Zymographic Analysis and Characterization of MMP-2 and -9 Forms in Human Sound Dentin

Mazzoni, Annalisa; Mannello, Ferdinando; Tay, FR; Tonti, Ga; Papa, Stefano; Mazzotti, Giovanni; Lenarda, Roberto Di; Pashley, D. H.; Breschi, Lorenzo

doi:10.1177/154405910708600509

Cited by 288 publications

(299 citation statements)

References 31 publications

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“…Following electrophoresis, the gel was divided into equal parts, and the individual strips were developed in different developing buffers containing 5 lg/ml CSE, 10 lg/ml CSE, or 1 mM EDTA as positive control, respectively (Albini et al 2001;Mazzoni et al 2007). …”

Section: Gelatin Zymographymentioning

confidence: 99%

Caesalpinia sappan extract inhibits IL1β-mediated overexpression of matrix metalloproteinases in human chondrocytes

Toegel

Otero

et al. 2011

Genes Nutr

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Exacerbated production of matrix metalloproteinases (MMPs) is a key event in the progression of osteoarthritis (OA) and represents a promising target for the management of OA with nutraceuticals. In this study, we sought to determine the MMP-inhibitory activity of an ethanolic Caesalpinia sappan extract (CSE) in human OA chondrocytes. Thus, human articular chondrocytes isolated from OA cartilage and SW1353 chondrocytes were stimulated with Interleukin-1beta (IL1b), without or with pretreatment with CSE. Following viability assays, the production of MMP-2 and MMP-13 was assessed using ELISA, whereas mRNA levels of MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-13 and TIMP-1, TIMP-2, TIMP-3 were quantified using RT-qPCR assays. Chondrocytes were co-transfected with a MMP-13 luciferase reporter construct and NF-kB p50 and p65 expression vectors in the presence or absence of CSE. In addition, the direct effect of CSE on the proteolytic activities of MMP-2 was evaluated using gelatin zymography. We found that CSE significantly suppressed IL1b-mediated upregulation of MMP-13 mRNA and protein levels via abrogation of the NF-kB(p65/p50)-driven MMP-13 promoter activation. We further observed that the levels of IL1b-induced MMP-1, MMP-3, MMP-7, and MMP-9 mRNA, but not TIMP mRNA levels, were down-regulated in chondrocytes in response to CSE. Zymographic results suggested that CSE did not directly interfere with the proteolytic activity of MMP-2. In summary, this study provides evidence for the MMP-inhibitory potential of CSE or CSE-derived compounds in human OA chondrocytes. The data indicate that the mechanism of this inhibition might, at least in part, involve targeting of NF-kB-mediated promoter activation.

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Section: Gelatin Zymographymentioning

confidence: 99%

Caesalpinia sappan extract inhibits IL1β-mediated overexpression of matrix metalloproteinases in human chondrocytes

Toegel

Otero

et al. 2011

Genes Nutr

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“…[1][2][3][4][5][6][7] A correlation between the presence of active endogenous matrix metalloproteinases (MMPs) in dentin matrices and premature degradation of hybrid layers has been demonstrated. 8,9 Different forms of MMPs have been identified in dentin matrices by zymographic analyses, with MMP-2 and MMP-9 being the most prevalent. 8,10,11 When exposed to an acidic environment, such as the one created by the application of acidic adhesive resins, these enzymes, initially secreted as pro-enzymes, become active proteinases.…”

Section: Introductionmentioning

confidence: 99%

“…8,9 Different forms of MMPs have been identified in dentin matrices by zymographic analyses, with MMP-2 and MMP-9 being the most prevalent. 8,10,11 When exposed to an acidic environment, such as the one created by the application of acidic adhesive resins, these enzymes, initially secreted as pro-enzymes, become active proteinases. 12 These host-derived MMPs have been shown to degrade sub-optimally infiltrated collagen fibers once they have been activated through bonding procedures.…”

Section: Introductionmentioning

confidence: 99%

Effect of a Chlorhexidine-containing Adhesive on Dentin Bond Strength Stability

Sabatini

2013

Operative Dentistry

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A novel chlorhexidine-containing adhesive may offer comparable dentin-resin bond stability to the use of 2.0% chlorhexidine digluconate applied as a therapeutic primer while also providing a simplified clinical application technique. SUMMARYPurpose: The present study aimed to investigate a novel adhesive system containing 0.2% chlorhexidine digluconate (CHX) for its ability to improve the stability of the adhesive interface compared with the use of 2% CHX as a therapeutic primer. Furthermore, the study aimed to confirm the inhibitory properties of these CHX concentrations (0.2% and 2.0%) on dentin matrix metalloproteinase activity by gelatin zymography.Methods: Superficial dentin substrate for bonding was obtained from 120 non-carious human molars. A conventional adhesive Peak LC Bond and a CHX-containing adhesive Peak Universal Bond were used either in combination with 35% phosphoric acid (etch-and-rinse approach) or with self-etching primer (selfetch approach) for evaluation of the variables CHX treatment (2.0% therapeutic primer and 0.2% adhesive), adhesive approach (etch-andrinse and self-etch), and storage time (24 hours and six months). A bonding jig was used to fabricate composite cylinders, which were stored for either 24 hours or six months, after which shear bond strength (SBS) was evaluated using a notched-edge testing device. A three-way analysis of variance and a Student t-test with a significance level of p,0.05 were used to analyze the data. Extracts from concentrated demineralized human dentin powder were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and incubated in the presence of 0.2% and 2.0% CHX.Results: No significant effect of CHX treatment, adhesive approach, storage time variables, or their interactions on mean SBS was

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“…More recently, MMPs were identified as being responsible for that activity (Tjäderhane et al 1998), and to date, the presence of gelatinases MMP-2 and -9 ( Fig. 1), collagenase MMP-8, stromelysin MMP-3, and MMP-20 have been reported (Martin-De Las Heras et al 2000;Sulkala et al 2002;Mazzoni et al 2007;Sulkala et al 2007;Boukpessi et al 2008;Mazzoni et al 2009;Santos et al 2009;Boushell et al 2011;.…”

mentioning

confidence: 99%

Role of Dentin MMPs in Caries Progression and Bond Stability

et al. 2014

Self Cite

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Dentin can be described as a biological composite with collagen matrix embedded with nanosized hydroxyapatite mineral crystallites. Matrix metalloproteinases (MMPs) and cysteine cathepsins are families of endopeptidases. Enzymes of both families are present in dentin and collectively capable of degrading virtually all extracellular matrix components. This review describes these enzymes and their presence in dentin, mainly focusing on their role in dentin caries pathogenesis and loss of collagen in the adhesive hybrid layer under composite restorations. MMPs and cysteine cathepsins present in saliva, mineralized dentin, and/or dentinal fluid may affect the dentin caries process at the early phases of demineralization. Changes in collagen and noncollagenous protein structure may participate in observed decreases in mechanical properties of caries-affected dentin and reduce the ability of caries-affected dentin to remineralize. These endogenous enzymes also remain entrapped within the hybrid layer during the resin infiltration process, and the acidic bonding agents themselves (irrespective of whether they are etch-and-rinse or self-etch) can activate these endogenous protease proforms. Since resin impregnation is frequently incomplete, denuded collagen matrices associated with free water (which serves as a collagen cleavage reagent for these endogenous hydrolase enzymes) can be enzymatically disrupted, finally contributing to the degradation of the hybrid layer. There are multiple in vitro and in vivo reports showing that the longevity of the adhesive interface is increased when nonspecific enzyme-inhibiting strategies are used. Different chemicals (i.e., chlorhexidine, galardin, and benzalkonium chloride) or collagen cross-linker agents have been successfully employed as therapeutic primers in the bonding procedure. In addition, the incorporation of enzyme inhibitors (i.e., quaternary ammonium methacrylates) into the resin blends has been recently promoted. This review will describe MMP functions in caries and hybrid layer degradation and explore the potential therapeutic role of MMP inhibitors for the development of improved intervention strategies for MMP-related oral diseases.

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Zymographic Analysis and Characterization of MMP-2 and -9 Forms in Human Sound Dentin

Cited by 288 publications

References 31 publications

Caesalpinia sappan extract inhibits IL1β-mediated overexpression of matrix metalloproteinases in human chondrocytes

Caesalpinia sappan extract inhibits IL1β-mediated overexpression of matrix metalloproteinases in human chondrocytes

Effect of a Chlorhexidine-containing Adhesive on Dentin Bond Strength Stability

Role of Dentin MMPs in Caries Progression and Bond Stability

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