Immunohistochemical and biochemical assay of MMP-3 in human dentine

Annalisa Mazzoni, Veronica Papa, Fernando Nato, Marcela Carrilho, Leo Tjäderhane, Alessandra Ruggeri, Pietro Gobbi, Giovanni Mazzotti, Franklin Chi Meng Tay, David Henry Pashley, Lorenzo Breschi

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Objective: The function of endogenous MMP-3 and its distribution within the human dentine is unclear. Thus, the aim of the present study was to assay the presence and distribution of MMP-3 within human sound dentine by means of biochemical and immunohistochemical assays. Methods: Powdered dentine from extracted human teeth was prepared and (1) partially demineralised with 1% H3PO4 for 10 min or (2) untreated (control). The presence of MMP-3 was measured using a colorimetric assay system (QuantiSir™, Epigentek, USA). Additional cryo-fractured dentine fragments were processed for immunohistochemical identification of MMP-3 under FEI-SEM. Casein-zymography was used to investigate MMP-3 activity. Results: MMP-3 detected level was 2.732 ng/μL in partially demineralised dentine powder, whilst it increased to 3.280 ng/μL in mineralised dentine. The FEI-SEM analysis revealed positive immunolabelling patterns for MMP-3, predominantly localized on the intertubular collagen fibrillar network showing MMP-3 directly or indirectly bound to the collagen fibrils. Casein-zymograms showed positive proteolytic activity for MMP-3 in demineralised dentine powder. Conclusion: The results of the study clearly revealed the presence and distribution of MMP3 in human sound dentine. Whilst the presence was verified, its role is still unclear. Future studies are needed to investigate the possible involvement of MMP-3 in physiological and pathological condition of the dentine-pulp complex.

Original languageEnglish (US)
Pages (from-to)231-237
Number of pages7
JournalJournal of Dentistry
Volume39
Issue number3
DOIs
StatePublished - Mar 1 2011

Fingerprint

Dentin
Matrix Metalloproteinases
Caseins
Powders
Fibrillar Collagens
Tooth
Collagen

Keywords

  • Biochemical assay
  • Dentine
  • Dentine bonding agent
  • Immunohistochemistry
  • MMP-3

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Mazzoni, A., Papa, V., Nato, F., Carrilho, M., Tjäderhane, L., Ruggeri, A., ... Breschi, L. (2011). Immunohistochemical and biochemical assay of MMP-3 in human dentine. Journal of Dentistry, 39(3), 231-237. https://doi.org/10.1016/j.jdent.2011.01.001

Immunohistochemical and biochemical assay of MMP-3 in human dentine. / Mazzoni, Annalisa; Papa, Veronica; Nato, Fernando; Carrilho, Marcela; Tjäderhane, Leo; Ruggeri, Alessandra; Gobbi, Pietro; Mazzotti, Giovanni; Tay, Franklin Chi Meng; Pashley, David Henry; Breschi, Lorenzo.

In: Journal of Dentistry, Vol. 39, No. 3, 01.03.2011, p. 231-237.

Research output: Contribution to journalArticle

Mazzoni, A, Papa, V, Nato, F, Carrilho, M, Tjäderhane, L, Ruggeri, A, Gobbi, P, Mazzotti, G, Tay, FCM, Pashley, DH & Breschi, L 2011, 'Immunohistochemical and biochemical assay of MMP-3 in human dentine', Journal of Dentistry, vol. 39, no. 3, pp. 231-237. https://doi.org/10.1016/j.jdent.2011.01.001
Mazzoni A, Papa V, Nato F, Carrilho M, Tjäderhane L, Ruggeri A et al. Immunohistochemical and biochemical assay of MMP-3 in human dentine. Journal of Dentistry. 2011 Mar 1;39(3):231-237. https://doi.org/10.1016/j.jdent.2011.01.001
Mazzoni, Annalisa ; Papa, Veronica ; Nato, Fernando ; Carrilho, Marcela ; Tjäderhane, Leo ; Ruggeri, Alessandra ; Gobbi, Pietro ; Mazzotti, Giovanni ; Tay, Franklin Chi Meng ; Pashley, David Henry ; Breschi, Lorenzo. / Immunohistochemical and biochemical assay of MMP-3 in human dentine. In: Journal of Dentistry. 2011 ; Vol. 39, No. 3. pp. 231-237.
@article{02c40951cd774ebf88575e780d7cd557,
title = "Immunohistochemical and biochemical assay of MMP-3 in human dentine",
abstract = "Objective: The function of endogenous MMP-3 and its distribution within the human dentine is unclear. Thus, the aim of the present study was to assay the presence and distribution of MMP-3 within human sound dentine by means of biochemical and immunohistochemical assays. Methods: Powdered dentine from extracted human teeth was prepared and (1) partially demineralised with 1{\%} H3PO4 for 10 min or (2) untreated (control). The presence of MMP-3 was measured using a colorimetric assay system (QuantiSir™, Epigentek, USA). Additional cryo-fractured dentine fragments were processed for immunohistochemical identification of MMP-3 under FEI-SEM. Casein-zymography was used to investigate MMP-3 activity. Results: MMP-3 detected level was 2.732 ng/μL in partially demineralised dentine powder, whilst it increased to 3.280 ng/μL in mineralised dentine. The FEI-SEM analysis revealed positive immunolabelling patterns for MMP-3, predominantly localized on the intertubular collagen fibrillar network showing MMP-3 directly or indirectly bound to the collagen fibrils. Casein-zymograms showed positive proteolytic activity for MMP-3 in demineralised dentine powder. Conclusion: The results of the study clearly revealed the presence and distribution of MMP3 in human sound dentine. Whilst the presence was verified, its role is still unclear. Future studies are needed to investigate the possible involvement of MMP-3 in physiological and pathological condition of the dentine-pulp complex.",
keywords = "Biochemical assay, Dentine, Dentine bonding agent, Immunohistochemistry, MMP-3",
author = "Annalisa Mazzoni and Veronica Papa and Fernando Nato and Marcela Carrilho and Leo Tj{\"a}derhane and Alessandra Ruggeri and Pietro Gobbi and Giovanni Mazzotti and Tay, {Franklin Chi Meng} and Pashley, {David Henry} and Lorenzo Breschi",
year = "2011",
month = "3",
day = "1",
doi = "10.1016/j.jdent.2011.01.001",
language = "English (US)",
volume = "39",
pages = "231--237",
journal = "Journal of Dentistry",
issn = "0300-5712",
publisher = "Elsevier BV",
number = "3",

}

TY - JOUR

T1 - Immunohistochemical and biochemical assay of MMP-3 in human dentine

AU - Mazzoni, Annalisa

AU - Papa, Veronica

AU - Nato, Fernando

AU - Carrilho, Marcela

AU - Tjäderhane, Leo

AU - Ruggeri, Alessandra

AU - Gobbi, Pietro

AU - Mazzotti, Giovanni

AU - Tay, Franklin Chi Meng

AU - Pashley, David Henry

AU - Breschi, Lorenzo

PY - 2011/3/1

Y1 - 2011/3/1

N2 - Objective: The function of endogenous MMP-3 and its distribution within the human dentine is unclear. Thus, the aim of the present study was to assay the presence and distribution of MMP-3 within human sound dentine by means of biochemical and immunohistochemical assays. Methods: Powdered dentine from extracted human teeth was prepared and (1) partially demineralised with 1% H3PO4 for 10 min or (2) untreated (control). The presence of MMP-3 was measured using a colorimetric assay system (QuantiSir™, Epigentek, USA). Additional cryo-fractured dentine fragments were processed for immunohistochemical identification of MMP-3 under FEI-SEM. Casein-zymography was used to investigate MMP-3 activity. Results: MMP-3 detected level was 2.732 ng/μL in partially demineralised dentine powder, whilst it increased to 3.280 ng/μL in mineralised dentine. The FEI-SEM analysis revealed positive immunolabelling patterns for MMP-3, predominantly localized on the intertubular collagen fibrillar network showing MMP-3 directly or indirectly bound to the collagen fibrils. Casein-zymograms showed positive proteolytic activity for MMP-3 in demineralised dentine powder. Conclusion: The results of the study clearly revealed the presence and distribution of MMP3 in human sound dentine. Whilst the presence was verified, its role is still unclear. Future studies are needed to investigate the possible involvement of MMP-3 in physiological and pathological condition of the dentine-pulp complex.

AB - Objective: The function of endogenous MMP-3 and its distribution within the human dentine is unclear. Thus, the aim of the present study was to assay the presence and distribution of MMP-3 within human sound dentine by means of biochemical and immunohistochemical assays. Methods: Powdered dentine from extracted human teeth was prepared and (1) partially demineralised with 1% H3PO4 for 10 min or (2) untreated (control). The presence of MMP-3 was measured using a colorimetric assay system (QuantiSir™, Epigentek, USA). Additional cryo-fractured dentine fragments were processed for immunohistochemical identification of MMP-3 under FEI-SEM. Casein-zymography was used to investigate MMP-3 activity. Results: MMP-3 detected level was 2.732 ng/μL in partially demineralised dentine powder, whilst it increased to 3.280 ng/μL in mineralised dentine. The FEI-SEM analysis revealed positive immunolabelling patterns for MMP-3, predominantly localized on the intertubular collagen fibrillar network showing MMP-3 directly or indirectly bound to the collagen fibrils. Casein-zymograms showed positive proteolytic activity for MMP-3 in demineralised dentine powder. Conclusion: The results of the study clearly revealed the presence and distribution of MMP3 in human sound dentine. Whilst the presence was verified, its role is still unclear. Future studies are needed to investigate the possible involvement of MMP-3 in physiological and pathological condition of the dentine-pulp complex.

KW - Biochemical assay

KW - Dentine

KW - Dentine bonding agent

KW - Immunohistochemistry

KW - MMP-3

UR - http://www.scopus.com/inward/record.url?scp=79952184778&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79952184778&partnerID=8YFLogxK

U2 - 10.1016/j.jdent.2011.01.001

DO - 10.1016/j.jdent.2011.01.001

M3 - Article

C2 - 21215789

AN - SCOPUS:79952184778

VL - 39

SP - 231

EP - 237

JO - Journal of Dentistry

JF - Journal of Dentistry

SN - 0300-5712

IS - 3

ER -