MMP-2 assay within the hybrid layer created by a two-step etch-and-rinse adhesive

Biochemical and immunohistochemical analysis

Annalisa Mazzoni, Marcela Carrilho, Veronica Papa, Leo Tjäderhane, Pietro Gobbi, Cesare Nucci, Roberto Di Lenarda, Giovanni Mazzotti, Franklin Chi Meng Tay, David Henry Pashley, Lorenzo Breschi

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Objective: Degradation of hybrid layers (HLs) within resin-infiltrated dentine results from multiple degradation factors, including collagenolytic activity of specific matrix metalloproteinases (MMPs). Inhibition of host-derived MMPs may, therefore, slow the degradation of HL. The null hypothesis tested is that the presence of MMP-2 is similar regardless of chlorhexidine (CHX) pre-treatment or the use of an adhesive. Methods: Powdered dentine prepared from extracted human teeth was divided into 4 groups: (G1) mineralised powder (control group); (G2) dentine powder treated with 1% phosphoric acid for 1 min; (G3) 1% phosphoric acid-etched dentine treated with Adper Scotchbond 1 XT (SB1XT; 3M ESPE); (G4) 1% phosphoric acid-etched dentine treated with 0.2% CHX followed by SB1XT. The concentration of detectable pro-MMP-2 and MMP-2 was assayed using a colorimetric assay system (QuantiSir). In addition, the presence of MMP-2 in the HL was assessed in 1 year-aged adhesive-dentine interfaces using an immunohistochemical approach under FEI-SEM/TEM. Results: In dentine powder treated with 1% phosphoric acid (G2), MMP-2 level decreased compared to controls (G1); the application of SB1XT (G3) resulted in an increase of MMP-2, whilst 0.2% CHX before SB1XT application (G4), reduced MMP-2. The FEI-SEM/TEM analysis revealed MMP-2 distribution within the HL of aged interfaces showing increase MMP-2 patterns in the control group and minor labelling in the CHX-pretreated specimens. Conclusion: The results of this study support the use of non-toxic MMPs inhibitors, such as CHX, as an appropriate additional step in bonding procedures in order to increase the longevity of the adhesive restorations.

Original languageEnglish (US)
Pages (from-to)470-477
Number of pages8
JournalJournal of Dentistry
Volume39
Issue number7
DOIs
StatePublished - Jul 1 2011

Fingerprint

Matrix Metalloproteinase 2
Adhesives
Dentin
Chlorhexidine
Powders
Matrix Metalloproteinases
Control Groups
Matrix Metalloproteinase Inhibitors
Tooth
phosphoric acid

Keywords

  • Biochemistry
  • Chlorhexidine
  • Dentine bonding agents
  • Immunohistochemistry
  • MMP-2

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

MMP-2 assay within the hybrid layer created by a two-step etch-and-rinse adhesive : Biochemical and immunohistochemical analysis. / Mazzoni, Annalisa; Carrilho, Marcela; Papa, Veronica; Tjäderhane, Leo; Gobbi, Pietro; Nucci, Cesare; Di Lenarda, Roberto; Mazzotti, Giovanni; Tay, Franklin Chi Meng; Pashley, David Henry; Breschi, Lorenzo.

In: Journal of Dentistry, Vol. 39, No. 7, 01.07.2011, p. 470-477.

Research output: Contribution to journalArticle

Mazzoni, A, Carrilho, M, Papa, V, Tjäderhane, L, Gobbi, P, Nucci, C, Di Lenarda, R, Mazzotti, G, Tay, FCM, Pashley, DH & Breschi, L 2011, 'MMP-2 assay within the hybrid layer created by a two-step etch-and-rinse adhesive: Biochemical and immunohistochemical analysis', Journal of Dentistry, vol. 39, no. 7, pp. 470-477. https://doi.org/10.1016/j.jdent.2011.04.004
Mazzoni, Annalisa ; Carrilho, Marcela ; Papa, Veronica ; Tjäderhane, Leo ; Gobbi, Pietro ; Nucci, Cesare ; Di Lenarda, Roberto ; Mazzotti, Giovanni ; Tay, Franklin Chi Meng ; Pashley, David Henry ; Breschi, Lorenzo. / MMP-2 assay within the hybrid layer created by a two-step etch-and-rinse adhesive : Biochemical and immunohistochemical analysis. In: Journal of Dentistry. 2011 ; Vol. 39, No. 7. pp. 470-477.
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abstract = "Objective: Degradation of hybrid layers (HLs) within resin-infiltrated dentine results from multiple degradation factors, including collagenolytic activity of specific matrix metalloproteinases (MMPs). Inhibition of host-derived MMPs may, therefore, slow the degradation of HL. The null hypothesis tested is that the presence of MMP-2 is similar regardless of chlorhexidine (CHX) pre-treatment or the use of an adhesive. Methods: Powdered dentine prepared from extracted human teeth was divided into 4 groups: (G1) mineralised powder (control group); (G2) dentine powder treated with 1{\%} phosphoric acid for 1 min; (G3) 1{\%} phosphoric acid-etched dentine treated with Adper Scotchbond 1 XT (SB1XT; 3M ESPE); (G4) 1{\%} phosphoric acid-etched dentine treated with 0.2{\%} CHX followed by SB1XT. The concentration of detectable pro-MMP-2 and MMP-2 was assayed using a colorimetric assay system (QuantiSir). In addition, the presence of MMP-2 in the HL was assessed in 1 year-aged adhesive-dentine interfaces using an immunohistochemical approach under FEI-SEM/TEM. Results: In dentine powder treated with 1{\%} phosphoric acid (G2), MMP-2 level decreased compared to controls (G1); the application of SB1XT (G3) resulted in an increase of MMP-2, whilst 0.2{\%} CHX before SB1XT application (G4), reduced MMP-2. The FEI-SEM/TEM analysis revealed MMP-2 distribution within the HL of aged interfaces showing increase MMP-2 patterns in the control group and minor labelling in the CHX-pretreated specimens. Conclusion: The results of this study support the use of non-toxic MMPs inhibitors, such as CHX, as an appropriate additional step in bonding procedures in order to increase the longevity of the adhesive restorations.",
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author = "Annalisa Mazzoni and Marcela Carrilho and Veronica Papa and Leo Tj{\"a}derhane and Pietro Gobbi and Cesare Nucci and {Di Lenarda}, Roberto and Giovanni Mazzotti and Tay, {Franklin Chi Meng} and Pashley, {David Henry} and Lorenzo Breschi",
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AU - Papa, Veronica

AU - Tjäderhane, Leo

AU - Gobbi, Pietro

AU - Nucci, Cesare

AU - Di Lenarda, Roberto

AU - Mazzotti, Giovanni

AU - Tay, Franklin Chi Meng

AU - Pashley, David Henry

AU - Breschi, Lorenzo

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N2 - Objective: Degradation of hybrid layers (HLs) within resin-infiltrated dentine results from multiple degradation factors, including collagenolytic activity of specific matrix metalloproteinases (MMPs). Inhibition of host-derived MMPs may, therefore, slow the degradation of HL. The null hypothesis tested is that the presence of MMP-2 is similar regardless of chlorhexidine (CHX) pre-treatment or the use of an adhesive. Methods: Powdered dentine prepared from extracted human teeth was divided into 4 groups: (G1) mineralised powder (control group); (G2) dentine powder treated with 1% phosphoric acid for 1 min; (G3) 1% phosphoric acid-etched dentine treated with Adper Scotchbond 1 XT (SB1XT; 3M ESPE); (G4) 1% phosphoric acid-etched dentine treated with 0.2% CHX followed by SB1XT. The concentration of detectable pro-MMP-2 and MMP-2 was assayed using a colorimetric assay system (QuantiSir). In addition, the presence of MMP-2 in the HL was assessed in 1 year-aged adhesive-dentine interfaces using an immunohistochemical approach under FEI-SEM/TEM. Results: In dentine powder treated with 1% phosphoric acid (G2), MMP-2 level decreased compared to controls (G1); the application of SB1XT (G3) resulted in an increase of MMP-2, whilst 0.2% CHX before SB1XT application (G4), reduced MMP-2. The FEI-SEM/TEM analysis revealed MMP-2 distribution within the HL of aged interfaces showing increase MMP-2 patterns in the control group and minor labelling in the CHX-pretreated specimens. Conclusion: The results of this study support the use of non-toxic MMPs inhibitors, such as CHX, as an appropriate additional step in bonding procedures in order to increase the longevity of the adhesive restorations.

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