Biochemical and immunohistochemical identification of MMP-7 in human dentin

Annalisa Mazzoni, Tatjana Maravić, Arzu Tezvergil-Mutluay, Leo Tjäderhane, Polliana Mendes Candia Scaffa, Roda Seseogullari-Dirihan, Alberto Bavelloni, Pietro Gobbi, David H. Pashley, Franklin R. Tay, Lorenzo Breschi

Research output: Contribution to journalArticle

Abstract

Objectives: Matrix metalloproteinases (MMPs) are dentinal endogenous enzymes claimed to have a vital role in dentin organic matrix breakdown. The aim of the study was to investigate presence, localization and distribution of MMP-7 in sound human dentin. Methods: Dentin was powdered, demineralized and dissolved in isoelectric focusing buffer. Resolved proteins were transferred to nitrocellulose membranes for western blotting (WB) analyses. For the zymographic analysis, aliquots of dentin protein were electrophoresed in 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing fluorescently labeled gelatin. Further, the concentrations of dentinal MMPs were measured using Fluorescent Microsphere Immunoassay with a human MMP-MAP multiplex kit. Pre- and post-embedding immunolabeling technique was used to investigate the localization and distribution of MMP-7 in dentin. Dentin was cryo-fractured, the fragments partially decalcified and labeled with a primary monoclonal anti-MMP-7 and a secondary antibody conjugated with gold nanoparticles. MMP-7 labelings were identified in the demineralized dentin matrix as highly electron-dense dispersed gold particles. Results: WB and zymographic analysis of extracted dentin proteins showed presence of MMP-7 (∼20–28 KDa). Further, MMP-7 was found in the supernatants of the incubated dentin beams using Fluorescent Microsphere Immunoassay. FEI-SEM and TEM analyses established MMP-7 as an intrinsic constituent of the human dentin organic matrix. Conclusion: This study demonstrated that MMP-7 is an endogenous component of the human dentin fibrillar network. Clinical significance: It is pivotal to understand the underlying processes behind dentin matrix remodeling and degradation in order to develop the most optimal clinical protocols and ensure the longevity of dental restorations.

Original languageEnglish (US)
Pages (from-to)90-95
Number of pages6
JournalJournal of Dentistry
Volume79
DOIs
StatePublished - Dec 2018

Fingerprint

Matrix Metalloproteinase 7
Dentin
Matrix Metalloproteinases
Microspheres
Immunoassay
Gold
Western Blotting
Proteins
Collodion
Isoelectric Focusing
Gelatin
Clinical Protocols
Sodium Dodecyl Sulfate
Nanoparticles
Polyacrylamide Gel Electrophoresis
Tooth
Buffers

Keywords

  • Biochemistry
  • Collagen
  • Dentine
  • Extra-cellular matrix
  • Immunohistochemistry
  • MMPs

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Mazzoni, A., Maravić, T., Tezvergil-Mutluay, A., Tjäderhane, L., Scaffa, P. M. C., Seseogullari-Dirihan, R., ... Breschi, L. (2018). Biochemical and immunohistochemical identification of MMP-7 in human dentin. Journal of Dentistry, 79, 90-95. https://doi.org/10.1016/j.jdent.2018.10.008

Biochemical and immunohistochemical identification of MMP-7 in human dentin. / Mazzoni, Annalisa; Maravić, Tatjana; Tezvergil-Mutluay, Arzu; Tjäderhane, Leo; Scaffa, Polliana Mendes Candia; Seseogullari-Dirihan, Roda; Bavelloni, Alberto; Gobbi, Pietro; Pashley, David H.; Tay, Franklin R.; Breschi, Lorenzo.

In: Journal of Dentistry, Vol. 79, 12.2018, p. 90-95.

Research output: Contribution to journalArticle

Mazzoni, A, Maravić, T, Tezvergil-Mutluay, A, Tjäderhane, L, Scaffa, PMC, Seseogullari-Dirihan, R, Bavelloni, A, Gobbi, P, Pashley, DH, Tay, FR & Breschi, L 2018, 'Biochemical and immunohistochemical identification of MMP-7 in human dentin', Journal of Dentistry, vol. 79, pp. 90-95. https://doi.org/10.1016/j.jdent.2018.10.008
Mazzoni A, Maravić T, Tezvergil-Mutluay A, Tjäderhane L, Scaffa PMC, Seseogullari-Dirihan R et al. Biochemical and immunohistochemical identification of MMP-7 in human dentin. Journal of Dentistry. 2018 Dec;79:90-95. https://doi.org/10.1016/j.jdent.2018.10.008
Mazzoni, Annalisa ; Maravić, Tatjana ; Tezvergil-Mutluay, Arzu ; Tjäderhane, Leo ; Scaffa, Polliana Mendes Candia ; Seseogullari-Dirihan, Roda ; Bavelloni, Alberto ; Gobbi, Pietro ; Pashley, David H. ; Tay, Franklin R. ; Breschi, Lorenzo. / Biochemical and immunohistochemical identification of MMP-7 in human dentin. In: Journal of Dentistry. 2018 ; Vol. 79. pp. 90-95.
@article{91d7062ffb1a405eabe9100bf5183773,
title = "Biochemical and immunohistochemical identification of MMP-7 in human dentin",
abstract = "Objectives: Matrix metalloproteinases (MMPs) are dentinal endogenous enzymes claimed to have a vital role in dentin organic matrix breakdown. The aim of the study was to investigate presence, localization and distribution of MMP-7 in sound human dentin. Methods: Dentin was powdered, demineralized and dissolved in isoelectric focusing buffer. Resolved proteins were transferred to nitrocellulose membranes for western blotting (WB) analyses. For the zymographic analysis, aliquots of dentin protein were electrophoresed in 12{\%} sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing fluorescently labeled gelatin. Further, the concentrations of dentinal MMPs were measured using Fluorescent Microsphere Immunoassay with a human MMP-MAP multiplex kit. Pre- and post-embedding immunolabeling technique was used to investigate the localization and distribution of MMP-7 in dentin. Dentin was cryo-fractured, the fragments partially decalcified and labeled with a primary monoclonal anti-MMP-7 and a secondary antibody conjugated with gold nanoparticles. MMP-7 labelings were identified in the demineralized dentin matrix as highly electron-dense dispersed gold particles. Results: WB and zymographic analysis of extracted dentin proteins showed presence of MMP-7 (∼20–28 KDa). Further, MMP-7 was found in the supernatants of the incubated dentin beams using Fluorescent Microsphere Immunoassay. FEI-SEM and TEM analyses established MMP-7 as an intrinsic constituent of the human dentin organic matrix. Conclusion: This study demonstrated that MMP-7 is an endogenous component of the human dentin fibrillar network. Clinical significance: It is pivotal to understand the underlying processes behind dentin matrix remodeling and degradation in order to develop the most optimal clinical protocols and ensure the longevity of dental restorations.",
keywords = "Biochemistry, Collagen, Dentine, Extra-cellular matrix, Immunohistochemistry, MMPs",
author = "Annalisa Mazzoni and Tatjana Maravić and Arzu Tezvergil-Mutluay and Leo Tj{\"a}derhane and Scaffa, {Polliana Mendes Candia} and Roda Seseogullari-Dirihan and Alberto Bavelloni and Pietro Gobbi and Pashley, {David H.} and Tay, {Franklin R.} and Lorenzo Breschi",
year = "2018",
month = "12",
doi = "10.1016/j.jdent.2018.10.008",
language = "English (US)",
volume = "79",
pages = "90--95",
journal = "Journal of Dentistry",
issn = "0300-5712",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Biochemical and immunohistochemical identification of MMP-7 in human dentin

AU - Mazzoni, Annalisa

AU - Maravić, Tatjana

AU - Tezvergil-Mutluay, Arzu

AU - Tjäderhane, Leo

AU - Scaffa, Polliana Mendes Candia

AU - Seseogullari-Dirihan, Roda

AU - Bavelloni, Alberto

AU - Gobbi, Pietro

AU - Pashley, David H.

AU - Tay, Franklin R.

AU - Breschi, Lorenzo

PY - 2018/12

Y1 - 2018/12

N2 - Objectives: Matrix metalloproteinases (MMPs) are dentinal endogenous enzymes claimed to have a vital role in dentin organic matrix breakdown. The aim of the study was to investigate presence, localization and distribution of MMP-7 in sound human dentin. Methods: Dentin was powdered, demineralized and dissolved in isoelectric focusing buffer. Resolved proteins were transferred to nitrocellulose membranes for western blotting (WB) analyses. For the zymographic analysis, aliquots of dentin protein were electrophoresed in 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing fluorescently labeled gelatin. Further, the concentrations of dentinal MMPs were measured using Fluorescent Microsphere Immunoassay with a human MMP-MAP multiplex kit. Pre- and post-embedding immunolabeling technique was used to investigate the localization and distribution of MMP-7 in dentin. Dentin was cryo-fractured, the fragments partially decalcified and labeled with a primary monoclonal anti-MMP-7 and a secondary antibody conjugated with gold nanoparticles. MMP-7 labelings were identified in the demineralized dentin matrix as highly electron-dense dispersed gold particles. Results: WB and zymographic analysis of extracted dentin proteins showed presence of MMP-7 (∼20–28 KDa). Further, MMP-7 was found in the supernatants of the incubated dentin beams using Fluorescent Microsphere Immunoassay. FEI-SEM and TEM analyses established MMP-7 as an intrinsic constituent of the human dentin organic matrix. Conclusion: This study demonstrated that MMP-7 is an endogenous component of the human dentin fibrillar network. Clinical significance: It is pivotal to understand the underlying processes behind dentin matrix remodeling and degradation in order to develop the most optimal clinical protocols and ensure the longevity of dental restorations.

AB - Objectives: Matrix metalloproteinases (MMPs) are dentinal endogenous enzymes claimed to have a vital role in dentin organic matrix breakdown. The aim of the study was to investigate presence, localization and distribution of MMP-7 in sound human dentin. Methods: Dentin was powdered, demineralized and dissolved in isoelectric focusing buffer. Resolved proteins were transferred to nitrocellulose membranes for western blotting (WB) analyses. For the zymographic analysis, aliquots of dentin protein were electrophoresed in 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing fluorescently labeled gelatin. Further, the concentrations of dentinal MMPs were measured using Fluorescent Microsphere Immunoassay with a human MMP-MAP multiplex kit. Pre- and post-embedding immunolabeling technique was used to investigate the localization and distribution of MMP-7 in dentin. Dentin was cryo-fractured, the fragments partially decalcified and labeled with a primary monoclonal anti-MMP-7 and a secondary antibody conjugated with gold nanoparticles. MMP-7 labelings were identified in the demineralized dentin matrix as highly electron-dense dispersed gold particles. Results: WB and zymographic analysis of extracted dentin proteins showed presence of MMP-7 (∼20–28 KDa). Further, MMP-7 was found in the supernatants of the incubated dentin beams using Fluorescent Microsphere Immunoassay. FEI-SEM and TEM analyses established MMP-7 as an intrinsic constituent of the human dentin organic matrix. Conclusion: This study demonstrated that MMP-7 is an endogenous component of the human dentin fibrillar network. Clinical significance: It is pivotal to understand the underlying processes behind dentin matrix remodeling and degradation in order to develop the most optimal clinical protocols and ensure the longevity of dental restorations.

KW - Biochemistry

KW - Collagen

KW - Dentine

KW - Extra-cellular matrix

KW - Immunohistochemistry

KW - MMPs

UR - http://www.scopus.com/inward/record.url?scp=85055963706&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055963706&partnerID=8YFLogxK

U2 - 10.1016/j.jdent.2018.10.008

DO - 10.1016/j.jdent.2018.10.008

M3 - Article

C2 - 30367893

AN - SCOPUS:85055963706

VL - 79

SP - 90

EP - 95

JO - Journal of Dentistry

JF - Journal of Dentistry

SN - 0300-5712

ER -