OBJECTIVE: To investigate the correlation between matrix metalloproteinase (MMP)-2/MMP-9 levels and childhood caries, and the saliva levels of MMP-2/MMP-9 among healthy children and those with different degrees of dental caries, both before and after treatment. METHODS: In the study, 368 children aged 3 to 5 years were separated into three groups: severe caries group (112 children), mild caries group (98 children) and caries free group (158 children). The children with severe caries were included in treatment group (83 children) after accepting a comprehensive treatment of caries. MMP-2 and MMP-9 levels were determined by enzyme-linked immunosorbent assay (ELISA) and the data were analyzed by the Statistics Package for Social Science (SPSS 13.0). The differences among severe caries group, mild caries group and caries free group were analyzed by SNK-q (Student Newman Keuls). The severe caries group and treatment group were compared by paired t test. The differences between each group were statistically analyzed. RESULTS: There was no significant difference of the age and gender composition among severe caries group, mild caries group, caries free group and treatment group. The MMP-2 level of severe caries group [(141.3±32.5) μg/L] was higher than those of mild caries group [(107.5±21.3) μg/L] and caries free group [(102.8±18.5) μg/L] (P<0.05). There was no significant difference between mild caries and caries free group (P>0.05). After analysis of 83 children in the treatment group, the level of MMP-2 [(120.1±24.8) μg/L] was lower than before [(144.6±30.3) μg/L] (P<0.05), but was higher than that of caries free group (P<0.05). The MMP-9 levels of severe caries group [(445.8±68.1) μg/L] and mild caries group [(428.6±59.2) μg/L] were higher than that of caries free group [(385.4±60.6) μg/L] (P<0.05), but the difference between severe caries group and mild caries group was not significant (P>0.05). After analysis of 83 children in the treatment group, the alteration of MMP-9 [(432.2±64.7) μg/L] was not significant either (P>0.05). CONCLUSION The saliva levels of MMP-2 and MMP-9 in children with severe caries were higher than those in caries free children, even if the treatment was implemented, which suggests that the MMP-2 and MMP-9 in saliva might be related to the caries in children.
Child, Preschool ; Dental Caries/enzymology* ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Matrix Metalloproteinase 2/analysis* ; Matrix Metalloproteinase 9/analysis* ; Matrix Metalloproteinases ; Saliva/chemistry*

Osteoarthritis is recognised to be an interactive pathological process involving the cartilage, subchondral bone and synovium. The signals from the synovium play an important role in cartilage metabolism, but little is known regarding the influence of the signalling from bone. Additionally, the collagenases and stromelysin-1 are involved in cartilage catabolism through mitogen-activated protein kinase (MAPK) signalling, but the role of the gelatinases has not been elucidated. Here, we studied the influence of osteoclastic signals on chondrocytes by characterising the expression of interleukin-1β (IL-1β)-induced gelatinases through MAPK signalling. We found that osteoclast-conditioned media attenuated the gelatinase activity in chondrocytes. However, IL-1β induced increased levels of gelatinase activity in the conditioned media group relative to the mono-cultured chondrocyte group. More specifically, IL-1β restored high levels of gelatinase activity in c-Jun N-terminal kinase inhibitor-pretreated chondrocytes in the conditioned media group and led to lower levels of gelatinase activity in extracellular signal-regulated kinase or p38 inhibitor-pretreated chondrocytes. Gene expression generally correlated with protein expression. Taken together, these results show for the first time that signals from osteoclasts can influence gelatinase activity in chondrocytes. Furthermore, these data show that IL-1β restores gelatinase activity through MAPK inhibitors; this information can help to increase the understanding of the gelatinase modulation in articular cartilage.
3T3 Cells ; Animals ; Cartilage, Articular ; cytology ; Cell Survival ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; enzymology ; Coculture Techniques ; Culture Media, Conditioned ; Gelatinases ; drug effects ; Interleukin-1beta ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; physiology ; Matrix Metalloproteinase 2 ; drug effects ; Matrix Metalloproteinase 9 ; drug effects ; Mice ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; drug effects ; Monocytes ; cytology ; NF-kappa B ; antagonists & inhibitors ; Osteoclasts ; physiology ; Protease Inhibitors ; analysis ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; Tissue Inhibitor of Metalloproteinase-2 ; drug effects ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

OBJECTIVETo investigate the changes of cytokines in induced sputum at different stages of silicosis patients.

METHODSA total of 200 workers from one of the Shandong Province gold mine were chosen as object of observation. Among which 40 patients at silicosis stage I and 40 patients at silicosis stage II were divided into silicosis observed object group, silicosis stage I group, silicosis stage II group, and another 80 workers exposed to silica dust without suffering from silicotic Clinical symptoms, however, were chosen as group of dust exposed, and 40 logistical workers without being exposed and history of silicosis's illness were chosen as control group. And ask their basic information by questionnaire. Then, spray-inhalation the induced sputum and apply the ELISA to assess the level of tumor necrosis factor (TNF), interleukin (IL), macrophage inflammatory protein-1 (MIP-1α), monocyte chemotactic factor-1 (MCP-1), metalloproteinases (MMP), transforming growth factor-β (TGF-β), platelet derived growth factor (PDGF) in induced sputum from subjects.

RESULTSThe level of TGF-β [(901.60 ± 30.09) ng/L] in the induced sputumof patients in silicosis stage I group is lower than that in the observed object group [(913.02 ± 20.51) ng/L], and the level of MMP-9 [(212.49 ± 5.97) ng/L], MCP-1 [(129.91 ± 4.30) ng/L] has various degrees of increase than that in control group, observed object group and dust exposed group. All the differences have statistical significances (P < 0.05). The level of TNF-α [(85.76 ± 3.78) ng/L] in the induced sputum of patients in silicosis stage I group reaches the maximum, there are significant differences comparing with that level in the silica dust exposure group and the control group, whose differences are statistically significant (P < 0.05). Compared with the control group, the level of MMP-2 (427.95 ± 23.64) in the induced sputum of patients in silicosis stage I group has increased, whose differences also have statically significant (P < 0.05). Compared with the control group, silica dust exposed group, the observation group of objects, the pneumosilicosis patients of IL-16 in induced sputum IL-16 (21.40 ± 9.24) decreased, the content of PDGF [(5.96 ± 0.51) ng/L], MMP-2 [(447.86 ± 27.10) ng/L], MMP-9 [(223.91 ± 12.28) ng/L], MCP-1 [(122.87 ± 6.08) ng/L] increased, the differences are statistically significant (P < 0.05).

CONCLUSIONAs silicosis biomarkers, TNF-alpha, TGF-beta, IL-16, PDGF, MMP-2, MMP-9 and MCP-1 have certain significance, further suggesting that early detection rate of patients with silicosis can be improved by employing the multiple indexes discriminate equation.

Biomarkers ; metabolism ; Case-Control Studies ; Chemokine CCL2 ; metabolism ; Chemokine CCL3 ; metabolism ; Cytokines ; metabolism ; Discriminant Analysis ; Dust ; Humans ; Interleukin-16 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Platelet-Derived Growth Factor ; metabolism ; Silicosis ; diagnosis ; Sputum ; chemistry ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe P38 mitogen-activated protein kinase (P38 MAPK) and matrix metalloproteinase-2 (MMP-2) mRNA expression level changes in neonatal rats with hyperoxia-induced lung injury,and to investigate the influence of P38 MAPK activation on MMP-2 mRNA expression.

METHODSThirty-six Sprague-Dawley (SD) rats were randomly divided into three groups: air control, hyperoxia and SB203580-treated hyperoxia (n=12). The rats were sacrificed on the 3rd and 7th days and the lungs were removed. Hematoxylin-eosine staining was used to observe the pathological changes in lung tissues.

RESULTSCompared with the air and SB203580-treated groups, levels of P38 MAPK and MMP-2 mRNA significantly increased in the hyperoxia group (P<0.01).

CONCLUSIONSExpression of P38 MAPK increases in neonatal rats with hyperoxia-induced acute lung injury and this may play a role in control of the expression of MMP-2 mRNA.

Animals ; Animals, Newborn ; Female ; Hyperoxia ; complications ; Lung ; pathology ; Lung Injury ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinases ; genetics ; p38 Mitogen-Activated Protein Kinases ; metabolism

Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Melanoma ; enzymology ; Skin Neoplasms ; enzymology

OBJECTIVETo observe the effect of neferine on Collagen-I, TIMP-1 and MMP-2 expressions and protein secretion of hepatic stellate cells.

METHODThe hepatic stellate cell line HSC-T6 was cultured in vitro, and then randomly divided into 5 groups: the control group, the platelet-derived growth factor (PDGF) group and PDGF + neferine (2, 6, 10 micromol x L(-1)) groups. All of the groups were cultured for 48 h, and their cells were collected to extract mRNA and detect Collagen-I, TIMP-1 and MMP-2 expressions with RT-PCR. Their cell supernatants were also collected to determine the protein content of three factors with ELISA.

RESULTCompared with the control group, PDGF could remarkably increase the Collagen-I, TIMP-1 and MMP-2 expressions and protein secretion of hepatic stellate cells. Compared with the PDGF group, PDGF + neferine (6, 10 micromol x L(-1)) groups showed a notable decrease in the Collagen-I and mRNA expression and protein secretion along with the increase in the concentration, whereas the PDGF + neferine (2 micromol x L(-1)) group showed no significant change in the Collagen-I and mRNA expression and protein secretion. Compared with the PDGF group, three PDGF + neferine groups showed no notable change in MMP-2 expression and protein secretion.

CONCLUSIONNeferine can inhibit the Collagen-I, TIMP-1 and mRNA protein expression and protein secretion of PDGF-induced HSCs along with the increase in the concentration, but with not remarkable effect on the MMP-2 expression and secretion.

Animals ; Benzylisoquinolines ; pharmacology ; Cells, Cultured ; Collagen Type I ; analysis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Hepatic Stellate Cells ; chemistry ; drug effects ; Matrix Metalloproteinase 2 ; analysis ; genetics ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; genetics

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.
Acid Phosphatase/analysis/blood ; Animals ; Arthritis, Experimental/enzymology/etiology/veterinary ; Biological Markers/*analysis/*blood ; Blotting, Western/veterinary ; Dislocations/complications/*veterinary ; Dog Diseases/*enzymology/etiology ; Dogs ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Isoenzymes/analysis/blood ; Male ; Matrix Metalloproteinase 2/analysis/blood ; Osteoarthritis/enzymology/etiology/*veterinary ; Spectrophotometry/veterinary ; Stifle/physiopathology ; Synovial Fluid/*enzymology ; Tissue Inhibitor of Metalloproteinase-2/analysis/blood

OBJECTIVES: To investigate expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in squamous cell carcinoma of the tonsil and to correlate expression profiles with clinicopathological characteristics. METHODS: Paraffin blocks were obtained from 45 tonsil squamous cell carcinoma (SCC) patients, who underwent surgery as an initial treatment between 1994 and 2004, and from 20 normal controls. Expressions of MMP-2, MMP-9, MMP-13, TIMP-1, and TIMP-2 were investigated immunohistochemically. RESULTS: The expressions of MMPs (except MMP-2) and TIMPs were found to be significantly different in tonsil SCC and normal control tissues. Furthermore, MMP-13 expression was found to be correlated with tumor invasion (P=0.05), and the expressions of MMP-9 and TIMP-1 with nodal metastasis (P=0.048, 0.031). No relation was found between MMP or TIMP expression and recurrence. However, MMP-9 expression was found to be significantly associated with 5-year survival in tonsil SCC patients by multivariate analysis (hazard ratio, 3.853; P=0.013). CONCLUSION: Significant overexpressions of multiple MMPs and TIMPs were found in tonsil SCC tissues. Furthermore, our findings suggest that MMP-9 expression might be a useful prognostic factor.
Carcinoma, Squamous Cell ; Humans ; Matrix Metalloproteinases ; Metalloproteases ; Multivariate Analysis ; Neoplasm Metastasis ; Palatine Tonsil ; Paraffin ; Prognosis ; Recurrence ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

The presence of matrix metalloproteinase-2 (MMP-2) in dentin has been reported, but its distribution and activity level in mature human coronal dentin are not well understood. The purpose of this study was to determine the MMP-2 distribution and relative activity in demineralized dentin. Crowns of twenty eight human molars were sectioned into inner (ID), middle (MD), and outer dentin (OD) regions and demineralized. MMP-2 was extracted with 0.33 mol x L(-1) EDTA/2 mol xL(-1) guanidine-HCl, pH 7.4, and MMP-2 concentration was estimated with enzyme-linked immunoabsorbant assay (ELISA). Further characterization was accomplished by Western blotting analysis and gelatin zymography. The mean concentrations of MMP-2 per mg dentin protein in the dentin regions were significantly different (P = 0.043): 0.9 ng (ID), 0.4 ng (MD), and 2.2 ng (OD), respectively. The pattern of MMP-2 concentration was OD > ID > MD. Western blotting analysis detected -.66 and -72 kDa immunopositive proteins corresponding to pro- and mature MMP-2, respectively, in the ID and MD, and a -66 kDa protein in the OD. Gelatinolytic activity consistent with MMP-2 was detected in all regions. Interestingly, the pattern of levels of Western blot immunodetection and gelatinolytic activity was MD > ID > OD. The concentration of MMP-2 in human coronal dentin was highest in the region of dentin that contains the dentinoenamel junction and least in the middle region of dentin. However, levels of Western blot immunodetection and gelatinolytic activity did not correlate with the estimated regional concentrations of MMP-2, potentially indicating region specific protein interactions.
Blotting, Western ; Dentin ; enzymology ; Electrophoresis, Polyacrylamide Gel ; Enzyme Precursors ; analysis ; Female ; Humans ; Male ; Matrix Metalloproteinase 2 ; analysis ; metabolism ; Molar, Third ; enzymology ; Tissue Distribution ; Tooth Crown ; enzymology ; Tooth Demineralization ; enzymology

OBJECTIVE: Supplementation with vitamin E is able to protect bone against free radical-induced elevation of bone-resorbing cytokines. We examined gene expression by microarray analysis during the differentiation of human mesenchymal stem cells treated with vitamin E into osteoblasts in vitro. METHODS: Human bone marrow stem cells were cultured in osteogenic differentiation medium and vitamin E was added. A colorimetric immunoassay for the quantification of cell proliferation was used to measure osteoblast differentiation. Gene expression was analyzed using a microarray technique. We also used a real time reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: It was found that vitamin E enhanced cell proliferation when compared to cells cultured in media without vitamin E. We focused on 68 genes which are related to osteogenesis and osteoclastogenesis. Alkaline phosphatase, transforming growth factor-beta 1, fibroblast growth factor receptor 1, matrix metalloproteinase 2, muscle segment homeobox 2, bone morphogenetic protein 1, biglycan, vascular endothelial growth factor B, dentin sialophosphoprotein, cartilage oligomeric matrix protein, runt-related transcription factor 2, fibroblast growth factor receptor 3, and SMAD2 were upregulated > 2-fold compared to the control. Conversely, osteopetrosis-associated transmembrane protein 1, microphthalmia-associated transcription factor, and epidermal growth factor receptor were downregulated > 2-fold compared to the control. Vitamin E produced a 1.5-fold increase in the expression of runt-related transcription factor 2 and transforming growth factor-beta 1 as determined by real time RT-PCR. CONCLUSION: Vitamin E had a positive effect on the gene expressions regarding osteogenic differentiation of mesenchymal stem cells.
Alkaline Phosphatase ; Biglycan ; Bone Marrow ; Bone Morphogenetic Protein 1 ; Cartilage ; Cell Proliferation ; Cytokines ; Dentin ; Durapatite ; Extracellular Matrix Proteins ; Gene Expression ; Genes, Homeobox ; Glycoproteins ; Humans ; Immunoassay ; Matrix Metalloproteinase 2 ; Mesenchymal Stromal Cells ; Microarray Analysis ; Microphthalmia-Associated Transcription Factor ; Muscles ; Osteoblasts ; Osteogenesis ; Phosphoproteins ; Receptor, Epidermal Growth Factor ; Receptor, Fibroblast Growth Factor, Type 1 ; Receptor, Fibroblast Growth Factor, Type 3 ; Sialoglycoproteins ; Stem Cells ; Transcription Factors ; Vascular Endothelial Growth Factor B ; Vitamin E ; Vitamins