Abstract

The aim of the present study was to determine the expression of MMP-9, MMP-13 and TIMP-3 after using the XP Bond ™ (Dentsply) dentin-bonding agent on 21 human teeth. Class I deep cavities were prepared and restored. After 01, 07, 14, 21, 30, 90 and 120 days, the teeth were extracted and processed for an immunohistochemical assessment (n=3). Immunohistochemical staining was performed using the monoclonal antibody anti-MMP-9, and polyclonal antibodies anti-MMP-13 and anti-TIMP-3. The immunoreactivity of metalloproteinases 9 and 13 was considered intense and moderate in the first storage intervals, before decreasing over time. In contrast, the immunoreactivity of TIMP-3 was considered absent in dentin and weak in pulp in the first time intervals, and completely absent in the subsequent intervals. MMP-9 expression in dentin and in pulp was prevalent in the dentinal tubules and odontoblastic layer, respectively. MMP-13 expression in dentin and in pulp was predominant in the pre-dentine region and odontoblastic layer, respectively. TIMP-3 expression was absent in dentin and predominantly located in the odontoblastic layer for pulp. In conclusion, the expression patterns of MMP-9, MMP-13 and TIMP-3 altered in the different time intervals studied. The increased expression of TIMP-3 in the first time intervals could suggest a synchronous response to the increased expression of MMPs in the same periods. The results confirmed the low performance of TIMP-3 in the physiological processes of the pulp-dentin complex for the conditions and materials tested herein.

Highlights

  • During the restorative procedure, the pulp responds to various forms of aggression, which result from cavity preparation and the toxic elements present in restorative materials, dentin bonding agents (Lovász et al 2021; Masioli et al 2007)

  • The expression patterns of more commonly known as metalloproteinases (MMPs)-9, MMP-13, and tissue inhibitors of MMPs (TIMPs)-3 were altered at different time intervals

  • Our results suggested that during a restorative procedure using the etch-and-rinse adhesive technique, there was a greater release of MMPs and TIMP studied in the first days after the restorative procedure, tending to decrease in the following days

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Summary

Introduction

The pulp responds to various forms of aggression, which result from cavity preparation and the toxic elements present in restorative materials, dentin bonding agents (Lovász et al 2021; Masioli et al 2007) Chemical mediators such as cytokines and tumor necrosis factor α (TNF-α), released during inflammatory responses, may activate the enzymes responsible for remodeling and degradation of the extracellular matrix (ECM). Evidence suggests that MMPs found in the dentin matrix or saliva can be responsible for the degradation of the exposed dentin organic matrix through acid demineralization produced by bacteria (Loreto et al 2014) They may be involved in the degradation of collagen fibrils (Li et al 2015) which are not completely infiltrated by resin monomers from the adhesive system used in the dentin bonding technique (Tahmiscija et al 2019; Nassar et al 2014; Pashley et al 2004). Recent studies have indicated the involvement of MMP-13 (collagenase-3) in the development and progression of dental tissue disorders; data regarding its presence and activity are still scarce and must be better elucidated (Loreto et al 2014)

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