Bisphosphonates have been reported to have chondroprotective activities in addition to its original functions. However, mechanisms for these just began to be elucidated. Under the hypothesis that bisphosphonates may regulate expression and activities of matrix enzymes during degradation of cartilage for bone formation, we administrated an alendronate (1 mg/kg) to newborn rats subcutaneously once a day for 4, 7, and 10 days. To identify the effects of alendronate on cartilage, thickness of cartilage layer was measured by histomorphometry on the proximal epiphysis of tibia. Immunofluorescence staining and RT-PCR were performed to investigate the expressions of matrix enzymes in both in vitro and in vivo. MTS assay revealed that at 10(-3) M in concentration, alendronate significantly reduced viability of chondrocytes. The mRNA expressions of MMP-1, MMP-9, EMMPRIN, and TIMP-3 in primary chondrocytes were decreased by the alendronate treatment. Interestingly, TIMP-1 mRNA expression was significantly increased, whereas a constitutive form, TIMP-2 was relatively unchanged by the treatment. The thickness of proliferating layer at postnatal day 7 was not significantly different, whereas thickness of hypertrophied layer was significantly thicker in the alendronate group than in the control (p<0.01). Immunofluorescence demonstrated that the expressions of MMP-9, TIMP-2 and -3 were reduced, whereas TIMP-1 expression was increased by the alendronate administration. These results suggest that the alendronate have chondroprotective properties by down-regulation of MMPs and up-regulation of TIMPs during endochondral ossification.
Alendronate ; Animals ; Antigens, CD147 ; Cartilage ; Chondrocytes ; Diphosphonates ; Down-Regulation ; Epiphyses ; Fluorescent Antibody Technique ; Humans ; Infant, Newborn ; Matrix Metalloproteinases ; Osteogenesis ; Rats ; RNA, Messenger ; Tibia ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2 ; Tissue Inhibitor of Metalloproteinase-3 ; Up-Regulation

No abstract available.
Matrix Metalloproteinases* ; Stomach Neoplasms ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

BACKGROUND: MMPs and TIMPs are important factors for abnormal remodeling the pulmonary parenchyme in idiopathic interstitial pneumonia(IIP) This study evaluated the expression of MMPs and TIMPs in the tissue of IPF, NSIP and normal control subjects. METHOD: The MMP-2 and -9 activity in the lung tissue was studied by gelatin zymography, and the expression of MMP-1, -2 ,-9, TIMP-1 and -2 in the lung tissue was measured by immunohistochemistry. Thirty five patients, who were diagnosed with IIP (UIP ; 22, NSIP ; 13), were enrolled in the immunohistochemical study. Thirteen patients with IIP (UIP ; 9, NSIP ; 4) and five patients with lung cancer were enrolled in the zymographic assay. RESULTS: (1) The immunohistochemistry for MMP-1,-2,-9, TIMP-1 and-2 ; MMP-1,-9 and TIMP-2 were stained stronger in the UIP subjects than NSIP and the normal control. TIMP-2 was strongly stained in the UIP tissue. particularly the fibroblasts in the fibroblastic foci. (2) Zymography for MMP-2 and MMP-9 revealed MMP-2 to have prominent expression in the UIP tissue than in the NSIP tissue. CONCLUSIONS: These results suggest that the overexpression of the TIMPs and gelatinases in UIP might be? important factors in the irreversible fibrosis of the lung parenchyme.
Fibroblasts ; Fibrosis ; Gelatin ; Gelatinases ; Humans ; Immunohistochemistry ; Lung ; Lung Neoplasms ; Matrix Metalloproteinases* ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

Matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of matrix metalloproteinases, TIMPs) play essential roles in the remodelling of the extracellular matrix. The balance between MMPs and TIMPs is altered in neoplasia, contributing to the invasive and metastatic properties of malignant tumors. Although MMP and TIMP are believed to play an important role in invasion and metastasis in malignant solid tumors, little is known about their involvement in malignant lymphoma. Immunohistochemical stains for MMP-1, MMP-2, MMP-9, TIMP-1 and TIMP-2 were performed using 56 paraffin blocks of the malignant lymphoma and the results were analyzed by using the tumor grade by Working Formulation. The expression of MMP-9 was noted in 45.5% of low grade, 86.1% of intermediate grade, and 100% of high grade malignant lymphoma. The incidence of MMP-9 expression in tumor cells was positively correlated with the grade of the malignant lymphoma (P<0.025). In nodal lymphomas, the incidence of the MMP-9 expression of the tumor cells was higher in malignant lymphoma with extracapsular invasion than those without extracapsular invasion (P=0.008). The incidence of TIMP-1 expression in the tumor cells and fibroblasts was positively correlated with the grade of the malignant lymphoma (P<0.025). In nodal lymphoma, the incidence of the TIMP-1 expression of the tumor cells was higher in malignant lymphoma with extracapsular invasion than those without extracapsular invasion (P=0.009). The incidences of the MMP-1, MMP-2, and TIMP-2 expression in malignant lymphoma were neither increased in the malignant lymphoma with extracapsular tumor invasion nor correlated with the grade by working formulation. There was no significant difference in the expression rate of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 in nodal- and extra-nodal malignant lymphoma. The above results suggest that the expressions of MMP-9 and TIMP-1 are positively correlated with the grade and the presence of extranodal tumor invasion in malignant lymphomas.
Coloring Agents ; Extracellular Matrix ; Fibroblasts ; Incidence ; Lymphoma* ; Matrix Metalloproteinases ; Neoplasm Metastasis ; Paraffin ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

OBJECTIVETo detect protein expression of MMPs and TIMPs in various salivary gland neoplasms and to investigate their roles in invasion and metastasis of the malignant salivary gland tumors.

METHODSImmunohistochemistry and Gelatin zymography analyses for MMP-2, MMP-9, MT1-MMP, TIMP-1 and TIMP-2 were performed in 26 malignant and 28 benign salivary gland tumors.

RESULTSThe expression of MMP-2 and MMP-9 was significantly higher in carcinomas than in adenomas (P < 0.05). The MMP-2/TIMP-1 and MMP-2/TIMP-2 was also significantly higher in carcinomas than in adenomas (P < 0.05). There was a cooperated effect among MMP-2, MT1-MMP and TIMP-2. The expression of active MMP-2, proMMP-9 and active MMP-9 was significantly higher in malignant tumors than in benign tumors (P < 0.05).

CONCLUSIONMMP-2 and MMP-9 may play important roles in invasion of malignant salivary gland tumors. A disturbed balance between MMP-2, MMP-9, TIMP-1 and TIMP-2 in malignant salivary gland tumors was detected. It was the absolute increase of MMP-2 and MMP-9 to induce the unbalance.

Enzyme Precursors ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; Salivary Gland Neoplasms ; Salivary Glands ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

BACKGROUND: In the idiopathic interstitial pneumonia (IIP), it has been known that imbalance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) is important factor for abnormal remodeling of lung parenchyme. This study was performed to compare the expression of MMPs and TIMPs in the normal subjects and patients with IIP. METHODS: Seventeen patients were diagnosed as IIP by open lung biopsy (male: female 7:10) and four patients as normal control were diagnosed as lung cancer and treated by lobectomy or pneumonectomy from March, 1999 to August 2001 at Gil medical center. IIP group divided into UIP (n=10) and NSIP (n=7). MMP-1 and TIMP-2 of their lung tissue were stained by immunohistochemical method and were graded 4 levels (grage 0-3) following stained status. RESULTS: MMP-1 was stained more strongly in the IIP than normal. But it had no differences between UIP and NSIP. TIMP-1 and-2 were stained more strongly in the UIP than NSIP but not stained in the normal. In the UIP, TIMP-2 was stained strongly in fibroblast foci. CONCLUSION: These results suggst that imbalance between MMPs and TIMPs may be important factor of pathogenesis of pulmonary fibrosis in the IIP. It is thought that major site of TIMP-2 is myofibroblast in the fibroblast foci.
Biopsy ; Female ; Fibroblasts ; Humans ; Idiopathic Interstitial Pneumonias* ; Lung ; Lung Neoplasms ; Matrix Metalloproteinase 1* ; Matrix Metalloproteinases ; Metalloproteases ; Myofibroblasts ; Pneumonectomy ; Pulmonary Fibrosis ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

PURPOSE: Although varicose veins are common, the primary cause predisposing to venous reflux of great saphenous vein is controversial. We compared histologic features and expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) in the great saphenous veins (GSVs) of normal and patients with varicose veins. METHOD: Ten normal proximal GSV and each twenty diseased proximal, mid, and distal above-knee GSV vein segments were used for this analysis. We compared venous architecture with histology and expression and localization of MMP and TIMP with immunohistochemistry. RESULT: Compared with normal veins, varicose veins showed thickening and disorganization of intimal and medial connective tissue elements. Elastic fibers of varicose vein were fragmented and degenerated. By the immunohistochemistry, MMP-1 and MMP-2 were more strongly expressed than MMP-9, TIMP-1 and TIMP-2 either normal and varicose veins. MMPs were localized to endothelial cells, medial connective tissues and adventitial microvessels in normal veins. Whereas they were more diffusely localized to subintimal and medial connective tissues in varicose veins. Compared with proximal varicose veins, TIMP-2 was more significantly expressed in distal varicose veins. CONCLUSION: In varicose veins, vein wall architectures are disorganized and MMPs are more prominently expressed in these degenerative subintimal and medial areas.
Connective Tissue ; Elastic Tissue ; Endothelial Cells ; Humans ; Immunohistochemistry ; Matrix Metalloproteinases* ; Microvessels ; Saphenous Vein* ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2 ; Varicose Veins* ; Veins

OBJECTIVE: Homeostasis of the extracellular matrix (ECM) is maintained by the action of a specific system of proteolytic enzymes known as matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP). The MMP/TIMP system regulates the composition and turnover of ECM to control the site and extent of connective tissue remodeling. In pathologic conditions, MMP play a key role in degradation of basement membrane and extracellular matrix, and is responsible for cancer invasion, progression and metastasis. The aim of this study is to evaluate the correlation between expressions of MMP/TIMP and clinicopathologic factors in endometrial cancer. METHODS: Expressions of MMP-2, MMP-9, TIMP-1, and TIMP-2 were assessed by immunohistochemistry in a total of 55 endometrial cancers and were analyzed by the correlation between expressions of MMP/TIMP and clinicopathologic factors in endometrial cancer. RESULTS: Expression rates of MMP-2,-9, TIMP-1, and TIMP-2 were 71.7%, 54.9%, 41.2%, and 76.5% respectively. Expression of MMP-2 was correlated with the group of positive lymph node metastasis in endometrial cancer (p=0.04). Specially, coexpression of MMP-2 and TIMP-2 was significantly more frequent in the group of positive lymph node metastasis (p<0.01) and the group of positive peritoneal CONCLUSION: The expressions of MMP and TIMP were not a significant difference in survival analysis, but this study was recognized that the coexpression MMP-2 and TIMP-2 is correlated with lymph node metastasis and positive peritoneal cytology.
Basement Membrane ; Connective Tissue ; Endometrial Neoplasms ; Extracellular Matrix ; Female ; Homeostasis ; Immunohistochemistry ; Lymph Nodes ; Matrix Metalloproteinases ; Metalloproteases ; Neoplasm Metastasis ; Peptide Hydrolases ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

PURPOSE: Kawasaki disease (KD) is a systemic vasculitis, a leading cause of pediatric acquired heart disease. Histopathological findings of coronary artery lesion (CAL) in KD indicate destruction of the coronary artery wall with diffuse vasculitis. Matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) might play central roles in this process. Special attention to MMP-9 has recently been emerging. This study was performed to investigate the clinical significance of MMP-9 and its inhibitors, TIMP-1 and TIMP-2, in KD. METHODS: We compared 47 KD patients with 14 febrile controls. Serum MMP-9 and TIMP-1, TIMP-2 were measured by ELISA and compared according to clinical stages and coronary involvement. RESULTS: In acute stage, MMP-9 and TIMP-1 were significantly higher, whereas TIMP-2 was lower, in KD than those in febrile controls (p<0.05). The elevated MMP-9 levels in acute phase significantly decreased during the subacute and convalescent phases (p<0.05). During acute phase, the MMP-9, TIMP-1, and MMP-9/TIMP-2 levels in the CAL group were lower than those in the non-CAL group, but they increased significantly in the subacute phase (p<0.05). MMP-9 has a positive correlation with TIMP-1 in the acute and subacute phases, and negative correlation with TIMP-2 in the subacute and convalescent phases (p<0.05). CONCLUSION: These results suggest that MMP-9, TIMP-1, and the imbalance in MMP-9 and TIMP-2 might play important roles on the pathophysiology of KD and especially on the development of CAL. However, further larger studies are needed.
Coronary Vessels ; Enzyme-Linked Immunosorbent Assay ; Heart Diseases ; Humans ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; Mucocutaneous Lymph Node Syndrome ; Systemic Vasculitis ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2 ; Tissue Inhibitor of Metalloproteinases ; Vasculitis

BACKGROUND: Matrix metalloproteinases (MMPs) are a large family of proteolytic enzymes, which are in volved in the degradation of many different components of the extracellular matrix. There is increasing evidence indicating that individual MMPs have important roles in tumor invasion and metastasis. A tissue inhibitor of metalloproteinase(TIMPs) has been reported to inhibit tumor invasion by inactivating the MMPs. In this study, the correlation between MMPs and TIMPs expression, and the clinical outcome was investigated. METHODS: Immunohistochemical staining of MMP-2,9 and TIMP-1,2 were performed on paraffin-embedded tumor sections from 74 resected primary non-small cell lung cancers. RESULTS: In 74 patients, MMP-2, MMP-9, TIMP-1, and TIMP-2 immunoreactivity was demonstrated in 24 (34%), 19(26%), 27(36%) and 32(43%) of the paraffin-embedded tumors, respectively. The median survival of the MMP-2 positive cases was significantly shorter than that of the negative cases(20 vs 34 months). The median survival of the TIMP-2 positive cases was also was significantly longer than that of negative case (34 vs 18 months). The MMP-2, and MMP-9 expression level had a positively correlation with a more advanced stage and lymph node metastasis. There was inverse correlation between TIMP-2 expression and tumor invasion. The median survival of the MMP-2 negative/TIMP-2 positive cases was higher than that of the other cases. CONCLUSION: These results suggest that tumor invasion and lymph node metastasis are closely related to MMP-2 and MMP-9 expression. There was an inverse correlation between TIMP-2 MMP-9 expression, and tumor invasion.
Carcinoma, Non-Small-Cell Lung* ; Extracellular Matrix ; Humans ; Lung Neoplasms ; Lymph Nodes ; Matrix Metalloproteinases ; Neoplasm Metastasis ; Peptide Hydrolases ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2