Carbodiimide Inactivation of MMPs and Effect on Dentin Bonding

Abstract

The use of protein cross-linking agents during bonding procedures has been recently proposed to improve bond durability. This study aimed to use zymography and in situ zymography techniques to evaluate the ability of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) cross-linker to inhibit matrix metalloproteinase (MMP) activity. The hypotheses tested were that: (1) bonding procedures increase dentin gelatinolytic activity and (2) EDC pre-treatment prevents this enzymatic activity. The zymographic assay was performed on protein extracts obtained from dentin powder treated with Optibond FL or Scotchbond 1XT with or without 0.3M EDC pre-treatment. For in situ zymography, adhesive/dentin interfaces were created with the same adhesives applied to acid-etched dentin slabs pre-treated or not with EDC conditioner. Zymograms revealed increased expression of dentin endogenous MMP-2 and -9 after adhesive application, while the use of EDC as a primer inactivated dentin gelatinases. Results of in situ zymograpy showed that hybrid layers of tested adhesives exhibited intense collagenolytic activity, while almost no fluorescence signal was detected when specimens were pre-treated with EDC. The correlative analysis used in this study demonstrated that EDC could contribute to inactivate endogenous dentin MMPs within the hybrid layer created by etch-and-rinse adhesives.

Transdentinal Cytotoxicity of Carbodiimide (EDC) and Glutaraldehyde on Odontoblast-like Cells

“…19,20,34 Additionally, EDC is able to preserve the resin-dentin bond over time. 35 In the medical field, EDC has been used to reinforce different structures for tissue engineering applications and has shown satisfactory biocompatibility.…”

Mechanisms of degradation of the hybrid layer in adhesive dentistry and therapeutic agents to improve bond durability—A literature review

“…Since lower biodegradation rates and high mechanical properties of collagen are desirable, the use of collagen cross-linking agents in adhesive procedures has gained increased popularity in recent years [63,66]. The use of glutaraldehyde, proanthrocyanidin and genipin [123], riboflavin [124], as well as tannic acid [122] and carbodiimide [70] has been proposed to enhance the mechanical and structural stability of dentin collagen, leading to a stable dentin matrix network that, after resin infiltration, should provide a durable hybrid layer [125][126][127][128][129]. Moreover, collagen cross-linking molecules have been reported to improve the resistance of uncross-linked or mildly cross-linked collagen matrices to degradation by bacterial collagenases [130,131], potentially contributing to the stabilization of the resin-dentin interface over time.…”

Role of Dentin MMPs in Caries Progression and Bond Stability

“…5A, B; Mazzoni et al 2012;Mazzoni et al 2014). Although previous studies demonstrated the presence and activity of endogenous proteolytic enzymes in human dentin, evidence of the activity of endogenous dentin MMPs within resin-bonded HLs was lacking.…”

“…The potential of cross-linkers is related to the possibility of improving the mechanical strength of the collagen network (Bedran-Russo et al 2014), improving the resistance to enzymatic degradation, and inactivating exposed MMPs bound to matrix collagen (Liu et al 2011;Tjäderhane et al 2013b). Together, this leads to a stable dentin matrix network, which ultimately could determine the formation of a stable HL (Cova et al 2011;Mazzoni, Angeloni, et al 2013;Tjäderhane et al 2013b;Bedran-Russo et al 2014;Mazzoni et al 2014). …”