the establishment of the metastatic process in primary breast tumors. Breast cancer samples (n=44) were analyzed by immunohistochemistry for MMP-2, ER, and PR. We observed that 90% of patients who had metastases and died showed positive staining for MMP-2 (p=0.0082 for both). Using Kaplan-Meier analysis, we found that negative ER patients who were also positive for MMP-2 had even worse disease-free survival (DFS) and overall survival (OS) (p= 0.012 and p=0.005, respectively). Similar results were found in PR-negative patients for DFS (a trend p=0.077) and OS (p=0.038). Regardless of our small sample size (n=44), the data obtained strongly suggest that MMP-2 in combination with already well-established markers could help to predict the emergence of metastases and death in patients with breast cancer.

Expression of SKA1 and MMP–9 in primary salivary adenoid cystic carcinoma: Correlation with tumor progression and patient prognosis

ABSTRACTConclusion The spindle and kinetochore-associated complex sub unit 1(SKA1) and matrix metalloproteinase-9 (MMP-9) are highly expressed in salivary adenoid cystic carcinoma (SACC), and SKA1 may be the novel, promising prognostic factor for clinical outcomes in head and neck adenoid cystic carcinoma patients. Objectives This study aimed to investigate the expression of SKA1 and MMP-9 in SACC tissues and the clinical significance. Methods SKA1 and MMP-9 expression in samples from 42 cases of SACC and 20 subjects with the normal tissue adjacent to carcinoma were detected by immunohistochemical analysis. Results The positive rate of SKA1 and MMP-9 staining was 78.6% and 66.7% in SACC, respectively, significantly higher than in normal salivary gland tissues (p < 0.001). Chi-square test showed that there was no significant correlation between SKA1 expression and MMP-9 expression in SACC tissues. However, SKA1 and MMP-9 expression was positively associated with advanced stage and solid histological pattern of SACC (p < 0.05). In addition, SKA1 and MMP-9 expression was positively associated with recurrence and perineural invasion, and survival time, respectively.

Treatment of hypertrophic scars and keloids by fractional carbon dioxide laser: a clinical, histological, and immunohistochemical study.

Treatment of keloids (K) and hypertrophic scars (HTS) is challenging. A few case reports reported good results in HTS treated by fractional CO2 laser. The aim of the present study was the assessment of the clinical response as well as histological changes in K and HTS treated by fractional CO2 laser and the role of matrix metalloproteinase 9 (MMP9) in the response. A randomized half of the scar was treated by fractional CO2 laser in 30 patients (18 K, 12 HTS) for a total of four sessions 6 weeks apart. Vancouver scar score (VSS) was done before and 1, 3, and 6 months after the last laser session by a blinded observer. Biopsies taken from normal skin, untreated scar, and treated scar tissue 1 and 3 months after the laser sessions were stained by HX & E for histological changes and Masson trichrome for collagen fiber arrangement. Immunohistochemical staining for MMP9 was done in before and 1 month after samples. Quantitative morphometric analysis was done for collagen and MMP9 by image analyzer. Nineteen patients completed the 6-month follow-up period (12 K, 7 HTS). VSS score was significantly lower in the treated compared to untreated areas after 3 and 6 months in both K and HTS but was mainly due to improved pliability of the scar. Histologically, dense inflammatory infiltrate and increased vascularity was apparent 1 month after laser sessions and disappeared at 3 months. Thinner better organized collagen bundle could be seen in 3 months after samples. MMP9 was significantly increased in after treatment samples but without significant correlation with VSS. Fractional CO2 resurfacing is safe but affects mainly pliability of K and HTS with collagen remodeling apparent 3 months after therapy. MMP9 may play a role in mechanism of action of CO2 laser in K and HTS.