Biochemical and immunohistochemical identification of MMP-7 in human dentin

Show full item record



Permalink

http://hdl.handle.net/10138/307702

Citation

Mazzoni , A , Maravic , T , Tezvergil-Mutluay , A , Tjäderhane , L , Scaffa , P M C , Seseogullari-Dirihan , R , Bavelloni , A , Gobbi , P , Pashley , D H , Tay , F R & 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

Title: Biochemical and immunohistochemical identification of MMP-7 in human dentin
Author: Mazzoni, Annalisa; Maravic, 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
Contributor: University of Helsinki, Department of Oral and Maxillofacial Diseases
Date: 2018-12
Language: eng
Number of pages: 6
Belongs to series: Journal of Dentistry
ISSN: 0300-5712
URI: http://hdl.handle.net/10138/307702
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 (similar to 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.
Subject: MMPs
Dentine
Collagen
Extra-cellular matrix
Immunohistochemistry
Biochemistry
MATRIX METALLOPROTEINASES
DEGRADATION
COLLAGEN
CARIES
MATRILYSIN
INVASION
TISSUE
STROMELYSIN-1
PROTEINASES
GELATINASES
313 Dentistry
3126 Surgery, anesthesiology, intensive care, radiology
Rights:


Files in this item

Total number of downloads: Loading...

Files Size Format View
1_s2.0_S0300571218302690_main.pdf 2.437Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record