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Detection of Functional Matrix Metalloproteinases by Zymography
Published on: November 8, 2010
Determination of matrix metalloproteinases in human radicular dentin
Juliana Santos1, Marcela Carrilho, Taina Tervahartiala
1Department of Restorative Dentistry, Endodontics Area, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil.
Abstract:
Matrix metalloproteinases (MMPs) are present in sound coronal dentin and may play a role in collagen network degradation in bonded restorations. We investigated whether these enzymes can also be detected in root dentin. Crown and root sections of human teeth were powderized, and dentin proteins were extracted by using guanidine-HCl and EDTA. Extracts were analyzed by zymography and Western blotting for matrix metalloproteinases detection. Zymography revealed gelatinolytic activities in both crown and root dentin samples, corresponding to MMP-2 and MMP-9. MMP-2 was more evident in demineralized root dentin matrix, whereas MMP-9 was mostly extracted from the mineralized compartment of dentin and presented overall lower levels. Western blot analysis detected MMP-8 equally distributed in crown and root dentin. Because MMPs are also present in radicular dentin, their contribution to the degradation of resin-dentin bonds should be addressed in the development of restorative strategies for the root substrate.

