In-situ zymography to assess the MMPs activity with different etching time and ethanol wet-bonding on radicular dentin

The aim of this in vitro study was to investigate the effect of different etching times and ethanol pre-treatment on metalloproteinasis (MMPs) gelatinolytic activity on root dentin. Twelve single root teeth, extracted for periodontal reasons, were selected and an endodontic treatment was performed. After seven days, an 8-mm post space was prepared with dedicated drills. Specimens were randomly divided into four groups according to different adhesive protocols, based on different etching time in phosphoric acid and pre-treatment application. Cementation of the fiber post was performed with a dual-curing cement (DC Core, Kuraray) polymerized for 40s. In situ zymographic analyses was performed to investigate endogenous MMPs activity within the dentin hybrid layer. Quantification analyses of the MMPs activity revealed that all tested groups activated enzymes. However, the ethanol wet-bonding pretreatment was able to reduce the MMPs activity, above all when radicular dentin was extensively etched. In conclusion, MMPs gelatinolytic activity was detected in all groups and in-situ zymography was able to measure it. Further investigations are needed to clinically validate the data obtained of the present study.