In vitro release and anti-herpetic activity of Cymbopogon citratus volatile oil-loaded nanogel

Abstract This study aimed to prepare hydrogel containing Cymbopogon citratus (DC.) Stapf, Poaceae, volatile oil encapsulated in poly (d,l-lactide-co-glycolide) nanoparticles and to evaluate its in vitro anti-herpetic activity. Polymeric nanoparticles were prepared by solvent emulsification-diffusion method and incorporated in carbomer hydrogels. In vitro release profiles for the nanogel, loaded nanoparticles and hydrogel containing free oil were evaluated by dialysis. Inhibitory activities against Herpes simplex for the formulations were investigated in Vero cells. Hydrogel was developed using nanoparticles with mean diameter of 217.1 nm and negative Zeta potential (−20.5 mV). Volatile oil release profile showed a biphasic pattern with an initial faster release and subsequent sustained phase in all formulations. Nanogel strongly inhibited virus in a non-cytotoxic concentration, 42.16 times lower than free oil, 8.76 and 2.23 times than loaded nanoparticles and hydrogel containing free oil, respectively. These results highlight the potential of nanogel to protect oil against volatilization, control release and improve its anti-herpetic activity.

Anti-cryptococcal activity of ethanol crude extract and hexane fraction from Ocimum basilicum var. Maria bonita: mechanisms of action and synergism with amphotericin B and Ocimum basilicum essential oil.

CONTEXT: Ocimum basilicum L. (Lamiaceae) has been used in folk medicine to treat headaches, kidney disorders, and intestinal worms. OBJECTIVE: This study evaluates the anti-cryptococcal activity of ethanol crude extract and hexane fraction obtained from O. basilicum var. Maria Bonita leaves. MATERIALS AND METHODS: The MIC values for Cryptococcus sp. were obtained according to Clinical and Laboratory Standards Institute in a range of 0.3-2500 µg/mL. The checkerboard assay evaluated the association of the substances tested (in a range of 0.099-2500 µg/mL) with amphotericin B and O. basilicum essential oil for 48 h. The ethanol extract, hexane fraction and associations in a range of 0.3-2500 µg/mL were tested for pigmentation inhibition after 7 days of treatment. The inhibition of ergosterol synthesis and reduction of capsule size were evaluated after the treatment with ethanol extract (312 µg/mL), hexane fraction (78 µg/mL) and the combinations of essential oil + ethanol extract (78 µg/mL + 19.5 µg/mL, respectively) and essential oil + hexane fraction (39.36 µg/mL + 10 µg/mL, respectively) for 24 and 48 h, respectively. RESULTS: The hexane fraction presented better results than the ethanol extract, with a low MIC (156 µg/mL against C. neoformans T444 and 312 µg/mL against C. neoformans H99 serotype A and C. gattii WM779 serotype C). The combination of the ethanol extract and hexane fraction with amphotericin B and essential oil enhanced their antifungal activity, reducing the concentration of each substance needed to kill 100% of the inoculum. The substances tested were able to reduce the pigmentation, capsule size and ergosterol synthesis, which suggest they have important mechanisms of action. CONCLUSIONS: These results provide further support for the use of ethanol extracts of O. basilicum as a potential source of antifungal agents.

O-glycosylation in cell wall proteins in Scedosporium prolificans is critical for phagocytosis and inflammatory cytokines production by macrophages.

In this study, we analyze the importance of O-linked oligosaccharides present in peptidorhamnomannan (PRM) from the cell wall of the fungus Scedosporium prolificans for recognition and phagocytosis of conidia by macrophages. Adding PRM led to a dose-dependent inhibition of conidia phagocytosis, whereas de-O-glycosylated PRM did not show any effect. PRM induced the release of macrophage-derived antimicrobial compounds. However, O-linked oligosaccharides do not appear to be required for such induction. The effect of PRM on conidia-induced macrophage killing was examined using latex beads coated with PRM or de-O-glycosylated PRM. A decrease in macrophage viability similar to that caused by conidia was detected. However, macrophage killing was unaffected when beads coated with de-O-glycosylated PRM were used, indicating the toxic effect of O-linked oligosaccharides on macrophages. In addition, PRM triggered TNF-α release by macrophages. Chemical removal of O-linked oligosaccharides from PRM abolished cytokine induction, suggesting that the O-linked oligosaccharidic chains are important moieties involved in inflammatory responses through the induction of TNF-α secretion. In summary, we show that O-glycosylation plays a role in the recognition and uptake of S. prolificans by macrophages, killing of macrophages and production of pro- inflammatory cytokines.

Antiherpetic activity of a flavonoid fraction from Ocotea notata leaves

This study describes the isolation of a flavonoid fraction from leaves of Ocotea notata (Nees & Mart.) Mez, Lauraceae, the identification of six major compounds (an A-type proanthocyanidin trimer [3], isoquercitrin [4], reynoutrin [5], miquelianin [6], quercitrin [7], afzelin [8]) and four minor compounds (catechin [1], epicatechin [2], quercetin [9], kaempferol [10]) present in the fraction and its activity against the Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). The 50% effective concentrations values (EC50) calculated from the dose-response curve and the selectivity indices (SI) against the virus were: EC50 35.8 µg/mL and SI 5.5 to HSV-1 and EC50 23.5 µg/mL and SI 8.5 to HSV-2. The flavonoid fraction was more active against HSV-2 than HSV-1. The mechanisms of antiviral action of the flavonoid fraction against the virus were also evaluated. The percentage inhibition (PI) obtained for HSV-2 was higher than 90% in the following assays: virucidal, pre-treatment of cells, treatment of cells after viral adsorption and treatment of cells after viral penetration. For HSV-1, the flavonoid fraction had no effect in pre-treatment of cells and showed 60% of inhibition in virucidal assay.

Evaluation of cytotoxicity and degree of conversion of glass ionomer cements reinforced with resin.

The objective of the present study was to evaluate the cytotoxicity and degree of monomer conversion of resin-reinforced glass ionomer cements (RGIC) over different time periods. Four RGICs: Fuji Ortho LC (FOLC), Fuji Ortho Band (FOB), Orthoglass (OGL), and Multicure Glass Ionomer (MCI) were evaluated for cytotoxicity in fibroblastic L929 cells and for their degree of monomer conversion over different time periods. Three control groups were also analysed: positive control (C+), consisting of Tween 80 cell detergent; negative control (C-), consisting of phosphate-buffered saline; and cell control (CC), consisting of cells exposed to any material. To evaluate the cytotoxicity, the dye-uptake technique was used and the degree of conversion was evaluated using infrared spectroscopy. The data obtained were analysed by analysis of variance and the Tukey's test. The results showed cytotoxicity of the RGICs at 1 and 24 hours; the viability values of these materials were statistically different from the C- and CC groups (P < 0.05). After 48 hours, the FOLC group was statistically similar to the CC and C- groups but different from the others. At 1 hour, there was no difference in the degree of conversion between the FOLC and OGL groups (P > 0.05) or between the FOB and MCI (P < 0.05) groups. However, at 48 hours, the FOLC group had greater conversion values than the other groups (P < 0.05). There is a direct relationship between the degree of conversion and RGIC cytotoxicity. Following initial polymerization, cytotoxicity decreases and, consequently, the degree of conversion of the material increases.

Glycolipids from macroalgae: potential biomolecules for marine biotechnology?

Brown, red and green algae from the Southeastern coast of Brazil were successively extracted with chloroform/methanol 2:1 and 1:2 (v/v). The crude lipid extract was partitioned according to Folch and the lower phase enriched in glycolipids was fractionated on a silica gel column chromatography eluted with chloroform, acetone and methanol. Three major orcinol-reactive bands present in the acetone and methanol fractions were detected by thin-layer chromatography with chromatographic mobilities corresponding to sulfoglycolipids and glycosyldiacylglycerols. These fractions exhibited potent antiviral activity against HSV-1-ACVs and HSV-1-ACVr and present low toxicity for cell cultures. Purification and identification of these bioactive glycolipids will be necessary in order to elucidate their primary structures and mechanism of action.