Biodegradable cross-linked chitosan nanoparticles improve anti-Candida and anti-biofilm activity of TistH, a peptide identified in the venom gland of the Tityus stigmurus scorpion.

Among several bioactive peptides identified from the venom glands of the Tityus stigmurus scorpion, one peptide with hypotensive action (TistH, Tityus stigmurus Hypotensin) showed multifunctional and biotechnological applications. The maximum efficacy of this class of compounds can be achieved by immobilizing it in specific and suitable biomaterials or suitable carriers. In this study, distinct entrapment methods of TistH in chitosan nanoparticles was tested using its incorporation (CN-TistH-Inc) or adsorption (CN-TistH-Ads) methods by ionotropic gelification. Physico-chemical properties as well as biocompatibility and antifungal efficacy were assessed for different samples. Atomic force microscopy and field emission gun scanning electronic microscopy images associated with particle size measurements demonstrated that the two methods induced cationic spherical, small (< 160 nm), and narrow-sized (PdI about 0.3) nanoparticles, even after peptide loading greater than 96.5%, which was confirmed using Fourier transform infrared spectroscopy. The colloidal suspensions showed to be stable for 8 weeks and were able to induce the desired slow in vitro peptide release. Cytotoxicity assays performed in normal cells originated from murine macrophages (RAW 264.7) and kidneys of African green monkeys (Vero E6) suggested biocompatibility of samples. The CN-TistH-Inc and CN-TistH-Ads showed a minimal inhibitory concentration of 89.2 µg.mL-1 against Candida albicans, 11.1 µg.mL-1 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. In this approach, biodegradable nanocarriers prepared using simple and reproducible methods demonstrated the ability to deliver the TistH peptide from T. stigmurus and improve its antifungal efficacy.

A biotechnological approach to immunotherapy: Antivenom against Crotalus durissus cascavella snake venom produced from biodegradable nanoparticles.

Snakebite envenoming is a tropical disease neglected worldwide. In Brazil, the Crotalus durissus cascavella (CDC) snake belongs to a genus with venom of highest lethality. A search for new immunoadjuvants aimed to expand the therapeutic alternatives to improve vaccines and antivenom. This approach proposed to produce small and narrow-sized cationic CDC venom-loaded chitosan nanoparticles (CHNP) able to induce antibody response against the CDC venom. The ionic gelation method induced the formation of stable and slightly smooth spherical nanoparticles (<160 nm) with protein loading efficiency superior to 90%. The interactions between venom proteins and CHNP assessed using FT-IR spectroscopy corroborated with the in vitro release behavior of proteins from nanoparticles. Finally, the immunization animal model using BALB/c mice demonstrated the higher effectiveness of CDC venom-loaded CHNP compared to aluminum hydroxide, a conventional immunoadjuvant. Thus, CHNPs loaded with CDC venom exhibited a promising biotechnological approach to immunotherapy.