RESUMO
Superparamagnetic iron oxide nanoparticles (SPIONs) are actively used as highly sensitive imaging probes to provide contrast in MRI. In this study, we propose the use of SPIONs encapsulated with antibody-conjugated poly(lactic-co-glycolic acid) (PLGA) as a potent theragnostic agent. The SPIONs were synthesized by a chemical co-precipitation method of ferric and ferrous ions, and subsequently encapsulated with PLGA by using an emulsification-diffusion method. Herceptin was chemically conjugated to the SPION-encapsulating PLGA nanoparticles to target the human epidermal growth factor receptor 2 (Her2/neu) overexpressing breast cancers. FACS and MR molecular imaging revealed that the Her2/neu overexpressing cell line showed a stronger contrast enhancement than the Her2/neu non-expressing cell lines, and the signal intensity of in vivo MR imaging decreased as the concentration of Herceptin increased. This strategy of encapsulating SPIONs with PLGA will be highly useful in functionalizing magnetic nanoparticles and improving the diagnostic and therapeutic efficacy of a wide array of cancer treatments.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Ácido Láctico , Imageamento por Ressonância Magnética , Nanopartículas de Magnetita , Ácido Poliglicólico , Linhagem Celular Tumoral , Meios de Contraste , Humanos , Nanosferas , Copolímero de Ácido Poliláctico e Ácido PoliglicólicoRESUMO
We developed Pyrene-Gadolinium (Py-Gd) nanoparticles as pH-sensitive magnetic resonance imaging (MRI) contrast agents capable of showing a high-Mr signal in cancer-specific environments, such as acidic conditions. Py-Gd nanoparticles were prepared by coating Py-Gd, which is a complex of gadolinium with pyrenyl molecules, with pyrenyl polyethyleneglycol PEG using a nano-emulsion method. These particles show better longitudinal relaxation time (T1) MR signals in acidic conditions than they do in neutral conditions. Furthermore, the particles exhibit biocompatibility and MR contrast effects in both in vitro and in vivo studies. From these results, we confirm that Py-Gd nanoparticles have the potential to be applied for accurate cancer diagnosis and therapy.
Assuntos
Meios de Contraste/síntese química , Gadolínio , Imageamento por Ressonância Magnética/instrumentação , Nanopartículas Metálicas , Neoplasias/diagnóstico , Animais , Células 3T3 BALB , Linhagem Celular Tumoral , Materiais Revestidos Biocompatíveis , Gadolínio/química , Humanos , Imageamento por Ressonância Magnética/métodos , Nanopartículas Metálicas/química , Camundongos , Polietilenoglicóis/química , Pirenos/químicaRESUMO
Well-designed nanoparticle-mediated, image-guided cancer therapy has attracted interest for increasing the efficacy of cancer treatment. A new class of smart theragnostic nanoprobes employing cetuximab (CET)-conjugated polyethylene glycol (PEG)ylated gold nanorods (CET-PGNRs) is presented; these nanoprobes target epithelial cancer cells using near-infrared light. The cetyltrimethylammonium bromide bilayer on GNRs is replaced with heterobifunctional PEG (COOH-PEG-SH) to serve as a biocompatible stabilizer and to increase specificity. The carboxylated GNRs are further functionalized with CET using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC-NHS) chemistry. To assess the potential of such GNRs, their optical properties, biocompatibility, colloidal stability, in vitro/in vivo binding affinities for cancer cells, absorption imaging, and photothermal therapy effects are investigated. CET-PGNRs exhibit excellent tumor targeting ability and strong potential for simultaneous absorption imaging and photothermal ablation of epithelial cancer cells.
Assuntos
Hipertermia Induzida/métodos , Nanopartículas Metálicas/química , Fototerapia/métodos , Absorção , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/química , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Linhagem Celular Tumoral , Cetuximab , Ouro , Humanos , Raios Infravermelhos , Nanopartículas Metálicas/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Microscopia de Fluorescência , Nanotubos , Polietilenoglicóis/químicaRESUMO
Heterogeneous stem-like populations within tumor tissues are the primary suspects in causing cancer recurrence and malignancy. It is essential to selectively kill these tumorigenic populations. We created a novel system for photothermally ablating specific cells from three-dimensional mammospheres. A CD44-positive subpopulation, with tumorigenic and self-renewal potential, spontaneously arises in MCF7 breast cancer cell-engineered mammospheres. Using anti-CD44 antibody-linked gold nanorods, which strongly absorb near infrared light and increase local temperature, we effectively targeted and photo-ablated atypical cells. This biomarker-specific photothermal ablation model, using a smart nanoplatform, is a promising new strategy for selectively killing cancer cells, while sparing normal tissues.
Assuntos
Técnicas de Ablação/métodos , Neoplasias da Mama/cirurgia , Ouro/química , Receptores de Hialuronatos/metabolismo , Nanotubos/química , Biomarcadores Tumorais/química , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Técnicas de Cultura de Células/métodos , Feminino , Citometria de Fluxo , Humanos , Células MCF-7 , Microscopia Confocal , Microscopia de Fluorescência , Microscopia de Contraste de Fase , Nanotecnologia , Polietilenoglicóis , Ligação ProteicaRESUMO
Amphiphilic surfactants have been used to disperse magnetic nanoparticles in biological media, because they exhibit a dual hydrophobic/hydrophilic affinity that facilitates the formation of a nanoemulsion, within which nanoparticle surfaces can be modified to achieve different physicochemical properties. For the investigation of the interactions of cells with charged magnetic nanoparticles in a biological medium, we selected the nanoemulsion method to prepare water-soluble magnetic nanoparticles using amphiphilic surfactant (polysorbate 80). The hydroxyl groups of polysorbate 80 were modified to carboxyl or amine groups. The chemical structures of carboxylated and aminated polysorbate 80 were confirmed, and water-soluble manganese ferrite nanoparticles (MFNPs) were synthesized with three types of polysorbate 80. Colloidal size, morphology, monodispersity, solubility and T2 relaxivity were found to be similar between the three types of MFNP. However, cationic MFNPs exhibited greater cytotoxicity in macrophages (RAW264.7 cells) and lower cellular membrane effective stiffness than anionic and non-ionic MFNPs. Moreover, cationic MFNPs exhibited large uptake efficiency for RAW264.7 cells compared with anionic or non-ionic MFNPs under the same conditions. Therefore, we propose that surface charge should be a key consideration factor in the design of magnetic nanoparticles for theragnostic applications.
Assuntos
Compostos Férricos/química , Compostos Férricos/toxicidade , Macrófagos/efeitos dos fármacos , Compostos de Manganês/química , Nanopartículas/química , Nanopartículas/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas , Íons/química , Íons/toxicidade , Macrófagos/citologia , Camundongos , Nanopartículas/ultraestrutura , Tamanho da Partícula , Polissorbatos/química , Solubilidade , Tensoativos/químicaRESUMO
Thiolated dextran-coated gold nanorods (DEX-GNRs) were synthesized for targeted delivery to inflammatory macrophages and their photothermal ablation under near-infrared (NIR) light irradiation. Successful synthesis of DEX-GNRs was achieved using thiolated dextran, generated by applying mercaptopropionic acid to transform a hydroxyl group of dextran into a thiol group which has strong binding affinity with surfaces of GNRs. We confirmed both the existence of a thiol group in the functionalized dextran using Ellman's reagent in a thiol group assay and the characteristic band of DEX-GNRs using FT-IR spectrum. Furthermore, a cellular uptake study revealed that dextran showed a superior ability to bind the GNRs surface against macrophages compared to those of PEGylated GNRs with various molecular weights of polyethyleneglycol (PEG). Consequently, an in vitro photothermal irradiation experiment using NIR light indicated that DEX-GNRs exhibited a significant cell-killing efficacy, even with a lower concentration of Au and a low-power light source.
Assuntos
Técnicas de Ablação/métodos , Dextranos/química , Dextranos/farmacologia , Ouro/química , Macrófagos/efeitos dos fármacos , Nanotubos/química , Animais , Transporte Biológico , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Dextranos/metabolismo , Dextranos/toxicidade , Inflamação/patologia , Inflamação/cirurgia , Raios Infravermelhos , Macrófagos/patologia , Macrófagos/efeitos da radiação , Peso Molecular , Polietilenoglicóis/química , TemperaturaRESUMO
We have successfully fabricated a nanocomposite, which is composed of polyaniline (PAni) and pyrene butyric acid (Pyba) via a solvent shift method, which was self-doped at a neutral pH value. This PAni nanocomposite can act as a fine nanoagent expressing absorbance, fluorescence, and Raman properties according to the surrounding pH values.
Assuntos
Compostos de Anilina/química , Compostos de Anilina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Nanocompostos/química , Compostos de Anilina/metabolismo , Ácido Butírico/química , Ácido Butírico/metabolismo , Ácido Butírico/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Humanos , Nanopartículas/química , Nanopartículas/metabolismo , Oxirredução/efeitos dos fármacos , Polissorbatos/química , Polissorbatos/metabolismo , Polissorbatos/farmacologiaRESUMO
We propose that nanogels (HLGs) prepared by simply blending an epidermal growth factor (EGF)-loaded hyaluronan (HA)-based nanoformulation and poloxamers can be efficient transdermal drug carriers. In particular, due to the thermogelling behavior of poloxamer, when the HLGs, which are liquid at room temperature, are applied to the skin's surface, they form a gel at skin temperature. First, lipid-based nanoformulations (EGF-LNs) were fabricated by the lipid thin film method and then chemically conjugated with HA on the surface of the films to prepare EGF-loaded HA-based nanoformulations (EGF-HLNs). Both EGF-LNs and EGF-HLNs exhibited a uniform size and spherical lamellar structure. The EGF-HLN was added to a poloxamer solution to form EGF-HLG, which is a liquid at room temperature and a gel at skin temperature. HLGs have been shown to be able to deliver and permeate EGF well into the skin using both in vitro and in vivo systems, thus serving as an effective transdermal delivery system. In addition, it has been confirmed that this system could be a possible implantable drug carrier. Therefore, HLGs, which are uncomplicated and easily prepared, are expected to be easily used not only in the pharmaceutical field but also in the cosmetic field.
Assuntos
Nanogéis , Cicatrização , Administração Cutânea , Portadores de Fármacos , Fator de Crescimento Epidérmico , PeleRESUMO
For the synthesis of biocompatible photothermal agents, gold nanorod-embedded polymeric nanoparticles (GPNs) were synthesized using a nanoprecipitation method. Uniform gold nanorods (GNRs), which are sensitive to a photothermal effect by near-infrared (NIR) light, with an aspect ratio of 4.0 were synthesized by a seed-mediated growth method. The hydroxyl groups of polycaprolactone diol (PCL diOH) were modified by esterification with mercaptopropionic acid to give a dithiol (polycaprolactone dithiol, PCL diSH) as a phase transfer and capping agent. Subsequently, hexadecyltrimethylammonium bromide (CTAB), a stabilizer of GNRs, was exchanged and/or removed by PCL diSH. PCL diSH-coated GNRs were further wrapped in a hydrophilic polymer, Pluronic F127, as a stabilizer. These newly formulated GPNs exhibit excellent stability in water and a maximum absorbance in the NIR region indicating a highly efficient surface plasmon resonance effect, phenomena useful for photothermal agents.
Assuntos
Ouro/química , Nanopartículas Metálicas/química , Nanotecnologia/métodos , Poliésteres/química , Materiais Biocompatíveis/química , Precipitação Química , Microscopia Eletrônica de Transmissão , Processos Fotoquímicos , Espectroscopia de Infravermelho com Transformada de Fourier , Espectroscopia de Luz Próxima ao Infravermelho , TemperaturaRESUMO
Cetuximab conjugated fluorescent magnetic nanohybrids (CET-FMNHs) were synthesized for detection of human epithelial cancer via magnetic resonance (MR) and optical imaging. Spherical FMNHs consist of MnFe(2)O(4) magnetic nanocrystals encapsulated in pyrene-labeled PCL-b-PMAA as a surfactant prepared by a nano-emulsion method. FMNHs demonstrated excellent colloidal stability and biocompatibility for biomedical application. Antibody conjugated fluorescent magnetic nanohybrids (CET-FMNHs) served as effective agents for both magnetic resonance (MR) and fluorescence optical imaging of cancer cell lines.
Assuntos
Anticorpos Monoclonais , Células Epiteliais/patologia , Corantes Fluorescentes , Nanopartículas , Anticorpos Monoclonais Humanizados , Linhagem Celular Tumoral , Cetuximab , Receptores ErbB , Citometria de Fluxo , Humanos , Imageamento por Ressonância Magnética , Microscopia de Fluorescência , Poliésteres/síntese química , Ácidos Polimetacrílicos/síntese química , Pirenos/síntese químicaRESUMO
Folate (FA) conjugated tri-block copolymers were prepared by bioconjugation of poly epsilon-caprolactonediol and various molecular weights of diamine polyethylene glycol. The synthetic tri-block copolymers were characterized by 1H-NMR. Three types of nanoparticles were prepared by nanoprecipitation. Their size and morphology were verified by laser scattering and transmission electron microscopy, respectively. The colloidal stability of the nanoparticles was evaluated by turbidity test. The anticancer drug doxorubicin (DOX) was encapsulated in the nanoparticles during preparation. Drug loading amounts and release behavior from prepared nanoparticles were investigated. Fluorescent-activated cell sorting analysis and epi-fluorescencic microscopic imaging of prepared nanoparticles exhibited good cellular uptake against target cells. FA receptor expressed OVCAR3 cells that showed higher mean fluorescence intensity than FA receptor defect A549 cells at specific polyethylene glycol chain lengths. The cell cytotoxicity of prepared nanoparticles was evaluated for receptor mediated drug delivery.
Assuntos
Sistemas de Liberação de Medicamentos , Nanopartículas/química , Nanopartículas/toxicidade , Polietilenoglicóis/química , Polietilenoglicóis/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fenômenos Químicos , Físico-Química , Humanos , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Transmissão , Estrutura Molecular , Peso Molecular , Nanopartículas/ultraestruturaRESUMO
We report the successful fabrication of the various types of water soluble PEGylated magnetic complexes (PMCs) for magnetism-related biomedical applications. Various types of PMCs were synthesized and tested to accomplish phase transfer from organic to aqueous phase using monomethoxy polyethylene glycol (mPEG)-fatty acid amphiphilic block copolymers (PFs) through conjugation of the hydroxyl group of mPEG with the carboxyl group of fatty acids. We also carefully investigate their colloidal stabilities in aqueous phase according to the ratio of hydrophilic and hydrophobic lengths relying on different types of fatty acids. Synthesized PMCs clearly demonstrated high magnetic sensitivity under magnetic field as magnetic resonance (MR) contrast agents. Furthermore, PMCs exhibited sufficient cell viabilities and excellent cell affinities in an in vitro model. Our results demonstrated that our PMCs possessed the potential for highly efficient magnetism-related biomedical applications such as MR image agents, drug delivery and tracking of cells.
Assuntos
Portadores de Fármacos/síntese química , Ácidos Graxos/química , Magnetismo , Polietilenoglicóis/química , Água , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/metabolismo , Portadores de Fármacos/metabolismo , Ácidos Graxos/metabolismo , Compostos Férricos/síntese química , Células HeLa , Humanos , Compostos de Manganês/síntese química , Nanopartículas/química , Polietilenoglicóis/metabolismo , Polímeros/síntese química , Polímeros/metabolismo , Solubilidade , Sais de Tetrazólio/química , Tiazóis/química , Aderências TeciduaisRESUMO
Oleyl dextran-coated magnetic nanoclusters (ODMCs) are fabricated for the accurate detection of macrophage-rich atherosclerotic plaques using magnetic resonance (MR) imaging. Dextran is introduced to the cluster surface of magnetic nanocrystals (MNCs) through self-assembly using amphiphilic oleic acid-conjugated dextran (ODex) to provide not only hydrophilicity but also a high affinity to macrophages. Enhanced magnetism of the ODMCs is engineered by optimizing the degree of substitution (DS) of the oleyl group in ODex and the concentration of ODex used for the synthesis of ODMC. Consequently, ODMCs exhibit significantly increased T2 relaxivity and excellent macrophage-targeting ability without cytotoxicity, in vitro. Moreover, in vivo T2-weighted MR imaging after intravenous injection of ODMCs into a rat artery balloon injury model demonstrates considerable MR contrast strength efficacy in the plaques of the injured carotid artery. These novel ODMCs may offer a highly efficient MR imaging nanoprobes for the selective diagnosis of atherosclerosis.
Assuntos
Imageamento por Ressonância Magnética , Nanopartículas de Magnetita/química , Nanotecnologia/métodos , Placa Aterosclerótica/diagnóstico , Polímeros/química , Animais , Linhagem Celular , Sobrevivência Celular , Dextranos/química , Fluorescência , Hidrodinâmica , Macrófagos/ultraestrutura , Nanopartículas de Magnetita/ultraestrutura , Masculino , Camundongos , Ácido Oleico/química , Tamanho da Partícula , Ratos Sprague-Dawley , Termogravimetria , Difração de Raios XRESUMO
Poly(ethylene glycol)-coated cross-linked iron oxide nanoparticles (PCIONs) are developed for therapeutic engineering of mesenchymal stem cells (MSCs) and their monitoring via magnetic resonance (MR) imaging at a time. PCIONs successfully combine with plasmid DNA (pDNA) via ionic interaction. Accordingly, PCION/pDNA complexes mediate superior translocations of vascular endothelial growth factor (VEGF) pDNA into intracellular regions of MSCs under external magnetic field, which significantly elevate production of VEGF from MSCs. Genetically engineered MSCs are also clearly visualized via MR imaging after administration to rat cerebrovascular ischemia models, which enable tracking of MSCs migration from injected sites to injured ischemic area.
Assuntos
Isquemia Encefálica/terapia , Rastreamento de Células/métodos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Células-Tronco Mesenquimais/metabolismo , Nanopartículas/química , Animais , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patologia , Engenharia Celular , Modelos Animais de Doenças , Compostos Férricos/química , Imageamento por Ressonância Magnética , Células-Tronco Mesenquimais/citologia , Plasmídeos/química , Polietilenoglicóis/química , Ratos , Ratos Sprague-Dawley , Transfecção , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Combined cancer treatment via co-delivery of siRNAs and an anticancer drug can be a promising strategy due to the synergistic effect of simultaneously minimizing gene/drug administration. In this study, Bcl-xL siRNA and doxorubicin (DOX) are encapsulated into designed methoxy-poly(ethylene glycol)-block-poly(D,L-lactic acid) (mPEG-b-PLA) block copolymer polymersomes (PSomes). A study of the cytotoxicity of Bcl-xL siRNA and DOX co-encapsulated PSomes (CPSomes) shows more inhibited proliferation of MKN-45 and MKN-28 human gastric cancer cell lines than only gene- and drug-loaded ones. Consequently, these results demonstrate that co-delivery of genes and drugs using PSomes results in a synergistic efficacy and indicates the potential of PSomes as efficient nanocarriers for combined cancer therapy.
Assuntos
Antineoplásicos/farmacologia , Materiais Biocompatíveis/química , Doxorrubicina/farmacologia , Sistemas de Liberação de Medicamentos , Poliésteres/química , Polietilenoglicóis/química , RNA Interferente Pequeno/metabolismo , Proteína bcl-X/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Microscopia Crioeletrônica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Humanos , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Hidrólise/efeitos dos fármacos , Concentração Inibidora 50 , Cinética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Tempo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Proteína bcl-X/genéticaAssuntos
Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Nanomedicina/métodos , Nanopartículas/uso terapêutico , Animais , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Imageamento por Ressonância Magnética , Magnetismo , Camundongos , Neoplasias Experimentais/tratamento farmacológico , Polímeros/uso terapêuticoRESUMO
Organic solvent-soluble nanocrystals suitable for magnetic resonance imaging are prepared by two routes, namely, a coprecipitation method and a multiple-step thermal decomposition method (seed-mediated growth). The size, shape, crystallinity, phase, and composition of the prepared nanocrystals are determined by various characterization techniques, including transmission electron microscopy, vibrating-sample magnetometer, X-ray diffraction, and inductively coupled plasma mass spectroscopy. Subsequently, the organic-soluble nanocrystals are rendered water-soluble by two methods, the microemulsion method and the ligand exchange method, for biomedical applications. Detailed protocols for the preparation of water-soluble nanocrystals, as well as procedures for drug-loading and antibody conjugation to the water-soluble nanocrystals are provided.
Assuntos
Desenho de Fármacos , Nanopartículas de Magnetita/química , Nanopartículas Metálicas/química , Nanotecnologia/métodos , Anticorpos/química , Precipitação Química , Emulsões , Interações Hidrofóbicas e Hidrofílicas , Tamanho da Partícula , Polímeros/química , TemperaturaRESUMO
To develop novel gene delivery carriers, aminated polysorbate 80 (P80-NH(2)) was synthesized with strong positively charged properties through the introduction of three amine groups. The resulting P80-NH(2) and DNA polyplex exhibited superb condensation abilities due to the high densities of positively charged amines groups. Size and surface charge of polyplex were shown to be well suited for cellular internalization. In addition, the P80-NH(2) /DNA polyplex demonstrated acceptable transfection efficiency in HeLa cells and was nontoxic relative to the conventional 25-kDa polyethyleneimine system.
Assuntos
Antineoplásicos/síntese química , DNA/química , Portadores de Fármacos/síntese química , Técnicas de Transferência de Genes , Polissorbatos/síntese química , Aminação , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Portadores de Fármacos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Proteínas de Fluorescência Verde/química , Células HeLa , Humanos , Teste de Materiais , Estrutura Molecular , Tamanho da Partícula , Plasmídeos/química , Polissorbatos/química , Polissorbatos/farmacologia , Relação Estrutura-Atividade , Propriedades de Superfície , TransfecçãoRESUMO
We investigated the synergism between shRNAs against Bcl-xL and doxorubicin (DOX) using aptamer-conjugated polyplexes (APs) in combination cancer therapy. Synergistic and selective cancer cell death was achieved by AP-mediated co-delivery of very small amounts of DOX and Bcl-xL-specific shRNA, which simultaneously activated an intrinsic apoptotic pathway. A branched polyethyleneimine (PEI) was grafted to polyethylene glycol (PEI-PEG) to serve as a vehicle for shRNA delivery, and its surface was further conjugated with an anti-PSMA aptamer (APT) for the selective delivery of APs to prostate cancer cells that express prostate-specific membrane antigens (PSMA) on their cell surface. The APs were finally obtained after intercalation of DOX to form shRNA/PEI-PEG-APT/DOX conjugates. Cell viability assays and FACS analysis of GFP expression against PC3 (PSMA deficient) and LNCaP (PSMA overexpressed) cells demonstrated that the synthesized APs inhibited the growth of PSMA-abundant prostate cancer cells with strong cell selectivity. Consequently, IC(50) values of APs loaded with both DOX and shRNA were approximately 17-fold less than those for the simple mixture of shRNA plus drug (shRNA/Lipofectamine + DOX). These results suggest that AP-mediated co-delivery of an anti-cancer drug and shRNA against Bcl-xL may widen the therapeutic window and allow for the selective destruction of cancer cells.
Assuntos
Antibióticos Antineoplásicos , Aptâmeros de Nucleotídeos , Morte Celular/efeitos dos fármacos , Doxorrubicina , Neoplasias da Próstata/tratamento farmacológico , RNA Interferente Pequeno , Proteína bcl-X , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacologia , Antibióticos Antineoplásicos/uso terapêutico , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/farmacologia , Aptâmeros de Nucleotídeos/uso terapêutico , Linhagem Celular Tumoral , Doxorrubicina/química , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Humanos , Masculino , Teste de Materiais , Polietilenoglicóis/química , Polietilenoglicóis/metabolismo , Polietilenoimina/química , Polietilenoimina/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/uso terapêuticoRESUMO
We fabricated multimode nanoprobes for acquisition of biological information at different object levels, i.e., in vivo detection and ex vivo validation for characterizing tumor angiogenesis. Fluorescent magnetic nanoprobes (FMNPs) were synthesized by using amphiphilic pyrenyl polyethylene glycol (Py-PEG) and superparamagnetic MnFe(2)O(4) nanocrystals (MNCs). Py-PEG, which is synthesized by conjugation of hydrophilic PEG with hydrophobic and fluorescent 1-pyrenebutyric acid through an esterification process, is capable of self-assembly and maintaining a high UV fluorescent intensity in aqueous phase. Py-PEG can be used as a fluorescent surfactant that simultaneously and efficiently encapsulates MNCs to exhibit fluorescent and magnetic properties as well as maintaining high water-solubility. Consequently, we proved that our biologically non-toxic FMNPs were prominent multimode imaging probes by showing not only excellent MR sensitivity but also high illumination intensity with strong signal strength under short exposure time of UV light from the extensive imaging studies of in vitro/vivo and ex vivo using orthotopic and xenograft mice models.