Natural products continue to be an invaluable resource of anticancer drug discovery in recent years. Propolis is known for its biological activities such as antimicrobial and antitumor effects. This study assessed the effects of Brazilian red propolis (BRP) on apoptosis and migration potential in human bladder cancer cells. The effect of BRP ethanolic extract (25, 50, and 100 μg/mL) on 5637 cells was determined by MTT, LIVE/DEAD, and migration (scratch assay) assays. Apoptosis induction was investigated through flow cytometry and gene expression profile was investigated by qRT‐PCR. Results showed cytotoxicity on MTT and LIVE/DEAD assays, with IC50 values of 95 μg/mL in 24 h of treatment. Cellular migration of 5637 cells was significantly inhibited through lower doses of BRP ethanolic extract (25 and 50 μg/mL). Flow cytometry analyses showed that BRP induced cytotoxicity through apoptosis‐like mechanisms in 5637 cells and qRT‐PCR revealed increased levels of Bax/Bcl‐2 ratio, p53, AIF, and antioxidant enzymes genes. Data suggest that BRP may be a potential source of drugs to bladder cancer treatment.
Most studies of Brazilian red propolis have explored the composition and biological properties of its ethanolic extracts. In this work, we chemically extracted and characterized the essential oil of Brazilian red propolis (EOP) and assessed its adjuvant, antiparasitic and cytotoxic activities. The chemical composition of EOP was analyzed using gas chromatography with mass spectrometry (GC-MS). EOP was tested for in vitro activity against Trichomonas vaginalis (ATCC 30236 isolate); trophozoites were treated with different concentrations of EOP (ranging from 25 to 500 μg/mL) in order to establish the MIC and IC50 values. A cytotoxicity assay was performed in CHO-K1 cells submitted to different EOP concentrations. BALB/c mice were used to test the adjuvant effect of EOP. The animals were divided in 3 groups and inoculated as follows: 0.4 ng/kg BW EOP (G1); 50 μg of rCP40 protein (G2); or a combination of 0.4 ng/kg BW EOP and 50 μg of rCP40 (G3). Total IgG, IgG1 and IgG2a levels were assessed by ELISA. The major constituent compounds of EOP were methyl eugenol (13.1%), (E)-β-farnesene (2.50%), and δ-amorphene (2.3%). Exposure to EOP inhibited the growth of T. vaginalis, with an IC50 value of 100 μg/mL of EOP. An EOP concentration of 500 μg/mL was able to kill 100% of the T. vaginalis trophozoites. The EOP kinetic growth curve showed a 36% decrease in trophozoite growth after a 12 h exposure to 500 μg/mL of EOP, while complete parasite death was induced at 24 h. With regard to CHO-K1 cells, the CC50 was 266 μg/mL, and 92% cytotoxicity was observed after exposure to 500 μg/mL of EOP. Otherwise, a concentration of 200 μg/mL of EOP was able to reduce parasite proliferation by 70% and was not cytotoxic to CHO-K1 cells. As an adjuvant, a synergistic effect was observed when EOP was combined with the rCP40 protein (G3) in comparison to the administration of each component alone (G1 and G2), resulting in higher concentrations of IgG, IgG1 and IgG2a. EOP is constituted by biologically active components with promising antiparasitic and immunostimulatory activities and can be investigated for the formulation of new vaccines or trichomonacidal drugs.
Wound infections are the main complication when treating skin wounds. This work reports a novel antimicrobial material using green synthesized zinc oxide nanoparticles (ZnONPs) incorporated in polymeric fibers for wound healing purposes. ZnONPs are a promising antimicrobial nanomaterial with high activity against a range of microorganisms, including drug-resistant bacteria. The electrospun fibers were obtained using polyacrylic acid (PAA) and polyallylamine hydrochloride (PAH) and were loaded with ZnONPs green synthesized from Ilex paraguariensis leaves with a spherical shape and ~18 nm diameter size. The fibers were produced using the electrospinning technique and SEM images showed a uniform morphology with a diameter of ~230 nm. EDS analysis proved a consistent dispersion of Zn in the fiber mat, however, particle agglomerates with varying sizes were observed. FTIR spectra confirmed the interaction of PAA carboxylic groups with the amine of PAH molecules. Although ZnONPs presented higher antimicrobial activity against S. aureus than E. coli, resazurin viability assay revealed that the PAA/PAH/ZnONPs composite successfully inhibited both bacteria strains growth. Photomicrographs support these results where bacteria clusters were observed only in the control samples. The PAA/PAH/ZnONPs composite developed presents antimicrobial activity and mimics the extracellular matrix morphology of skin tissue, showing potential for wound healing treatments.