Preparation of nanocapsules of active ingredients of Garcinia hanburyi and evaluation of their antidiabetic activity / 中国药房

OBJECTIVE To prepare Gambogic acid （GA） nanocapsules （GA-LNCs） and Neogambogic acid （NGA） nanocapsules（NGA-LNCs），and to evaluate their antidiabetic activities. METHODS Using water as the aqueous phase ，medium- chain triglyceride as the oil phase and polyethylene glycol monostearate as the surfactant ，GA-LNCs and NGA-LNCs were prepared by phase inversion method. Using entrapment efficiency and drug-loading amount as index ，the formulation technologies of above 2 nanocapsules were optimized by simplex lattice design. Its physical and chemical properties were investigated. The diabetic mice model was established. GA-LNCs and NGA-LNCs （1.92 and 2.42 mg/kg respectively ）were given intragastrically ，once a day ，for consecutive 6 weeks. The fasting blood glucose of mice ，the activities of superoxide dismutase （SOD）and glutathione peroxidase （GSH-Px），the contents of malondialdehyde （MDA），total cholesterol （TC），triglyceride （TG），high-density lipoprotein cholesterol（HDL-C）and low-density lipoprotein cholesterol （LDL-C）were all detected. RESULTS The optimal formulation of 2 kinds of nanocapsules included 60％ water，10％ medium-chain triglyceride ，30％ polyethylene glycol monostearate （total amount of the three was 2 g）and 35 mg GA or NGA . The encapsulation efficiencies of GA-LNCs and NGA-LNC obtained by the optimal formulation were （92.01±0.68）％ and（93.12±2.11）％；the drug-loading amount were （0.99±0.21）％ and（1.21±0.22）％， respectively. GA-LNCs and NGA-LNCs were yellow ，homogeneous and transparent liquid without precipitation. They were spherical in microscopic shape ， and had obvious shell- Δ 基金项目：吉林省科技发展计划项目（No.20200404090YY）；吉 membrane structure. The particle sizes were （28.11 ± 9.76） 林省教育厅科学技术研究项目（No.JJKH20210372KJ） ＊硕士研究生 。研究方向 ：植物药 。E-mail：zhanhe0108@163. and（22.06±6.84）nm；Zeta potential were （－4.09±1.00） com and（－17.40±1.32）mV，and polydispersity were 0.93±0.06 # 通信作者：讲师，博士。研究方向：中药有效成分治疗疾病的作 and 0.74±0.12. The results of animal experiments showed that 用机制。E-mail：chenweijia_jlau@163.com both GA-LNCs and NGA-LNCs could sig nificantly increase 中国药房 2022年第33卷第9期 China Pharmacy 2022Vol. 33 No. 9 ·1075· the activities of SOD and GSH-Px and the seru m content of HDL-C （P＜0.05 or P＜0.01）in model mice ，and significantly decreased the fasting blood glucose and the serum contents of MDA ， TC， TG and LDL-C （P＜0.05 or P＜0.01）. CONCLUSIONS GA-LNCs，NGA-LNCs prepared in this study are good in physical and chemical properties and have good anti-diabetes activity.

Docosahexaenoic fatty acid nanoencapsulated with Anti-PECAM-1 as strategy to increase atherosclerotic plaque stability / Ácido graxo docosahexaenoico nanoencapsulado com anti-PECAM-1 como uma estratégia para aumentar a estabilidade de placas ateroscleróticas

Cardiovascular diseases (CVDs) are the main cause of mortality worldwide, being the ischemic heart disease responsible for 85% of deaths. Atherosclerosis is a chronic inflammation of the arteries that underlies ischemic forms of CVD and involves the innate and adaptive immune systems, from initial fatty streak formation to atherosclerotic plaque ruptures, which defines the beginning and end stages of disease, respectively. Recent research on the reduction of systemic inflammation in order to treat CVD is controversial, since results show that this reduced inflammation can also increase patient susceptibility to general infection. Therefore, new tissue-targeting strategies are necessary. Docosahexaenoic fatty acid (DHA) is a natural bioactive precursor of pro-resolving oxylipins that can reduce inflammation. Based on these factors, the objective of this study was to develop a nanocapsule containing algae oil as a DHA source and apply anti-PECAM-1 on its surface to drive it to the inflamed endothelium. Initially, a surface-functionalized metal-complex multi-wall nanocapsule containing algae oil in its nucleus (MLNC-DHA-a1) was developed. This nanocapsules presented a mean diameter of 163 ± 5 nm, was spherical in shape, showed 94.80% conjugation efficiency using 200 µg/mL of anti-PECAM-1 on the surface, and did not show significant toxicity toward HUVECs at concentrations from 0.14 to 2.90x1011 nanocapsules/mL. The nanocapsules were also stable for 2 h, sufficient time to allow for clinical applications. In cell viability assays, concentrations of 0.14 to 1.40x1011 nanocapsules/mL did not significantly affect the viability of immortalized murine macrophages (RAW 264.7) and U-937 cells after 24, 48, and 72 h of treatment. Finally, macrophages were incubated with 0.75x1011 MLNC-DHA-a1 nanocapsules/mL for 4 h and showed a significant uptake, observed using dark-field hyperspectral microscopy (CytoViva®). Once inside murine macrophages (RAW 264.7), MLNC-DHA-a1 nanocapsules promoted a strong increase in M2 phenotype polarization compared to non-treated control cells. Our results suggest that DHA-enriched algae oil, as part of a lipid core nanocapsules, does not reduce cell viability and improves macrophage phenotype, making it a promising potential therapy for controlling chronic inflammation and healing or stabilizing atherosclerotic plaques

As doenças cardiovasculares (DCVs) são a principal causa de mortalidade no mundo, sendo os eventos isquêmicos responsáveis por 85% das mortes. A aterosclerose é uma inflamação crônica das artérias associada aos eventos isquêmicos das DCVs, na qual o sistema imunológico inato e adaptativo estão envolvidos desde a formação inicial das estrias gordurosas até a ruptura das placas ateroscleróticas. Pesquisas recentes direcionadas à redução da inflamação sistêmica têm mostrado resultados controversos, pois essa abordagem pode aumentar a susceptibilidade do paciente a infecções. Nesse sentido, novas estratégias direcionadas ao tecido lesionado são necessárias. No que se refere a medicamentos anti-inflamatórios ou suplementos alimentares, o ácido docosaexaenóico (DHA) tem sido relatado como um precursor natural de oxilipinas pró- resolutivas. Baseado nesse contexto, o objetivo deste estudo foi desenvolver nanocápsulas contendo óleo de alga como fonte de DHA e vetorizar essas nanopartículas com o anticorpo antiPECAM-1 em sua superfície, visando direcioná-las ao endotélio inflamado. Inicialmente, a nanocápsula multiparede metal-complexa funcionalizada contendo óleo de alga em seu núcleo (MLNC-DHA-a1) foi desenvolvida, apresentando um diâmetro médio de 163 ± 5 nm, formato esférico, onde a eficiência de conjugação do anti-PECAM-1 (200 µg/mL) foi de 94,80% sem toxicidade significativa em HUVECs nas concentrações de 1.14 a 2.9 x 1011 nanocápsulas/mL. As nanocápsulas apresentaram uma estabilidade de 2h, o que representa tempo suficiente para a sua aplicação clínica. A seguir, ensaios de viabilidade celular foram realizados em outras linhagens de células para avaliar a toxicidade das nanocápsulas. As concentrações de 0.14 a 1.40 x 1011 de nanocápsulas/mL não afetaram significativamente a viabilidade celular de macrófagos murinos imortalizados (RAW 264.7) e U-937 após 24, 48 e 72 h de tratamento. Por fim, os macrófagos (RAW 264.7) foram incubados com 0.75 x 1011 MLNC-DHA-a1/mL durante 4 h e apresentam uma captação significativa das nanocápsulas, observada por microscopia hiperespectral de campo escuro (CytoViva®). Uma vez captadas pelos macrófagos murinos imortalizados (RAW 264.7), as nanoformulações MLNC-DHA-a1 promoveram um forte aumento da polarização do fenótipo M2 em comparação com as células controle não tratadas. Nossos resultados sugerem que o óleo de alga rico em DHA presente no núcleo lipídico das nanocápsulas, não reduziu a viabilidade celular e estimulou uma maior polarização de macrófagos para o tipo M2, sendo assim uma terapia potencial para controlar a inflamação crônica e cicatrizar ou estabilizar placas ateroscleróticas

Stability and pharmacokinetics of hyaluronic acid-modified asparaginase self-assembled biomimetic nanocapsules / 中国药科大学学报

@#The stability and pharmacokinetic properties of hyaluronic acid-modified asparaginase (Asp) self-assembled bionic nanocapsules (ASNCs) were preliminarily investigated. ASNCs were prepared by molecular self-assembly method to investigate their morphology, particle size, zeta potential and antitrypsin stability. After intravenous injection of free Asp and ASNCs, rat plasma samples at different times were taken to determine Asp activity. Pharmacokinetic parameters were calculated by DAS pharmacokinetic software. The particle size of ASNCs was (99.17 ± 0.21) nm and the potential was -(13.13 ± 0.60) mV. In trypsin solution, ASNCs showed more excellent stability. The area under the activity-time curve (AUC0-48 h) of ASNCs was about 2 times higher than that of Asp; the mean residence time (MRT0-48 h) was about 1.7 times higher than that of Asp, and the bioavailability was 195% of Asp. The results showed that ASNCs could improve the stability and bioavailability of Asp against trypsin and prolong the circulation time of Asp in vivo.

PEG-PCL Nanocapsules Containing Curcumin: Validation of HPLC Method for Analyzing Drug-loading Efficiency, Stability Testing and Cytotoxicity on NIH-3T3 Cell Line

Abstract Curcumin (CUR) shows potential use for treating cancer. However, CUR has low solubility and reduced bioavailability, which limit its clinical effect. Therefore, the development of nanocarriers can overcome these problems and can ensure the desired pharmacological effect. In addition, it is mandatory to prove the quality, the efficacy, and the safety for a novel nanomedicine to be approved. In that sense, this paper aimed (a) to prepare CUR-loaded polyethylene glycol-poly(ε-caprolactone) nanocapsules; (b) to validate an analytical method by high performance liquid chromatography (HPLC) for quantifying CUR in these nanoformulations; (c) to evaluate the physicochemical stability of these formulations; and to investigate their cytotoxicity on NIH-3T3 mouse fibroblast cells. The HPLC method was specific to CUR in the loaded nanocapsules, linear (r = 0.9994) in a range of 10.0 to 90.0 µg.mL-1 with limits of detection and quantification of 0.160 and 0.480 µg.mL-1, respectively. Precision was demonstrated by a relative standard deviation lower than 5%. Suitable accuracy (102.37 ± 0.92%) was obtained. Values of pH, particle size, polydispersity index, and zeta potential presented no statistical difference (p > 0.05) for CUR-loaded nanoparticles. No cytotoxicity was observed against NIH-3T3 mouse embryo fibroblast cell line using both the tetrazolium salt and sulforhodamine B assays. In conclusion, a simple and inexpensive HPLC method was validated for the CUR quantification in the suspensions of nanocapsules. The obtained polymeric nanocapsules containing CUR showed suitable results for all the performed assays and can be further investigated as a feasible novel approach for cancer treatment.

Effect of Cilostazol-Loaded PCL/PEG Nanocapsules on Abdominal Aortic Tunics and Lipid Profile of Wistar Rats

Abstract Cilostazol (CLZ) is a phosphodiesterase III inhibitor with antiplatelet and vasodilator properties. It has been recently verified that CLZ plays a significant role in the arteries by inhibiting the proliferation and growth of muscle cells, increasing the release of nitric oxide by the endothelium and promoting angiogenesis. Considering these promising effects, the use of nanocapsules may be an interesting strategy to optimize its pharmacokinetics and pharmacodynamics at the vascular level for preventing atherosclerosis. The aim of this study was to evaluate the effect of cilostazol-loaded nanocapsules in the abdominal aortic tunics and on the lipid profile of Wistar rats in order to investigate its potential role in the prevention of atherosclerosis. Thirty-two animals were divided into four groups of eight animals, with 30-day treatment. Group 1 received nanoencapsulated CLZ; Group 2, control nanocapsules with no drug; Group 3, propylene glycol and water; and Group 4, a solution of CLZ in propylene glycol and water. After 30 days, there was no statistically significant difference between the groups regarding the cellularity and thickness of the arterial tunics of the abdominal aorta. However, the group that received nanoencapsulated CLZ (Group 1) had an improvement in HDL-c and triglyceride values compared to unloaded nanocapsules (Group 2).

Study on influence of polymer and surfactant on in vitro performance of biodegradable aqueous-core nanocapsules of tenofovirdisoproxil fumarate by response surface methodology

The major objective of this study was to investigate the effect of biodegradable polymer type and surfactant concentration on various characteristics viz. particle size, entrapment efficiency and drug release rate constant of aqueous core nanocapsules (ACNs) containing tenofovirdisoproxil fumarate. In this study, the nanocapsules were prepared by modified multiple emulsion technique with biodegradable polymers viz. poly(lactide-co-glycolide) of two different grades (PLGA RG502H and PLGA RG503H) and poly lactic acid (PLA R203H); and the surfactant employed was span 80. The experiments were designed under response surface methodology by employing the Design Expert software. Entrapment efficiency, particle size and drug release rate constant were taken as response variables. The prepared nanocapsules were subjected to characterization studies and the obtained results were statistically analyzed by Analysis of Variance (ANOVA) for response surface 2-Factorial Interaction model. ANOVA studies showed that the influence of both factors on all the response variables were significant at p<0.05. The optimized formulation was found to have the entrapment efficiency of 71.58%, particle size of 252.41 nm and the drug release rate constant of 0.045 h-1; thus, indicating that the ACNs were obtained with finest characteristics. SEM studies showed that the particles were spherical.

Sistemas de nanopartículas poliméricas II: estructura, métodos de elaboración, características, propiedades, biofuncionalización y tecnologías de auto-ensamblaje capa por capa (layer-by-layer self-assembly) / Polymeric nanoparticle systems: structure, elaboration methods, characteristics, properties, biofunctionalization and self-assembly layer by layer technologies

Los materiales poliméricos han sido ampliamente investigados para aplicaciones biomédicas, teniendo especial relevancia cuando se encuentran en forma de micro- y nano-partículas. Últimamente se ha ampliado su campo de aplicación al ser conjugados con péptidos y ácidos nucleicos, por lo tanto, el interés en el estudio de este tipo de materiales, así como también en la formulación de nanoestructuras funcionalizadas como materiales, dispositivos y vehículos de transporte de agentes terapéuticos ha aumentado. Las recientes investigaciones en nanosistemas se inspiran en fenómenos naturales que estimulan la integración de señales moleculares y la mimetización de procesos a nivel celular, de tejidos y órganos. Tecnológicamente, la capacidad de obtener nanoestructuras esféricas mediante la combinación de materiales que presenten propiedades distintas a las que ningún otro material individual posee por sí solo, es lo que hace que las nanocápsulas sean particularmente atractivas. Las potenciales ventajas de los sistemas de nanopartículas de tipo polimérico se destacan a lo largo de cada parte de este artículo de revisión. El presente artículo aborda los aspectos más relevantes sobre la estructura, composición y algunos métodos de elaboración de los sistemas nanoparticulados. Además, expone algunos de los trabajos más recientes, centrados en sistemas de nanopartículas basados en polímeros dirigidos a la administración de agentes, publicados en artículos especializados de investigación y revisiones durante los últimos años.

Polymeric materials have been extensively investigated for biomedical applications including micro- and nanoparticles. Modern advances have broadened horizons for application with peptides and nucleic acids. Therefore, interests increased in the formulation of materials, devices and vehicles for transporting therapeutic agents in functionalized nanostructures. Recent nano-systems are inspired by natural phenomena that stimulate the integration of molecular signals and the mimicking of natural cellular processes, at tissue and organ levels. Technologically, the ability to obtain spherical nanostructures, which combine different properties, that no other single material possesses on its own, makes nanocapsules particularly attractive. Potential advantages over polymer nanoparticulate systems are highlighted throughout each part of this review article. Here, we address the most relevant aspects of structure, composition and methods of formulation of nanoparticulate systems. In addition, we outline some of the more recent works focusing on nanosized preparations, based on agent-directed polymers, found in specialized research articles that have emerged in the recent years.

Preparation,Characterization and Biocompatibility Evaluation in vitro of DiR-PEG-PLGA Fluorescent Nanocapsules / 中国药房

OBJECTIVE:To prepare and characterize Fluorescent dye 1,1′-octacosyl-3,3,3′,3′-tetramethylindocarbocyanine iodide(DiR)-loading polyethylene glycol-poly lactic-co-glycolic acid(DiR-PEG-PLGA)nanocapsules,and to evaluate its biocompatibility in vitro. METHODS:Using PLGA and PEG-PLGA as carrier,DiR-PEG-PLGA nanocapsules were prepared by modified ultrasonic emulsification method. The particle size,Zeta potential,morphology,stability and fluorescence in vitro of nanocapsules were detected respectively. MTT assay was used to evaluate cytotoxicity in vitro of nanocapsules to human-derived HL7702 hepatocytes,and hemolysis test was carried out to investigate its hemolysis effects. RESULTS:Prepared DiR-PEG-PLGA nanocapsules were spherical with a clear core-shell structure. The average particle size was(507.53 ± 7.87)nm,polydispersity coetficient of particle size was 0.306 1±0.001 5 and Zeta potential was(-35.20±0.92)mV with good stability within 6 months under 4℃. Fluorescence signal intensity(y)of nanocapsules was increased linearly with DiR mass concentration(x)in vitro. The linear eguation was y=0.345 2x+0.433 4(R2=0.997 3).The toxicity of nanocapsules to HL7702 cells was between 0-1 degree,and no hemolytic effect was observed. CONCLUSIONS:The study successfully prepare fluorescent DiR-PEG-PLGA nanocapsules with high biocompatibility in vitro,which is further expected to become a safe optical drug carrier.

Production and characterization of nanocapsules encapsulated linalool by ionic gelation method using chitosan as wall material

Abstract Linalool has been extensively applied in various fields, such as flavoring agent, perfumes, cosmetics and medical science. However, linalool is unstable, volatile and readily oxidizable. A sensitive substance can be encapsulated in a capsule, so encapsulation technology can solve these problems. In this paper, linalool-loaded nanocapsules (Lin-nanocapsules) were prepared via the ionic gelation method and Lin-nanocapsules were characterized. The results of Fourier transformation infrared spectroscopy (FTIR) showed that linalool was successfully encapsulated in the wall materials. Scanning electron microscopy (SEM) results demonstrated that the shapes of Lin-nanocapsules, with smooth surfaces, were nearly spherical. Lin-nanocapsule average particle size was 352 nm and its polydispersity index (PDI) was proved to be 0.214 by the results of dynamic light scattering (DLC). Thermogravimetric results indicated that linalool loading capacity (LC) was 15.17%, and encapsulation could decrease linalool release and increase linalool retaining time under the high temperature. Oscillatory shear and steady-state shear measurements of Lin-nanocapsule emulsions were systematically investigated. The results of steady-state shear showed that Lin-nanocapsule emulsion, which was Newtonian only for high shear rate, was non-Newtonian. It was proved by oscillatory shear that when oscillation frequency changed from low to high, Lin-nanocapsules emulsion changed from viscous into elastic.

Permeating perfluoropentane drops-encapsulated mesoporous silica nanocapsules collaborative high-intensity focused ultrasound surface ablation of bovine liver in vitro / 中国介入影像与治疗学

Objective To discusse the influence of permeating perfluoropentane drops-encapsulated mesoporous silica nanocapsules (MSNC-PFP) on HIFU surface ablation in vitro bovine liver.Methods Fifity bovine liver tissues were divided into 5 groups:Control group,0.25 mg/ml group,0.50 mg/ml group,1.00 mg/ml group,2.00 mg/ml group according to the concentration of MSNC-PFP.The synergistic agent was injected along the ablation line path under the guiding of ultrasound.The surface ablation model was structured by 6 coagulation necrosis side by using HIFU linear scanning,inner region without melting,and the ultrasonographic changes were observed in the melt area.The coagulation necrosis range was assessed by TTC staining,and necrosis degree was observed by HE staining.The coagulation necrosis volume was measured,the coverage index (CI),external volume index (EI),energy efficiency factor (EEF) were used to evaluate the ablation effect in each group.Results The MSNC-PFP more than 1.00 mg/ml was shown as strong echo in ultrasonic image,fade after 3-5 min.The grey value was changed in HIFU ablation area related to the concentration of MSNC-PFP.The total ablation volume of 0.50 mg/ml group,1.00 mg/ml group,2.00 mg/ml group were more than control group,the EEF was lower than that of the control group (all P＜0.01).The CI and EI of 1.00 mg/ml group and 2.00 mg/ml group were higher than those of the other group (all P＜0.01).The coagulation necrosis became complete when the concentration of MSNC-PFP above 0.50 mg/ml.Conclusion MSNC-PFP can increase the volume of coagulation necrosis,decrease the EEF and increase the efficiency of HIFU surface ablation of bovine liver in vitro.

Preparation of mupirocin-loaded polymeric nanocapsules using essential oil of rosemary

Abstract The purpose of this study was to prepare and characterize mupirocin-loaded polymeric nanocapsules using two different oils and to develop and validate an analytical method for quantitative determination by high performance liquid chromatography. The mean size of the nanoparticles was 233.05 nm and 275.03 nm for nanocapsules with a rosemary oil like oily core and caprylic/capric triglyceride, respectively, and a good polydispersity index below 0.25 for both formulations. The nanocapsules showed good stability when stored at 40 ºC and room temperature for 30 days. The quantitative method was performed with a mobile phase consisting of ammonium ammonium acetate (0.05 M adjusted to pH 5.0 with acetic acid) and acetonitrile 60:40 (v/v); the flow rate was 0.8 mL/min, UV detection at 230 nm. The analytical method was linear in the range of 5.0-15.0 µg/mL, specific for both oils, accurate, precise (intermediate precision RSD = 1.68% and repeatability RSD = 0.81%) and robust under the evaluated conditions. Therefore, this method can be performed for quantification of mupirocin in polymeric nanocapsules containing both oils.

Pullulan as a stabilizer agent of polymeric nanocapsules for drug delivery

ABSTRACT Polymeric stabilizers have received attention in the preparation of nanostructured systems due to their ability to enhance formulation stability. Considering this, the objective of this work was to prepare poly(ε-caprolactone) nanocapsules using the pullulan as a polymeric stabilizer. The nanocapsules were prepared using the interfacial deposition method of preformed polymers and they were characterized in terms of pH, average diameter, polydispersity index, zeta potential, beclomethasone dipropionate content, encapsulation efficiency, photostability and drug release profiles. The formulations showed physicochemical characteristics consistent with nanocarriers for drug delivery such as: average diameter lower than 270 nm, polydispersity indexes lower than 0.2, negative zeta potential (-22.7 to -26.3 mV) and encapsulation efficiencies close to 100%. In addition, the nanocapsules were able to delay the beclomethasone dipropionate photodegradation under UVC radiation and by the dialysis bag diffusion technique, the nanocapsules were able to prolong the drug release. Thus, pullulan could be considered an interesting excipient to formulate polymeric nanocapsules.

Uso de antimicrobiano nanoparticulado para o tratamento da mastite subclínica de ovelhas de corte no período seco / Use of nanoparticulated antimicrobians to treat subclinical mastitis of ewes during the dry period

A inflamação da glândula mamária é uma das principais causas de prejuízo na ovinocultura. Este estudo teve como objetivo investigar as taxas de cura do tratamento da mastite subclínica após infusão intramamária de princípio ativo antimicrobiano no momento da secagem, em formulações convencional e nanoparticulada. Os rebanhos estavam localizados em São Carlos, São Paulo, Brasil. Analisou-se um total de 584 glândulas mamárias de 307 ovelhas de aptidão para produção de carne. Triagem prévia dos casos subclínicos de mastite foi efetuada por meio do California Mastitis Test (CMT) e/ou da contagem de células somáticas (CCS). Análises microbiológicas foram realizadas para confirmação da etiologia infecciosa. As glândulas mamárias com mastite subclínica foram distribuídas em três grupos: G1 (Controle; glândulas mamárias que não receberam tratamento antimicrobiano); G2 (glândulas mamárias em que foi administrado 100 mg de cloxacilina benzatina em estrutura convencional) e G3 (glândulas mamárias em que foi administrado 50 mg de cloxacilina benzatina em estrutura nanoencapsulada). O tratamento aplicado ao G3 mostrou-se mais eficiente (P=0,047) na cura de glândulas mamárias com mastite subclínica. O uso da cloxacilina nanoencapsulada no momento da secagem de ovelhas de corte auxilia no controle da mastite subclínica infecciosa e reduz os prejuízos consequentes.(AU)

Inflammation of the mammary gland is one of the main causes of losses in sheep-rearing. This study aimed to investigate the cure rates from treating subclinical mastitis after intramammary infusion of active antimicrobial agents as conventional formulations or as nanoparticles, at the time when the ewes are being dried off. A total of 584 mammary glands in 307 ewes in meat-producing herds located in São Carlos, São Paulo, Brazil, were analyzed. Prescreening of subclinical mastitis cases was done using the California mastitis test (CMT) and/or the somatic cell count (SCC). Microbiological analyses were performed to confirm the infectious etiology. The mammary glands with subclinical mastitis were distributed into three groups: G1 (control; mammary glands that did not receive any antimicrobial treatment); G2 (mammary glands to which 100mg of benzathine cloxacillin in conventional form were administered); and G3 (mammary glands to which 50mg of benzathine cloxacillin in nanoparticulate form were administered). The treatment applied to G3 was more efficient (P=0.047) in curing mammary glands with subclinical mastitis. Use of cloxacillin nanoparticles at the time when the ewes are being dried off helps to control infectious subclinical mastitis and reduces consequential losses among meat-producing herds.(AU)

Quinine-Loaded Polymeric Nanoparticles: Validation of a simple HPLC-PDA Method to Determine Drug Entrapment and Evaluation of its Photostability.

Liquid chromatographic method was developed and validated for quantitative determination of quinine in polymeric nanoparticles. The method was performed using a Waters RP-18 column using a mobile phase consisting of acetonitrile:water:triethylamine (60:40:0.01 v/v/v, and pH aqueous phase adjusted to 3.0 with phosphoric acid). The flow rate was 1.0 mL min-1 and the detection was achieved with a UV-PDA set at 232 nm. The response was linear over a range of 12.0 to 24.0 μg.mL-1 (r = 0.9995). The relative standard deviation values for intra-day and inter-day precision studies were less than 2% and the accuracy was 98.8% to Nc1 and 97.3% to Nc2. The samples free of quinine and quinine-loaded polymeric nanoparticles were subjected to photodegradation conditions. A considerable reduction of degradation of quinine occurred in polymeric nanoparticles. Through these results, it was clear that the nanoencapsulation of quinine protects the drug from degradation by exposure to UV-A light. The analytical method was validated according to International Conference on Harmonization Guidelines and Center for Drug Evaluation and Research.

Pharmacokinetics and bioequivalence of self-assembly nanocapsules loaded with asparaginase / 第二军医大学学报

Objective To study the pharmacokinetics and bioequivalence of asparaginase (Asp) hyaluronic acid-graft-poly (ethylene glycol)/dimethyl-β-cyclodextrin (DCD) nanocapsules (AHDPs) in male Sprague-Dawley (SD) rats. Methods AHDPs were observed under the transmission electron microscope. The size, zeta potential and entrapment efficiency of AHDPs were examined. Asp activities were assayed after intravenous injection of AHDPs or free Asp in rats. Pharmacokinetic parameters were calculated by software DAS 2.1.1. Then the bioequivalence of AHDPs and free Asp were evaluated. Results The average particle size of AHDPs was (439.63±8.49) nm, zeta potential was (-20.43±2.20) mV, and entrapment efficiency was (55.75±4.11)% (n=3). AUC(0-48 h) of AHDPs and free Asp were (138.93±0.89) U•mL-1•h and (46.38±1.98) U•mL-1•h, the AUC0-∞ were (175.22±13.59) U•mL-1•h and (51.44±3.01) U•mL-1•h, and t1/2 was (4.46±1.04) h and (1.86±0.38) h, respectively. Compared with free Asp, the AUC0-48 h, AUC0-∞ and t1/2 of AHDPs were increased by 3.00, 3.40 and 2.40 times, respectively. The 90% confidential intervals of AUC0-48 h, AUC0-∞ and Cmax were 76.9%-78.3%, 76.9%-78.3% and 92.8%-94.4%, respectively. Conclusion AHDPs can prolong the biological half-life and improve the bioavailability of Asp in rats. AHDPs and free Asp are not bioequivalent.

Caracterização farmacêutica de nanocápsulas de timol e avaliação da permeação cutânea e da atividade repelente contra Aedes aegypti

A dengue é uma doença infecciosa que pode se manifestar de forma grave, com quadros hemorrágicos. O agente etiológico é um arbovirus transmitido pela picada do mosquito Aedes aegypti, principal vetor da doença. As principais medidas de combate são controle do mosquito e proteção individual, que pode dar-se com o uso de repelentes. Quase a totalidade dos repelentes no mercado possui DEET como ativo, que possui restrições de uso. Nestes entido, o timol (monoterpeno) tem-se mostrado promissor como repelente de inseto (Ae.aegypti), sendo a agregação de tecnologias essencial para a viabilidade de um produto repelente à base de timol. Diante do exposto, o objetivo do presente trabalho foi a preparação e caracterização farmacêutica de nanocápsulas de timol, com avaliação da permeação cutânea, citotoxicidade e atividade repelente de inseto (Ae. aegypti). Para tanto, foi desenvolvido evalidado método analítico para identificação e quantificação do timol por CLAE-DAD nas nanocápsulas de timol (NCT – 1%), que apresentou um teor de ativo em torno de 85%.Prosseguindo a caracterização das NCT, foram determinados o diâmetro médio, índice depolidispersão (PDI), potencial zeta (PZ), eficiência de encapsulação e pH, além do estudo de estabilidade. Análises das NCT mostraram um diâmetro médio de aproximadamente 150 nm,PZ negativo (-27,83 ± 2,60), PDI abaixo de 0,2 e uma eficiência de encapsulação de 98 %...

Dengue is an infectious disease that may also happen in a severe form with hemorrhagicevents. The etiological agent of the disease is an arbovirus wich is transmitted by themosquito Aedes aegypti, the primary vector of the disease. The main actions to combat thedisease are mosquito control and personal protection that can take place using repellents.Almost all repellents have DEET as active substance, which has use restrictions. In thiscontext, thymol (monoterpene) has become a potential insect repellent against Ae. aegypti,and the use of technologies is essential to the viability of a repellent thymol based product.Given the above, the aim of this study was the preparation and pharmaceuticalcharacterization of thymol nanocapsules and evaluation of skin permeation, in vitrocytotoxicity and mosquito repellence (Ae. aegypti). For this purpose, we developed andvalidated analytical method for identification and quantification of thymol by HPLC-PDA.Nanocapsules thymol (NCT - 1%) showed an active content of 85%. Continuing thecharacterization, NCT were analyzed for the mean diameter (D), polydispersity index (PDI),potential zeta (PZ), encapsulation efficiency, pH and product stability . Results shared showedan average diameter of approximately 150 nm, negative PZ (-27,83 ± 2,60), PDI less than 0.2and an encapsulation efficiency of 98%...

Validation of high performance liquid chromatography method for determination of meloxicam loaded PEGylated nanocapsules

abstract A method to ensure that an analytical method will produce reliable and interpretable information about the sample must first be validated, making sure that the results can be trusted and traced. In this study, we propose to validate an analytical high performance liquid chromatography (HPLC) method for the quantitation of meloxicam loaded PEGylated nanocapsules(M-PEGNC). We performed a validation study, evaluated parameters including specificity, linearity, quantification limit, detection limit, accuracy, precision and robustness. PEGylated nanocapsules were prepared by interfacial deposition of preformed polymer, and the particle size, polydispersity index, zeta potential, pH value and encapsulation efficiency were characterized. The proposed HPLC method provides selective, linear results in the range of 1.0-40.0 μg/mL; quantification and detection limits were 1.78 μg/mL and 0.59 μg/mL, respectively; relative standard deviation for repeatability was 1.35% and intermediate precision was 0.41% and 0.61% for analyst 1 and analyst 2, respectively; accuracy between 99.23 and 101.79%; robustness between 97.13 and 98.45% for the quantification of M-PEGNC. Mean particle diameters were 261 ± 13 nm and 249 ± 20 nm, polydispersity index was 0.15 ± 0.07 and 0.17 ± 0.06, pH values were 5.0 ± 0.2 and 5.2 ± 0.1, and zeta-potential values were -37.9 ± 3.2 mV e -31.8 ± 2.8 mV for M-PEGNC and placebo(B-PEGNC), respectively. In conclusion, the proposed analytical method is suitable for the quality control of M-PEGNC. Moreover, suspensions showed monomodal size distributions and low polydispersity index indicating high homogeneity of formulations with narrow size distributions, and appropriate pH and zeta potential. The extraction process was efficient for release of meloxicam from nanostructured systems.

resumo Para se assegurar que um método analítico produzirá informação confiável e interpretável sobre a amostra este deve ser inicialmente validado, tornando claro que os resultados podem ser confiados e rastreados. Neste estudo, propomos validar um método de cromatografia líquida de alta eficiência (CLAE) para a quantificação do meloxicam encapsulado em nanocápsulas PEGuiladas (M-PEGNC). Efetuamos a validação, avaliando parâmetros de especificidade, linearidade, limite de quantificação, limite de detecção, exatidão, precisão e robustez. As nanocápsulas PEGuiladas foram preparadas por deposição interfacial do polímero pré-formado e caracterizaram-se o tamanho da partícula, índice de polidispersão, potencial zeta, pH e eficiência de encapsulação. O método de CLAE proposto fornece resultados seletivos e lineares na faixa de 1,0-40,0 mg/mL; limites de quantificação e detecção de 1,78 mg/mL e 0,59 mg/mL, respectivamente; desvio padrão relativo para a repetibilidade de 1,35% e precisão intermediária de 0,41% e 0,61% para o analista 1 e analista 2, respectivamente; exatidão entre 99,23 e 101,79%; robustez entre 97,13 e 98,45% para a quantificação de M-PEGNC. Os diâmetros médios das partículas foram 261 ± 13 nm e 249 ± 20 nm; índice de polidispersão de 0,15 ± 0,07 e 0,17 ± 0,06, valores de pH de 5,0 ± 0,2 e 5,2 ± 0,1 e valores do potencial zeta de -37,9 ± 3,2 mV e -31,8 ± 2,8 mV para o M-PEGNC e o placebo(B-PEGNC), respectivamente. Concluindo, o método analítico proposto é adequado para o controle de qualidade do M-PEGNC. Além disso, suspensões mostraram distribuição de tamanho monomodal e baixo índice de polidispersão, indicando alta homogeneidade das formulações com distribuição estreita de tamanho, pH e potencial zeta apropriados. O processo de extração foi eficiente para a liberação do meloxicam dos sistemas nanoestruturados.

Evaluation of gastroprotective activity of Passiflora alata

AbstractPassiflora alata Curtis, Passifloraceae, is a liana popularly known in Brazil as ‘maracujá-doce’ that has been used for treating different illnesses. Its leaves are described in the Brazilian Pharmacopoeia, but the gastroprotective activity has never been investigated. In the present study a freeze-dried crude 60% ethanol–water extract of P. alata aerial parts was prepared. Total flavonoid content, expressed as vitexin, was 0.67% ± 0.01. The hemolytic activity was 32 units for P. alata, using Saponin (Merck®) as reference. P. alata presented EC50 of 1061.2 ± 8.5 µg/ml in the 2,2-diphenyl-1-picryhydrazyl assay and 1076 ± 85 µmol Trolox/g in the Oxygen Radical Absorbance Capacity assay. P. alata, its solvent fractions and a P. alatananopreparation were investigated for gastroprotective activity. The test samples exhibited gastroprotective activity on HCl/ethanol induced gastric mucosal lesions in rats. P. alata at doses of 100, 200 and 400 mg/kg, using the necrotizing agent at 150 mmol/l, inhibited 100% of ulcer formation (compared to the negative control), while lansoprazole (30 mg/kg) 77%. When tested against a more concentrated necrotizing agent (300 mmol/l), fractions of P. alata at 100 mg/kg reduced 57% (n-hexane), 34% (ethyl acetate) and 72% (aqueous fraction) the ulcer formation. In this assay, lansoprazole (30 mg/kg) inhibited 47%. When encapsulated, P. alata inhibited ulcer formation at 55%, 94% and 90% for dosages of 25, 50 and 100 mg/kg. These results suggest the potential use of P. alata as a gastroprotective herbal medicine.

Efeitos das nanocápsulas de núcleo lipídico contendo acetileugenol em melanomas: estudos in vivo e in vitro / Effects of lipid-core nanocapsules with acetyleugenol in melanomas: in vivo and in vitro studies

O melanoma é uma neoplasia de pele invasivo, com maior taxa de morte, sem tratamento efetivo. Nanocápsulas poliméricas de núcleo lipídico (LNC) tem sido empregadas com sucesso como carreadores de fármacos hidrofóbicos. Como o eugenol é um composto hidrofóbico com atividades antiproliferativas e pró-apoptóticas em células cancerosas, visamos avaliar os efeitos dos tratamentos com acetileugenol (AC), LNC ou LNC contendo acetileugenol (LNC-AC) em modelo de melanoma in vivo em camundongos C57B6, e a citotoxicidade dos mesmos em células endoteliais (HUVEC) e de melanoma (SK-Mel-28) in vitro. Os resultados obtidos mostraram que: 1) tratamentos i.p. com as LNC ou com LNC-AC (50 mg/kg, 3-10 dia de indução do tumor) induziram toxicidade sistêmica e, somente o tratamento com LNC inibiu o desenvolvimento do melanoma. O tratamento com LNC, mas não com a mistura de triglicerídeos de cadeia média, por via oral, inibiu o desenvolvimento tumoral, sem toxicidade. Adicionalmente, os tratamentos com AC, LNC ou LNC-AC não foram eficazes quando administrados em fase tardia de evolução tumoral (50 mg/kg, 7-17 dia de indução do tumor, via oral); 2) os tratamentos agudos com AC, LNC ou LNC-AC (20 mg/kg, 200 µL, e.v.) não alteraram o número de leucócitos circulantes, mas os tratamentos com LNC ou com LNC-AC reduziram o comportamento de rolling dos leucócitos em vênulas póscapilares do músculo cremaster e causaram hemólise, sendo que este último efeito também foi observado após tratamento in vitro em hemácias murinas; 3) Os estudos in vitro mostraram que as LNC e LNC-AC foram captadas pelas células HUVEC e SK-Mel-28 após 1 hora de incubação; que a incubação com LNC-AC induziu apoptose tardia e necrose com maior eficácia em SK-Mel-28 do que em HUVEC; que as incubações com LNC ou LNC-AC exerceram efeitos antiproliferativos, induzindo parada na fase G2/M do ciclo celular das duas linhagens de células avaliadas; que somente a incubação com AC ou LNC-AC inibiu a adesão ao Matrigel® com maior eficácia na linhagem SK-Mel-28 do que HUVEC; que somente a incubação com as LNC reduziram a expressão de VCAM-1 em HUVEC e que as incubações com LNC ou LNC-AC reduziram a expressão de ß3 integrina em SK-Mel- 28; que nenhum dos tratamentos alterou a migração celular das HUVEC ou SK-Mel- 28; que somente a incubação com LNC-AC reduziu os níveis de espécies reativas de oxigênio em HUVEC e SK-Mel-28; que a incubação com LNC ou LNC-AC aumentou a produção de óxido nítrico (NO) pelas duas linhagens de células avaliadas; que o tratamento com L-NAME reverteu os níveis de NO e a inibição sobre a proliferação celular induzida pela incubação com LNC ou LNC-AC e; que o tratamento de células de melanoma murino com LNC ou LNC-AC parece alterar a polarizar os neutrófilos para o fenótipo N1. Associados, os resultados obtidos mostram o tratamento oral com LNC inibe o crescimento do melanoma sem induzir efeitos tóxicos, e que este efeito benéfico pode ser dependente, pelo menos em parte, da nanoencapsulação dos triglicerídios de cadeia média e da supraestrutura da formulação, com toxicidade direta sobre as células de melanoma e possível modulação do microambiente tumoral

Melanoma is the most invasive skin cancer, with high rates of death without effective treatment. Polymeric lipid-core nanocapsules (LNC) has been successfully used as carriers of hydrophobic drugs. As eugenol is an hydrophobic compound with antiproliferative and pro-apoptotic activity in cancer cells, here we aimed to evaluate the effects of treatments with acetyleugenol (AC), LNC or LNC containing acetyleugenol (LNC-AC) in an in vivo melanoma model in C57BL6 mice and the cytotoxicity of the treatments in vitro, using endothelial (HUVEC) and melanoma (SK-Mel- 28) cells. The results obtained showed that: 1) i.p. treatments with LNC or LNCAC (50 mg/kg, 3-10 days of tumor injection) induced systemic toxicity and, only the treatment with LNC inhibited the melanoma development. Treatment with LNC, but not with mix of triglycerides of medium chain, by oral route, inhibited the tumor development, without toxicity. In addition, the treatments with AC, LNC or LNC-AC were not effective when administered in the late stage of tumor evolution (50 mg/kg, 10-20 days of tumor induction, oral route); 2) the acute treatments with AC, LNC or LNC-AC (20 mg/kg, 200 µL, intravenous route) did not altered the number of circulating leukocytes, but the treatments with LNC or LNC-AC reduced the rolling behavior of leukocytes in postcapillary venules of the cremaster muscle and induced hemolysis. The latter effect was also observed after in vitro treatment using murine erythrocytes; 3) In vitro studies showed that the LNC and LNC-AC suffered uptake by HUVEC and SK-Mel-28 cells after 1 hour of incubation; that the incubation with LNC-AC induced late apoptosis and necrosis more effectively in SK-Mel-28 than in HUVEC cells; that the incubation with LNC or LNC-AC presented antiproliferative effects, by inducing G2M arrest in cell cycle in both cells lines evaluated; that only the incubation with AC or LNC-AC inhibited the adhesion in Matrigel® with more efficaccy in SK-Mel-28 than in HUVEC cells; that only incubtion with LNC reduced the VCAM-1 expression in HUVEC and the incubation with LNC or LNC-AC reduced the ß3 integrin expression in SK-Mel-28 cells; that any treatment affected the HUVEC or SK-Mel- 28 migration; that only the incubation with LNC-AC reduced the levels of reactive species of oxygen in HUVEC and SK-Mel-28 cells; that the incubation with LNC or LNC-AC increased the nitric oxide (NO) production by both cell lines used; that the treatment with L-NAME reversed the NO levels and the inhibition on cell proliferation induced by incubation with LNC or LNC-AC and; that the in vitro treatment of murine with LNC or LNC-AC altered the neutrophil polarization to N1 phenotype. Together, results obtained show that the oral treatment with LNC inhibit the melanoma growth without any toxic effect, and that the beneficial effect could be dependent, at least in part, of nanoencapsulation of medium chain triglycerides and the supraestrucuture of the formulation, with direct toxicity on melanoma cells and possible modulation of tumor microenvironment

Toxicity of lidocaine solid lipid nanoparticles in human neurons / 中华麻醉学杂志

Objective To investigate the toxicity of lidocaine solid lipid nanoparticles (SLNs) in human neurons.Methods Lidocaine-loaded SLNs were prepared using high pressure homogenization.SHSY5Y cells were cultured in vitro and inoculated on 96-well plates (100 μl/well) at a density of 5× 105 cells/ml.SH-SY5Y cells were randomized into 10 groups (n =30 each) using a random number table:control group (group C), different concentrations of lidocaine groups (L1-4 groups), different concentrations of lidocaine SLN groups (L-SLN1-4 groups), and blank SLN group (group SLN).The cells were cultured routinely in group C.The cells were incubated with the culture medium containing lidocaine with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.In LSLN1-4 groups, the cells were incubated with the culture medium containing lidocaine SLNs with the final concentrations of 1.000％, 0.500％, 0.250％ and 0.125％ in L1-4 groups, respectively.Before incubation (at the corresponding time points in group C), and at 1, 12 and 24 h of culture or incubation (T0-3) , 6 wells in each group were selected for measurement of the cell survival rate (using methyl thiazolyl tetrazolium assay).The cell morphology was examined with optical microscope at T3.Results Compared with that at T0, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in L-SLN3 group, and at T3 in L-SLN4 group (P＜0.05) , and no significant change was found in SLN and C groups (P＞0.05).The cell survival rate was significantly lower at T2,3 in L1-4 and L-SLN1-3 groups, and at T3 in group L-SLN4 than that at T1, and at T3 in L1-4 and L-SLN1-4 groups than that at T2 (P＜0.05).Compared with group C, the cell survival rate was significantly decreased at each time point in L1-4 and L-SLN1,2 groups, at T2,3 in group L-SLN3, and at T3 in group L-SLN4 (P＜0.05) , and no significant change was found in group SLN (P＞0.05).Compared with group L-SLN at the corresponding concentration, the cell survival rate was significantly decreased at each time point in group L1-4 (P＜0.05).Conclusion Lidocaine SLNs have toxic effect on human neurons, but the effect is weaker than that caused by Iidocaine solution.