Os autores reconhecem a bolsa Arábia Saudita-financiado (I821) para fazer a pesquisa possível. Os autores agradecem ao Dr Xianwei Liu de apoio especializado em análise de TEM da Universidade de Cranfield.

1. Preparação de emulsão de óleo-em-água <ol><li> Pesar palmitoestearato de glicerilo (120 mg), miristato de isopropilo (60 mg), d-α-tocoferol succinato de polietileno-glicol (30 mg) e a lecitina de soja (30 mg), e adicione-os a 12,5 ml de solventes orgânicos (6,25 ml de acetona e 6,25 ml de etanol). dissolvem-se rapidamente a mistura à temperatura de 70 ° C (5 ° C acima do ponto de fusão do lípido sólido) em banho de água. </li><li> Adicionar tanto 125, 250 ou 375 mg de Poloxamer188 em 12,5 ml de ddH2O para atingir 1-3% (respectivamente) de solução de Poloxâmero 188, o qual é sujeito a aquecimento à mesma temperatura como acima. </li><li> Adicionar a solução da fase aquosa do passo 1.2 para a solução de fase oleosa do passo 1.1, gota a gota, para formar emulsões, sob agitação magnética a 400 rpm. Agita-se a emulsão a 400 rpm durante mais 4 h, para permitir a evaporação dos solventes orgânicos. </li></ol> 2. Solidificação e homogeneização <ol><li> deixar tele emulsão numa sala fria (4 ° C) durante 2 horas para solidificar / cristalizar. </li><li> Para obter o NLC, submeter a emulsão de alta dispersão pura (HSD) com um homogeneizador a 10.000-15.000 rpm para 10-40 min. </li></ol> 3. Optimização da Preparação NLC <ol><li> Retirar amostras a partir do passo 2.2 com uma concentração de surfactante de 1, 2 e 3% HSD submetido a uma velocidade de 10.000, 15.000 e 20.000 rpm, respectivamente, e intervalos de tempo de 10, 20, 30 e 40 min, respectivamente, </li><li> Examinar as amostras para o tamanho de partículas (PS), poli índice de dispersão (PDI), morfologia e ultra-estrutura 20. Nota: Os parâmetros que produzem partículas com o menor tamanho (155 nm) e o valor PDI (0,2) são determinadas como ideal. </li></ol> 4. Preparação de NLC Dac-carregado (NLC-Dac) <ol><li> Preparar a solução de fase oleosa, tal como descrito na etapa 1.1, com a adição da DAC (70 mg) antes de se dissolver a mistura a 70 ° C em banho de água. </li><li> Prepare uma solução de fase aquosa, tal como descrito na etapa 1.2, com 1% de tensioactivo, e adicionar esta solução ao preparado no passo 4.1, gota a gota, para formar uma emulsão sob agitação magnética a 400 rpm. Agita-se a emulsão durante mais 4 horas para evaporar os solventes orgânicos. </li><li> Deixar a emulsão numa sala fria (4 ° C) durante 2 horas para solidificar / cristalizar, tal como descrito no passo 2, e finalmente, sujeitar a emulsão de alta dispersão pura (HSD), utilizando os parâmetros óptimos determinados no passo 3. </li></ol>

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</ol>

Os autores não têm nada a revelar.

nanoestruturados partículas à base de lípidos têm sido utilizados para proporcionar um suporte altamente lipófilo para a entrega de fármacos hidrofóbicos. Um NLC é a segunda geração de transportador nanoestruturada lipídicas sólidas, que são sólidos à temperatura ambiente e corporal. A incorporação de um lípido sólido num lípido líquido num resultados NLC em uma cristalização menos perfeita, aumentando assim a eficiência de carga de droga e também reduzindo a expulsão de fármacos encapsulados ...

Dacarbazina (CAD) é um agente alquilante que exerce uma actividade anti-tumor através de metilação de ácidos nucleicos ou danos directa de ADN, levando à paragem do ciclo celular e morte celular 1. Como um primeiro agente quimioterapêutico linha, Dac foi usado sozinho ou em combinação com outras drogas de quimioterapia para o tratamento de vários cancros 2-6. É o agente mais ativo até agora utilizada no tratamento do melanoma cutâneo e metastático, que é a forma mais agressiva de câncer de pele 3,7,8. A taxa de resposta, no entanto, é de apenas 20% no máximo, e os efeitos terapêuticos são muitas vezes acompanhada com graves efeitos secundários sistémicos. Na sua forma natural, Dac é hidrofílico e é instável devido à sua fotossensibilidade 9. A única fórmula disponíveis para uso clínico, é actualmente um pó estéril para ser utilizado na suspensão para 7,8 infusão intravenosa. A taxa de resposta de baixa e alta toxicidade sistémica rComeram do medicamento é em grande parte atribuível à sua fraca solubilidade em água, por conseguinte, de baixa disponibilidade no local alvo, e uma distribuição elevada nos sítios não-alvo, o que limita a dose máxima do medicamento 10. A rápida degradação e metabolismo após a admissão por via intravenosa juntamente com o desenvolvimento de resistência aos fármacos limita a aplicação clínica e efeito terapêutico do fármaco 11. Por conseguinte, existe uma necessidade urgente de desenvolver formulações alternativas do CAD para o tratamento de melanoma maligno. Sistemas coloidais contendo lipossomas, micelas ou partículas nanoestruturados têm sido intensamente investigado por seu uso na entrega da droga como revisto por Marilene et al. 12 partículas nanoestruturados como portadores potenciais da droga têm atraído cada vez mais atenção na última década devido à sua capacidade de aumentar a carga de droga eficiência, liberação de drogas de controle, melhorar a farmacocinética e biodistribuição e, portanto, reduce droga toxicidade sistêmica 13. Apenas alguns nanoformulações, no entanto, têm sido investigados até agora para entrega Dac, que mostra a protecção da droga a partir de degeneração foto, o aumento da solubilidade do fármaco, melhorado e efeito terapêutico 10,14,15. No entanto, estas formulações sofreram de baixa eficiência de encapsulação enquanto alguns também usando nanopartículas de polímeros sintéticos que não são rentáveis. Transportadores lipídicos nanoestruturados (NLC), constituídos por uma mistura de lípidos sólidas e líquidas, têm sido desenvolvidos para entrega de drogas 16,17. Os medicamentos a serem encapsuladas são muitas vezes solúveis em ambos os lípidos líquidos e lípidos fases sólidas 18, resultando numa carga elevada e de libertação controlada 19. Este estudo tem como objetivo desenvolver uma nova formulação Dac baseado em NLC-encapsulamento usando palmitoestearato glicerilo e miristato de isopropilo como lipídios. A preparação envolvidos óleo-em-água, a evaporação, a solidificação, e homogenization. As preparações foram caracterizadas para o tamanho do NLC, forma, ultra-estrutura, e da distribuição, eficiência de encapsulação de drogas e de carga de droga 20.

<table><tbody><tr><td>Dacarbazine (DAC)</td><td>Sigma Aldrich (Gillingham Dorset, UK)</td><td>D2390-100MG</td><td>drug used for uploading</td></tr><tr><td>glyceryl palmitostearate&amp;nbsp;</td><td>Gattefoss&amp;eacute; (Saint_Priest_c&amp;eacute;dex, France)</td><td>85251-77-0</td><td>solid lipid&amp;nbsp;</td></tr><tr><td>d-&amp;alpha;- Tocopherol polyethylene glycol succinate (TPGS)</td><td>Sigma Aldrich (Gillingham Dorset, UK)</td><td>57668</td><td>lipid phase surfactant</td></tr><tr><td>Poloxamer 188</td><td>Sigma Aldrich (Gillingham Dorset, UK)</td><td>15759-1KG</td><td>liqiud phase surfactant</td></tr><tr><td>Acetone&amp;nbsp;</td><td>Sigma Aldrich (Gillingham Dorset, UK)</td><td>650501-1L</td><td>organic solvent</td></tr><tr><td>Ethanol&amp;nbsp;</td><td>Sigma Aldrich (Gillingham Dorset, UK)</td><td>459836-1L</td><td>organic solvent</td></tr><tr><td>Soybean lecithin (SL)</td><td>Cuisine Innovation (Dijon, France)</td><td>SLL1402</td><td>lipid phase surfactant</td></tr><tr><td>Double-distilled water was collected in our laboratory from</td><td>Millipore-Q Gradient A10 ultra-pure water system (Millipore, France)</td><td>SAS - 67120&amp;nbsp;</td><td>aqueous phase&amp;nbsp;</td></tr><tr><td>T 25 digital ULTRA-TURRAX</td><td>IKA</td><td>3725000</td><td>as high shear disperser</td></tr><tr><td>Hotplate Magnetic Stirrer</td><td>Scientific Support, Inc</td><td>1454&amp;nbsp;</td><td>emulsion homogenization</td></tr></tbody></table>

encapsulation of cancer therapeutic agent dacarbazine using nanostructured lipid carrier

A única fórmula de dacarbazina (DAC) em utilização clínica é a infusão intravenosa, que apresenta um perfil terapêutico pobres, devido ao baixo dispersibilidade do fármaco na solução aquosa. Para superar isto, um veículo lipídico nanoestruturada (NLC) consistindo de palmitoestearato de glicerilo e miristato de isopropilo foi desenvolvido para encapsular Dac. NLCs com tamanho controlado foram alcançados usando a dispersão de alto cisalhamento (HSD) na sequência de solidificação da emulsão de óleo-em-água. Os parâmetros de síntese, incluindo a concentração de tensioactivo, a velocidade e tempo de HSD foram optimizados para conseguir o menor NLC com dimensão, o índice de polidispersão e potencial zeta de 155 ± 10 nM, 0,2 ± 0,01, e -43,4 ± 2 mV, respectivamente. Os parâmetros óptimos também foram empregues para a preparação NLC DAC-carregado. A NLC resultante carregado com Dac possuía tamanho, índice de polidispersão e o potencial zeta de 190 ± 10 nm, 0,2 ± 0,01 e -43,5 ± 1,2 mV, respectivamente. O ef encapsulamento de drogaseficiên- e carga de fármaco atingiu 98% e 14%, respectivamente. Este é o primeiro relatório sobre encapsulamento de Dac usando NLC, o que implica que NLC poderia ser um novo candidato potencial como transportadora de drogas para melhorar o perfil terapêutico de Dac.

As preparações da NLC e NLC-Dac usando palmitoestearato glicerilo e miristato de isopropilo com diferentes parâmetros foram caracterizados por PS, PDI, morfologia e ultra-estrutura 20. O PS e PDI dos NLCs foram concentração de surfactante, velocidade HSD e dependente duração. Tal como avaliado pelo PS e PDI dos NLCs, os melhores resultados foram alcançados com 1% de agente tensioactivo e uma velocidade de dispersão pura de 15.000 rpm durante 30 min (Figura 1A,...

Watch this Scientific Journal Video about Encapsulation of Cancer Therapeutic Agent Dacarbazine Using Nanostructured Lipid Carrier at JoVE.com

Encapsulation of Cancer Therapeutic Agent Dacarbazine Using Nanostructured Lipid Carrier

O método mais vulgarmente utilizado para a síntese transportador lipídico nanoestruturada (NLC) envolve emulsão de óleo-em-água, de homogeneização e de solidificação. Este foi modificado aqui pela aplicação de dispersão de elevado corte, após a solidificação para alcançar um NLC com tamanho desejável, uma melhor encapsulação de drogas e a eficiência de carga de fármaco como um transportador potencial para entrega dacarbazina.

Encapsulation of Cancer Terapêutica Agente dacarbazina Usando Nanoestruturados Lipid Transportador

The only formula of dacarbazine (Dac) in clinical use is intravenous infusion, presenting a poor therapeutic profile due to the low dispersity of the drug ...

encapsulation-cancer-therapeutic-agent-dacarbazine-using

Research

JoVE Journal

Chemistry

11.1K visualizações.  Cranfield University.  O método mais vulgarmente utilizado para a síntese transportador lipídico nanoestruturada (NLC) envolve emulsão de óleo-em-água, de homogeneização e de solidificação. Este foi modificado aqui pela aplicação de dispersão de elevado corte, após a solidificação para alcançar um NLC com tamanho desejável, uma melhor encapsulação de drogas e a eficiência de carga de fármaco como um transportador potencial para entrega dacarbazina. 

Vídeo: Encapsulation of Cancer Therapeutic Agent Dacarbazine Using Nanostructured Lipid Carrier

Anticancer Efficacy of Photodynamic Therapy with Lung Cancer-Targeted Nanoparticles

Photodynamic therapy (PDT) is a non-invasive and non-surgical method representing an attractive alternative choice for lung cancer treatment. Photosensitizers selectively accumulate in tumor tissue and lead to tumor cell death in the presence of oxygen and the proper wavelength of light.
To increase the therapeutic effect of PDT, we developed both photosensitizer- and anticancer agent-loaded lung cancer-targeted nanoparticles. Both enhanced permeability and retention (EPR) effect-based passive targeting and hyaluronic-acid-CD44 interaction-based active targeting were applied. CD44 is a well-known hyaluronic acid receptor that is often introduced as a biomarker of non-small cell lung cancer. 
In addition, a combination of PDT and chemotherapy is adopted in the present study. This combination concept may increase anticancer therapeutic effects and reduce adverse reactions. 
We chose hypocrellin B (HB) as a novel photosensitizer in this study. It has been reported that HB causes higher anticancer efficacy of PDT compared to hematoporphyrin derivatives1. Paclitaxel was selected as the anticancer drug since it has proven to be a potential treatment for lung cancer2. 
The antitumor efficacies of photosensitizer (HB) solution, photosensitizer encapsulated hyaluronic acid-ceramide nanoparticles (HB-NPs), and both photosensitizer- and anticancer agent (paclitaxel)-encapsulated hyaluronic acid-ceramide nanoparticles (HB-P-NPs) after PDT were compared both in vitro and in vivo. The in vitro phototoxicity in A549 (human lung adenocarcinoma) cells and the in vivo antitumor efficacy in A549 tumor-bearing mice were evaluated.
The HB-P-NP treatment group showed the most effective anticancer effect after PDT. In conclusion, the HB-P-NPs prepared in the present study represent a potential and novel photosensitizer delivery system in treating lung cancer with PDT.

Photodynamic therapy (PDT) is an alternative choice for lung cancer treatment. To increase the therapeutic effect of PDT, lung cancer-targeted nanoparticles combined with chemotherapy were developed. Both in vitro and in vivo anticancer efficacies of PDT with prepared nanoparticles were evaluated.

Anticancer eficácia da terapia fotodinâmica com Nanopartículas Lung Cancer segmentadas

Photodynamic therapy (PDT) is a non-invasive and non-surgical method representing an attractive alternative choice for lung cancer treatment. ...

Pesquisa do câncer

Facile Preparation of Internally Self-assembled Lipid Particles Stabilized by Carbon Nanotubes

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled (functionalized) CNTs are used as stabilizers to produce Pickering type oil-in-water (O/W) emulsions. Lipids namely, Dimodan U and Phytantriol are used as emulsifiers, which in excess water self-assemble into the bicontinuous cubic Pn3m phase. This highly viscous phase is fragmented into smaller particles using a probe ultrasonicator in presence of conventional surfactant stabilizers or CNTs as done here. Initially, the CNTs (powder form) are dispersed in water followed by further ultrasonication with the molten lipid to form the final emulsion. During this process the CNTs get coated with lipid molecules, which in turn are presumed to surround the lipid droplets to form a particulate emulsion that is stable for months. The average size of CNT-stabilized nanostructured lipid particles is in the submicron range, which compares well with the particles stabilized using conventional surfactants. Small angle X-ray scattering data confirms the retention of the original Pn3m cubic phase in the CNT-stabilized lipid dispersions as compared to the pure lipid phase (bulk state). Blue shift and lowering of the intensities in characteristic G and G&#39; bands of CNTs observed in Raman spectroscopy characterize the interaction between CNT surface and lipid molecules. These results suggest that the interactions between the CNTs and lipids are responsible for their mutual stabilization in aqueous solutions. As the concentrations of CNTs employed for stabilization are very low and lipid molecules are able to functionalize the CNTs, the toxicity of CNTs is expected to be insignificant while their biocompatibility is greatly enhanced. Hence the present approach finds a great potential in various biomedical applications, for instance, for developing hybrid nanocarrier systems for the delivery of multiple functional molecules as in combination therapy or polytherapy.

We report on a smart application of carbon nanotubes for kinetic stabilization of lipid particles that contain self-assembled nanostructures in their cores. The preparation of lipid particles requires rather low concentrations of carbon nanotubes permitting their use in biomedical applications such as drug delivery.

Preparação Facile da Internamente auto-montados Lipid partículas estabilizada por nanotubos de carbono

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled ...

Química

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

Customizado aparelho de aquecimento à base de laser para lançamento acionado de cisplatina de termossensível lipossomas com Ressonância Magnética Orientação Imagem

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Bioengenharia

Preparation and Characterization of Lipophilic Doxorubicin Pro-drug Micelles

Micelles have been successfully used for the delivery of anticancer drugs. Amphiphilic polymers form core-shell structured micelles in an aqueous environment through self-assembly. The hydrophobic core of micelles functions as a drug reservoir and encapsulates hydrophobic drugs. The hydrophilic shell prevents the aggregation of micelles and also prolongs their systemic circulation in vivo. In this protocol, we describe a method to synthesize a doxorubicin lipophilic pro-drug, doxorubicin-palmitic acid (DOX-PA), which will enhance drug loading into micelles. A pH-sensitive hydrazone linker was used to conjugate doxorubicin with the lipid, which facilitates the release of free doxorubicin inside cancer cells. Synthesized DOX-PA was purified with a silica gel column using dichloromethane/methanol as the eluent. Purified DOX-PA was analyzed with thin layer chromatography (TLC) and 1H-Nuclear Magnetic Resonance Spectroscopy (1H-NMR). A film dispersion method was used to prepare DOX-PA loaded DSPE-PEG micelles. In addition, several methods for characterizing micelle formulations are described, including determination of DOX-PA concentration and encapsulation efficiency, measurement of particle size and distribution, and assessment of in vitro anticancer activities. This protocol provides useful information regarding the preparation and characterization of drug-loaded micelles and thus will facilitate the research and development of novel micelle-based cancer nanomedicines.

A protocol for the preparation and characterization of lipophilic doxorubicin pro-drug loaded 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG) micelles is described.

Preparação e caracterização de Lipofílica Doxorrubicina micelas Pro-drogas

Micelles have been successfully used for the delivery of anticancer drugs. Amphiphilic polymers form core-shell structured micelles in an aqueous ...

Phase Change Dimethyldioctadecylammonium-Shelled Microdroplets as a Promising Drug Delivery System: Results on 3D Spheroids of Mammalian Tumor Cells

Significant improvement of phase-change perfluorocarbon microdroplets (MDs) in the vast theranostic scenario passes through the optimization of the MDs composition with respect to synthesis efficiency, stability, and drug delivery capability. To this aim, decafluoropentane (DFP) MDs stabilized by a shell of dimethyldioctadecylammonium bromide (DDAB) cationic surfactant were designed. A high concentration of DDAB-MDs was readily obtained within a few seconds by pulsed high-power insonation, resulting in low polydisperse 1 &#181;m size droplets. Highly positive &#950;-potential, together with a long, saturated hydrocarbon chains of the DDAB shell, are key factors to stabilize the droplet and the drug cargo therein. The high affinity of the DDAB shell with cell plasma membrane allows for localized chemotherapeutics delivery by increasing the drug concentration at the tumor cell interface and boosting the uptake. This would turn DDAB-MDs into a relevant drug delivery tool exhibiting high antitumor activity at very low drug doses.
In this work, the efficacy of such an approach is shown to dramatically improve the effect of doxorubicin against 3D spheroids of mammalian tumor cells, MDA-MB-231. The use of&#160;three-dimensional (3D) cell cultures developed in the form of multicellular tumor spheroids (i.e., densely packed cells in a spherical shape) has numerous advantages compared to 2D cell cultures: in addition to have the potential to bridge the gap between conventional in vitro studies and animal testing, it will improve the ability to perform more predictive in vitro screening assays for preclinical drug development or evaluate the potential of off-label drugs and new co-targeting strategies.

<meta charset="utf8"></head><body>Microgotículas com casca de dimetildioctadecilamônio de mudança de fase como um sistema promissor de entrega de medicamentos: resultados em esferóides 3D de célula

Decafluoropentane microdroplets developed with a shell of dimethyldioctadecylammonium bromide exhibited an exceptional colloidal stability and an actractive biointerface. DDAB-MDs proved to be efficient drug reservoirs characterized by a high affinity to plasma membranes together with enhanced uptake and antitumor activity of Doxorubicin against human triple-negative breast cancer (MDA-MB-231) 3D model.

Microgotículas com casca de dimetildioctadecilamônio de mudança de fase como um sistema promissor de entrega de medicamentos: resultados em esferóides 3D de células tumorais de mamíferos

Significant improvement of phase-change perfluorocarbon microdroplets (MDs) in the vast theranostic scenario passes through the optimization of the MDs ...

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier

The controlled delivery of drug/imaging agents to cells is critical for the development of therapeutics and for the study of cellular signaling processes. Recently, nanoparticles (NPs) have shown significant promise in the development of such delivery systems. Here, a liquid crystal NP (LCNP)-based delivery system has been employed for the controlled delivery of a water-insoluble dye, 3,3&#8242;-dioctadecyloxacarbocyanine perchlorate (DiO), from within the NP core to the hydrophobic region of a plasma membrane bilayer. During the synthesis of the NPs, the dye was efficiently incorporated into the hydrophobic LCNP core, as confirmed by multiple spectroscopic analyses. Conjugation of a PEGylated cholesterol derivative to the NP surface (DiO-LCNP-PEG-Chol) enabled the binding of the dye-loaded NPs to the plasma membrane in HEK 293T/17 cells. Time-resolved laser scanning confocal microscopy and F&#246;rster resonance energy transfer (FRET) imaging confirmed the passive efflux of DiO from the LCNP core and its insertion into the plasma membrane bilayer. Finally, the delivery of DiO as a LCNP-PEG-Chol attenuated the cytotoxicity of DiO; the NP form of DiO exhibited ~30-40% less toxicity compared to DiOfree delivered from bulk solution. This approach demonstrates the utility of the LCNP platform as an efficient modality for the membrane-specific delivery and modulation of hydrophobic molecular cargos.

A liquid crystal nanoparticle (LCNP) nanocarrier is exploited as a vehicle for the controlled delivery of a hydrophobic cargo to the plasma membrane of living cells.

Targeted Plasma Membrane entrega de um hidrofóbica Carga encapsulado em um cristal líquido nanopartículas Transportador

The controlled delivery of drug/imaging agents to cells is critical for the development of therapeutics and for the study of cellular signaling processes. ...

Formulation and Characterization of Bioactive Agent Containing Nanodisks

The term nanodisk refers to a discrete type of nanoparticle comprised of a bilayer forming lipid, a scaffold protein, and an integrated bioactive agent. Nanodisks are organized as a disk-shaped lipid bilayer whose perimeter is circumscribed by the scaffold protein, usually a member of the exchangeable apolipoprotein family. Numerous hydrophobic bioactive agents have been efficiently solubilized in nanodisks by their integration into the hydrophobic milieu of the particle&#39;s lipid bilayer, yielding a largely homogenous population of particles in the range of 10-20 nm in diameter. The formulation of nanodisks requires a precise ratio of individual components, an appropriate sequential addition of each component, followed by bath sonication of the formulation mixture. The amphipathic scaffold protein spontaneously contacts and reorganizes the dispersed bilayer forming lipid/bioactive agent mixture to form a discrete, homogeneous population of nanodisk&#160;particles. During this process, the reaction mixture transitions from an opaque, turbid appearance to a clarified sample that, when fully optimized, yields no precipitate upon centrifugation. Characterization studies involve the determination of bioactive agent solubilization efficiency, electron microscopy, gel filtration chromatography, ultraviolet visible (UV/Vis) absorbance spectroscopy, and/or fluorescence spectroscopy. This is normally followed by an investigation of biological activity using cultured cells or mice. In the case of nanodisks harboring an antibiotic (i.e., the macrolide polyene antibiotic amphotericin B), their ability to inhibit the growth of yeast or fungi as a function of concentration or time can be measured. The relative ease of formulation, versatility with respect to component parts, nanoscale particle size, inherent stability, and aqueous solubility permits myriad in vitro and in vivo applications of nanodisk technology. In the present article, we describe a general methodology to formulate and characterize nanodisks containing amphotericin&#160;B as the hydrophobic bioactive agent.

Here, we describe the production and characterization of bioactive agents containing nanodisks. Amphotericin B nanodisks are taken as an example to describe the protocol in a stepwise manner.

Formulação e caracterização de agentes bioativos contendo nanodiscos

The term nanodisk refers to a discrete type of nanoparticle comprised of a bilayer forming lipid, a scaffold protein, and an integrated bioactive agent. ...

Bioquímica

Uptake of New Lipid-coated Nanoparticles Containing Falcarindiol by Human Mesenchymal Stem Cells

Nanoparticles are the focus of an increased interest in drug delivery systems for cancer therapy. Lipid-coated nanoparticles are inspired in structure and size by low-density lipoproteins (LDLs) because cancer cells have an increased need for cholesterol to proliferate, and this has been exploited as a mechanism for delivering anticancer drugs to cancer cells. Moreover, depending on drug chemistry, encapsulating the drug can be advantageous to avoid degradation of the drug during circulation in vivo. Therefore, in this&#160;study, this design is used to fabricate lipid-coated nanoparticles of the anticancer drug falcarindiol, providing a potential new delivery system of falcarindiol in order to stabilize its chemical structure against degradation and improve its uptake by tumors. Falcarindiol nanoparticles, with a phospholipid and cholesterol monolayer encapsulating the purified drug core of the particle, were designed. The lipid monolayer coating consists of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol (Chol), and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE PEG 2000) along with the fluorescent label DiI (molar ratios of 43:50:5:2). The nanoparticles are fabricated using the rapid injection method, which is a fast and simple technique to precipitate nanoparticles by good-solvent for anti-solvent exchange. It consists of a rapid injection of an ethanol solution containing the nanoparticle components into an aqueous phase. The size of the fluorescent nanoparticles is measured using dynamic light scattering (DLS) at 74.1 &#177; 6.7 nm. The uptake of the nanoparticles is tested in human mesenchymal stem cells (hMSCs) and imaged using fluorescence and confocal microscopy. The uptake of the nanoparticles is observed in hMSCs, suggesting the potential for such a stable drug delivery system for falcarindiol.

This article describes the encapsulation of falcarindiol in lipid-coated 74 nm nanoparticles. The cellular uptake of the nanoparticles&#160;by human stem cells into lipid droplets is monitored by fluorescent and confocal imaging. Nanoparticles are fabricated by the rapid injection method of solvent shifting, and their size is measured with the dynamic light scattering technique.

Captação de novas nanopartículas revestidos de lipídeos contendo Falcarindiol por células-tronco mesenquimais humanas

Nanoparticles are the focus of an increased interest in drug delivery systems for cancer therapy. Lipid-coated nanoparticles are inspired in structure and ...

Preparation and Characterization of Nanoliposomes for the Entrapment of Bioactive Hydrophilic Globular Proteins

Liposome nanocapsules have been applied for many purposes in the pharmaceutical, cosmetic, and food industries. Attributes of liposomes include their biocompatibility, biodegradability, non-immunogenicity, non-toxicity, and ability to entrap both hydrophilic and hydrophobic compounds. The classical hydration of thin lipid films in an organic solvent is applied herein as a technique to encapsulate tarin, a plant lectin, in nanoliposomes. Nanoliposome size, stability, entrapment efficiency, and morphological characterization are described in detail. The nanoliposomes are prepared using 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (ammonium salt; DSPE-MPEG 2000), and cholesterylhemisuccinate (CHEMS) as the main constituents. Lipids are first dissolved in chloroform to obtain a thin lipid film that is subsequently rehydrated in ammonium sulfate solution containing the protein to be entrapped and incubated overnight. Then, sonication and extrusion techniques are applied to generate nanosized unilamellar vesicles. The size and polydispersity index of the nanovesicles are determined by dynamic light scattering, while nanovesicle morphology is assessed by scanning electron microscopy. Entrapment efficiency is determined by the ratio of the amount of unencapsulated protein to original amount of initially loaded protein. Homogeneous liposomes are obtained with an average size of 155 nm and polydispersity index value of 0.168. A high entrapment efficiency of 83% is achieved.

This study describes classical hydration using the thin lipid film method for nanoliposome preparation followed by nanoparticle characterization. A 47 kDa-hydrophilic and globular protein, tarin, is successfully encapsulated as a strategy to improve stability, avoid fast clearance, and promote controlled release. The method can be adapted to hydrophobic molecules encapsulation.

Preparação e caracterização de Nanolipossomas para a aprisionamento de proteínas globulares hidrofílicas bioativas

Liposome nanocapsules have been applied for many purposes in the pharmaceutical, cosmetic, and food industries. Attributes of liposomes include their ...

Encapsulation of Cancer Terapêutica Agente dacarbazina Usando Nanoestruturados Lipid Transportador

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