Nanocarried antioxidants in freezing extenders for boar spermatozoa.

Post-thawing cryoinjuries in boar spermatozoa due to oxidative stress may be reduced by adding nanoencapsulated antioxidants to freezing extenders. This study evaluated post-thawing kinetics, structural and biochemical functions of boar spermatozoa frozen with extenders including resveratrol and vitamin E loaded into polymeric nanocapsules. Resveratrol was added at 0 (control), 5, 10, 20, 40 and 80 µg/ml, whereas Vitamin E was added at 0 (control), 50, 100, 200 and 400 µg/ml. Both antioxidants were tested in free and nanoencapsulated presentations. In contact with empty nanocapsules, some sperm kinetics parameters were impaired compared to the control (p < .05), whereas lipoperoxidation declined (p < .05). With inclusion of 40 µg/ml nanoencapsulated resveratrol, some sperm kinetics parameters were improved (p < .01), but sperm motility, structural and biochemical functions did not differ from the control (p > .05). No improvement in sperm quality occurred with inclusion of vitamin E, although sperm kinetics with 400 µg/ml nanoencapsulated vitamin E was reduced compared to the control (p < .01). Inclusion of 40 µg/ml nanoencapsulated resveratrol benefitted boar sperm kinetics after thawing, but no improvement resulted from inclusion of vitamin E.

Safety assessment of poly(N-vinylcaprolactam) as a potential drug carrier in extenders for boar sperm cryopreservation.

Polymers may be used to deliver compounds in freezing extenders to minimize injuries in spermatozoa during cryopreservation, although their activity and toxicity for boar sperm are unknown. This study investigated the effects of the polymer (N-vinylcaprolactam) (PNVCL), when included in extenders for boar sperm cryopreservation. In Experiment 1, sperm was exposed to PNVCL at: 0 (control); 39.1; 78.1; 156.3; and 312.5 µg/mL. Spermatozoa structure, kinetics and biochemical functions were unaltered in contact with PNVCL at 38 °C (P > .05) but declined with prolonged exposure (10, 60 and 120 min) in all treatments (P > .05). In Experiment 2, after inclusion of PNVCL in the freezing extender at the same concentrations, post-thawing sperm quality did not differ compared to the control (P > .05). Lipid peroxidation and the production of reactive oxygen species were the only parameters of sperm quality that were unaffected in both experiments, even after contact with PNVCL for 120 min (P > .05). As no negative effects were observed in post-thawing boar sperm quality, PNVCL did not incur in cytotoxicity and may be a potential carrier for antioxidants in freezing extenders.

Inhibitory effects of a standardized extract of Justicia pectoralis in an experimental rat model of airway hyper-responsiveness.

OBJECTIVE: Justicia pectoralis is a plant useful for the treatment of respiratory diseases. Here, we studied the antiasthmatic properties of a standardized extract of J. pectoralis (Jp). METHODS: Ovalbumin (OVA)-sensitized rats were actively challenged with saline or OVA to study airway hyper-responsiveness after oral treatment with saline or Jp. The ability of Jp to inhibit hyper-reactivity was evaluated in isolated trachea mounted in isolated organ bath chamber. KEY FINDINGS: Using KCl or carbachol as contractile agents, tracheal rings of OVA-challenged rats contracted with higher magnitude than trachea of rats challenged with saline. Such hyper-responsive phenotype of OVA-challenged tissues decreased with Jp administration. In Ca+ -free medium, Jp or its major constituent coumarin inhibited preferentially the contractions induced by Ca2+ addition in tissues of OVA-challenged rats stimulated with KCl or acetylcholine. In tissues depleted of their internal Ca+ stores in the presence of thapsigargin, Jp inhibited the contraction induced by capacitative Ca2+ entry. By gavage, Jp abolished the increase caused by challenge with OVA on the levels of IL-1ß and TNF-α in the bronchoalveolar fluid and also impaired the changes in gene expression of canonical transient receptor proteins. CONCLUSIONS: Jp has antiasthmatic properties in an experimental model that reproduces tracheal hyper-reactivity.

Lipid-Core Nanocapsules Improved Antiedematogenic Activity of Tacrolimus in Adjuvant-Induced Arthritis Model.

Despite significant technological advances, rheumatoid arthritis remains an incurable disease with great impact on the life quality of patients. We studied the encapsulation of tacrolimus in lipidcore nanocapsules (TAC-LNC) as a strategy to enhance its systemic anti-arthritic properties. TAC-LNC presented unimodal distribution of particles with z-average diameter of 212 +/- 11, drug content close to the theoretical value (0.80 mg mL(-1)), and 99.43% of encapsulation efficiency. An in vitro sustained release was determined for TAC-LNC with anomalous transport mechanism (n = 0.61). In vivo studies using an arthritis model induced by Complete Freund's Adjuvant demonstrated that the animals treated with TAC-LNC presented a significantly greater inhibition of paw oedema after intraperitoneal administration. Furthermore, the encapsulation of TAC in lipid-core nanocapsules was potentially able to prevent hyperglycemia in the animals. In conclusion, TAC-LNC was prepared with 100% yield of nanoscopic particles having satisfactory characteristics for systemic use. This formulation represents a promising strategy to the treatment of rheumatoid arthritis in the near future.

Co-encapsulation of imiquimod and copaiba oil in novel nanostructured systems: promising formulations against skin carcinoma.

In this study, two types of cutaneous-directed nanoparticles are proposed for the co-encapsulation of imiquimod (a drug approved for the treatment of basal cell carcinoma) and copaiba oil (oil that exhibits anti-proliferative properties). Nanostructured copaiba capsules (NCCImq) were prepared using the interfacial deposition method, and nanostructured Brazilian lipids (NBLImq) were prepared by high-pressure homogenization. The formulations exhibited average diameter, zeta potential, pH and drug content of approximately 200nm, -12mV, 6 and 1mgmL(-1), respectively. In addition, the formulations exhibited homogeneity regarding particle size, high encapsulation efficiency and stability. Both nanocarriers controlled imiquimod release, and NBLImq exhibited slower drug release (p < 0.05), likely due to increased interaction of the drug with the solid lipid (cupuaçu seed butter). The in vitro evaluation of the imiquimod-loaded nanocarriers was performed using healthy skin cells (keratinocytes, HaCaT); no alteration was observed, suggesting the biocompatibility of the nanocarriers. In addition, in vitro skin permeation/penetration using pig skin was performed, and NCCImq led to increased drug retention in the skin layers and reduced amounts of drug found in the receiver solution. Thus, NCCImq is considered the most promising nanoformulation for the treatment of skin carcinoma.

Mucoadhesive Amphiphilic Methacrylic Copolymer-Functionalized Poly(ε-caprolactone) Nanocapsules for Nose-to-Brain Delivery of Olanzapine.

Nose-to-brain drug delivery has been proposed to overcome the low absorption of drugs in central nervous system due to the absence of brain-blood barrier in the olfactory nerve pathway. However, the presence of a mucus layer and quick clearance limit the use of this route. Herein, amphiphilic methacrylic copolymer-functionalized poly(ε-caprolactone) nanocapsules were proposed as a mucoadhesive system to deliver olanzapine after intranasal administration. In vitro evaluations showed that these nanocapsules were able to interact with mucin (up to 17% of increment in particle size and 30% of reduction of particle concentration) and nasal mucosa (2-fold higher force for detaching), as well as to increase the retention of olanzapine (about 40%) on the nasal mucosa after continuous wash. The olanzapine-loaded amphiphilic methacrylic copolymer-functionalized PCL nanocapsules enhanced the amount of drug in the brain of rats (1.5-fold higher compared to the drug solution). In accordance with this finding, this formulation improved the prepulse inhibition impairment induced by apomorphine, which is considered as an operational measure of pre-attentive sensorimotor gating impairment present in schizophrenia. Besides, nanoencapsulated olanzapine did not affect the nasal mucosa integrity after repeated doses. These data evidenced that the designed nanocapsules are a promising mucoadhesive system for nose-to-brain delivery of drugs.

A novel approach to arthritis treatment based on resveratrol and curcumin co-encapsulated in lipid-core nanocapsules: In vivo studies.

Resveratrol and curcumin are two natural polyphenols extensively used due to their remarkable anti-inflammatory activity. The present work presents an inedited study of the in vivo antioedematogenic activity of these polyphenols co-encapsulated in lipid-core nanocapsules on Complete Freund's adjuvant (CFA)-induced arthritis in rats. Lipid-core nanocapsules were prepared by interfacial deposition of preformed polymer. Animals received a single subplantar injection of CFA in the right paw. Fourteen days after arthritis induction, they were treated with resveratrol, curcumin, or both in solution or loaded in lipid-core nanocapsules (1.75 mg/kg/twice daily, i.p.), for 8 days. At the doses used, the polyphenols in solution were not able to decrease paw oedema. However, nanoencapsulation improved the antioedematogenic activity of polyphenols at the same doses. In addition, the treatment with co-encapsulated polyphenols showed the most pronounced effects, where an inhibition of 37-55% was observed between day 16 and 22 after arthritis induction. This treatment minimized most of the histological changes observed, like fibrosis in synovial tissue, cartilage and bone loss. In addition, unlike conventionally arthritis treatment, resveratrol and curcumin co-encapsulated in lipid-core nanocapsules did not alter important hepatic biochemical markers (ALP, AST, and ALT). In conclusion, the strategy of co-encapsulating resveratrol and curcumin in lipid-core nanocapsules improves their efficacy as oedematogenic agents, with no evidence of hepatotoxic effects. This is a promising strategy for the development of new schemes for treatment of chronic inflammation diseases, like arthritis.

Justicia pectoralis Jacq: , Acanthaceae: preparation and characterisation of the plant drug including chromatographic analysis by HPLC-PDA

Justicia pectoralis Jacq., Acanthaceae, é uma erva conhecida popularmente no Nordeste como chambá e, utilizada tradicionalmente no tratamento de doenças do trato respiratório, como a asma, tosse e bronquite. Essa espécie encontra-se na Relação Nacional de Plantas Medicinais de Interesse para o SUS. O objetivo do presente estudo foi elaborar protocolo para a preparação da droga vegetal a partir de J. pectoralis e realizar a sua caracterização visando seu emprego como matéria-prima farmacêutica. A parte aérea de J. pectoralis, após secagem em estufa com circulação e renovação de ar (35 °C) durante diferentes períodos de tempo (1-5 dias) mostrou a partir de 24 h de secagem um teor de umidade abaixo do valor máximo permitido para drogas vegetais. O pó da droga vegetal foi classificado como pó moderadamente grosso e, caracterizado quanto aos teores de cinzas totais, extrativos solúveis em água e etanol. Análise do extrato hidroalcoólico (etanol 20 por cento) de J. pectoralis por Cromatografia Líquida de Alta Eficiência (CLAE-DAD) determinou um teor de cumarina e umbeliferona de 16,2 e 0,81 mg/g da droga vegetal, respectivamente. As condições de preparação da droga vegetal e os parâmetros de controle de qualidade determinados para J. pectoralis no presente estudo são de interesse no desenvolvimento de fitoterápicos que empreguem esse matéria-prima ativa.

Justicia pectoralis Jacq., Acanthaceae, is a herb popularly known in Brazilian northeast as "chambá" and used in folk medicine for the treatment of respiratory tract conditions such as asthma, cough and bronchitis. This species is included in the National Register of Plants of Interest to the National Health System. The aim of the present study was to develop a protocol for the preparation of the plant drug from J. pectoralis and to characterise the plant drug for its use as a pharmaceutical raw material. The aerial parts of J. pectoralis, after drying chamber with forced air circulation (35 °C) for different periods of time (1-5 days), presented after one day a moisture content below the maximum allowed for plant drugs. The powder of the plant drug was classified as moderately coarse, and the total ashes content and the water- or ethanol-soluble extractives were determined. Analysis of hydroalcoholic (ethanol 20 percent) extract of J. pectoralis by high performance liquid chromatography-photo diode array (HPLC-PDA) determined the content of coumarin and umbelliferone (16.2 and 0.81 mg/g plant drug, respectively). The preparation conditions of the plant drug and the quality control parameters established for J. pectoralis in this study are of interest for the development of phytomedicines which use this active raw material.