Avaliação da aplicação de uma solução experimental inibidora de metaloproteinase na resistência de união mediata de sistemas adesivos

Abstract

This study aimed to evaluate, in vitro, the mediate microtensile bond strength- μTBS (12 months) of adhesive systems Adper Scotchbond SE / 3M/ESPE (Adper SE) and Adper Scotchbond Multi-Purpose / 3M/ESPE (Adper SMP) to sound dentin surface, treated or not with an experimental acid solution inhibitor of metalloproteinases (MMPs), as well as the predominant pattern of failure by fractography; and evaluate, in vitro, the inhibition of gelatinolytic activity by an experimental acid solution for dentin surface etching containing tetracycline compared with an experimental solution containing tetracycline. For mediate μTBS evaluation, in Adper SE Control and Adper SMP Control groups, the adhesive system was applied according to the manufacturer's instructions. In Adper SE Experimental group was applied the experimental solution prior to the adhesive system, and in the Adper Experimental SMP group, the etching with 37% phosphoric acid was replaced by applying the experimental solution. After applying the adhesive systems, the teeth were prepared for μTBS test, and the sticks were stored in an oven at 37°C. After 12 months μTBS test was realized and the sticks were evaluated by SEM. For enzymatic activity evaluation, saliva samples were collected from healthy donors, centrifuged and the supernatant was used by technique of zymography. This technique was based on the use of electrophoresis, in which a protein substrate (gelatin) was co-polymerized with acrylamide. The enzymes were separated under non-reducing conditions, renatured and then incubated in suitable buffer, based on the association between the substrate digestion and verification of the molecular weight of proteases. To quantify the relative inhibition by the two solutions was used the Image J software (NIH, Bethesda, MD, USA) and compared with the control. For mediate μTBS evaluation, according to the Tukey test (α = 0.05), no difference between the control and experimental groups was found for each adhesive system. The predominant pattern of failure for the system Adper SE was cohesive in composite while for Adper SMP was cohesive in hybrid layer. In the enzymatic activity evaluation, the enzymes were characterized as matrix metalloproteinases, because its expression was inhibited by EDTA (a known inhibitor of MMPs), while NEM (an inhibitor of serine proteases) had no effect on enzymatic activity. The MMP identified in the study, was characterized as MMP-2, with a molecular weight of 62KDa. The experimental acid solution presented an inhibition of the gelatinolytic activity of 93.66% (concentration 0.05%) and 58.58% (0.01% concentration), while the experimental solution had a percentage inhibition of 31.28% and 6.83%, respectively. Based on these results we conclude that after 12 months of storage, the adhesive system Adper SE showed better performance that Adper SMP in the microtensile bond strength test, also seeming to be more effective in maintaining the integrity of the adhesive interface, under fractographic analysis; and that the experimental acid solution for dentin surface etching was capable of almost completely inhibits the enzymatic activity of MMP-2 in a dose dependent manner.