PRODUÇÃO E CARACTERIZAÇÃO DE NANOCÁPSULAS DE NÚCLEO AQUOSO CONTENDO O PEPTÍDEO SSIEFARL DO HERPES SIMPLEX VÍRUS-1 PARA APLICAÇÃO VACINAL

• Autor(a) principal: Melo, Larissa Daiane Willrich de
• Data de Publicação: 2013
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional Universidade Franciscana
• Texto Completo: http://tede.universidadefranciscana.edu.br:8080/handle/UFN-BDTD/208; http://www.tede.universidadefranciscana.edu.br:8080/handle/UFN-BDTD/305
• topic: Nanocápsulas; Herpes virus simplex; SSIEFARL; Vacina; Nanocapsules; Herpes simplex virus; SSIEFARL; Vaccine; CNPQ::ENGENHARIAS

Efforts to develop an effective vaccine against Herpes Simplex Virus-1 (HSV-1) has been intense enough, however, there is no effective vaccine yet. The using of nanocapsules (Ncs) for production of viral vaccines may contribute to the enhancement and effectiveness of the already developed vaccines models. The main properties of the nanostructured system, which may contribute to the improvement of the immune response generated by vaccine is the slow and gradual release of the encapsulated content, protection of the encapsulated molecule or protein and adjuvanticity, inherent in every Ncs. The objective of this study was to produce and characterize nanocapsules with aqueous core to encapsulate the SSIEFARL peptide from HSV-1. This peptide has hydrophilic characteristics which impossibility the encapsulation in conventional nanocapsules with oil core. For the standardization of the tunnel several adaptations where modified in the system previously developed by Lambert et al. Among these, the stirring of the suspension, temperature, amount of surfactant, amount of peptide and settling time were adapted. All protocols two aqueous suspensions were tested, one with the organic solvent ethanol in the aqueous phase or only water. The results showed that the production with magnetic stirring in the presence of ethanol in the aqueous phase lead to Ncs with better physicochemical characteristics. Empty Ncs presented an average of particle size of 285 nm (+ 19,4) and zeta potential of -40.1 mV (+ 1,7) and the Ncs containing SSIEFARL peptide presented the size of 315 nm (+ 20,7) and zeta potential of -35,5 mV (+ 2,4). Other condition of production tested was the water temperature added during the production. When water was used at 40 °C the physico-chemical parameters were better than when room temperature water was used. Additionally, we tested the stability of the suspensions after storage at room temperature or 4 °C. The Ncs were stable for longer periods of time under refrigeration. For the characterization of morphostructure, Ncs were analyzed by transmission electron microscopy (TEM). This analysis revealed that Ncs present as spherical structures, homogeneous and uniform as standards in nanostructure for biological application. The analysis of Ncs by confocal microscopy was applied to investigate the encapsulation. Three amounts of SSIEFARL-6 FAM peptide were used (20, 30 or 50 μL), and the 20 μL showed better encapsulation. The analysis of the release profile of the peptide from Ncs showed that of the suspension produced with 20 μL was more linear, demonstrating slow release for at least 24 h. These data indicate that the method of production under magnetic stirring with ethanol and 20 μL of peptide in the aqueous phase is more suitable for production of aqueous core Ncs. Furthermore, empty Ncs at low concentrations showed no cytotoxicity to lymphoid cells from BALB-c in cell culture. Thus, we suggest that this approach has promising potential for developing a vaccine against HSV-1.