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Effect of a new cystatin derived from "Maqui Berry" on acquired pellicle thickness and viscoelastic properties, as well as early colonizers, microcosm biofilm profile and viability, and prevention of enamel demineralization

Abstract

Despite being easily preventable, carie is the most prevalent chronic disease worldwide. In the thematic project associated with this proposal (FAPESP 2019/26070-1), we proposed a completely new concept for the prevention of dental caries, through engineering of acquired enamel pellicle (AEP). Modification of the protein profile of this integument, by incorporating acid-fast proteins, such as cystatin derived from sugar cane (CaneCPI-5), interferes with the initial colonization of dental biofilm, reducing counts of mutans streptococcus and lactobacillus, as well as enamel demineralization. Recently, our research group carried out the heterologous cloning and production (E. coli) of several cystatins derived from the leaves of a Chilean plant called maqui (Aristotelia chinensis). One of them, called Maqui-3, showed a high yield (~ 110 mg/L), 15 times higher than that of CaneCPI-5, making it attractive for commercial applications. Previous experiments by our group, using acquired pellicle engineering procedures and initial dental erosion protocols, revealed that the protective effect of Maqui-3 was greater than that of CaneCPI-5 (unpublished data). However, the anticaries potential of Maqui-3 has never been evaluated, which will be the objective of the present study. We intend to evaluate the effect of Maqui-3: 1) on the thickness and viscoelastic properties of PAE, using the QCM-D (quartz crystal microbalance with dissipation monitoring); 2) in the profile (count of colony-forming units) and viability (resazurin) of microcosm biofilm, as well as in enamel demineralization (cross-sectional microradiography - TMR); 3) changes in the initial colonizing microorganisms of the biofilm, through metagenomic analyses. (AU)

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