Metalloproteinases (MMPs) have been implicated with metabolism of collagen in physiological and pathological processes in human dentine. As bovine teeth have been used as a substitute for human teeth in laboratory analysis, this study evaluated the activity of MMP-2 and -9 in bovine versus human dentine. Bovine and human dentine fragments, from crowns and roots, were powderized. Protein extraction was performed by two protocols: a neutral extraction with guanidine-HCl/EDTA (pH 7.4) and an acidic extraction with citric acid (pH 2.3). Gelatinolytic activities of extracts were revealed by zymography. MMP-2 and -9 were detected in crown and root dentine from bovine and human teeth. Total activities of MMP-2 were 11.4 ± 2.2, 14.6 ± 2.0, 9.7 ± 1.2 and 12.4 ± 0.9 ng/ml for bovine root, human root, bovine crown and human crown dentine, respectively. Corresponding activities for MMP-9 were 14.9 ± 2.0, 15.3 ± 1.3, 15.4 ± 1.3 and 15.5 ± 1.3 ng/ml, respectively. Bovine dentine was found to be a reliable substrate for studies involving the activity of MMP-2 and -9.

Keywords:
Bovine tooth, Dentine, Gelatinase, Human tooth, Matrix metalloproteinases
1.
Caterina J, Shi J, Sun X, Qian Q, Yamada S, Liu Y, Krakora S, Bartlett JD, Yamada Y, Engler JA, Birkedal-Hansen H, Simmer JP: Cloning, characterization, and expression analysis of mouse enamelysin. J Dent Res 2000;79:1697–1703.
2.
DenBesten PK, Heffernan LM, Treadwell BV, Awbrey BJ: The presence and possible functions of the matrix metalloproteinase collagenase activator protein in developing enamel matrix. Biochem J 1989;264:917–920.
3.
Fonseca RB, Haiter-Neto F, Carlo HL, Soares CJ, Sinhoreti MA, Puppin-Rontani RM, Correr-Sobrinho L: Radiodensity and hardness of enamel and dentin of human and bovine teeth, varying bovine teeth age. Arch Oral Biol 2008;53:1023–1029.
4.
Francisconi LF, Honório HM, Rios D, Magalhães AC, Machado MA, Buzalaf MA: Effect of erosive pH cycling on different restorative materials and on enamel restored with these materials. Oper Dent 2008;33:203–208.
5.
Hannas AR, Pereira JC, Granjeiro JM, Tjäderhane L: The role of matrix metalloproteinases in the oral environment. Acta Odontol Scand 2007;65:1–13.
6.
Hara AT, Queiroz CS, Paes Leme AF, Serra MC, Cury JA: Caries progression and inhibition in human and bovine root dentine in situ. Caries Res 2003;37:339–344.
7.
Kato MT, Leite AL, Hannas AR, Buzalaf MA: Gels containing MMP inhibitors prevent dental erosion in situ. J Dent Res 2010a;89:468–472.
8.
Kato MT, Leite AL, Hannas AR, Oliveira RC, Pereira JC, Tjäderhane L, Buzalaf MA: Effect of iron on matrix metalloproteinase inhibition and on the prevention of dentine erosion. Caries Res 2010b;44:309–316.
9.
Kato MT, Magalhães AC, Rios D, Hannas AR, Attin T, Buzalaf MA: Protective effect of green tea on dentin erosion and abrasion. J Appl Oral Sci 2009;17:560–564.
10.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265–275.
11.
Magalhães AC, Wiegand A, Rios D, Hannas A, Attin T, Buzalaf MA: Chlorhexidine and green tea extract reduce dentin erosion and abrasion in situ. J Dent 2009;37:994–998.
12.
Martin-De Las Heras S, Valenzuela A, Overall CM: The matrix metalloproteinase gelatinase A in human dentine. Arch Oral Biol 2000;45:757–765.
13.
Mazzoni A, Mannello F, Tay FR, Tonti GA, Papa S, Mazzotti G, Di Lenarda R, Pashley DH, Breschi L: Zymographic analysis and characterization of MMP-2 and -9 forms in human sound dentin. J Dent Res 2007;86:436–440.
14.
Ogata Y, Enghild JJ, Nagase H: Matrix metalloproteinase 3 (stromelysin) activates the precursor for the human matrix metalloproteinase 9. J Biol Chem 1992;267:3581–3584.
15.
Reis AF, Giannini M, Kavaguchi A, Soares CJ, Line SR: Comparison of microtensile bond strength to enamel and dentin of human, bovine, and porcine teeth. J Adhes Dent 2004;6:117–121.
16.
Rios D, Honório HM, Magalhães AC, Delbem AC, Machado MA, Silva SM, Buzalaf MA: Effect of salivary stimulation on erosion of human and bovine enamel subjected or not to subsequent abrasion: an in situ/ex vivo study. Caries Res 2006;40:218–223.
17.
Santos J, Carrilho M, Tervahartiala T, Sorsa T, Breschi L, Mazzoni A, Pashley D, Tay F, Ferraz C, Tjaderhane L: Determination of matrix metalloproteinases in human radicular dentin. J Endod 2009;35:686–689.
18.
Schilke R, Lisson JA, Bauss O, Geurtsen W: Comparison of the number and diameter of dentinal tubules in human and bovine dentine by scanning electron microscopic investigation. Arch Oral Biol 2000;45:355–361.
19.
Schmalz G, Hiller KA, Nunez LJ, Stoll J, Weis K: Permeability characteristics of bovine and human dentin under different pretreatment conditions. J Endod 2001;27:23–30.
20.
Sorsa T, Tjäderhane L, Salo T: Matrix metalloproteinases (MMPs) in oral diseases. Oral Dis 2004;10:311–318.
21.
Sulkala M, Larmas M, Sorsa T, Salo T, Tjäderhane L: The localization of matrix metalloproteinase-20 (MMP-20, enamelysin) in mature human teeth. J Dent Res 2002;81:603–607.
22.
Sulkala M, Tervahartiala T, Sorsa T, Larmas M, Salo T, Tjäderhane L: Matrix metalloproteinase-8 (MMP-8) is the major collagenase in human dentin. Arch Oral Biol 2007;52:121–127.
23.
Tersariol IL, Geraldeli S, Minciotti CL, Nascimento FD, Paakkonen V, Martins MT, Carrilho MR, Pashley DH, Tay FR, Salo T, Tjäderhane L: Cysteine cathepsins in human dentin-pulp complex. J Endod 2010;36:475–481.
24.
Tjäderhane L, Larjava H, Sorsa T, Uitto V-J, Larmas M, Salo T: The activation and function of host matrix metalloproteinases in dentin matrix breakdown in caries lesions. J Dent Res 1998;77:1622–1629.
25.
van Strijp AJ, Jansen DC, DeGroot J, ten Cate JM, Everts V: Host-derived proteinases and degradation of dentine collagen in situ. Caries Res 2003;37:58–65.
26.
Vieira A, Lugtenborg M, Ruben JL, Huysmans MC: Brushing abrasion of eroded bovine enamel pretreated with topical fluorides. Caries Res 2006;40:224–230.
27.
Visse R, Nagase H: Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 2003;92:827–839.
28.
Wegehaupt F, Gries D, Wiegand A, Attin T: Is bovine dentine an appropriate substitute for human dentine in erosion/abrasion tests? J Oral Rehabil 2008;35:390–394.
29.
Zhu L, Tanimoto K, Robinsin S, Chen J, Witkowska E, Hall S, Le T, Denbesten PK, Li W: Comparative properties of recombinant human and bovine matrix metalloproteinase-20. Arch Oral Biol 2008;53:785–790.
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2011
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