Desenvolvimento de duas formulações de enxaguante bucal à base de Cloramina T: um estudo in vitro

Abstract

Certain oral pathologies can lead to tooth loss, particularly dental caries, a multifactorial disease that affects the hard tissues of the teeth, causing demineralization and gingival inflammation. The development of dental biofilm is directly related to the occurrence of caries. Chlorhexidine has long been established as the gold standard for treatment; however, research suggests concerns regarding bacterial resistance to this medication, highlighting the need for alternatives. It is essential to develop new compositions and evaluate their characteristics. This study aimed to develop two chloramine T-based mouthwash formulations, characterize them, and assess their cytotoxic, antibacterial, and antibiofilm properties, as well as their stability. Was developed two formulations containing chloramineT, one designed for children and the other for adults. The formulations were characterized using FT-IR spectroscopy, thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). Their stability was assessed by measuring pH over 60 days under three different conditions. Antimicrobial activity was determined by broth microdilution followed by a kill curve analysis. The antibiofilma ctivity of the formulations was evaluated using crystal violet quantification, totalcarbohydrate assay, and atomic force microscopy (AFM). Cytotoxicity was assessed through hemolytic assays, MTT, and neutral red uptake assays. Results indicated that both formulations remained stable at 4ºC. Characterization suggested that chloramine T was preserved after formulation preparation. Both formulations demonstrated effective antibacterial activity, with MIC and MBC values of 3.125 mM for Streptococcus mutans and 0.875 mM for Staphylococcus aureus, indicating bactericidal properties. The bacterial kill curve corroborated the activity against the tested strains. Antibiofilm assays showed activity against all formed biofilms, with reduced crystal violet staining and carbohydrate content when the formulations were applied. The antibiofilm activity was further supported by AFM results. High cytotoxicity was observed in all assays, with mean IC50 values of 0.849 mM (F1) and 1.106 mM (F2). Conclusion: Despite demonstrating high cytotoxicity, both formulations showed promising in vitro results regarding antibacterial and antibiofilm activities against S. aureus and S. mutans. They also exhibited certain stability, and characterization indicated the stable presence of chloramine T after formulation synthesis. Therefore, these formulations suggest a potential alternative treatment to those currently available commercially.