Desenvolvimento, avaliação clínica e microbiológica de verniz dentário de Copaifera Langsdorffii Desf

Abstract

The use of biologically active natural products in different types of formulations and forms of applications has been researched to prevent oral diseases. Studies have demonstrated the therapeutic potential of Copaifera langsdorffi (copaiba) in several functions, especially the anti-inflammatory and antimicrobial properties. The varnishes represent a good prevention strategy, given the great adhesion to the tooth and slow, continuous and prolonged release of the active principle. Objective: The aim of the study was to do a prospective study of patents for copaiba products, develop and evaluate longitudinally the clinical and antimicrobial efficacy of a 1% Copaifera langsdorffi DESF (C. langsdorffii) dental varnish to prevent dental caries in high-risk children. Materials and Methods: Initially, a prospective study of copaiba dental product patents was carried out in intellectual property banks in Brazil and worldwide. In the pilot study, 24 children, aged between 36 and 71 months, caries-free (ICDAS II = 0) and with good systemic health were recruited. The children were divided into 4 groups for saliva dose-response evaluation after varnish application at different concentrations (1%, 5%, 10% and 20%). Each patient received the varnish application on the occlusal face of the all second deciduous molars and saliva was collected before and after the varnish application to verify the dose-response. The third stage of the study is a longitudinal, randomized, controlled study. A total of 90 participants were selected with the same inclusion and exclusion criteria from the pilot study and were randomized into 3 groups with 30 patients each: Group I - C. langsdorffii varnish 1%; Group II - 1% chlorhexidine varnish; Group III- Fluoride Varnish (5% NaF). At this stage, each patient received the varnish application on the occlusal face of the all deciduous second molars. Varnishes were applied to occlusal surfaces of second deciduous molars on the baseline (D0), 3 months after (D90), and 6 months after (D180). Saliva was collected before starting treatment (D0), 90 days after day 0 (D90), 6 months (D180) and 1 year after initiation of treatment (D360) to evaluate the S.mutans reduction. Microbiological analysis was performed in duplicates (1:10 and 1:100 mL dilutions). S. mutans isolates were identified by its characteristic colony morphology and the values were expressed as log10 (CFU/mL). Statistics were carried out by applying repeated measures analysis of variance, Tukey’s multiple comparisons test, and paired t test. Results: In the prospective study, 9 patents were found using copaiba resin oil resin in dental products. The National Institute of Industrial Property (INPI-Brazil) had the largest number of deposits (5), followed by Espacenet (2) and Free Patents (2). C. Langsdorffii has been highlighted as the most widely used species in vehicle products and deposits in formulations (3). In the pilot study all concentrations showed antimicrobial activity, with no statistical difference between them. In the pre and post treatment comparative analysis, the 1% concentration obtained the best dose-response (p = 0.0026). In the longitudinal study we found the MS levels (mean± standard deviation) were as follows: Chlorhexine (D0: 0.58 ± 0.43; D90: 0.38 ± 0.23; D180:0.33 ± 0.14; D360: 0.55 ±0.52), Fluoride (D0:0.86 ± 0.37; D90:0.51 ± 0.33; D180:0.41 ± 0.24; D360:0.53 ± 0.44), Copaiba (D0:1.32 ± 0.61; D90:0.99 ± 0.57; D180:0.39 ± 0.22; D360:0.12 ± 0.19). Copaiba demonstrated significant S.mutans reduction: D360 versus D0 (p < 0.0001), D180 versus D0 (p < 0.001), D360versus D90 (p < 0.001), D180 versus D90 (p < 0.001), D360 versus D180 (p < 0.05). Chlorhexidine varnish significantly reduced S.mutans: D180 versus D0 (p < 0.05). Fluoride reduced S.mutans at D180 versus D0 (p < 0.001). Copaiba consistently reduced S.mutans throughout the 360-day period. At D360, copaiba produced lower S.mutans levels compared to fluoride (p < 0.001) and chlorhexidine (p < 0.01). Conclusions: Although biological activity has been documented for centuries, few patents for dental purposes have been found with copaiba. The 1% concentration obtained better responses. Three annual applications of the aqueous extract of Copaifera langsdorffii dental varnish showed substantial antimicrobial activity against S.mutans for up to 12 months in high-risk caries-free children. Further studies to identify the anticaries effect of this varnish are required to establish its use in caries prevention.