Osteoarthritis (OA) is a complex chronic progressive disease attacked by biological and mechanical factors and a result from the anabolic and catabolic imbalance in chondrocyte, subchondral bone and extracellular matrix(ECM). Etiology and pathological of OA are not yet entirely clear. The degradation and destruction of collagen II caused by matrix metalloproteinase -13 (MMP-13) is considered the core factor in the occurrence and development of OA. The research of MMP-13 inhibitor provide ideas and methods for the treatment of OA. In this article,the role and determination of MMP-13 in OA and the development prospect of MMP-13 inhibitor in the treatment of OA research progress were reviewed.
Animals ; Collagen ; metabolism ; Humans ; Matrix Metalloproteinase 13 ; analysis ; physiology ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Osteoarthritis ; drug therapy ; etiology

OBJECTIVETo study the changes of metalloproteinases 1, 2, 13 in granulation wound after the treatment of vacuum-assisted closure (VAC).

METHODSThe chronic wounds in 5 patients were treated with VAC. The expressions of MMP-1, 2, 13 in the granulation tissues of the chronic wounds were determined and quantified using RT-PCR technique before and at 1, 4, 7 days after the treatment.

RESULTSThe MMP-1, 13 mRNA showed obvious decrease, with the steepest variation of MMP-13. The MMP-2 mRNA also showed a decreased tendency, though in an undulatory fashion.

CONCLUSIONVAC can promote healing of chronic wounds through depressing the expressions of MMP-1, 2, 13 mRNA and protein synthesis, depressing the degradations of collagen and gelatin.

Collagen ; metabolism ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 13 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Negative-Pressure Wound Therapy ; Polymerase Chain Reaction ; Time Factors ; Wound Healing

OBJECTIVE: To investigate the effect of pure Chinese herbal extract Mangiferin on the malignant biological behaviors of multiple myeloma (MM) cells, and to analyze the molecular mechanism of the anti-myeloma effect of Mangiferin, so as to provide experimental basis for MM replacement therapy. METHODS: U266 and RPMI8226 of human MM cell lines were intervened with different concentrations of Mangiferin. Cell proliferation was detected by CCK-8 method. Annexin V/PI double staining flow cytometry was used to detect cell apoptosis. Western blot was used to detect the expression of apoptosis and related signaling pathway proteins, and real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of matrix metalloproteinase (MMP) and CXC chemokine receptor (CXCR) family. RESULTS: Mangiferin could inhibit the proliferation activity of U266 and RPMI8226 cells and induce cells apoptosis. After Mangiferin intervened in U266, RPMI8226 cells for 48 h, the expression of Bcl-2 family pro-apoptotic protein Bax was up-regulated, while the expression of survivin and Bcl-xL proteins was down-regulated and caspase-3 was hydrolyzed and activated to promote cell apoptosis, besides, the expression of Bcl-2 protein in U266 cells was also significantly down-regulated to induce apoptosis (P<0.05). After Mangiferin intervenes in MM cells, it can not only increase the expression level of tumor suppressor p53, but also induce programmed cell death of MM cells by inhibiting the expression of anti-apoptotic molecules and down-regulating the phosphorylation levels of AKT and NF-κB. In addition, after the intervention of Mangiferin, the expressions of CXCR4, MMP2 and MMP9 in U266 cells were down-regulated (P<0.05), while there is no effect on the expressions of CXCR2, CXCR7 and MMP13 (P>0.05). However, the expressions of CXCR4, MMP9, and MMP13 in RPMI8226 cells were down-regulated (P<0.01), the expression of MMP2 was weakly affected, and the expression of CXCR2 and CXCR7 was basically not affected (P>0.05). CONCLUSION Mangiferin can inhibit the proliferation and induce apoptosis of MM cells, and its mechanism may be related to inhibiting the activation of NF-κB signaling pathway, affecting the expression of Bcl-2 family proteins, and inhibiting the expression of core members of MMP and CXCR family.
Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinase 13 ; Cell Line, Tumor ; NF-kappa B ; Multiple Myeloma/pathology* ; Cell Proliferation ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2

OBJECTIVETo study the effect of hyperthermia on anti-invasion of Tca8113 and the expression change of matrix metalloproteinase-13 (MMP-13) and calcium-binding protein S100A4 (S100A4).

METHODSTca8113 cell pools were incubated at 43 degrees C for 0, 40, 80, 120 min, respectively, and at 37, 41, 43, 45 degrees C respectively for 80 min. The effect of high temperatures on the invasion ability of Tca8113 was measured in vitro. The slides of cells were made and incubated at 43 degrees C for 0, 40, 80, 120 min, respectively. Immunocytochemical method was employed for detecting the expression change of MMP-13 and S100A4 protein. Tca8113 cells were incubated at 43 degrees C for 0, 40, 80, 120 min respectively and at 37, 41, 43, 45 degrees C respectively for 80 min. Western blot method was conducted for detecting the expressionchange of MMP-13 and S100A4 protein.

RESULTSAs incubating time at higher temperature lasted, the proportion of the cells with invasion ability decreased. Except groups of 40 min and 80 min at 43 degrees C and 41, 43 degrees C for 80 min, the rest groups show significant statistic differences (P < 0.05). The expression intensity of MMP-13 and S100A4 proteins in Tca8113 cells would decrease as incubating time at higher temperature lasted. The content of MMP-13 and S100A4 proteins would decrease as incubating time at higher temperature lasted or incubating temperature increased. Except the groups of 40, 80 min at 43 degrees C and 41, 43 degrees C for 80 min, statistic differences were identified (P < 0.05).

CONCLUSIONThe invasion of Tca8113 could be inhibited by hyperthermia. The mechanism of this effect may be due to protein expression inhibition of MMP-13 and S100A4.

Calcium-Binding Proteins ; Cell Line, Tumor ; Humans ; Hyperthermia, Induced ; Matrix Metalloproteinase 13 ; S100 Proteins

INTRODUCTION: Middle ear cholesteatoma is notorious for its capacity to absorb surrounding bone, but the exact mechanism of bone destruction is not fully understood until now. Recently, a group of matrix metalloproteinases (MMPs) were found to dissolve extracellular matrices. Among them, collagenase-3 (MMP-13), a newly found collagenase, is thought to be a strong enzyme to dissolve bone and cartilage. So we tried to find out whether Collagenase-3 is found in cholesteatoma and plays some role in bone destruction by cholesteatoma. MATERIALS AND METHOD: We performed immunohistochemical stains on 5 cholesteatomas, 5 middle ear granulations, 5 normal middle ear mucosa and 5 deep meatal skin specimens with anticollagenase-3 antibody and analysed the staining patterns. RESULTS: All the cholesteatomas showed strong immunoreactivity with collagenase-3. Two out of 5 granulations showed somewhat weaker and more disseminated patterns of immunoreactivity, but the rest 3 granulations showed no immunoreactivity. Normal middle ear mucosa and deep meatal skin specimens showed no immunoreactivity at all. CONCLUSIONS: Collagenase-3 may play an active role in the process of bone destruction by cholesteatoma; however, a further study using zymography or blotting method is needed to make this clear.
Cartilage ; Cholesteatoma ; Cholesteatoma, Middle Ear* ; Collagenases ; Coloring Agents ; Ear, Middle* ; Extracellular Matrix ; Humans* ; Matrix Metalloproteinase 13* ; Matrix Metalloproteinases ; Mucous Membrane ; Skin

BACKGROUND: Various topical cosmeceuticals and lasers have been used to treat photo-aged skin that has wrinkles, acne scars and dilated pores. The microneedle therapy system (MTS) that mechanically makes multiple holes on the skin has come into the limelight to treat these skin problems via stimulating collagen remodeling. The automicroneedle therapy system (AMTS) is a developed version of MTS and it has several advantages compared with conventional MTS. AMTS can achieve regular treatment results because of its automatically punching method. In addition, AMTS can treat smaller area and it has cost advantages due to the inexpensive disposable needle head. OBJECTIVE: This study was designed to determine the dermal proliferative effects and safety of AMTS on the skin compared with that of the conventional MTS roller. METHODS: Twelve hairless mice were divided into two groups; one group was treated with a 0.25 mm needle and the other group was treated with a 2 mm needle. The first group was subdivided into the AMTS-H and the MTS groups and the no treatment group as a control, while the second group was subdivided into the AMTS-H, AMTS, MTS and control groups. Each treated group underwent four procedures every other day. The dermal proliferative efficacies of the treatment were evaluated by the histology, including the dermal thickness and the densities of the collagen fibers. Western blot was also performed for the evaluation of the protein expression of procollagen type I and matrix metalloproteinase-13. For safety profiles, we performed gross observation, basal skin barrier function testing and histologic examination. RESULTS: Treatment by AMTS significantly increased dermal collagen synthesis and the dermal thickness in the hairless mice. In addition, the expression of procollagen type I protein was increased, which accounted for the increased dermal collagen density. There was no specific safety problem related to the treatment. CONCLUSION: These results indicated that AMTS is an effective, safe modality for treating skin problems that require dermal proliferation. We anticipate that AMTS could be a new therapeutic option for inducing dermal proliferation or regeneration.
Acne Vulgaris ; Animals ; Blotting, Western ; Cicatrix ; Collagen ; Collagen Type I ; Head ; Matrix Metalloproteinase 13 ; Mice ; Mice, Hairless ; Needles ; Regeneration ; Skin

OBJECTIVE: To prepare a novel hyaluronic acid methacrylate (HAMA) hydrogel microspheres loaded polyhedral oligomeric silsesquioxane-diclofenac sodium (POSS-DS) patricles, then investigate its physicochemical characteristics and in vitro and in vivo biological properties. METHODS: Using sulfhydryl POSS (POSS-SH) as a nano-construction platform, polyethylene glycol and DS were chemically linked through the "click chemistry" method to construct functional nanoparticle POSS-DS. The composition was analyzed by nuclear magnetic resonance spectroscopy and the morphology was characterized by transmission electron microscopy. In order to achieve drug sustained release, POSS-DS was encapsulated in HAMA, and hybrid hydrogel microspheres were prepared by microfluidic technology, namely HAMA@POSS-DS. The morphology of the hybrid hydrogel microspheres was characterized by optical microscope and scanning electron microscope. The in vitro degradation and drug release efficiency were observed. Cell counting kit 8 (CCK-8) and live/dead staining were used to detect the effect on chondrocyte proliferation. Moreover, a chondrocyte inflammation model was constructed and cultured with HAMA@POSS-DS. The relevant inflammatory indicators, including collagen type Ⅱ, aggrecan (AGG), matrix metalloproteinase 13 (MMP-13), recombinant A disintegrin and metalloproteinase with thrombospondin 5 (Adamts5), and recombinant tachykinin precursor 1 (TAC1) were detected by immunofluorescence staining and real-time fluorescence quantitative PCR, with normal cultured chondrocytes and the chondrocyte inflammation model without treatment as control group and blank group respectively to further evaluate their anti-inflammatory activity. Finally, by constructing a rat model of knee osteoarthritis, the effectiveness of HAMA@POSS-DS on osteoarthritis was evaluated by X-ray film and Micro-CT examination. RESULTS: The overall particle size of POSS-DS nanoparticles was uniform with a diameter of about 100 nm. HAMA@POSS-DS hydrogel microspheres were opaque spheres with a diameter of about 100 μm and a spherical porous structure. The degradation period was 9 weeks, during which the loaded POSS-DS nanoparticles were slowly released. CCK-8 and live/dead staining showed no obvious cytotoxicity at HAMA@POSS-DS, and POSS-DS released by HAMA@POSS-DS significantly promoted cell proliferation (P<0.05). In the chondrocyte anti-inflammatory experiment, the relative expression of collagen type Ⅱ mRNA in HAMA@POSS-DS group was significantly higher than that in control group and blank group (P<0.05). The relative expression level of AGG mRNA was significantly higher than that of blank group (P<0.05). The relative expressions of MMP-13, Adamts5, and TAC1 mRNA in HAMA@POSS-DS group were significantly lower than those in blank group (P<0.05). In vivo experiments showed that the joint space width decreased after operation in rats with osteoarthritis, but HAMA@POSS-DS delayed the process of joint space narrowing and significantly improved the periarticular osteophytosis (P<0.05). CONCLUSION HAMA@POSS-DS can effectively regulate the local inflammatory microenvironment and significantly promote chondrocyte proliferation, which is conducive to promoting cartilage regeneration and repair in osteoarthritis.
Animals ; Rats ; Matrix Metalloproteinase 13 ; Microspheres ; Hydrogels ; Collagen Type II ; Diclofenac ; Inflammation ; Osteoarthritis, Knee/drug therapy* ; Hyaluronic Acid ; Aggrecans

OBJECTIVE: To explore the effect of miR-143 regulating matrix metalloproteinase(MMP)-13 expression on migration and invasion of osteosarcoma cells. METHODS: The mouse osteosarcoma cell line 143B cells were cultured in 96-well plates, and blank group, negative group, positive group, and intervention group were set up. Then, the blank group did no treatment 50 μg miR-143 mimic was added to positive group, negative group added equal mimic NC (control sequence of miR-143 mimic), the intervention group was added 50 μg miR-143 mimic and 10 μg MMP-13 protein, all groups continued to culture for 3 to 6 hours, and finally the serum was aspirated to treat for half an hour. The protein expressions of miR-143 and MMP-13 in each group were measured by fluorescence quantitative PCR experiment and Western blot experiment, respectively, and the invasion and migration abilities of cells were measured by Transwell and scratch experiments. RESULTS: The expression of MMP-13 protein in the positive group and the intervention group was significantly lower than that in the blank group, and the positive group was lower than the intervention group (P<0.05);The mean numbers of invasive cells in blank group, negative group, positive group and intervention group were (1 000.01±44.77), (959.25±46.32), (245.04±4.33), (634.06±33.78) cells/field, respectively;the scratch healing rate of the positive group and the intervention group was significantly lower than that of the blank group, and the positive group was lower than the intervention group (P<0.05). CONCLUSION MMP-13 is a target of miR-143, which can reduce the migration and invasion ability of osteosarcoma cells by inhibiting the expression of MMP-13.
Osteosarcoma/pathology* ; MicroRNAs/genetics* ; Matrix Metalloproteinase 13/genetics* ; Neoplasm Invasiveness ; Animals ; Mice ; Cell Line, Tumor ; Cell Movement

AIMTo explore the possible impact of endogenous hydrogen sulfide (H2S) on the content and metabolism of collagen in rats with high pulmonary blood flow.

METHODSThirty-two male SD rats, weighing 120-140 g, were randomly divided into 4 groups (n = 8), shunt group, shunt + PPG (propargylglycine, an antagonist of endogenous H2S producing enzyme) group, sham group and sham + PPG group. Rats in shunt group and shunt + PPG group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. In the sham group and sham + PPG group, rats experienced the same experimental processes except the shunting procedure. After 4 weeks of experiment, lung tissue H2S content of rat was determined by a modified sulfide electrode method. Pulmonary artery collagen I, collagen III, MMP-13 and TIMP-1 protein expressions of rat were investigated by immunohistochemistry.

RESULTSAfter 4 weeks of experiment, lung tissue H2S content increased significantly in rats of shunt group as compared with that of sham group (P < 0.05). Pulmonary artery collagen I and collagen III protein expression increased obviously in rats of shunt group as compared with that of sham group (P < 0.01). After administration of PPG for 4 weeks, lung tissue H2S content decreased significantly in rats of shunt + PPG group as compared with that of shunt group (P < 0.05). In contrast to rats in shunt group, collagen I and collagen III protein expression in pulmonary arteries of shunt + PPG group increased significantly, respectively (P < 0.05). Compared with rats of shunt group, pulmonary artery MMP-13, TIMP-1 and the ratio of MMP-13/TIMP-1 in shunt + PPG group down-regulated significantly (P < 0.05).

CONCLUSIONEndogenous H2S might play a protective regulatory role in the development of pulmonary hypertension and pulmonary vascular structural remodelling in rats by decreasing the content of pulmonary artery collagen resulting from catabolism of collagen.

Animals ; Collagen ; metabolism ; Hydrogen Sulfide ; metabolism ; Lung ; blood supply ; metabolism ; Male ; Matrix Metalloproteinase 13 ; metabolism ; Pulmonary Artery ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo investigate whether the functional polymorphisms in the promoter region of MMP-12 (-82A/G) and MMP-13(-77A/G) are associated with epithelial ovarian carcinoma (EOC).

METHODSThe MMP-12 -82A/G and MMP-13 -77A/G were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 300 epithelial ovarian carcinoma patients and 300 control women.

RESULTSThe A/G genotype frequency of the MMP-12 gene was significantly higher in the patients than in the controls (P= 0.003); similarly, the frequency of MMP-12 -82G allele was higher in the patient group (P= 0.004). Compared with the A/A genotype, the A/G genotype carriers significantly increased the risk of EOC development (OR= 2.81, 95%CI: 1.38-5.74). No overall association between the MMP-13 -77A/G polymorphism and EOC(P= 0.15) was observed. However, the A/A genotype carriers in the MMP-13 -77A/G locus had significantly higher risk of developing serous-papillary and mucinous ovarian cancer (OR= 1.93, 95% CI: 1.05-3.53; OR= 5.16, 95% CI: 1.62-16.44, respectively), comparing with the G/G genotype carriers. Combining the two SNPs, the haplotype distributions in patients were not significantly different from that in control women (P= 0.06).

CONCLUSIONThese results suggested that individuals with MMP-12 -82A/G and MMP-13 -77A/A might have higher risk of overall or special histological type of EOC development.

Adult ; Aged ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Matrix Metalloproteinase 12 ; genetics ; Matrix Metalloproteinase 13 ; genetics ; Middle Aged ; Neoplasms, Glandular and Epithelial ; genetics ; Ovarian Neoplasms ; genetics ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics ; Young Adult