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Abstract

The co-expression of MMPs and cysteine cathepsins in the human dentin-pulp complex indicates that both classes of enzymes can contribute to the endogenous proteolytic activity of dentin. Chlorhexidine (CHX) is an efficient inhibitor of MMP activity. This study investigated whether CHX could also inhibit cysteine cathepsins present in dentin. The inhibitory profile of CHX on the activity of dentin-extracted and recombinant cysteine cathepsins (B, K, and L) was monitored in fluorogenic substrates. The rate of substrate hydrolysis was spectrofluorimetrically measured, and inhibitory constants were calculated. Molecular docking was performed to predict the binding affinity between CHX and cysteine cathepsins. The results showed that CHX inhibited the proteolytic activity of dentin-extracted cysteine cathepsins in a dose-dependent manner. The proteolytic activity of human recombinant cathepsins was also inhibited by CHX. Molecular docking analysis suggested that CHX strongly interacts with the subsites S2 to S2′ of cysteine cathepsins B, K, and L in a very similar manner. Taken together, these results clearly showed that CHX is a potent inhibitor of the cysteine cathepsins-proteolytic enzymes present in the dentin-pulp complex.

Keywords

cysteine cathepsins chlorhexidine proteolytic activity dentin collagen degradation

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References

Barrett

(1980). Fluorimetric assays for cathepsin B and cathepsin H with methyl coumaryl amide substrates. Biochem J 187:909-912.

Berman

Bhat

Bourne

Feng

Gilliland

Weissig

. (2000). The Protein Data Bank and the challenge of structural genomics. Nat Struct Biol 7(Suppl):957-959.

Bradford

(1976). A rapid and sensitive for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254.

Breschi

Martin

Mazzoni

Nato

Carrilho

Tjäderhane

. (2010). Use of a specific MMP inhibitor (Galardin) for preservation of hybrid layer. Dent Mater 26:571-578.

Carrilho

Geraldeli

Tay

de Goes

Carvalho

Tjäderhane

. (2007). In vivo preservation of the hybrid layer by chlorhexidine. J Dent Res 86:529-533.

Carrilho

Carvalho

Sousa

Nicolau

Breschi

Mazzoni

. (2010). Substantivity of chlorhexidine to human dentin. Dent Mater 26:779-785.

Chowdhury

Sivaraman

Wang

Devanathan

Lachance

. (2002). Design of noncovalent inhibitors of human cathepsin L. From the 96-residue proregion to optimized tripeptides. J Med Chem 45:5321-5329.

Dickinson

(2002). Cysteine peptidases of mammals: their biological roles and potential effects in the oral cavity and other tissues in health and disease. C.rit Rev Oral Biol Med 13:238-275

Gendron

Grenier

Sorsa

Mayrand

(1999). Inhibition of the activities of matrix metalloproteinases 2, 8, and 9 by chlorhexidine. Clin Diagn Lab Immunol 6:437-439.

10.

Gesteira

Coulson-Thomas

Taunay-Rodrigues

Oliveira

Thacker

Juliano

. (2011). Inhibitory peptides of the sulfotransferase domain of the heparan sulfate enzyme, N-deacetylase-N-sulfotransferase-1. J Biol Chem 286:5338-5346.

11.

Gordon

Schmidt

(2005). Advances in electronic structure theory: GAMESS a decade later. In: Theory and applications of computational chemistry: the first forty years. Dykstra

Frenking

Kim

Scuseria

, editors. Amsterdam: Elsevier, pp. 1167-1189.

12.

Greenspan

Clark

Tommasi

Cowen

McQuire

Farley

. (2001). Identification of dipeptidyl nitriles as potent and selective inhibitors of cathepsin B through structure-based drug design. J Med Chem 44:4524-4534.

13.

Halgren

(1999). MMFF VI. MMFF94s option for energy minimization studies. J Comput Chem 20:720-729.

14.

Hebling

Pashley

Tjäderhane

Tay

(2005). Chlorhexidine arrests subclinical degradation of dentin hybrid layers in vivo. J Dent Res 84:741-746.

15.

Kim

Mai

Carrilho

Yiu

Pashley

Tay

(2010). An all-in-one adhesive does not etch beyond hybrid layers. J Dent Res 89: 482-487.

16.

Lessa

Nogueira

Vargas Fda

Spolidorio

Hebling

García-Godoy

. (2010). Direct and transdentinal antibacterial activity of chlorhexidine. Am J Dent 23:255-259.

17.

Lessa

Nogueira

Huck

Hebling

Costa

(2010). Transdentinal cytotoxic effects of different concentrations of chlorhexidine gel applied on acid-conditioned dentin substrate. J Biomed Mater Res B Appl Biomater 92:40-47.

18.

Yasuda

Bogyo

Katz

Gordon

. (2004). Regulation of collagenase activities of human cathepsins by glycosaminoglycans. J Biol Chem 279:5470-5479.

19.

Machado

Rioli

Dalio

Castro

Juliano

Tersariol

. (2007). The role of Tyr⁶⁰⁵ and Ala⁶⁰⁷ of thimet oligopeptidase and Tyr⁶⁰⁶ and Gly⁶⁰⁸ of neurolysin in substrate hydrolysis and inhibitor binding. Biochem J 404:279-288.

20.

Martin-De Las Heras

Valenzuela

Overall

(2000). The matrix metalloproteinase gelatinase A in human dentine. Arch Oral Biol 45:757-765.

21.

Mazzoni

Mannello

Tay

Tonti

Papa

Mazzotti

. (2007). Zymographic analysis and characterization of MMP-2 and -9 forms in human sound dentin. J Dent Res 86:436-440.

22.

Musil

Zucic

Turk

Engh

Mayr

Huber

. (1991). The refined 2.15Å x-ray crystal structure of human liver cathepsin B: the structural basis for its specificity. EMBO J 10:2321-2330.

23.

Nägler

Storer

Portaro

Carmona

Juliano

Ménard

(1997). Major increase in endopeptidase activity of human cathepsin B upon removal of occluding loop contacts. Biochemistry 36: 12608-12615.

24.

Nascimento

Minciotti

Geraldeli

Carrilho

Pashley

Tay

. (2011). Cysteine cathepsins in human carious dentin. J Dent Res 90:506-511.

25.

Nerurkar

Zentner

Rytting

(1995). Effect of chloride in the releasing of chlorhexidine salts from methyl methacrylate: 2-hydroxyethyl methacrylate copolymer reservoir devices. J Control Release 33:357-363.

26.

Pashley

Tay

Yiu

Hashimoto

Breschi

Carvalho

. (2004). Collagen degradation by host-derived enzymes during aging. J Dent Res 83:216-221.

27.

Pashley

Tay

Breschi

Tjäderhane

Carvalho

Carrilho

. (2011). State of the art etch-and-rinse adhesives. Dent Mater 27: 1-16.

28.

Quraishi

Nägler

Fox

Sivaraman

Cygler

Mort

Storer

(1999). The occluding loop in cathepsin B defines the pH dependence of inhibition by its propeptide. Biochemistry 38:5017-5023.

29.

Sulkala

Tervahartiala

Sorsa

Larmas

Salo

Tjäderhane

(2007). Matrix metalloproteinase-8 (MMP-8) is the major collagenase in human dentin. Arch Oral Biol 52:121-127.

30.

Tersariol

Geraldeli

Minciotti

Nascimento

Pääkkönen

Martins

. (2010). Cysteine cathepsins in human dentin-pulp complex. J Endod 36:475-481.

31.

Thompson

Halbert

Bossard

Tomaszek

Levy

Zhao

. (1997). Design of potent and selective human cathepsin K inhibitors that span the active site. Proc Natl Acad Sci USA 94:14249-14254.

32.

Tjäderhane

Larjava

Sorsa

Uitto

Larmas

Salo

(1998). The activation and function of host matrix metalloproteinases in dentin matrix breakdown in caries lesions. J Dent Res 77:1622-1629.

33.

Turk

(1997). Structural and functional aspects of papain-like cysteine proteinases and their protein inhibitors. Biol Chem 378: 141-150.

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Chlorhexidine Inhibits the Activity of Dental Cysteine Cathepsins

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