RESUMO
Nanotechnology provides a powerful tool to overcome many disadvantages of small-molecule photosensitizers for photodynamic cancer therapy, such as hydrophobicity, rapid blood clearance, low accumulation in tumor tissue and low cell penetration, etc. The occurrence of quench in photosensitizer-loaded nanoparticle greatly downregulates the ability to generate singlet oxygen with light irradiation. Stimuli-responsive nanocarriers can improve the efficacy of PDT to a certain extent. However, insufficient release of photosensitizer from either endogenous or exogenous stimuli responsive nanocarriers in the short period of light irradiation restricts full usage of the photosensitizer delivered into cancer cells. We here report a dual-step light irradiation strategy to enhance the efficacy of cancer PDT. Ce6 as a photosensitizer is loaded in singlet oxygen-sensitive micelles (Ce6-M) via self-assembly of amphiphilic polymer mPEG2000-TK-C16. After co-incubation of Ce6-M with cancer cells or i.v. injection of Ce6-M, cancer cells or tumor tissues are irradiated with light for a short time to trigger Ce6 release, and 2 h later, re-irradiated for relatively long time. The sufficient release of Ce6 in the period between twice light irradiation significantly improves the generation of singlet oxygen, leading to more efficient cancer therapeutic effects of dual-step irradiation than that of single-step irradiation for the same total irradiation time.
Assuntos
Nanopartículas , Neoplasias , Fotoquimioterapia , Porfirinas , Linhagem Celular Tumoral , Micelas , Neoplasias/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Polímeros , Oxigênio SingleteRESUMO
Excessive amounts of Al3+ in the human body can cause adverse effects on immune function and induce several neurodegenerative disorders. So far, most of the reported fluorescent probes for Al3+ present some common drawbacks, such as low sensitivity and poor water solubility. In addition, a number of traditional fluorescent probes failed to image Al3+ in tumor cells due to the lack of tumor cell targeting capacity and cell penetrating abilities. To overcome these shortcomings, we constructed tumor-targeting fluorescent mixed nano-micelles (mPEG-Dye-Biotin) with an average particle size of 21 nm from an amphiphilic polymer containing a Schiff-base fluorescent unit (mPEG-Dye) and another amphiphilic polymer containing a tumor cell recognition ligand (DSPE-PEG-Biotin), through the co-self-assembly of both amphiphilic polymers in water using the film rehydration method. The as-prepared nanoprobe showed a highly sensitive and selective turn-on fluorescence response to Al3+ in aqueous solution with a low detection limit. MTT assay revealed the negligible cytotoxicity of the mPEG-Dye-Biotin nanoprobe to both HeLa cells and COS-7 cells, indicating the safety of mPEG-Dye-Biotin for biological applications. More importantly, the biotinylated nanoprobe showed better ability to enter biotin receptor-positive HeLa cells than that of the non-biotinylated micelle mPEG-Dye, which made it more suitable for imaging Al3+ in biotin receptor-positive tumor cells. This work provides a simple and general strategy to design a highly sensitive and tumor cell-specific metal ion nanoprobe, which can not only be applied in Al3+ imaging, but can also be extended to other ions or biomolecules by changing the incorporated fluorescent unit in the amphiphilic polymer.
Assuntos
Alumínio/análise , Biotina/química , Corantes Fluorescentes/química , Imagem Óptica , Polímeros/química , Neoplasias do Colo do Útero/diagnóstico por imagem , Animais , Células COS , Chlorocebus aethiops , Feminino , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Ligantes , Micelas , Estrutura Molecular , Tamanho da Partícula , Bases de Schiff/química , Propriedades de SuperfícieRESUMO
The effects of poly(ethylene glycol) (PEG) on improving the biological compatibility and circulation time of nanocarriers are determined by the surface density of PEG on nanoparticles. PEG with high surface density on nanocarriers has greater accumulation in tumor tissues. However, this impairs the release of drugs loaded in the nanoparticles in the tumor tissues. The relations and internal regularities between the controlled stripping of PEG of nanoparticles and its fate and antitumor efficacy in vivo remain unsolved. Redox-sensitive hybrid nanoparticles coated with varied PEG densities were prepared by blending a redox-sensitive polymer of DLPE-SS-MPEG. To keep identical nanoproperties, these nanoparticles were prepared with a similar size distribution of around 100 nm. The effects of controlled stripping of PEG on antitumor activities of nanoparticles were then investigated. As the PEG surface density increased, lower cellular internalization by tumor cells was observed. However, nanoparticles with higher controlled stripping of PEG showed greater accumulation in tumor tissues and advanced antitumor activities in vivo.
Assuntos
Nanopartículas , Polietilenoglicóis , Linhagem Celular Tumoral , Oxirredução , Polietilenoglicóis/metabolismo , PolímerosRESUMO
Polyaspartamide-based oligo-ethylenimine brushes (PASP-EDA, PASP-TEPA, PASP-PEHA, and PASP-PEI 423) were synthesized from polysuccinimide (PSI) via a ring-opening reaction with N-Boc protected ethylenediamine, tetraethylenepentamine, pentaethylenehexamine, and linear polyethylenimine (Mn 423), respectively. PASP-TEPA, PASP-PEHA, and PASP-PEI 423 possess high buffer capacity between pH 5 and pH 7, which is comparable to that of branched PEI 25000. The cytotoxicity assay indicated that they all are less toxic than PEI 25000. At an N/P ratio of above 2, all of the four synthetic polycation brushes can condense plasmid DNA to form small sized (160-400 nm) polyelectrolyte complexes with positive surface charge. The transfection of HEK 293 cells with oligo-ethylenimine brush/pRE Luc polyplexes indicated that the transfection efficiencies increased with increasing the length of oligo-ethylenimine side chains. The luciferase expression with PASP-PEHA and PASP-PEI 423 were as high as or even a little higher than that of PEI 25000. The results demonstrate that polyaspartamide-based oligo-ethylenimine brushes are a very promising class of novel polycations for highly efficient and less toxic gene delivery.
Assuntos
DNA/administração & dosagem , Poliaminas/química , Polietilenoimina/química , Proteínas/química , Transfecção , Aziridinas/química , Aziridinas/metabolismo , Linhagem Celular , Sobrevivência Celular , Humanos , Tamanho da Partícula , Plasmídeos/administração & dosagem , Poliaminas/metabolismo , Polieletrólitos , Polietilenoimina/metabolismo , Proteínas/metabolismoRESUMO
Three hydrolytically degradable poly(beta-aminoester)s containing ester bonds in the main chain and primary amines in the side chain, synthesized by Michael polyaddition, were applied to deliver foreign DNA into cells in vitro. These linear polycations can condense DNA into small-sized particles with positive surface charge at high N/P ratios. Their high buffer capacity at pH 5-7 facilitated the escape of DNA from the endosome and resulted in efficient gene expression. Under the optimal conditions, poly(beta-aminoester)s with a pendant aminoethyl group showed higher transfection efficiencies than branched poly(ethylenimine) (PEI) 25KDa in 293T cells. The effect of side chain structure of the poly(beta-aminoester) on transfection efficiency has been investigated, which indicated that the poly(beta-aminoester) containing the pendant aminoethyl group was the most efficient carrier for both of 293T cells and COS-7 cells. The combination of hydrolytical degradation, high buffer capacity, relatively low cytotoxicity, and high transfection efficiency suggested that this kind of poly(beta-aminoester)s are novel promising nonviral gene carriers.
Assuntos
Aminas/química , Portadores de Fármacos/química , Técnicas de Transferência de Genes , Polímeros/química , Animais , Células COS , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , DNA/química , Relação Dose-Resposta a Droga , Portadores de Fármacos/síntese química , Portadores de Fármacos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Humanos , Concentração de Íons de Hidrogênio , Tamanho da Partícula , Polímeros/síntese química , Polímeros/farmacologia , Propriedades de SuperfícieRESUMO
Photodynamic therapy (PDT) as an alternative choice of cancer treatment method has attracted increasing attention in the past few decades. A sufficient amount of oxygen is essential for the production of singlet oxygen (1O2) in successful PDT; however, hypoxia is a typical hallmark of cancer, which is one of the most important limitation factors of PDT. To overcome the hypoxic tumour microenvironment and achieve highly efficient photodynamic cancer therapy, herein, a photosensitizer Ce6-loaded fluorinated polymeric micelle (Ce6-PFOC-PEI-M) was constructed via the self-assembly of an amphiphilic polymer prepared from perfluorooctanoic acid and branched polyethyleneimine (10 kDa). The introduction of perfluoroalkyl groups in the polymeric micelle Ce6-PFOC-PEI-M retained the oxygen-carrying capacity similar to perfluorocarbon, increased the oxygen level and overcame the hypoxia in C6 glioma cells under oxygen-deficient conditions. As a control, Ce6-OC-PEI-M without a perfluoroalkyl group could not increase the oxygen level in C6 glioma cells under the same conditions. With laser irradiation, Ce6-PFOC-PEI-M generated much more reactive oxygen species (ROS) in C6 glioma cells than Ce6-OC-PEI-M, leading to a higher phototoxicity in vitro and photodynamic tumour growth inhibition in vivo than Ce6-OC-PEI-M. Furthermore, there were no differences in the contents of Ce6 in tumour tissue between Ce6-PFOC-PEI-M and Ce6-OC-PEI-M. The higher efficacy of Ce6-PFOC-PEI-M in PDT is ascribed to its oxygen-carrying ability rather than higher content of Ce6 in the tumour. The presented fluorinated polymeric micelle could provide a new platform in the delivery of various photosensitizers and has great potential to improve the efficacy of PDT cancer therapy.
Assuntos
Flúor/química , Fotoquimioterapia/métodos , Polímeros/química , Porfirinas/química , Hipóxia Tumoral , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Clorofilídeos , Portadores de Fármacos , Liberação Controlada de Fármacos , Feminino , Xenoenxertos , Humanos , Concentração de Íons de Hidrogênio , Lasers , Luz , Camundongos Endogâmicos BALB C , Camundongos Nus , Micelas , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/química , Porfirinas/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Distribuição TecidualRESUMO
The controllable and sustained release of DNA from the surfaces of biomaterials or biomedical devices represents a new method for localized gene delivery. We report the synthesis of a novel polycation containing disulfide bonds in its backbone and the fabrication of polycation/plasmid DNA multilayered thin films by layer-by-layer assembly. The films are very stable during preparation and in storage, however, they gradually degrade and release the incorporated DNA when incubated in PBS buffer containing dithiothreitol (DTT), which results from the degradation of a disulfide-contained polymer under reductive conditions. The film degradation rate and DNA release rate can be tuned by the concentration of reducing agent. This approach will be useful in gene therapy and tissue engineering by controlled administration of therapeutic DNA deposited on the surface of implantable biomedical devices or tissue engineering scaffolds.
Assuntos
DNA/química , Plasmídeos/química , Poliaminas/química , Materiais Biocompatíveis/química , Cromatografia/métodos , Ditiotreitol/química , Etídio/química , Concentração de Íons de Hidrogênio , Luz , Microscopia de Força Atômica , Polieletrólitos , Polímeros/química , Espalhamento de Radiação , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Engenharia Tecidual/métodosRESUMO
Codelivery is a promising strategy to overcome the limitations of single chemotherapeutic agents in cancer treatment. Despite progress, codelivery of two or more different functional drugs to increase anticancer efficiency still remains a challenge. Here, reduction-sensitive lipid-polymer hybrid nanoparticles (LPNPs) drug delivery system composed of monomethoxy-poly(ethylene glycol)-S-S-hexadecyl (mPEG-S-S-C16), soybean lecithin, and poly(D,L-lactide-co-glycolide) (PLGA) was used for codelivery of doxorubicin (DOX) and a Chinese herb extract triptolide (TPL). Hydrophobic DOX and TPL could be successfully loaded in LPNPs by self-assembly. More importantly, drug release and cellular uptake experiments demonstrated that the two drugs were reduction sensitive, released simultaneously from LPNPs, and taken up effectively by the tumor cells. DOX/TPL-coloaded LPNPs (DOX/TPL-LPNPs) exhibited a high level of synergistic activation with low combination index (CI) in vitro and in vivo. Moreover, the highest synergistic therapeutic effect was achieved at the ratio of 1:0.2 DOX/TPL. Further experiments showed that TPL enhanced the uptake of DOX by human oral cavity squamous cell carcinoma cells (KB cells). Overall, DOX/TPL-coencapsulated reduction-sensitive nanoparticles will be a promising strategy for cancer treatment.
Assuntos
Diterpenos/administração & dosagem , Diterpenos/uso terapêutico , Doxorrubicina/administração & dosagem , Doxorrubicina/uso terapêutico , Sistemas de Liberação de Medicamentos , Lipídeos/química , Nanopartículas/química , Neoplasias/tratamento farmacológico , Fenantrenos/administração & dosagem , Fenantrenos/uso terapêutico , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Doxorrubicina/química , Sinergismo Farmacológico , Compostos de Epóxi/administração & dosagem , Compostos de Epóxi/uso terapêutico , Citometria de Fluxo , Humanos , Ácido Láctico , Camundongos Endogâmicos BALB C , Camundongos Nus , Nanopartículas/ultraestrutura , Oxirredução/efeitos dos fármacos , Tamanho da Partícula , Ácido Poliglicólico , Copolímero de Ácido Poliláctico e Ácido PoliglicólicoRESUMO
The major challenge in current clinic contrast agents (CAs) and chemotherapy is the poor tumor selectivity and response. Based on the self-quench property of IR820 at high concentrations, and different contrast effect ability of Gd-DOTA between inner and outer of liposome, we developed "bomb-like" light-triggered CAs (LTCAs) for enhanced CT/MRI/FI multimodal imaging, which can improve the signal-to-noise ratio of tumor tissue specifically. IR820, Iohexol and Gd-chelates were firstly encapsulated into the thermal-sensitive nanocarrier with a high concentration. This will result in protection and fluorescence quenching. Then, the release of CAs was triggered by near-infrared (NIR) light laser irradiation, which will lead to fluorescence and MRI activation and enable imaging of inflammation. In vitro and in vivo experiments demonstrated that LTCAs with 808 nm laser irradiation have shorter T1 relaxation time in MRI and stronger intensity in FI compared to those without irradiation. Additionally, due to the high photothermal conversion efficiency of IR820, the injection of LTCAs was demonstrated to completely inhibit C6 tumor growth in nude mice up to 17 days after NIR laser irradiation. The results indicate that the LTCAs can serve as a promising platform for NIR-activated multimodal imaging and photothermal therapy.
Assuntos
Meios de Contraste/química , Imagem Multimodal/métodos , Neoplasias Experimentais/diagnóstico por imagem , Animais , Feminino , Compostos Heterocíclicos/química , Humanos , Verde de Indocianina/análogos & derivados , Verde de Indocianina/química , Raios Infravermelhos , Lipossomos/química , Imageamento por Ressonância Magnética/métodos , Camundongos Endogâmicos BALB C , Camundongos Nus , Imagem Multimodal/instrumentação , Neoplasias/patologia , Neoplasias Experimentais/terapia , Compostos Organometálicos/química , Fototerapia/métodos , Razão Sinal-Ruído , Nanomedicina Teranóstica/instrumentação , Nanomedicina Teranóstica/métodosRESUMO
The importance of mitochondrial delivery of an anticancer drug to cancer cells has been recognized to improve therapeutic efficacy. The introduction of lipophilic cations, such as triphenylphosphonium (TPP), onto the surface of nanocarriers was utilized to target mitochondria via strong electrostatic interactions between positively charged TPP and the negatively charged mitochondrial membrane. However, the highly positive charge nature of TPP leads to rapid clearance from the blood, decrease of circulation lifetime, and nonspecific targeting of mitochondria of cells. Here, we report a strategy for improving the anticancer efficacy of paclitaxel via redox triggered intracellular activation of mitochondria-targeting. The lipid-polymer hybrid nanoparticles (LPNPs) are composed of poly(d,l-lactide-co-glycolide) (PLGA), a TPP-containing amphiphilic polymer (C18-PEG2000-TPP) and a reduction-responsive amphiphilic polymer (DLPE-S-S-mPEG4000). The charges of TPP in LPNPs were almost completely shielded by surface coating of a PEG4000 layer, ensuring high tumor accumulation. After uptake by cancer cells, the surface charges of LPNPs were recovered due to the detachment of PEG4000 under intracellular reductive conditions, resulting in rapid and precise localization in mitochondria. This kind of simple, easy and practicable mitochondria-targeting nanoplatform showed high anticancer activity, and the activatable strategy is valuable for developing a variety of nanocarriers for application in the delivery of other drugs.
Assuntos
Antineoplásicos/administração & dosagem , Portadores de Fármacos , Mitocôndrias/efeitos dos fármacos , Nanopartículas , Neoplasias/tratamento farmacológico , Oxirredução , Animais , Feminino , Humanos , Lipossomos , Células MCF-7 , Camundongos Endogâmicos BALB C , Camundongos Nus , Compostos Organofosforados , Polietilenoglicóis , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In recent years, there has been increasing interest in developing a multifunctional nanoscale platform for cancer monitoring and chemotherapy. However, there is still a big challenge for current clinic contrast agents to improve their poor tumor selectivity and response. Herein, we report a new kind of Gd complex and folate-coated redox-sensitive lipid-polymer hybrid nanoparticle (Gd-FLPNP) for tumor-targeted magnetic resonance imaging and therapy. Gd-FLPNPs can simultaneously accomplish diagnostic imaging, and specific targeting and controlled release of doxorubicin (DOX). They exhibit good monodispersity, excellent size stability, and a well-defined core-shell structure. Paramagnetic nanoparticles based on gadolinium-diethylenetriaminepentaacetic acid-bis-cetylamine have paramagnetic properties with an approximately two-fold enhancement in the longitudinal relaxivity compared to clinical used Magnevist. For targeted and reduction-sensitive drug delivery, Gd-FLPNPs released DOX faster and enhanced cell uptake in vitro, and exhibited better antitumor effect both in vitro and in vivo.
Assuntos
Doxorrubicina/farmacologia , Sistemas de Liberação de Medicamentos , Lipídeos/química , Imageamento por Ressonância Magnética , Nanopartículas/química , Polímeros/química , Animais , Linhagem Celular Tumoral , Doxorrubicina/química , Endocitose/efeitos dos fármacos , Feminino , Ácido Fólico/química , Ácido Fólico/farmacologia , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , OxirreduçãoRESUMO
An amphiphilic polymer DLPE-S-S-MPEG was synthesized and employed with PCL to prepare two-component reduction-sensitive lipid-polymer hybrid nanoparticles (SLPNPs) for in vitro and in vivo delivery of a hydrophobic anticancer drug (Doxorubicin, DOX). Insensitive lipid-polymer hybrid nanoparticles (ILPNPs) were prepared as a control. The mean sizes of the LPNPs ranged from 100 nm to 120 nm. The TEM observations showed that the LPNPs have spherical morphologies with homogeneous distribution. The disulfide bond of DLPE-S-S-MPEG was cleaved by dithiothreitol (DTT), which resulted in the disassembly of SLPNPs and triggered the release of encapsulated DOX. The in vitro cytotoxicities of DOX/LPNPs against HeLa cells, HepG2 cells and COS-7 cells were studied. It was demonstrated that DOX/SLPNPs showed higher cytotoxicity against HeLa cells and HepG2 cells than DOX/ILPNPs, but showed a slight difference in the case of COS-7 cells. CLSM observation and FCM measurement further confirmed that the introduction of S-S bonds caused fast intracellular release of DOX from SLPNPs. Moreover, compared with DOX/ILPNPs and free DOX, DOX/SLPNPs exhibited higher antitumor activity. Both DOX/SLPNPs and DOX/ILPNPs showed lower cardiac toxicity and kidney toxicity than free DOX, which were confirmed by histological and immunohistochemical analyses. The tissue distribution of DOX in mice exhibited that two kinds of DOX/LPNPs accumulated extensively in the liver and spleen, while free DOX accumulated mainly in the heart and kidney 12 h after injection. Two-component SLPNPs may be a promising drug delivery carrier for reduction-triggered delivery of DOX.
Assuntos
Antineoplásicos/administração & dosagem , Antineoplásicos/química , Nanopartículas/química , Polímeros/farmacologia , Animais , Antineoplásicos/farmacologia , Células COS , Chlorocebus aethiops , Doxorrubicina/administração & dosagem , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Células HeLa , Coração/efeitos dos fármacos , Células Hep G2 , Humanos , Rim/efeitos dos fármacos , Camundongos , Polímeros/química , Distribuição TecidualRESUMO
The model drugs ibuprofen (IBU) and tegafur (T-Fu) were loaded into poly[N-isopropylacrylamide-co-(acryloyl beta-cyclodextrin)] [P(NIPA-co-A-CD)] and PNIPA hydrogels by immersing dried gels in IBU or T-Fu alcohol solutions until they reached equilibrium. Drug release studies were carried out in water at 25 degrees C. In contrast to the release time of conventional PNIPA hydrogel, that of IBU from the beta-CD incorporated hydrogel was significantly prolonged and the drug loading was also greatly increased, which may be the result of the formation of inclusion complexes between CD and ibuprofen. However, another hydrophilic drug, tegafur, did not display these properties because it could not form a complex with the CD groups. [diagram in text].
Assuntos
Resinas Acrílicas/química , Ciclodextrinas/química , Preparações de Ação Retardada , Hidrogéis/química , Temperatura , Preparações de Ação Retardada/química , Ibuprofeno/administração & dosagem , Ibuprofeno/química , Tegafur/administração & dosagem , Tegafur/químicaRESUMO
The development and evaluation of folate-targeted and reduction-triggered biodegradable nanoparticles are introduced to the research on targeted delivery of doxorubicin (DOX). This type of folate-targeted lipid-polymer hybrid nanoparticles (FLPNPs) is comprised of a poly(D,L-lactide-co-glycolide) (PLGA) core, a soybean lecithin monolayer, a monomethoxy-poly(ethylene glycol)-S-S-hexadecyl (mPEG-S-S-C16) reduction-sensitive shell, and a folic acid-targeted ligand. FLPNPs exhibited high size stability but fast disassembly in a simulated cancer cell reductive environment. The experiments on the release process in vitro revealed that as a reduction-sensitive drug delivery system, FLPNPs released DOX faster in the presence of 10 mM dithiothreitol (DTT). Results from flow cytometry, confocal image and in vitro cytotoxicity assays revealed that FLPNPs further enhanced cell uptake and generated higher cytotoxicity against human epidermoid carcinoma in the oral cavity than non-targeted redox-sensitive and targeted redox-insensitive controls. Furthermore, in vivo animal experiments demonstrated that systemic administration of DOX-loaded FLPNPs remarkably reduced tumor growth. Experiments on biodistribution of DOX-loaded FLPNPs showed that an increasing amount of DOX accumulated in the tumor. Therefore, FLPNPs formulations have proved to be a stable, controllable and targeted anticancer drug delivery system.
Assuntos
Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Ácido Fólico/química , Ácido Láctico/química , Lipídeos/química , Nanopartículas/química , Polietilenoglicóis/química , Ácido Poliglicólico/química , Antineoplásicos/química , Linhagem Celular Tumoral , Doxorrubicina/química , Humanos , Ligantes , Micelas , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Distribuição TecidualRESUMO
A new type of shell-sheddable micelles with disulfide linkages between the hydrophobic polyester core and hydrophilic poly(ethylene glycol) (PEG) shell was developed based on Y-shaped amphiphilic polymers mPEG-S-S-(PCL)2. The micelles were then used for the glutathione-mediated intracellular delivery of the anticancer drug doxorubicin (DOX) into tumor cells. The polymer could self-assemble into micelles with an average diameter of 135nm in aqueous solution and load DOX at a total content of 3.6%. The hydrophilic PEG shell of these micelles could be shed in the presence of reducing agent dithiothreitol (DTT), which resulted in size change of the micelles. In vitro release studies revealed that DOX-loaded mPEG-S-S-(PCL)2 micelles exhibited faster DOX release in the presence of DTT. MTT assay demonstrated that DOX-loaded mPEG-S-S-(PCL)2 micelles showed higher cytotoxicity against 10mM of glutathione monoester (GSH-OEt) pretreated HeLa cells than that of the non-pretreated ones. Confocal laser scanning microscopy and flow cytometry analyses indicated that DOX-loaded mPEG-S-S-(PCL)2 micelles were efficiently internalized into HeLa cells and exhibited faster DOX release in GSH-OEt-pretreated cells than in cells with no pretreatment. Endocytosis inhibition results proved that mPEG-S-S-(PCL)2 micelles entered the cells mainly through the clathrin-mediated endocytosis pathway, and caveolae-mediated endocytosis was involved to a small extent. These results indicate the great potential of the proposed Y-shaped reduction-sensitive polymer for application in effective intracellular anticancer drug delivery.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Micelas , Polietilenoglicóis/química , Polímeros/química , Sobrevivência Celular/efeitos dos fármacos , Citoplasma/metabolismo , Endocitose/efeitos dos fármacos , Células HeLa , Humanos , Interações Hidrofóbicas e Hidrofílicas , Tamanho da Partícula , Poliésteres/químicaRESUMO
Cationic polyphosphoesters (PPEs) with different side-chain charge groups were designed and synthesized as biodegradable gene carriers. Poly(N-methyl-2-aminoethyl propylene phosphate) (PPE-MEA), with a secondary amino group (-CH(2)CH(2)NHCH3) side chain released DNA in several hours at N/P (amino group of polymer to phosphate group of DNA) ratios from 0.5 to 5; whereas PPE-HA, bearing -CH(2)(CH2)(4)CH(2)NH(2) groups in the side chain, did not release DNA at the same ratio range for 30 days. Hydrolytic degradation and DNA binding results suggested that side chain cleavage, besides the polymer degradation, was the predominant factor affected the DNA release and transfection efficiencies. The side chain of PPE-MEA was cleaved faster than that of PPE-HA, resulting poor cellular uptake and no transgene expression for PPE-MEA/DNA complexes in COS-7 cells at charge ratios from 4 to 12. In contrast, PPE-HA/DNA complexes were stable enough to be internalized by cells and effected gene transfection (3400 folds higher than background at a charge ratio of 12). Interestingly, gene expression levels mediated by PPE-MEA and PPE-HA in mouse muscle following intramuscular injection of complexes showed a reversed order: PPE-MEA/DNA complexes mediated a 1.5-2-fold higher luciferase expression in mouse muscle as compared with naked DNA injection, while PPE-HA/DNA complexes induced delayed and lowered luciferase expression than naked DNA. These results suggested that the side chain structure is a crucial factor determining the mechanism and kinetics of hydrolytic degradation of PPE carriers, which in turn influenced the kinetics of DNA release from PPE/DNA complexes and their transfection abilities in vitro and in vivo.
Assuntos
Técnicas de Transferência de Genes , Compostos Organofosforados/química , Plasmídeos/administração & dosagem , Polímeros , Animais , Materiais Biocompatíveis/química , Células COS , Cátions/química , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , DNA/administração & dosagem , Ésteres , Feminino , Expressão Gênica , Injeções Intramusculares , Luciferases/genética , Camundongos , Camundongos Endogâmicos BALB C , Músculo Esquelético/metabolismo , Compostos Organofosforados/toxicidadeRESUMO
Novel temperature-sensitive poly(N-isopropylacrylamide)/amine-terminated polyamidoamine dendrimer G6-NH2 hydrogels with fast responsive properties were synthesized by forming semi-interpenetrating polymeric networks. In contrast to the conventional PNIPA hydrogel, these new gels showed rapid shrinking rate at the temperature above lower critical solution temperature (LCST), and exhibited higher equilibrium swelling ratio at room temperature. All these properties might be attributed to the incorporation of polyamidoamine dendrimer G6-NH2, which forms water-releasing channels and increases the hydrophilicity of PNIPA network. The novel hydrogels have potential applications in drug and gene delivery.
Assuntos
Resinas Acrílicas , Aminas , Hidrogéis , Resinas Acrílicas/química , Aminas/química , Hidrogéis/química , Cinética , Temperatura , Água/químicaRESUMO
Poly(vinyl alcohol) (PVA) and polyamidoamine (PAMAM) dendrimers are water-soluble, biocompatible and biodegradable polymers, which have been widely applied in biomedical fields. In this paper, novel physically cross-linked hydrogels composed of PVA and amine-terminated PAMAM dendrimer G6-NH(2) were prepared by cyclic freezing/thawing treatment of aqueous solutions containing PVA and G6-NH(2). The FT-IR analysis and elemental analysis indicated that PAMAM dendrimer G6-NH(2) was successfully introduced into the formed hydrogels, possibly via hydrogen bonds among hydroxyl groups, amide groups and amino groups in PVA and PAMAM dendrimer in the process of freezing-thawing cycle. Compared with physically cross-linked PVA hydrogel, PVA/G6-NH(2) hydrogels show higher swelling ratios and faster re-swelling rate due to the higher hydrophilicity of PAMAM dendrimer G6-NH(2). Higher contents of G6-NH(2) in PVA/G6-NH(2) hydrogels resulted in higher swelling ratios and faster re-swelling rates. With increasing freezing/thawing cyclic times, the swelling ratios and re-swelling rates of PVA/G6-NH(2) hydrogels decreased, which is similar to that of physically cross-linked PVA hydrogel. Combining the special host property of polyamidoamine dendrimer, these novel physically cross-linked hydrogels are expected to have potential use in drug delivery, including improving drug-loading amounts in hydrogels and prolonging drug release time. Swelling ratios of physically cross-linked PVA/G6-NH(2)-50 hydrogels prepared by three, six, nine freezing/thawing cycles. The swelling equilibrium experiments were carried out in distilled water at 25 degrees C.
Assuntos
Hidrogéis/química , Hidrogéis/síntese química , Nylons/química , Poliaminas/química , Álcool de Polivinil/química , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/química , Reagentes de Ligações Cruzadas/síntese química , Reagentes de Ligações Cruzadas/química , Concentração de Íons de Hidrogênio , Teste de MateriaisRESUMO
Reduction-sensitive micelles were prepared from monomethoxy-poly(ethylene glycol)-S-S-hexadecyl (mPEG-S-S-C16), an amphiphilic poly(ethylene glycol) derivative containing a disulfide bond. The micelles were then used for the intracellular delivery of the anticancer drug doxorubicin (DOX) into tumor cells, and the cellular uptake mechanisms of the micelles were determined. To serve as a control, monomethoxy-poly(ethylene glycol)-C-C-hexadecyl (mPEG-C-C-C16) with an analogous structure but without a disulfide bond was also prepared. The polymer could self-assemble into micelles in an aqueous solution and be loaded with high-content DOX. In vitro release studies revealed that DOX-loaded mPEG-S-S-C16 micelles released DOX faster than DOX-loaded mPEG-C-C-C16 micelles in the presence of dithiothreitol (DTT), but showed similar release rates in the absence of DTT. MTT assay demonstrated significantly enhanced cytotoxicity of DOX-loaded mPEG-S-S-C16 micelles against the human cervical cancer cells (HeLa) compared with DOX-loaded mPEG-C-C-C16 micelles, but there was no significant difference in the cytotoxicity between the two DOX-loaded micelles against the african green monkey SV40-transformed kidney fibroblast cells (COS-7). Confocal laser scanning microscopy observation and flow cytometry analyses indicated that DOX-loaded mPEG-S-S-C16 micelles were efficiently internalized into HeLa cells, released DOX into the cytoplasm, and entered the nuclei. By contrast, in the case of DOX-loaded mPEG-C-C-C16 micelles, little DOX was found in the nuclei. Endocytosis inhibition results proved that both mPEG-S-S-C16 and mPEG-C-C-C16 micelles entered the HeLa cells mainly through the clathrin-mediated endocytosis pathway, and caveolae-mediated endocytosis was involved to a small extent. These results indicated that the different behaviors of cell uptake between reduction-sensitive and -insensitive micelles may occur after the micelles were internalized into the cells, but not during endocytosis, and the potential of this reduction-sensitive polymer for the effective intracellular delivery of anticancer drugs.
Assuntos
Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Endocitose/efeitos dos fármacos , Espaço Intracelular/metabolismo , Micelas , Animais , Transporte Biológico/efeitos dos fármacos , Células COS , Morte Celular/efeitos dos fármacos , Chlorocebus aethiops , Citometria de Fluxo , Células HeLa , Humanos , Espaço Intracelular/efeitos dos fármacos , Luz , Espectroscopia de Ressonância Magnética , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Oxirredução/efeitos dos fármacos , Tamanho da Partícula , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Espalhamento de Radiação , Tensoativos/síntese química , Tensoativos/química , Resultado do TratamentoRESUMO
Three poly(amidoamine)s with pendant primary amines were synthesized by Michael polyaddition of a diamine to N,N-methylenebis(acrylamide). The poly(amidoamine)s with primary and tertiary amines possess a high buffer capacity between pH = 5 and 7, which facilitates the escape of polymer/DNA complexes from endosomes according to the 'proton sponge hypothesis'. In vitro cytotoxicity of poly(amidoamine)s depends on the side-chain structure of the polymer. Polymers 1a-1c can condense plasmid DNA by electrostatic interactions to form nanosized polyelectrolyte complexes with a positive surface charge. In vitro transfection results indicate that the transfection efficiency of polymer 1b under optimized conditions is comparable to that of commercial branched PEI 25,000.