NaF Inhibits Matrix-Bound Cathepsin-Mediated Dentin Matrix Degradation

2016 ◽  
Vol 50 (2) ◽  
pp. 124-132 ◽  
Author(s):  
Pinar Altinci ◽  
Murat Mutluay ◽  
Roda Seseogullari-Dirihan ◽  
David Pashley ◽  
Leo Tjäderhane ◽  
...  
Keyword(s):  
Mass Loss ◽  
Artificial Saliva ◽  
Gelatin Zymography ◽  
Cathepsin K ◽  
Dry Mass ◽  
Direct Inhibition ◽  
Cysteine Cathepsins ◽  
Matrix Bound ◽  
Dentin Matrix ◽  
Time Point

Matrix metalloproteinases (MMPs) and cysteine cathepsins (CCs) degrade the collagen fibrils of demineralized dentin. Sodium fluoride (NaF) has previously been shown to inhibit recombinant MMP-2 and MMP-9. This study aimed to evaluate the efficacy of NaF on the inhibition of dentin-bound MMPs and CCs. Dentin beams were completely demineralized in 10% phosphoric acid. The baseline total MMP activity and dry masses were measured. Beams were assigned to test groups based on similar MMP activity and dry mass (n = 10/group), and incubated in artificial saliva (control) or artificial saliva with NaF containing 6-238 mM fluoride for 1, 7 and 21 days. The dry mass loss and MMP activities were reassessed at each time point. The proteolytic activity was screened by gelatin zymography. ICTP and CTX released to the incubation medium were analyzed as indices of MMP and cathepsin K activity, respectively. The beams were examined under scanning electron microscopy. All NaF doses reduced the dry mass loss after 21 days (p < 0.05). NaF inhibition of the total MMP activity ranged between 5 and 80%. In gelatin zymography, the bands of MMP-2 and MMP-9 became less prominent with increasing NaF levels. NaF did not decrease the released ICTP (p > 0.05). Less CTX release was detected with F ≥179 mM (p < 0.05). CaF2-like minerals were observed on the beams. High levels of NaF may slow the degradation of the dentin matrix due to the inhibition of cathepsin K. Fluoride does not seem effective in the direct inhibition of proteolysis by dentin matrix-bound MMPs.

Effect of EGCG Application on Collagen Degradation in Dentine Caries

2013 ◽  
Vol 455 ◽  
pp. 112-116 ◽  
Author(s):  
Huang Qin Chen ◽  
Bin Huang
Keyword(s):  
Mass Loss ◽  
Storage Time ◽  
Artificial Saliva ◽  
Acetic Acid Solution ◽  
Dry Weight ◽  
Collagen Matrix ◽  
Dry Mass ◽  
Collagen Degradation ◽  
Experimental Media ◽  
Dentin Matrix

Caries progress might be controlled when collagen matrix could be preserved after demineralization. The aim of this study was to investigate the effect of EGCG, a major polyphenol of green tea, on changes in loss of dry weight or solubilization of collagen from demineralized dentin sections. Dentin sections (1mm×5mm×5mm) were completely exposed to the acetic acid solution to form dentine caries. After baseline measurements of dry mass, the samples were divided into 2 groups and incubated at 37°C in either media containing EGCG or artificial saliva only for 24h, 3 days, 1 week, 2 week and 3 weeks. Following hydrolyzed with HCl, hydroxyproline (HOP) as an index of solubilization of collagen was analysed quantitatively. Both the storage medium and the storage time showed significant effects on mass loss and HOP release (p < 0.05). The incubation in EGCG resulted in relatively rapid and significant (p < 0.05) decrease in the amounts of mass loss. The HOP content of the experimental media also increased with incubation time but was significantly lower (p < 0.05) than in the control artificial saliva medium. It can be concluded that EGCG can inhibit collagen degradation and affects the further demineralization in dentin matrix, which consequently inhibits the advanced dentine caries.

Zinc Inhibits Collagenolysis by Cathepsin K and Matrix Metalloproteinases in Demineralized Dentin Matrix

2017 ◽  
Vol 51 (6) ◽  
pp. 576-581 ◽  
Author(s):  
Pinar Altinci ◽  
Roda Seseogullari-Dirihan ◽  
Gulsen Can ◽  
David Pashley ◽  
Arzu Tezvergil-Mutluay
Keyword(s):  
Matrix Metalloproteinases ◽  
Mass Loss ◽  
Type I Collagen ◽  
Cathepsin K ◽  
Collagen Degradation ◽  
Type I ◽  
Level Control ◽  
Physiological Level ◽  
Cysteine Cathepsins ◽  
Demineralized Dentin

The enzymatic degradation of dentin organic matrix occurs via both the action of matrix metalloproteinases (MMPs) and cysteine cathepsins (CCs). Zinc can prevent collagen hydrolysis by MMPs. However, its effect on the activity of dentin-bound CCs is not known. The aim of this study was to investigate the effect of zinc on matrix-bound cathepsin K and MMP activity in dentin. Completely demineralized dentin beams were divided into test groups (n = 9) and incubated at 37°C in an incubation media (1 mL) containing ZnCl2 of 0.02 (physiological level, control), 0.2, 0.5, 1, 5, 10, 20, 30, or 40 mM. The dry mass changes of the beams were determined, and incubation media were analyzed for cathepsin K- and MMP-specific collagen degradation end products - CTX (C-terminal cross-linked telopeptide of type I collagen) and ICTP (cross-linked carboxy-terminal telopeptide of type I collagen) - at 1, 3, and 7 days of incubation. The mass loss of the beams decreased when the zinc level in the incubation media was ≥5 mM (p < 0.05). The release of liberated collagen degradation telopeptides decreased in accordance with the decrease in the mass loss rates of the beams. Cathepsin K-induced dentin collagen degradation can be strongly inhibited by zinc. Zinc levels of ≥5 mM can be considered as a reliable threshold for the stabilization of dentin matrices.

Inhibition of dentin matrix-bound cysteine cathepsins by potassium fluoride

2018 ◽  
Vol 127 (1) ◽  
pp. 1-9
Author(s):  
Pinar Altinci ◽  
Murat Mutluay ◽  
Leo Tjäderhane ◽  
Arzu Tezvergil-Mutluay
Keyword(s):  
Potassium Fluoride ◽  
Cysteine Cathepsins ◽  
Matrix Bound ◽  
Dentin Matrix

Pharmacological inhibitors to identify roles of cathepsin K in cell-based studies: a comparison of available tools

2009 ◽  
Vol 390 (9) ◽  
Author(s):  
Sylvie Desmarais ◽  
Frédéric Massé ◽  
M. David Percival
Keyword(s):  
Human Cell ◽  
Type I Collagen ◽  
Cathepsin K ◽  
The Other ◽  
Enzyme Assays ◽  
Type I ◽  
Treatment Of Osteoporosis ◽  
Cysteine Cathepsins ◽  
Rodent Cell ◽  
Enzyme Selectivity

Abstract Cathepsin K (Cat K) degrades bone type I collagen and is a target for the pharmacological treatment of osteoporosis. Further roles for Cat K have been recently described, some of which are supported by the use of purportedly selective Cat K inhibitors in human and rodent cell-based assays. Twelve commercial and non-commercial Cat K inhibitors were profiled against a panel of purified human, rat, and mouse cysteine cathepsins and in two cell-based enzyme occupancy assays for activity against Cat K, B, and L. Ten inhibitors, including the carbohydrazide Cat K inhibitor II (Boc-Phe-Leu-NHNH-CO-NHNH-Leu-Z), the non-covalent K4b, and the epoxide NC-2300, have either little Cat K selectivity, or appear poorly cell penetrant. The amino-acetonitrile-containing inhibitors L-873724 and odanacatib show greater than 100-fold human Cat K enzyme selectivity and have similar IC50 values against each cathepsin in cell-based and enzyme assays. The basic inhibitor balicatib has greater cellular potencies than expected on the basis of purified enzyme assays. The accumulation of [14C]-balicatib in fibroblasts is blocked by prior treatment of the cells with NH4Cl, consistent with balicatib having lysosomotropic properties. These results support the use of L-873724 and odanacatib as tools to identify novel roles for Cat K using human cell-based systems, but suggest using caution in the interpretation of studies employing the other compounds.

Seed production in Cardiocrium cordatum: effects of nitrogen and dry mass availability of a plant

2006 ◽  
Vol 84 (5) ◽  
pp. 805-812
Author(s):  
Satoki Sakai ◽  
Akiko Sakai ◽  
Kohta Fujioka
Keyword(s):  
Mass Loss ◽  
Seed Production ◽  
Plant Size ◽  
Dry Mass ◽  
Stem Diameter ◽  
N Content ◽  
N Concentration ◽  
The Mean ◽  
Number Of Seeds

To clarify how seed production depends on the relative N and dry mass availability, we examined the number and size of seeds, and the loss of dry mass in Cardiocrium cordatum (Thunb.) Makaino plants that differed naturally in overall size and stem N content. After adjusting for plant size (basal stem diameter was used as a criterion of plant size), the amount of dry mass lost because of respiration in a plant decreased with an increase in the stem N content of the plant, whereas the total dry mass of seeds of a plant increased with an increase in the stem N content. Plants with a high stem N content relative to dry mass status used dry mass resources more efficiently in seed production by reducing the loss of dry mass. Plants with a higher stem N content produced a greater number of seeds, and the amount of dry mass lost decreased with an increase in the number of seeds of the plant, possibly because the resources allocated to seed production are consumed rapidly, if numerous seeds simultaneously absorb those resources. However, the stem N content of a plant had no influence on the mean seed dry mass, seed N content, and the seed N concentration, and the latter three parameters did not affect respiration loss of dry mass. We concluded that plants undergoing seed production reduce dry mass loss if their relative N to dry mass availability is high, through an increase in the numbers of seeds they produce.

scholarly journals In Vitro Evaluation of Benzalkonium Chloride in the Preservation of Adhesive Interfaces

2014 ◽  
Vol 39 (3) ◽  
pp. 283-290 ◽  
Author(s):  
C Sabatini ◽  
JH Kim ◽  
P Ortiz Alias
Keyword(s):  
Bond Strength ◽  
Mass Loss ◽  
Benzalkonium Chloride ◽  
Artificial Saliva ◽  
In Vitro Study ◽  
Control Group ◽  
Significance Level ◽  
Bond Stability ◽  
Etch And Rinse

SUMMARY Inhibition of endogenous dentin matrix metalloproteinases (MMPs) by benzalkonium chloride (BAC) decreases collagen solubilization and may help improve resin-dentin bond stability. Objective: This study evaluated the resin-dentin bond stability of experimental adhesive blends containing BAC and the stability of dentin matrices by assessing the mass loss and collagen solubilization from dentin beams pretreated with BAC. Materials and Methods Twenty-five healthy molars were used for the bond strength evaluation of a two-step etch-and-rinse adhesive (Adper Single Bond Plus, SB) modified with BAC or not. The following groups were tested: 1) SB with no inhibitor (control); 2) topical 2.0% chlorhexidine + SB; 3) 1.0% BAC etchant + SB; 4) 0.5% BAC-SB; and 5) 1.0% BAC-SB. Microtensile bond strength (μTBS) and failure mode distribution under standard error of the mean were evaluated after 24 hours and six months of storage in artificial saliva (AS). A two-way analysis of variance and Tukey test with a significance level of p&lt;0.05 was used for data analysis. In addition, 30 completely demineralized dentin beams from human molars were either dipped in deionized water (DW, control) or dipped in 0.5% and 1.0% BAC for 60 seconds, and then incubated in AS. Collagen solubilization was assessed by evaluating the dry mass loss and quantifying the amount of hydroxyproline (HYP) released from hydrolyzed specimens after four weeks of incubation. Results The control group demonstrated lower μTBS than some of the experimental groups containing BAC at 24 hours and six months (p&lt;0.05). When BAC was incorporated into the adhesive blend in concentrations of 0.5% and 1.0%, no reduction in dentin bond strength was observed after six months (p&lt;0.05). Less mass loss and HYP release was seen for dentin matrices pretreated with BAC relative to the control pretreated with DW (p&lt;0.05). Conclusion This in vitro study demonstrates that BAC contributes to the preservation of resin-dentin bonds by reducing collagen degradation.

scholarly journals Effects of Composites Containing Bioactive Glasses on Demineralized Dentin

2017 ◽  
Vol 96 (9) ◽  
pp. 999-1005 ◽  
Author(s):  
A. Tezvergil-Mutluay ◽  
R. Seseogullari-Dirihan ◽  
V.P. Feitosa ◽  
G. Cama ◽  
D.S. Brauer ◽  
...  
Keyword(s):  
Bioactive Glass ◽  
Direct Contact ◽  
Artificial Saliva ◽  
Dry Mass ◽  
Sem Analysis ◽  
Dental Composites ◽  
Bioactive Glasses ◽  
Beneficial Effects ◽  
Before And After ◽  
Demineralized Dentin

The aim of this study was to evaluate the degradation of completely demineralized dentin specimens in contact with a filler-free or 2 ion-releasing resins containing micrometer-sized particles of Bioglass 45S5 (BAG) or fluoride-containing phosphate-rich bioactive glass (BAG-F). Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) were also used to evaluate the remineralization induced by the experimental ion-releasing resin-based materials. Dentin beams were totally demineralized in H3PO4 (10%) and placed in direct contact with a filler-free (RESIN) or 2 experimental ion-releasing resins (BAG or BAG-F) and immersed in artificial saliva (AS) up to 30 d. Further specimens were also processed and submitted to FTIR and SEM analysis to evaluate the remineralization induced by such ion-releasing resins before and after AS immersion. BAG and BAG-F alkalinized the incubation media. A significant decrease of the dry mass was observed between the specimens of all groups stored for 3 and 30 d in AS. However, the fluoride-containing phosphate-rich bioactive glass incorporated into a resin-based material (BAG-F) showed greater ability in reducing the solubilization of C-terminal cross-linked telopeptide (ICTP) and C-terminal telopeptide (CTX) after prolonged AS storage. Moreover, after 30 d of AS storage, BAG-F showed the greatest remineralizing effect on the stiffness of the completely demineralized dentin matrices. In conclusion, fluoride-containing phosphate-rich bioactive glass incorporated as micrometer-sized filler in dental composites may offer greater beneficial effects than Bioglass 45S5 in reducing the enzyme-mediated degradation and remineralization of demineralized dentin.

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

2007 ◽  
Vol 86 (5) ◽  
pp. 436-440 ◽  
Author(s):  
A. Mazzoni ◽  
F. Mannello ◽  
F.R. Tay ◽  
G.A.M. Tonti ◽  
S. Papa ◽  
...  
Keyword(s):  
Gelatin Zymography ◽  
Molecular Forms ◽  
Zymographic Analysis ◽  
Caries Progression ◽  
Dentin Matrix ◽  
Demineralized Dentin Matrix ◽  
And Function ◽  
Role And Function ◽  
Demineralized Dentin

The role and function of dentin matrix metalloproteinases (MMPs) are not well-understood, but they may play a key role in dentinal caries and the degradation of resin-bonded dentin matrices. To test the null hypothesis that MMP-9 is not found in dentin matrix, we used gelatin zymography to extract and isolate all molecular forms of gelatinolytic MMPs in demineralized mature sound dentin powder obtained from extracted human molars, characterizing and identifying the enzymes by Western blotting. Gelatinolytic MMPs were detected in extracts of demineralized dentin matrix and identified as MMP-2 and MMP-9. Acidic extracts (pH 2.3) yielded 3–8 times more MMP activity than did EDTA (pH 7.4). Their activation may contribute to dentin matrix degradation, which occurs during caries progression and following resin bonding. Inhibition of MMP-2 and -9 proteolytic activity may slow caries progression and increase the durability of resin-dentin bonds.

Metalloproteinases and their role in the degradation of bonding systems. Part 2

2018 ◽  
Vol 23 (4) ◽  
Author(s):  
Aneta Zduniak ◽  
Sylwia Olszewska ◽  
Agnieszka Mielczarek
Keyword(s):  
Collagen Matrix ◽  
Bacterial Biofilm ◽  
Medical Database ◽  
Hybrid Layer ◽  
Cysteine Cathepsins ◽  
Dentin Matrix ◽  
Endogenous Proteases ◽  
The Stability ◽  
Bonding Systems

The stability of the hybrid layer is crucial for ensuring the durability of fillings made of composite materials. Factors which weaken the bond strength are related to, among others, the presence of bacteria and their enzymes in the structure of the bacterial biofilm. Chronic damage the hybrid layer is also a result of hydrolysis and leaching of adhesive monomers which infiltrated the demineralised dentin matrix. Nanoleakage is also among the factors contributing to degradation. Many studies examine the effect of endogenous proteases on the degradation of the hybrid layer. Endogenous collagenolytic enzymes: metalloproteinases (MMPs) and cysteine cathepsins, are responsible for the degradation of the collagen matrix in the hybrid layer. Inhibition of endogenous proteases is therefore necessary to slow the degradation of fillings. The enzyme activity in dentine and in the hybrid layer can be regulated by endo- and exogenous inhibitors. The paper is a review of the available literature published in the PubMed medical database, as well as in Polish dental journals in the years 2002-2017. Its aim is to assess the role of metalloproteinases and cysteine cathepsins in the degradation of the hybrid layer and to review the compounds with inhibitory properties in relation to these enzyme groups.

Regulation of cathepsin K activity by hydrogen peroxide

2008 ◽  
Vol 389 (8) ◽  
Author(s):  
Emmanuel Godat ◽  
Virginie Hervé-Grépinet ◽  
Florian Veillard ◽  
Fabien Lecaille ◽  
Maya Belghazi ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Lung Inflammation ◽  
Cathepsin K ◽  
Initial Activity ◽  
Acute Lung Inflammation ◽  
Sulfenic Acid ◽  
Cysteine Cathepsins ◽  
Dose Dependent ◽  
Insight Into ◽  
K Activity

Abstract Although cysteine cathepsins, including cathepsin K, are sensitive to oxidation, proteolytically active forms are found at inflammatory sites. Regulation of cathepsin K activity was analyzed in the presence of H2O2 to gain an insight into these puzzling observations. H2O2 impaired processing of procathepsin K and inactivated its mature form in a time- and dose-dependent mode. However, as a result of the formation of a sulfenic acid, as confirmed by trapping in the presence of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazol, approximately one-third of its initial activity was restored by dithiothreitol. This incomplete inactivation may partially explain why active cysteine cathepsins are still found during acute lung inflammation.

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