RESUMO
In recent years, we have discovered Esc(1-21) and its diastereomer (Esc peptides) as valuable candidates for the treatment of Pseudomonas lung infection, especially in patients with cystic fibrosis (CF). Furthermore, engineered poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) were revealed to be a promising pulmonary delivery system of antimicrobial peptides. However, the "ad hoc" development of novel therapeutics requires consideration of their stability, tolerability, and safety. Hence, by means of electrophysiology experiments and preclinical studies on healthy mice, we demonstrated that neither Esc peptides or Esc-peptide-loaded PLGA NPs significantly affect the integrity of the lung epithelium, nor change the global gene expression profile of lungs of treated animals compared to those of vehicle-treated animals. Noteworthy, the Esc diastereomer endowed with the highest antimicrobial activity did not provoke any pulmonary pro-inflammatory response, even at a concentration 15-fold higher than the efficacy dosage 24 h after administration in the free or encapsulated form. The therapeutic index was ≥70, and the peptide was found to remain available in the bronchoalveolar lavage of mice, after two days of incubation. Overall, these studies should open an avenue for a new up-and-coming pharmacological approach, likely based on inhalable peptide-loaded NPs, to address CF lung disease.
RESUMO
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 (< 160â¯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 8â¯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⯵g.mL-1 against Candida albicans, 11.1⯵g.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.
Assuntos
Antifúngicos , Proteínas de Artrópodes , Candida/crescimento & desenvolvimento , Quitosana , Nanopartículas/química , Peptídeos , Venenos de Escorpião/química , Escorpiões/química , Animais , Antifúngicos/química , Antifúngicos/farmacologia , Proteínas de Artrópodes/química , Proteínas de Artrópodes/farmacologia , Quitosana/química , Quitosana/farmacologia , Chlorocebus aethiops , Camundongos , Peptídeos/química , Peptídeos/farmacologia , Células RAW 264.7 , Células VeroRESUMO
Photodynamic therapy (PDT), which enables the localized therapeutic effect by light irradiation, provides an alternative and complementary modality for the treatment of tumor. However, the aggregation of photosensitizers in acidic microenvironment of tumor and the non-targeted distribution of photosensitizers in normal tissues significantly affect the PDT efficiency. In this study, we developed a biodegradable nanocomplex HA-Arg-PEA from hyaluronic acid (HA) and arginine based poly(ester amide)s (Arg-PEA) as the nanocarrier for chlorin e6 (Ce6). HA enhanced the tumor-specific endocytosis mediated by the overexpression of CD44 receptor. Arg-PEA not only provide electrostatic interaction with HA to form self-assembled nanostructure, but also improve the monomerization of Ce6 at physiological pH as well as mildly acidic pH. The biodegradable characteristic of HA-Arg-PEA nanocomplex enabled the intracellular delivery of Ce6, in which its release and generation of singlet oxygen can be accelerated by enzymatic degradation of the carrier. The in vitro PDT efficiency of Ce6-loaded HA-Arg-PEA nanocomplex was examined in CD44 positive MDA-MB-435/MDR multidrug resistant melanoma cells. CD44-mediated uptake of Ce6-loaded HA-Arg-PEA nanocomplex significantly improved Ce6 level in MDA-MB-435/MDR cells within short incubation time, and the PDT efficiency in inhibiting multidrug resistant tumor cells was also enhanced at higher Ce6 concentrations. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1487-1499, 2017.