Obtenção, caracterização e avaliação da atividade antifúngica de nanopartículas de quitosana contendo o peptídio TistH identificado na peçonha do escorpião Tityus stigmurus
The Tityus stigmurus scorpion venom has been studied in recent years and revealed many toxins (peptides) with high potential for biotechnological applications. The transcriptomics approach from the venom glands of T. stigmurus, realized by our scientific group, identified several bioactive peptid...
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| Formato: | doctoralThesis |
| Idioma: | pt_BR |
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| Endereço do item: | https://repositorio.ufrn.br/jspui/handle/123456789/27580 |
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| Resumo: | The Tityus stigmurus scorpion venom has been studied in recent years and revealed many
toxins (peptides) with high potential for biotechnological applications. The transcriptomics
approach from the venom glands of T. stigmurus, realized by our scientific group,
identified several bioactive peptides with pharmacological potential. Among the molecules
identified, the activity of peptide like-hypotensins class (TistH, Tityus stigmurus
hypotensin), reported in the literature, evidenced the TistH with bradykinin-potentiating
peptide, in addition exert antifungal effect. The maximum efficacy of this class of
compounds can be achieved by immobilizing it in specific and suitable biomaterials or
suitable carriers. This study objective to obtained and characterized physiochemically
TistH-loading cross-linked chitosan nanoparticles for incorporation and adsorption,
evaluate the biocompatibility as well as improve of antifungal effect. The nanossystems
containing the TistH by incorporation (CN-TistH-Inc) and absorption (CN-TistH-Ads)
methods at concentration of 0.5 and 1.0% were obtained by ionic gelation technique. The
nanosystems at 0.5 and 1.0% were analyzed by dynamic light scattering showed positive
zeta potential, with size particles less than 160 nm and polydispersity index less than 0.3.
The encapsulation efficiency were greater than 96.5% and the success was confirmed by
polyacrylamide gel electrophoresis and Fourier transform infrared spectroscopy. The CNTistH-Inc and CN-TistH-Ads at 0.5 and 1.0% were analyzed by atomic force microscopy
and scanning electronic microscopy evidenced nanometric particles, spherical shape with
smooth surface and homogeneous aspects. The stability was evaluated by particle size and
polydispersity index can possible observed stable colloidal dispersions for 4 months.
According in vitro release profile was observed for all formulations prolonged and
sustained release for 24 hours, with kinetic parabolic diffusion mechanism. The
biocompatibility was evaluated in erythrocytes, normal cells originated from murine
macrophage (RAW 264.7) and kidney of African green monkeys (Vero E6) suggested
biocompatibility that both nanosystems, in concentration used for antifungal assay, not
demonstrated be hemolytic and cytotoxic, respectively. The antifungal activity was
evaluated through of determination of the Minimum inhibitory concentration (MIC),
minimum fungicidal concentration (MFC) and antibiofilm activity used yeast of genus
Candida. The CN-TistH-Inc and CN-TistH-Ads showed a minimal inhibitory
concentration of 89.2 µg/mL against Candida albicans, 11.1 µg/mL for C. parapsilosis and
C. tropicalis, confirmed by minimum fungicidal concentrations assay. Moreover, the
TistH-loaded cross-linked chitosan nanoparticles significantly reduced the biofilm
formation of clinical yeast sepsis of C. tropicalis and C. krusei, as well as clinical yeasts of
vulvovaginal candidiasis of C. albicans. Thus, this study demonstrated that chitosan
nanoparticles are promising for carrying and delivering the peptides TistH from Tityus
stigmurus, with improve antifungal activity against genus Candida spp. may eventually be
applied in the future as a possible antifungal agent. |
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