In Vitro Activity of Melaleuca alternifolia (Tea Tree) in Its Free Oil and Nanoemulsion Formulations Against Pythium insidiosum.

Pythium insidiosum is an important aquatic oomycete which can cause pythiosis in both animals and humans. This microorganism shows low susceptibility to antifungal drugs available. This study analyzed the in vitro antimicrobial activity of Melaleuca alternifolia in its free oil (FO) and nanoemulsion (NE) formulations against Brazilian P. insidiosum isolates. The antimicrobial activity evaluation was performed by the broth microdilution method according to CSLI M38-A2 document adapted to phytopharmaceuticals. Twenty-six P. insidiosum isolates were evaluated, and the minimum inhibitory concentration was determined at 100 % growth inhibition. Melaleuca alternifolia essential oil or FO was obtained commercially. The NE containing 1 % M. alternifolia essential oil was prepared by the spontaneous emulsification method. All P. insidiosum isolates evaluated showed minimum inhibitory concentrations (MIC) ranging from 531.5 to 2125 µg/mL for the FO formulation; MIC50 and MIC90 showed values between 1062.5 and 2125 µg/mL, respectively. When the NE formulation was evaluated, MIC values ranged from 132.7 to 2125 µg/mL and both MIC50 and MIC90 corresponded to 1062.5 µg/mL. FO and NE formulations of M. alternifolia showed antimicrobial activity against P. insidiosum. This study demonstrated that M. alternifolia oil can be an additional therapy in pythiosis treatment; however, further studies are needed to evaluate the applicability of the plant essential oils in the treatment of clinical pythiosis.

In vitro and in vivo evaluation of a desonide gel-cream photostabilized with benzophenone-3.

CONTEXT: Our group previously reported the photoinstability of some desonide topical commercial formulations under direct exposure to UVA radiation. OBJECTIVE: This study aimed to prepare and characterize a gel-cream containing desonide, with greater photostability than the commercial gel-cream (C-GC). Benzophenone-3 (BP-3) was used as a photostabilizing agent. METHODS: The gel-cream developed (D-GC) containing BP-3 at 0.1% was prepared and characterized regarding its pH, drug content, spreadability, viscosity, in vitro drug release and in vitro permeation. The in vivo anti-inflammatory effect was assessed by ear edema measurement, croton oil-induced acute skin inflammation and myeloperoxidase assay. RESULTS AND DISCUSSION: D-GC presented characteristics compatible with topical application, appropriate drug content and good spreadability, and non-Newtonian behavior with pseudoplastic flow. D-GC showed a good photostability profile, presenting a desonide content of 95.70% after 48 h of exposure to UVA radiation, and stability under room conditions during 60 days. The amount of desonide released from D-GC and C-GC was 57.8 and 51.7 µg/cm2, respectively, measured using the vertical Franz cell. The in vitro skin permeation showed that desonide reached the site of action of the topical corticosteroids, from both formulations; however, the desonide amount retained in the dermis was lower with D-GC. The in vivo evaluation of topical anti-inflammatory activity indicated that D-GC presented the same biological effect as C-GC. CONCLUSION: D-GC represents a promising approach to treat dermatological disorders, since it presented satisfactory physicochemical characteristics, the same biological activity as C-GC and superior photostability, conferred by the addition of BP-3 at 0.1%.

Genotoxic effects of Campomanesia xanthocarpa extracts on Allium cepa vegetal system.

UNLABELLED: CONTEXT. Campomanesia xanthocarpa Berg. (Myrtaceae), popularly known in Brazil as guabiroba, is a plant used as antidiarrheic, anti-inflammatory and antirheumatic agents, and in stomach and hepatic disorders. OBJECTIVE: The antiproliferative and genotoxic effects of aqueous extracts and essential oil of C. xanthocarpa were evaluated. MATERIALS AND METHODS: Cytotoxicity and genotoxicity of the aqueous extracts (6 and 30 mg/mL) and essential oil (0.25%, v/v) obtained from leaves of C. xanthocarpa were evaluated using the Allium cepa L. (Amaryllidaceae) assay. Mitotic index was calculated as the percentage of dividing cells of total cells observed; chromosome abnormalities were observed and counted during cell division. Additionally, the composition of the essential oil and the quantification of the main compounds of the extracts were determined by gas chromatography/mass spectrometry and high performance liquid chromatography coupled with diode array detector, respectively. RESULTS AND DISCUSSION: Aqueous extracts (6 and 30 mg/mL) led to a reduction of 67.7% and 34.1% of the mitotic index, respectively, whereas the treatment with essential oil caused a 48.2% reduction in the mitotic index, when compared with negative control. Chromosomal mutations were observed and included anaphase bridges, delay chromosome, break chromosome, as well as metaphase with disorganized chromosomal and binuclear cells. The main compounds of the essential oil were ß-caryophyllene (8.87%), viridiflorol (6.40%), spathulenol (5.16%), δ-cadinene (4.92%), linalool (4.46%) and α-cadinol (4.25%). Gallic acid (3.19%), chlorogenic acid (1.04%), quercetin (2.97%) and rutin (4.82%) were identified in an aqueous extract (30 mg/mL). CONCLUSION: Our results demonstrated that genotoxic and antiproliferative activities are present in C. xanthocarpa infusions using the in vivo onion root-tip cell test.

Spray-dried powders improve the controlled release of antifungal tioconazole-loaded polymeric nanocapsules compared to with lyophilized products.

This work aimed to obtain solid formulations from polymeric nanocapsules and nanoemulsions containing tioconazole, a broad spectrum antifungal drug. Two dehydration methods were used: spray-drying and freeze drying, using lactose as adjuvant (10%, w/v). The liquid formulations had a mean particle size around 206 nm and 182 nm for nanocapsules and nanoemulsions, respectively, and an adequate polydispersity index. Tioconazole content was close to the theoretical amount (1.0 mg/mL). After drying, the content ranged between 98 and 102%with a mean nanometric size of the dried products after redispersion. Scanning electron microscopy showed that the particles are rounded, sphere-shaped for the dried products obtained by spray-drying, and shapeless and irregular shapes for those obtained by freeze-drying. In the microbiological evaluation, all dried products remained active against the yeast Candida albicans when compared to the original systems. The dried products obtained by spray-drying from nanocapsules presented better control of the tioconazole release when compared to the freeze-drying products.

Antinociceptive and anti-inflammatory effects of Aloe saponaria Haw on thermal injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: In Brazil, the plant Aloe saponaria Haw, popularly known as "babosa pintadinha", has been empirically used for its potential effect on thermal injury. Because there are no scientific data confirming its popular use, the aim of the present study was to investigate the effects of Aloe saponaria on nociceptive and inflammatory parameters in a rat model of thermal injury. MATERIALS AND METHODS: Adult male Wistar rats were subjected to a thermal injury or sham procedure (immersion in water at 70 or 37°C, respectively, for 5 or 8s). Burned animals were topically treated with vehicle (base cream), sulfadiazine 1% (positive control) or Aloe saponaria cream (0.3%-30%) once a day for 2 or 6 days. Each day, 30min before the treatment, we measured nociceptive (static and dynamic mechanical allodynia, thermal allodynia and spontaneous pain) and inflammatory (paw edema) parameters. Moreover, enzymatic indicators of leukocyte infiltration into burned tissue were also determined 2 or 6 days after the thermal injury. RESULTS: The thermal injury (fist and second-degree) procedure, but not the sham procedure, induced nociception and inflammation from 1 to 6 days after the injury. The topical treatment with Aloe saponaria cream (10%) reduced nociceptive behaviors from day 1 to 6 (peak at day 2), edema at days 5 and 6 (peak at day 6) and myeloperoxidase, N-acetyl-glucosaminidase and eosinoperoxidase activities at day 6. The antinociceptive and anti-inflammatory effects of Aloe saponaria were obtained with doses of 3%-30%, with maximal inhibition obtained with a dose of 10% (reductions of 39±9%, 41±9%, 31±7%, 83±7% and 23±2% for static and dynamic mechanical allodynia, thermal allodynia, spontaneous pain and paw edema, respectively). CONCLUSION: Our results demonstrate that topically applied Aloe saponaria presented antinociceptive and anti-inflammatory effects in rats subjected to a thermal injury, which supports its traditional use for burn injuries.

Haloperidol-loaded polysorbate-coated polymeric nanocapsules decrease its adverse motor side effects and oxidative stress markers in rats.

Haloperidol is the most widely used antipsychotic drug in the treatment of psychiatric disorders. Despite its satisfactory therapeutic effect, its chronic use is related to severe motor side effects. Here, we investigate the incidence of motor side effects of haloperidol-loaded nanocapsules when compared to free haloperidol and the relation with oxidative stress (OS) development. Both vehicle (B-NcFO) and haloperidol loaded polysorbate-coated nanocapsules suspension (H-NcFO) prepared with fish oil as core showed uniform and rounded particles, nanometric size, negative zeta potential, low polydispersity indices and high encapsulation efficiency. Wistar rats received a single dose of free haloperidol (FH), B-NcFO or H-NcFO (0.2 mg/kg ip) and were submitted to acute motor side effects evaluation 1 h after the injection. Lower catalepsy time and oral dyskinesia were observed in H-NcFO-treated group than in FH group; however, both formulations decreased animals' locomotor activity. In a experiment performed subchronically, rats injected daily with H-NcFO (0.2 mg/kg-ip) for 28 days showed decreased oral dyskinesia frequency and catalepsy time and no impairment on locomotor activity as compared to FH group (0.2 mg/kg-ip). FH group showed higher OS, as observed by increased lipid peroxidation and reduced glutathione levels and catalase activity in extrapyramidal region. Our findings showed that nanocapsules may be an efficient form to prevent or minimize haloperidol motor side effects, which are related to OS development, ameliorating psychiatric patients' quality of life.

HPLC determination of bezafibrate in human plasma and its application to pharmacokinetics studies.

An isocratic high-performance liquid chromatographic (HPLC) method was developed and validated for the determination of bezafibrate in biological fluids. Bezafibrate was separated on a C(18) analytical column (150 x 4.6 mm i.d., 5 microm particle size) with 0.01 M phosphate buffer (pH 3.5)-acetonitrile-methanol (50:40:10) as mobile phase at a flow rate of 1.0 mL/min. The UV detector was set to 230 nm. Bezafibrate was extracted from human plasma using a simple liquid-liquid extraction with tert-butyl methyl ether. Parameters such as linearity, precision, accuracy, recovery, specificity, and stability were evaluated by method validation studies. All the parameters remained within acceptable limits. The validated procedure was linear in the concentration range of 0.2-50 microg/mL. The proposed method used for individual drug determinations is applicable for therapeutic monitoring purposes as well as for use in pharmacokinetic investigations. As an example, the practical quantification limit for bezafibrate in plasma was about 0.05 microg/mL with precision of 10.2% and accuracy of 112.6%. The method was applied in a study of the pharmacokinetics of bezafibrate in six healthy volunteers, who ingested a single oral dose of 200 mg.