Immunoexpression Of MMP-9 Research Articles

CXCR4 and MMP-9 Immunoexpression are Associated with Metastasis in Infiltrating Urothelial Carcinoma of The Bladder

Background: Infiltrating urothelial carcinoma of the bladder (IUCB) is an aggressive urinary tract cancer. Most of the patients were diagnosed with advanced stages, and many have metastasized. Metastasis is responsible for 90% of cancer deaths compared to the primary tumor itself. In previous research, the expression of CXCR4 and MMP-9 has been suggested to correlate with metastasis and prognosis in several cancers. This study aimed to analyze the association of CXCR4 and MMP-9 expression with metastasis in infiltrating urothelial carcinoma of the bladder. Methods: This cross-sectional study included 40 selected paraffin-embedded tissue blocks from histopathologically diagnosed IUCB patients, consisting of 20 cases of non-metastasizing and 20 cases of metastasizing IUCB. Immunohistochemical staining for CXCR4 and MMP-9 was performed in all cases. All data were analyzed using the Chi-Square test with p < 0.05 of a significant level and then processed using SPSS 24.0 for Windows.Results: High CXCR4 immunoexpression was found in 18 (45%) of all cases, and high MMP-9 immunoexpression was found in 19 (47.5%) cases. High immunoexpression of CXCR4 and MMP9 in infiltrating urothelial carcinoma of the bladder showed significant association with metastasis (p < 0.05). The stepwise logistic regression analysis revealed that both CXCR-4 and MMP-9 immunoexpressions were independent prognostic factors for metastasis.Conclusions: CXCR4 and MMP-9 play a role in the metastatic process in infiltrating urothelial carcinoma of the bladder.

Immunoexpression of MMP-8 and MMP-9 in chronic subdural hematoma.

To determine the possible role of matrix metallopeptidase (MMP)-8 and MMP-9 in the development of chronic subdural hematoma (CSDH), we investigated their expression in CSDH. In our previous study, we analyzed hematoma fluid and peripheral blood of 83 patients with CSDH, including 17 postoperative patients. Based on these results, we included 50 people in the normal group and analyzed 20 markers in the peripheral blood of each person. In order to identify representative markers, it was assessed by using overall differential gene expression. The concentration of MMP-8 was significantly higher in the normal group than that in the preoperative and postoperative groups. The concentration of MMP-9 was significantly lower in the normal group than in both preoperative and postoperative groups. Immunohistochemistry confirmed the expression of MMP-8 and MMP-9 in CSDH membranes. In conclusion, our results provide evidence of the expression of MMP-8 and MMP-9 in CSDH. In addition, the expression of MMP-8 and MMP-9 suggests angiogenesis in CSDH formation.

IMMUNOHISTOCHEMICAL EXPRESSION OF METALLOPROTEINASES AND TFN-Α BY MACROPHAGES AND LANGERHANS CELLS IN RADICULAR CYSTS

Objectives This research aimed to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMPs-9, -13), and tumor necrosis factor (TNF)-α in the lining epithelium of cystic lesions to elucidate the cyst expansion. Study Design A sectional and retrospective study was performed with 20 radicular cysts (RCs) and 20 residual radicular cysts (RRCs) by anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. Results The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared to RRCs (P = .011 and P = .012, respectively). An intense inflammatory infiltrate was significantly correlated with CD68 immunoexpression in RCs (P = .025). Macrophages demonstrated a significant positive correlation with MMP-13 (P = .015). A moderate correlation was observed between MMP-9 and MMP-13 (P = .01). TNF-α expression was more common in RCs (P = .001). Langerhans cells were more frequently expressed in atrophic epithelium (P = .041) and significantly correlated with TNF-α (P = .014) in RCs. Conclusions Macrophages induce a greater release of TNF-α. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared to RRCs. The microorganisms are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these cysts. This research aimed to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMPs-9, -13), and tumor necrosis factor (TNF)-α in the lining epithelium of cystic lesions to elucidate the cyst expansion. A sectional and retrospective study was performed with 20 radicular cysts (RCs) and 20 residual radicular cysts (RRCs) by anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared to RRCs (P = .011 and P = .012, respectively). An intense inflammatory infiltrate was significantly correlated with CD68 immunoexpression in RCs (P = .025). Macrophages demonstrated a significant positive correlation with MMP-13 (P = .015). A moderate correlation was observed between MMP-9 and MMP-13 (P = .01). TNF-α expression was more common in RCs (P = .001). Langerhans cells were more frequently expressed in atrophic epithelium (P = .041) and significantly correlated with TNF-α (P = .014) in RCs. Macrophages induce a greater release of TNF-α. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared to RRCs. The microorganisms are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these cysts.

Release of Matrix Metalloproteinases by Macrophages in Radicular Cysts and Residual Radicular Cysts.

Radicular cysts (RCs) and residual radicular cysts (RRCs) are the sequelae of dental caries and that leads to proliferation of epithelial rests of Malassez in periapical tissues. The aim was to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMP-9, MMP-13), and tumor necrosis factor-alpha (TNF-α) in the capsule and lining epithelium of cystic lesions. Twenty RCs and 20 RRCs were submitted to immunohistochemical analysis with anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. The Mann-Whitney test and the Spearman correlation test were used for analysis of the data (P<0.05). The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared with RRCs (P=0.011 and 0.012, respectively). The presence of an intense inflammatory infiltrate was significantly correlated with the immunoexpression of CD68 in RCs (P=0.025). Expression of CD68 showed a significant positive correlation with MMP-13 (P=0.015). A moderate correlation was observed between MMP-9 and MMP-13 (P=0.010). TNF-α expression was more common in RCs (P=0.001). CD1a was more frequently expressed in atrophic epithelium (P=0.041) and was significantly correlated with TNF-α (P=0.014). Langerhans cells induce a greater release of TNF-α which, in turn, is responsible for the stimulation of M1 macrophages. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared with RRCs. Therefore, the toxins of microorganisms present in highly inflamed RCs are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these lesions.

Evaluation of the Connective Tissue Wall in Sporadic Cases of Keratocystic Odontogenic Tumor (KCOT) Using MMP-9 and Confocal Microscopy: A Retrospective Study

Background: Keratocystic odontogenic tumor (KCOT) is an odontogenic lesion which manifests distinct biological behavior. Predominant studies in KCOT attribute this behavior to high epithelial proliferative capacity. Besides, a few studies facet loosely arranged collagen can contribute to the behavior of KCOT. Matrix metalloproteinases (MMP) are enzymes that degrade extracellular matrix components under both physiologic and pathologic conditions. The loosely arranged collagen in connective tissue wall of KCOT could be related to the degree of MMP-9 expression. Aim: To evaluate the arrangement of collagen fibers along with immunoexpression of MMP-9 and to relate to its neoplastic biologic behavior in sporadic cases of KCOT. Materials and Methods: KCOT ( n = 23) and dentigerous cyst (DC) ( n = 15) samples were processed for the following techniques: Masson’s trichrome stain for light microscopy, PMA-PSR stain for confocal microscopy, and MMP-9 for immunohistochemistry. Results: In Masson’s trichrome analysis, correlation of collagen fiber arrangement in the deeper regions with color intensity for KCOT was found to be statistically significant ( P = .033). In confocal microscopy, there was no difference between intensities in KCOT ( P = .990) and DC ( P = .233), respectively. The immunoexpression of MMP-9 in the connective tissue wall of DC (73.3%) was relatively higher than that of KCOT (60.8%). However, on comparison between KCOT and DC in the presence of inflammation, the immunoexpression of MMP-9 was higher in DC (100%) than KCOT (69.9%) and was statistically significant ( P = .028). Conclusion: It was concluded that the loose connective tissue wall in KCOT is because of the inherent nature of the lesion that could facilitate its biologic behavior. If inflammation is present, this could further aggravate the tumorigenic behavior.

Immunoexpression of bone resorption biomarkers in apical periodontitis in diabetics and normoglycaemics.

To compare the immunoexpression of RANK, MMP-9 and PTHrP in apical periodontitis lesions of diabetic and normoglycaemic individuals. Primary chronic apical periodontitis lesions associated with teeth indicated for extraction in 13 type 2 diabetic individuals and 13 normoglycaemic individuals who were screened for the glycaemic index and glycated haemoglobin (HbA1c) were analysed. Individuals with other systemic diseases and users of anti-inflammatories and/or antibiotics in the previous 3months were excluded. Silanized slides with paraffin sections were used for immunohistochemical reactions and stained with haematoxylin and eosin for histopathological classification. The images were analysed with an optical microscope, and the slides were subdivided into five large fields assigning scores (0-2), according to the number of positive markings for each antibody. Fisher's exact test evaluated the parameters: gender, type of lesion, location and position in the arch. Nonparametric Mann-Whitney test was used for age, HbA1c values and comparison of marker expression. The chi-squared test was used to associate the expression of the markers. And the Spearman's coefficient correlated the markers with the size of the periapical lesion. The samples consisted of 69% periapical granulomas and 31% periapical cysts in each group. RANK expression was considered weak/moderate and strong in, respectively, 62% and 38% of the cases in both groups. MMP-9 expression was weak/moderate and strong in, respectively, 38% and 62% of the cases from the diabetic group, in comparison with 38% and 38% in the normoglycaemics (24% cases from this group were negative). In contrast, PTHrP expression was negative, weak/moderate and strong in, respectively, 46%, 46% and 8% of the cases from the diabetic group, in comparison with 38% negative and 62% weak/moderate in normoglycaemics. Quantitative analysis revealed that there were no significant differences in the immunoexpression of RANK (P=0.26), MMP-9 (P=0.17) and PTHrP (P=0.43) between the groups. There was no significant correlation between the expression of bone resorption markers and the macroscopic size of the periapical lesions (P>0.05). The bone resorption mediators analysed had similar immunoexpression in the periapical lesions of diabetic and normoglycaemic individuals.

Immunoexpression of metalloproteinases 9 (MMP-9) and 2 (MMP-2) and their inhibitors (TIMP-1 and TIMP-2) in normal and neoplastic canine mammary tissue

Abstract The aim of this study was to perform the immunostaining of MMP-9 and MMP-2 and its inhibitors, TIMP-1 and TIMP-2, on normal and neoplastic canine mammary tissue in order to evaluate the behavior of these proteins in extracellular matrix (ECM) remodeling in different neoplastic mammary types. Thus, 48 samples of canine mammary tissue were analyzed, 14 of which complex carcinomas, 13 tubulopapillary carcinomas, six single adenomas and 15 normal mammary tissue. There were differences in MMP-9, TIMP-1 and TIMP-2 according to mammary histomorphology, and MMP-9 presented increased immunoexpression in epithelial and stromal cells in tubulopapillary and complex carcinomas. TIMP-1 exhibited reduced immunostaining in the stromal cells of the complex carcinomas and TIMP-2 enhanced immunostaining in the epithelial cells of tubulopapillary carcinomas. There was a positive correlation between MMP-9 and TIMP-1 in epithelial and stromal cells regarding immunostaining intensity and number of labeled cells in the normal breast. There was a positive correlation between MMP-9 and TIMP-2 in the epithelial cells of tubulopapillary carcinomas. It is concluded that balanced activity between MMP-9, MMP-2, TIMP-1 and TIMP-2 maintains normal canine mammary tissue homeostasis while increased immunoexpression of MMP-9 and TIMP-2 and reduced TIMP- 1 in carcinomas suggest a favorable condition for tumor evolution.

Immunoexpression of Matrix Metalloproteinase-9 in Colorectal Adenocarcinoma

Colorectal carcinoma is one of the most common malignant tumors of gastrointestinal tract and is a contributing factor of cancer mortality in Myanmar. Matrix metalloproteinase-9 is a proteolytic enzyme which breaks down extracellular matrix leading to tumor progression, contri-buting as a potential prognostic marker nowadays. The study was aimed to determine MMP-9 immunoexpression in colorectal adenocarcinoma and its association with Astler-Coller staging. A cross-sectional descrip-tive study was done on 42 specimens of colorectal adenocarcinoma. All tissue specimens were studied with haematoxylin and eosin to categorize histological types and grades. Out of 42 cases, 21.4% were well differentiated, 57.2% were moderately differentiated and 21.4% were poorly differentiated. According to Astler-Coller staging, 11.9% were found to be in stage B1, 40.5% in stage B2, 7.1% in stage C1, 33.3% in stage C2 and 7.1% in stage D. 81% (34/42) of colorectal carcinoma showed positive MMP-9 immunoexpression. Positive MMP-9 immuno-expression was seen in 91% of conventional adenocarcinoma, 40% of mucinous carcinoma and no cases signet ring carcinoma. The findings of the study pointed out that MMP-9 immunoexpression was positively associated with histological types of colorectal adenocarcinoma (p=0.001). MMP-9 immunoexpression was positive in 88.9% of well differentiated adenocarcinoma, 91.7% of moderately differentiated adenocarcinoma and 44.4% of poorly-differentiated adenocarcinoma (p=0.007). Regarding the immunoexpression of MMP-9 in different Astler-Coller staging, positive MMP-9 immunoexpression was seen in 60% of the cases in stage B1, 76.5% in stage B2, 66.7% in stage C1, 92.9% in stage C2 and 100% in stage D. The findings of the study can be extrapolated to predict prognosis and help in better management of colorectal adenocarcinoma by introducing targeted therapy against MMP-9 in future.

Presence of metalloproteinases 2 and 9 and 8-OHdG in the fibrotic process in skeletal muscle of Mdx mice

Inflammation and oxidative stress occurs in muscle of Duchenne muscular dystrophy (DMD). The relationship between a panel of biomarkers and the DMD outcome is necessary to indicate of disease progression and response to rehabilitation programs. The aim was to analyze the connective tissue of muscle of Mdx mice and immunoexpression of MMP-2, MMP-9, and 8-OHdG, which signalizes oxidative stress related to DNA damage. Biceps brachii of male C57BL/10 and C57BL/10-Dmdmdx mice was submitted to Hematoxylin-Eosin, Sirius red and immunohistochemistry (MMP-2, MMP-9 and 8-OHdG) analysis. Mdx showed focal lesions with intense inflammation and fibrosis related to immunoexpression of MMP-2 and MMP-9, proving the hypothesis that these MMPs are linked to muscular tissue degeneration, which can be regenerated by their inhibition, improving the treatment of DMD carriers. Histopathological findings related to centralized nuclei increase were related to higher 8-OHdG immunomarked nuclei in Mdx, which signalizes oxidative stress associated with DNA damage provoked by DMD. Such result shows that the evaluation of 8-OHdG during the evolution of the disease could be a method to evaluate DMD disease progression.

Analysis of tryptase-positive mast cells and immunoexpression of MMP-9 and MMP-13 in periapical lesions.

To evaluate the immunoexpression of tryptase, MMP-9 and MMP-13 in periapical lesions, correlating them with the type of lesion, intensity of the inflammatory infiltrate and thickness of the epithelial lining. Twenty periapical granulomas (PGs), twenty radicular cysts (RCs) and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using antitryptase, anti-MMP-9 and anti-MMP-13 antibodies. Immunoexpression of MMP-9 and MMP-13 was quantitatively evaluated both in the connective tissue of all lesions and in the epithelial lining of RCs and RRCs. Tryptase-positive mast cells were counted only in the connective tissue. The data were analysed using Kruskal-Wallis and Fisher's exact tests, as well as Spearman's correlation test (P<0.05). In comparison with RCs and RRCs, PGs exhibited higher immunoexpression of tryptase, MMP-9 and MMP-13 (P=0.002, P = < 0.001 and P<0.001, respectively). In comparison with lesions with inflammatory infiltrates grades I and II, lesions with inflammatory infiltrate grade III had higher median percentages of MMP-13-positive cells (P=0.003) and a tendency for higher expression of MMP-9 (P=0.059). No significant difference was observed between the expression of the studied markers and epithelial thickness (P>0.05). There were positive correlations between the number of tryptase-positive mast cells and the immunoexpression of MMP-9, as well as between the immunoexpression of MMP-9 and MMP-13. A larger number of tryptase-positive mast cells and greater enzymatic activity of MMP-9 and MMP-13 were found in PGs compared to RCs and RRCs. These findings are a characteristic of the dynamics of periapical diseases.

Metalloproteinases 2 and 9 Immunoexpression in Periapical Lesions from Primary Endodontic Infection: Possible Relationship with the Histopathological Diagnosis and the Presence of Pain

IntroductionThe aim of this study was to evaluate the possible associations among the histopathological diagnosis, the inflammatory infiltrate profile, the presence of pain, and the immunoexpression of matrix metalloproteinases MMP-2 and MMP-9 in periapical lesions from primary endodontic infection. MethodsFifty-one primary periapical lesions obtained from extracted teeth were selected for this study. Patients were previously evaluated for the presence of pain and sinus tract related to the tooth to be extracted. Tissues were processed for microscopic examination and MMP-2 and MMP-9 immunoexpression. Microscopically, samples were classified as periapical granulomas or periapical cysts and the inflammatory infiltrate as chronic or mixed. The percentage of immunopositive cells for MMP-2 and MMP-9 of each case was performed based on 10 consecutive microscopic fields. The Student t or chi-square tests were used in the statistical analysis. ResultsOf the total, 28 cases were classified as periapical granulomas (54.90%) and 23 cases as periapical cysts (45.10%). Seventeen patients (33.33%) reported pain associated with the extracted tooth, with 12 cases of periapical granulomas (70.58%) and 5 cases of periapical cysts (29.42%). All cases showed immunopositivity for MMP-2 and MMP-9 in a high percentage of cells, mainly in the cytoplasm of the leukocytes. MMP-2 was expressed more in periapical granulomas than periapical cysts (P < .05) and in symptomatic cases (P < .05). ConclusionsAccording to the results, we may conclude that MMP-2 and MMP-9 are highly expressed in periapical lesions from a primary endodontic infection. Moreover, we may suggest MMP-2 is expressed more in periapical granuloma and in cases associated with pain.

Immunohistochemical expression of matrix metalloproteinase-1, matrix metalloproteinase-2 and matrix metalloproteinase-9, myofibroblasts and Ki-67 in actinic cheilitis and lip squamous cell carcinoma.

Matrix metalloproteinases (MMPs), myofibroblasts (MFs) and epithelial proliferation have key roles in neoplastic progression. In this study immunoexpression of MMP-1, MMP-2 and MMP-9, presence of MFs and the epithelial proliferation index were investigated in actinic cheilitis (AC), lip squamous cell carcinoma (LSCC) and mucocele (MUC). Thirty cases of AC, thirty cases of LSCC and twenty cases of MUC were selected for immunohistochemical investigation of the proteins MMP-1, MMP-2, MMP-9, α-smooth muscle actin (α-SMA) and Ki-67. The MMP-1 expression in the epithelial component was higher in the AC than the MUC and LSCC. In the connective tissue, the expression was higher in the LSCC. MMP-2 showed lower epithelial and stromal immunostaining in the LSCC when compared to the AC and MUC. The epithelial staining for MMP-9 was higher in the AC when compared to the LSCC. However, in the connective tissue, the expression was lower in the AC compared to other lesions. The cell proliferation rate was increased in proportion to the severity of dysplasia in the AC, while in the LSCC it was higher in well-differentiated lesions compared to moderately differentiated. There were no statistically significant differences in number of MFs present in the lesions studied. The results suggest that MMPs could affect the biological behaviour of ACs and LSCCs inasmuch as they could participate in the development and progression from premalignant lesions to malignant lesions.

Comparação da imunoexpressão de VEGF, TGF-β e MMP-9 em ameloblastoma e tumor odontogênico adenomatóide

Purpose: Studies on odontogenic tumors have identified various molecular dysfunctions involved on their development, and some mechanisms such as angiogenesis and matrix modulation, are useful means of investigating the differences in biologic behavior of these tumors. Some important markers to identify tumor aggressiveness by immunohistochemistry are VEGF, TGF-β and MMP-9 proteins. This study aimed to compare the immunohistochemical expression of VEGF, TGF-β, and MMP-9 in ameloblastoma and adenomatoid odontogenic tumor (AOT). Methods: Immunoexpression of VEGF, TGF-β, and MMP-9 was studied in 15 solid ameloblastomas, and 15 AOTs. A semi-quantitative analysis of the immunostained cells was performed, and the statistical analysis was made using the Mann-Whitney nonparametric and Spearman correlation test, with significance level at .05 (P .05). Conclusion: The results suggest the involvement of angiogenesis in tumor progression of ameloblastomas, and the inductive effect of stromal cells in AOT, hence justifying its lower growth potential.

The relationships between the immunoexpression of KAI1, MMP-2, MMP-9 and steroid receptors expression in endometrial cancer.

The role of the parallel expression of the KAI1 protein and metalloproteinases (MMP-2 and MMP-9) in respect to the status of steroid receptors in endometrial cancer is still incompletely understood. The aim of this study was to evaluate the expression of and correlation between KAI1 on one hand and MMP-2 and MMP-9 on the other hand in terms of the status of the estrogen (ER) and progesterone receptors (PR) in 100 patients with endometrial cancer. The expressions of KAI1, MMP-2, MMP-9, ER and PR were assessed immunohistochemically on paraffin-embedded tissues. No correlations were found between these biomarkers and the clinical and pathological parameters of the endometrial cancer. However, in KAI1-positive cases, the expression was limited to a small area of tumor tissue in FIGO stages III-IV. A tendency towards the high expression of MMP-9 and MMP-2 was observed in the advanced stages of endometrial cancer (FIGO IIIA-IV). Positive correlations between the presence of KAI1 and PR and between the presence of MMP-9 and PR were found in endometrial cancer. A positive correlation was also observed between KAI1 and MMP-2 expression, and a borderline one between KAI1 and MMP-9 expression in endometrial cancer. KAI1+/PR+ and KAI1+/ER- immunophenotypes were observed more frequently in FIGO low stages and with well-differentiated tumor grade. However, the KAI1-/ER+ and KAI1-/PR+ immunophenotypes were mainly observed in advanced stages of endometrial cancer. KAI1+/MMP-2+ and KAI1+/MMP-9+ immunophenotypes were observed in FIGO Istage and with well-differentiated tumors. KAI1-/MMP-2+ and KAI1-/MMP-9+ phenotypes were more often observed in FIGO stage II. Our study showed that KAI1 protein, as well as steroid receptors, might modulate MMP-2 and MMP-9 expression in endometrial cancer. Our study revealed that the overlapping expression of the biomarkers investigated here suggests that cooperation between these molecules exists, even at the early stages of endometrial cancer growth, and may determine the speed of tumor cell dissemination and might characterize the biological behavior of endometrial cancer.

Immunoexpression of ROCK-1 and MMP-9 as prognostic markers in breast cancer

Breast cancer is the most common tumor in women and it has high mortality mainly due to the occurrence of tumor metastasis. Both the processes of cell migration and anchorage to the substrate are essential for the development of metastasis. These processes occur by rearrangements of the actin cytoskeleton, regulated by Rho-associated protein kinase 1 (ROCK-1). The degradation of the extracellular matrix, influenced by metalloproteinase 9 (MMP-9) also exerts greater cell invasiveness. The present study evaluated the ROCK-1 and MMP-9 proteins using an immunohistochemical method through the selection of invasive ductal breast carcinoma. The protein expression was correlated to clinicopathological parameters and overall survival of the patients. High expression of the ROCK-1 protein was correlated statistically to the status of lymph nodes (p=0.007) and showed variable expression in different clinical stages of the tumor. MMP-9 showed a strong immunostaining in patients with metastasis that had died, whereas there was no marker in normal breast tissues. In addition, 46.6% of patients classified as poor prognosis showed high expression of ROCK-1 and MMP-9 protein and another 40.0% just showed high expression of MMP-9. Thus, the differential expression of ROCK-1 and MMP-9 proteins suggests their potential use as prognostic markers in breast cancer.

Matrix metalloproteinase expression and activity in trophoblast-decidual tissues at organogenesis in CF-1 mouse

During early placentation, matrix metalloproteinases (MMPs) play important roles in decidualization, trophoblast migration, invasion, angiogenesis, vascularization and extracellular matrix (ECM) remodeling of the endometrium. The aim of our study was to analyze the localization, distribution and differential expression of MMP-2 and -9 in the organogenic implantation site and to evaluate in vivo and in vitro decidual MMP-2 and -9 activities on day 10 of gestation in CF-1 mouse. Whole extracts for Western blotting of organogenic E10-decidua expressed MMP-2 and -9 isoforms. MMP-2 immunoreactivity was found in a granular and discrete pattern in ECM of mesometrial decidua (MD) near maternal blood vessels and slightly in non-decidualized endometrium (NDE). Immunoexpression of MMP-9 was also detected in NDE, in cytoplasm of decidual cells and ECM of vascular MD, in trophoblastic area and in growing antimesometrial deciduum. Gelatin zymography showed that MMP-9 activity was significantly lower in CM compared to the active form of direct (not cultured) and cultured decidua. The decidual active MMP-9 was significantly higher than the active MMP-2. These results show differential localization, protein expression and enzymatic activation of MMPs, suggesting specific roles for MMP-2 and MMP-9 in decidual and trophoblast tissues related to organogenic ECM remodeling and vascularization during early establishment of mouse placentation.

Matrix metalloproteinases and their role in the renal epithelial mesenchymal transition.

Tubular cell epithelial-mesenchymal transition (EMT) is a fundamental contributor to renal fibrosis. The aim of this study was to investigate the activity of different matrix metalloproteinases by immunohistochemistry and gel-zymography in a model of chronic canine kidney disease. Immunohistochemistry for antibodies against MMP-9, MMP-2, MMP-13, MMP-14 and TIMP-2 was performed on 28 renal biopsy specimens. Selected cases were chosen for gelatin zymography. In moderate and severe tubulo-interstitial damage, increased expression of MMP-2 was noted. A peculiar staining pattern for MMP-2 in variable-sized vesicles, corresponding to the area of basement membrane splitting, was observed. The immunoexpression of MMP-9 and TIMP-2 was reduced in the same cases, compared to control dogs. The splitting of the membrane suggests an active role of this gelatinase in the disruption of type-IV collagen, the main basement membrane component, confirmed by MMP2 gelatinolytic activity by gel-zymography. These data could provide the basis for clinical trials examining the potential benefits of selective MMP-2 inhibitors in dogs with chronic kidney disease.

Mast cells and MMP‐9 in the lamina propria during eruption of rat molars: quantitative and immunohistochemical evaluation

During the active tooth eruption process, structural changes in the lamina propria are necessary to provide extracellular matrix remodelling and for the establishment of the eruptive pathway. A large number of resident cells, recruited cells and proteases have been demonstrated in the eruptive process, but the participation of MMP-9 and mast cells has not yet been demonstrated. In this study, we set out to evaluate the intensity of MMP-9 immunoexpression, the frequency of mast cells and the correlation between the incidence of mast cells and bone resorption in different phases of tooth eruption. Fragments of maxilla containing first molars, obtained from 9-, 11-, 13- and 16-day-old rats, were fixed in 4% formaldehyde, decalcified and embedded in paraffin. Sagittal sections were stained with Masson's trichrome or submitted to the tartrate-resistant acid phosphatase method for quantification of osteoclasts. Sections stained by 1% toluidine blue were used for quantification of metachromatic mast cells mm(-2) of lamina propria. The expression of MMP-9 in the lamina propria was evaluated by immunohistochemistry. In the 9-day-old rats, the lamina propria contained few mast cells and occasional osteoclasts were found in the bone surface overlying the occlusal portion of the tooth germs. Otherwise, a significant increase in the number of mast cells was observed in the intra-osseous phase of tooth eruption (11-day-old rats), period in which numerous TRAP-positive osteoclasts were found in the bone surface. MMP-9 immunolabelling was detected in fibroblasts, mast cells and macrophage-like cells of the lamina propria in all ages studied. However, an enhanced immunolabelling was evident in the advanced phase of tooth eruption (16-day-old rats). During the intra-osseous phase, the parallel between the high frequency of both mast cells and osteoclasts suggests that mast cells could exert a paracrine function on the osteoclasts and then stimulate bone resorption. The immunoexpression of MMP-9 in different cells of lamina propria, including mast cells, indicates that this enzyme participates in the degradation of ECM, mainly during late phase of mucosal penetration. Thus mast cells and MMP-9 are involved in the complex process of degradation of the eruptive pathway extracellular matrix.

Differential glomerular immunoexpression of matrix metalloproteinases MMP-2 and MMP-9 in idiopathic IgA nephropathy and Schoenlein-Henoch nephritis.

Both idiopathic IgA nephropathy (IgAN) and Schoenlein-Henoch nephritis (SHN) are characterized by cell proliferation and abnormal extracellular matrix (ECM) remodeling by mesangial cells leading to fibrosis, sclerosis and end-stage renal disease. Matrix metalloproteinases MMP-2 and MMP-9 are reported as the most important proteolytic enzymes involved in remodeling of ECM. Therefore, the aim of the present study was to determine glomerular immunoexpression of MMP-2 and MMP-9 in IgAN and SHN. Another purpose of this study was to examine the relationship between expression of MMPs and mesangial cells, a-smooth muscle actin (alpha-SMA) staining, and monocytes/macrophages. Fifteen patients with idiopathic IgAN and 12 with SHN were examined by percutaneous renal biopsy. Glomerular staining intensity of MMP-2 and MMP-9 was recorded semiquantitatively, whereas mesangial cells, glomerular alpha-SMA staining and glomerular CD 68+ cells were assessed quantitatively using computer image analysis system. Our study revealed that the mean values of glomerular immunoexpression of MMP-2, mesangial cells, alpha-SMA staining and glomerular CD 68+ cells were in SHN patients significantly increased as compared to IgAN cases whereas glomerular staining for MMP-9 did not differ in these groups. Moreover, a glomerular staining of MMP-2 was significantly positively correlated with mesangial cells as well as glomerular alpha-SMA staining in both SHN and IgAN. A positive significant correlation between glomerular MMP-2 staining and glomerular CD68+ cells was noted only in SHN group. The correlations of glomerular MMP-9 and these parameters were weak and not significant. In conclusion, our results confirm increased glomerular staining of MMP-2 but not MMP-9 in SHN patients. A suggestion that augmented mesangial cells proliferation in these cases depends on MMP-2, alpha-SMA and monocytes/macrophages needs further investigations including double staining study.

Specific matrix metalloproteinase expression in focal myositis: an immunopathological study

The aim of our study was to investigate immunoexpression of matrix metalloproteinases MMP2, MMP7 and MMP9 in muscle specimens from patients with focal myositis (FM) vs polymyositis (PM) and dermatomyositis. We studied muscle biopsy samples from seven patients affected by FM; samples from five patients each with PM and dermatomyositis were studied as disease controls. MMP2 immunoreactivity was present in PM and dermatomyositis, MMP7 only in PM, and MMP9 in PM, dermatomyositis and FM. Our results confirm that increased MMP9 immunoreactivity in muscle fibres is a common feature of all inflammatory myopathies and suggest that MMP2 and MMP7 cannot be implicated in the inflammatory events of FM.