Avaliação da atividade antimicrobiana da lactoferricina B (LfcinB) e da anfotericina B (AMB) em sistemas de liberação controlada em biofilmes polimicrobianos in vitro relacionados à periodontite associada à apneia obstrutiva do sono

Abstract

Periodontitis is a highly prevalent disease, being one of the six most common human diseases. Due to its capacity to create biofilms, a dysbiotic microbiota is considered as a predisposing factor since it might result in a localized, chronic inflammatory response. The degree of involvement may vary between individuals due to genetic susceptibility and lifestyle. Some systemic diseases, such as obstructive sleep apnea (OSA), can increase the risk of development and progression of periodontitis. OSA is a sleep disorder that obstructs the airway, preventing the passage of air. The increase in periodontitis in individuals with OSA may be caused by a variety of factors, including genetic susceptibility, an inflammatory response that both conditions share, and/or pathophysiological features of OSA such oral dryness, hypoxia, and hypercapnia. All these factors can be considered a significant risk in the persistence of periodontitis in individuals with OSA. In order to propose a therapeutic alternative for the treatment of OSA-related periodontitis, the first objective of this study was based on establishing the association between both conditions, through the role of the microbiota and the local inflammatory response, based on the identification of cultivable microorganisms and quantification of five proinflammatory cytokines in samples from the oral cavity of 94 individuals over 30 years of age, with and without Periodontitis, with and without OSA, from the Sleep Clinic of the San Ignacio University Hospital and the School of Dentistry from the Pontificia Universidad Javeriana. From the microbial identification by MALDI-TOF and the quantification of cytokines by the Luminex assay, it was determined that the culturable oral microbiota of patients with periodontitis and OSA, was altered in terms of diversity, associated with the presence of bacteria of orange complexes, such as Prevotella spp. and yeasts of the genus Candida spp., with C. albicans being the most prevalent species, and the levels of IL-1β, IL-6, IL-17A and IL-33 increased, with the presence of Candida spp. with increased levels of IL-17A and IL-33. From the microbial profile of individuals with Periodontitis and OSA, nine bacteria and C. albicans were selected, isolated and cultured to establish an in vitro polymicrobial biofilm model, with which the anti-biofilm effect of nanofibers loaded with Amphotericin B(AMB), a palindromic antimicrobial peptide; Lactoferricin B (LfcinB (21-25)pal) and/or Zinc Oxide (ZnO) were evaluated. In the same way, the cytotoxic effect and the physicochemical characteristics of the nanofibers were evaluated. It was determined that the nanofibers were biocompatible showing a cell viability greater than 50% of human periodontal ligament fibroblasts. The nanofiber with AMB, LfcinB (21-25)pal and ZnO (Nf 6) showed greater inhibition, time-dependent, of the polymicrobial biofilm in vitro, demonstrating the ability of AMB to inhibit bacteria, together with LfcinB (21-25)pal and ZnO increasing the anti-biofilm activity. The Nf 6 nanofiber exhibited good mechanical and thermal stability, was easy to handle, had a homogeneous fiber diameter distribution, and was hydrophilic. Finally, a technological survey of the use of these nanofibers was carried out. This technology prospection study demonstrated that this nanofiber may be a promising therapeutic alternative for the treatment of OSA-associated periodontitis, therefore, this nanofiber is proposed as a possibly patentable technology.