Gelatinolytic activity after dentin pretreatment with dimethyl sulfoxide (DMSO) combined to dental bonding systems: Perspectives for biological responses

Abstract

Endogeneous proteolytic responses in dentin bonding interface have addressing to strategies to preventive and therapeutic approaches of clinical use of dentin bonding systems (DBSs), but still present limitations. The aim of this study was to examine the gelatinolytic profile by means of in situ zymography regarding the use of 1% dimethyl sulfoxide (DMSO) as an aprotic solvent. Sound human third molars were prepared and randomized in 10 groups, following the factors 1- DBS: Adper™ Scotchbond Multipurpose [MP], Adper™ Single Bond 2 [SB], Clearfil™ SE Bond [CSE] and Adper™ Scotchbond Universal – Etch-and-rinse [SU-ER] mode and self-etch mode [SU-SE], 2- dentin pretreatment: Control – Water [C], 2% CHX and 1% DMSO and 3- time: Initial-24 h [I], 6 months [6M] and 30 months [30M]. Pretreatments were applied before primer application for 30s. After restoration, specimens were cut into slices, in which one third were incubated with fluorescein-conjugated gelatin for 24h at 37 °C and analyzed by confocal laser scanning microscopy. The other two-thirds were stored for 6 or 30 months at 37 °C. Fluorescence was quantified using Image J and data was subjected for two-way ANOVA followed by Tukey test (p<0.05). Neither DMSO nor CHX affected initial analyses for any tested conditions. After 6 months, it was observed increased fluorescence for MP using both pretreatments and for SB using only DMSO. Regardless time and pretreatment, CSE and SU-SE showed stabilized gelatinolytic pattern. For SU-ER, both CHX and DMSO were able to maintain a lower fluorescence compared to control group after 6 months. 30-month performance states the susceptibility of degradation for all etched-dentin systems. DMSO pretreatment can be promising to reduce gelatinolytic activity combined with an universal adhesive system under etch-and-rinse mode. For self-etching strategies, DMSO was successful to stabilize the gelatinolytic reactions.