RESUMO
Iron oxide nanoparticles coated with dextran were synthesized via four variations on the co-precipitation method. The methods ranged from in situ formation of the nanoparticles within the dextran solution to the adsorption of dextran to the nanoparticle surface following nucleation and extensive washing. The timing of the addition of dextran into the reaction mixture was found to greatly influence the physical and chemical properties of the magnetic nanoparticles. Batches of dextran coated iron oxide nanoparticles were synthesized by each method in triplicate, and the nanoparticles were further crosslinked with epichlorohydrin. The properties of the nanoparticles such as size, percentage of dextran coating, stability in solution, crystallinity, and magnetic properties were evaluated. The simultaneous semi-two-step method injected the reducing agent and the dextran solution into the reaction vessel at the same time. This method resulted in the greatest batch-to-batch reproducibility of nanoparticle properties and the least variation in nanoparticles synthesized in the same batch. The two-step method resulted in the greatest variation of the characteristics examined between batches. The one-step method was synthesized with both five grams and one gram of dextran to investigate the effects of solution viscosity on the resulting nanoparticle characteristics. The one-step method with five grams of dextran resulted in nanoparticles with significantly smaller crystal sizes (5.4 ± 1.9 nm) and lower specific adsorption rate (SAR) values (138.4 ± 13.6 W/g) in an alternating magnetic field (58 kA/m, 292 kHz). However, this method resulted in nanoparticles that were very stable in PBS over 12 hours, which is most likely due to the greater dextran coating (60.0 ± 2.7 weight percent). For comparison, the simultaneous semi-two-step method generated nanoparticles 179.2 ± 18.3 nm in diameter (crystal size 12.1 ± 0.2 nm) containing 18.3 ± 1.2 weight percent dextran with a SAR value of 321.1 ± 137.3 W/g.
RESUMO
Inhalable lung surfactant-based carriers composed of synthetic phospholipids, dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG), along with paclitaxel (PTX), were designed and optimized as respirable dry powders using organic solution co-spray-drying particle engineering design. These materials can be used to deliver and treat a wide variety of pulmonary diseases with this current work focusing on lung cancer. In particular, this is the first time dry powder lung surfactant-based particles have been developed and characterized for this purpose. Comprehensive physicochemical characterization was carried out to analyze the particle morphology, surface structure, solid-state transitions, amorphous character, residual water content, and phospholipid bilayer structure. The particle chemical composition was confirmed using attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) spectroscopy. PTX loading was high, as quantified using UV-VIS spectroscopy, and sustained PTX release was measured over weeks. In vitro cellular characterization on lung cancer cells demonstrated the enhanced chemotherapeutic cytotoxic activity of paclitaxel from co-spray-dried DPPC/DPPG (co-SD DPPC/DPPG) lung surfactant-based carrier particles and the cytotoxicity of the particles via pulmonary cell viability analysis, fluorescent microscopy imaging, and transepithelial electrical resistance (TEER) testing at air-interface conditions. In vitro aerosol performance using a Next Generation Impactor™ (NGI™) showed measurable powder deposition on all stages of the NGI and was relatively high on the lower stages (nanometer aerodynamic size). Aerosol dispersion analysis of these high-performing DPIs showed mass median diameters (MMADs) that ranged from 1.9 to 2.3 µm with excellent aerosol dispersion performance as exemplified by high values of emitted dose, fine particle fractions, and respirable fractions.
Assuntos
1,2-Dipalmitoilfosfatidilcolina/química , Antineoplásicos/administração & dosagem , Portadores de Fármacos , Inaladores de Pó Seco , Neoplasias Pulmonares/tratamento farmacológico , Paclitaxel/administração & dosagem , Fosfatidilgliceróis/química , Administração por Inalação , Aerossóis , Antineoplásicos/química , Varredura Diferencial de Calorimetria , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Química Farmacêutica , Cristalografia por Raios X , Preparações de Ação Retardada , Relação Dose-Resposta a Droga , Impedância Elétrica , Desenho de Equipamento , Humanos , Neoplasias Pulmonares/patologia , Microscopia de Fluorescência , Paclitaxel/química , Tamanho da Partícula , Difração de Pó , Pós , Solubilidade , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Tecnologia Farmacêutica/métodos , Fatores de TempoRESUMO
Past work has shown that Treponema pallidum, the causative agent of syphilis, binds host fibronectin (FN). FN and other host proteins are believed to bind to rare outer membrane proteins (OMPs) of T. pallidum, and it is postulated that this interaction may facilitate cell attachment and mask antigenic targets on the surface. This research seeks to prepare a surface capable of mimicking the FN binding ability of T. pallidum in order to investigate the impact of FN binding with adsorbed Tp0483 on the host response to the surface. By understanding this interaction, it may be possible to develop more effective treatments for infection and possibly mimic the stealth properties of the bacteria. Functionalized self-assembled monolayers (SAMs) on gold were used to investigate rTp0483 and FN adsorption. Using a quartz crystal microbalance (QCM), rTp0483 adsorption and subsequent FN adsorption onto rTp0483 were determined to be higher on negatively charged carboxylate-terminated self-assembled monolayers (-COO(-) SAMs) compared to the other surfaces analyzed. Kinetic analysis of rTp0483 adsorption using surface plasmon resonance (SPR) supported this finding. Kinetic analysis of FN adsorption using SPR revealed a multistep event, where the concentration of immobilized rTp0483 plays a role in FN binding. An examination of relative QCM dissipation energy compared to the shift in frequency showed a correlation between the physical properties of adsorbed rTp0483 and SAM surface chemistry. In addition, AFM images of rTp0483 on selected SAMs illustrated a preference of rTp0483 to bind as aggregates. Adsorption on -COO(-) SAMs was more uniform across the surface, which may help further explain why FN bound more strongly. rTp0483 antibody studies suggested the involvement of amino acids 274-289 and 316-333 in binding between rTp0483 to FN, while a peptide blocking study only showed inhibition of binding with amino acids 316-333. Finally, surface adsorbed rTp0483 with FN bound significantly less anti-RGD and gelatin compared to FN adsorbed directly to -COO(-) SAMs, indicating that one or both binding regions may play a role in binding between rTp0483 and FN.
Assuntos
Adesinas Bacterianas/metabolismo , Fibronectinas/metabolismo , Membranas Artificiais , Adsorção , Sítios de Ligação , Fibronectinas/sangue , Ouro/química , Humanos , Cinética , Técnicas de Microbalança de Cristal de Quartzo , Propriedades de SuperfícieRESUMO
Molecular switches are designer molecules that combine the functionality of two individual proteins into one, capable of manifesting an "on/off" signal in response to a stimulus. These switches have unique properties and functionalities and thus, can be employed as nanosensors in a variety of applications. To that end, we have developed a bioluminescent molecular switch for cyclic AMP. Bioluminescence offers many advantages over fluorescence and other detection methods including the fact that there is essentially zero background signal in physiological fluids, allowing for more sensitive detection and monitoring. The switch was created by combining the properties of the cyclic AMP receptor protein (CRP), a transcriptional regulatory protein from E. Coli that binds selectively to cAMP with those of aequorin, a bioluminescent photoprotein native of the jellyfish Aequorea victoria . Genetic manipulation to split the genetic coding sequence of aequorin in two and genetically attach the fragments to the N and C termini of CRP resulted in a hybrid protein molecular switch. The conformational change experienced by CRP upon the binding of cyclic AMP is suspected to result in the observed loss of the bioluminescent signal from aequorin. The "on/off" bioluminescence can be modulated by cyclic AMP over a range of several orders of magnitude in a linear fashion in addition to the capacity to detect changes in cellular cyclic AMP of intact cells exposed to different external stimuli without the need to lyse the cells. We envision that the molecular switch could find applications in vitro as well as In Vivo cyclic AMP detection and/or imaging.
Assuntos
Equorina/genética , Técnicas Biossensoriais/métodos , AMP Cíclico/metabolismo , Engenharia de Proteínas/métodos , Receptores de AMP Cíclico/genética , Proteínas Recombinantes de Fusão/genética , Equorina/química , Equorina/metabolismo , Animais , AMP Cíclico/farmacologia , Relação Dose-Resposta a Droga , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Hidrozoários , Medições Luminescentes , Modelos Moleculares , Conformação Proteica , Receptores de AMP Cíclico/química , Receptores de AMP Cíclico/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismoRESUMO
Angiogenesis plays cardinal role in normal developmental processes as well as in numerous pathologies. Multiple cytokines are released and act simultaneously to activate endothelial cells in vivo. The present study investigated the relative ability of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), tumor necrosis factor-alpha (TNF-alpha) in modulating cell monolayer permeability, migration, proliferation and tube formation individually and in combination. While the common methods for assaying angiogenesis were conducted for studying cell migration, proliferation and differentiation, endothelial cell monolayer permeability studies were carried out using a whole cell-based biosensor. The biosensor, consisting of a confluent monolayer of human umbilical vein endothelial cells (HUVECs) on a potassium ion-selective electrode, takes advantage of cell monolayer permeability dysfunction to detect the presence of small quantities of cytokines. When a confluent monolayer of cells was formed on the membrane surface, the response of the electrode toward the marker ion, potassium, was inhibited. The response obtained after exposing this sensor to different cytokines for 1 and 3h, can be attributed to the modulation of monolayer permeability by these cytokines. The present study demonstrated that at the concentrations experimented with, the relative change in permeability assay in the presence of cytokines compared to the control was much higher than that observed in other assays, thereby bolstering the potential of the biosensor to act as a quick screening tool for angiogenesis.
Assuntos
Bioensaio/instrumentação , Técnicas Biossensoriais/instrumentação , Permeabilidade da Membrana Celular/fisiologia , Eletroquímica/instrumentação , Células Endoteliais/fisiologia , Neovascularização Fisiológica/fisiologia , Bioensaio/métodos , Técnicas Biossensoriais/métodos , Células Cultivadas , Eletroquímica/métodos , Células Endoteliais/citologia , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Angiogenesis occurs during the early phase of cancer. Recruitment of new blood vessels by existing cancer cells leads to the release of higher concentrations of cytokines as compared to cells in healthy individuals. Some of the common cytokines observed at higher concentrations, such as vascular endothelial growth factor, basic fibroblast growth factor, hepatocyte growth factor and tumor necrosis factor-alpha, are also known to induce increased permeability across an endothelial cell monolayer. A whole-cell-based biosensor has been developed that can detect the presence of small quantities of the abovementioned cytokines individually and in different combinations. It was observed that the biosensor could differentiate between the cytokine concentrations observed in the sera of healthy individuals and cancer patients. The biosensor was also evaluated by exposing it to actual serum. These results demonstrated that the sensor can distinguish between healthy individuals and cancer patients and that the corresponding biosensor responses correlate with the stages of cancer.
Assuntos
Proteínas Angiogênicas/sangue , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Eletrodos Seletivos de Íons , Programas de Rastreamento , Estadiamento de Neoplasias , Neoplasias/diagnóstico , Biomarcadores Tumorais/sangue , Neoplasias da Mama/diagnóstico , Membrana Celular/fisiologia , Feminino , Humanos , Neoplasias Hepáticas/diagnóstico , Masculino , Neoplasias Ovarianas/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Permeabilidade , Padrões de Referência , Sensibilidade e EspecificidadeRESUMO
AIM: In the present study, we examine the effects of internalized peptide-conjugated iron oxide nanoparticles and their ability to locally convert alternating magnetic field (AMF) energy into other forms of energy (e.g., heat and rotational work). MATERIALS & METHODS: Dextran-coated iron oxide nanoparticles were functionalized with a cell penetrating peptide and after internalization by A549 and H358 cells were activated by an AMF. RESULTS: TAT-functionalized nanoparticles and AMF exposure increased reactive oxygen species generation compared with the nanoparticle system alone. The TAT-functionalized nanoparticles induced lysosomal membrane permeability and mitochondrial membrane depolarization, but these effects were not further enhanced by AMF treatment. Although not statistically significant, there are trends suggesting an increase in apoptosis via the Caspase 3/7 pathways when cells are exposed to TAT-functionalized nanoparticles combined with AMF. CONCLUSION: Our results indicate that internalized TAT-functionalized iron oxide nanoparticles activated by an AMF elicit cellular responses without a measurable temperature rise.
Assuntos
Neoplasias Pulmonares/terapia , Nanopartículas de Magnetita/uso terapêutico , Fragmentos de Peptídeos/uso terapêutico , Produtos do Gene tat do Vírus da Imunodeficiência Humana/uso terapêutico , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Caspase 7/metabolismo , Linhagem Celular Tumoral , Humanos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Lisossomos/metabolismo , Lisossomos/patologia , Campos Magnéticos , Magnetismo , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/ultraestrutura , Fragmentos de Peptídeos/química , Espécies Reativas de Oxigênio/metabolismo , Produtos do Gene tat do Vírus da Imunodeficiência Humana/químicaRESUMO
Cancer and the inflammatory system share a complex intertwined relationship. For instance, in response to an injury or stress, vascular endothelial cells will express cell adhesion molecules as a means of recruiting leukocytes. However, circulating tumor cells (CTCs) have been shown to highjack this expression for the adhesion and invasion during the metastatic cascade. As such, the initiation of endothelial cell inflammation, either by surgical procedures (cancer resection) or chemotherapy can inadvertently increase the metastatic potential of CTCs. Yet, systemic delivery of anti-inflammatories, which weaken the entire immune system, may not be preferred in some treatment settings. In this work, we demonstrate that a long-term releasing flavone-based polymer and subsequent nanoparticle delivery system can inhibit tumor cell adhesion, through the suppression of endothelial cell adhesion molecule expression. The degradation of a this anti-inflammatory polymer provides longer term, localized release profile of active therapeutic drug in nanoparticle form as compared with that of the free drug, permitting more targeted anti-metastatic therapies. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1438-1447, 2016.
Assuntos
Apigenina/farmacologia , Plásticos Biodegradáveis/farmacologia , Neoplasias da Mama/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Apigenina/química , Plásticos Biodegradáveis/química , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Metástase NeoplásicaRESUMO
Three-dimensional (3D) lung multicellular spheroids (MCS) in liquid-covered culture (LCC) and air-interface culture (AIC) conditions have both been developed for the evaluation of aerosol anticancer therapeutics in solution and aerosols, respectively. The MCS were formed by seeding lung cancer cells on top of collagen where they formed spheroids due to the prevalence of cell-to-cell interactions. LCC MCS were exposed to paclitaxel (PTX) in media whereas AIC MCS were exposed to dry powder PEGylated phospholipid aerosol microparticles containing paclitaxel. The difference in viability for 2D versus 3D culture for both LCC and AIC was evaluated along with the effects of the particles on lung epithelium via transepithelial electrical resistance (TEER) measurements. For LCC and AIC conditions, the 3D spheroids were more resistant to treatment with higher IC50 values for A549 and H358 cell lines. TEER results initially indicated a decrease in resistance upon drug or particle exposure, however, these values increased over the course of several days indicating the ability of the cells to recover. Overall, these studies offer a comprehensive in vitro evaluation of aerosol particles used in the treatment of lung cancer while introducing a new method for culturing lung cancer MCS in both LCC and AIC conditions.
Assuntos
Técnicas de Cultura de Células/métodos , Neoplasias Pulmonares/patologia , Esferoides Celulares/patologia , Aerossóis , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Avaliação de Medicamentos , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Paclitaxel/farmacologia , Esferoides Celulares/efeitos dos fármacosRESUMO
To increase the efficacy of radiation, iron oxide nanoparticles can be utilized for their ability to produce reactive oxygen species (ROS). Radiation therapy promotes leakage of electrons from the electron transport chain and leads to an increase in mitochondrial production of the superoxide anion which is converted to hydrogen peroxide by superoxide dismutase. Iron oxide nanoparticles can then catalyze the reaction from hydrogen peroxide to the highly reactive hydroxyl radical. Therefore, the overall aim of this project was to utilize iron oxide nanoparticles conjugated to a cell penetrating peptide, TAT, to escape lysosomal encapsulation after internalization by cancer cells and catalyze hydroxyl radical formation. It was determined that TAT functionalized iron oxide nanoparticles and uncoated iron oxide nanoparticles resulted in permeabilization of the lysosomal membranes. Additionally, mitochondrial integrity was compromised when A549 cells were treated with both TAT-functionalized nanoparticles and radiation. Pre-treatment with TAT-functionalized nanoparticles also significantly increased the ROS generation associated with radiation. A long term viability study showed that TAT-functionalized nanoparticles combined with radiation resulted in a synergistic combination treatment. This is likely due to the TAT-functionalized nanoparticles sensitizing the cells to subsequent radiation therapy, because the nanoparticles alone did not result in significant toxicities.
Assuntos
Compostos Férricos/administração & dosagem , Óxido Ferroso-Férrico/administração & dosagem , Produtos do Gene tat/farmacocinética , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/radioterapia , Radiossensibilizantes/administração & dosagem , Radioterapia Conformacional/métodos , Células A549 , Sobrevivência Celular/efeitos da radiação , Produtos do Gene tat/administração & dosagem , Humanos , Terapia de Alvo Molecular/métodos , Nanocápsulas/administração & dosagem , Nanocápsulas/química , Neoplasias Experimentais/patologia , Tolerância a Radiação , Espécies Reativas de Oxigênio/metabolismo , Resultado do TratamentoRESUMO
This review highlights the state-of-the-art in the application of magnetic nanoparticles (MNPs) and their composites for remote controlled therapies. Novel macro- to nano-scale systems that utilize remote controlled drug release due to actuation of MNPs by static or alternating magnetic fields and magnetic field guidance of MNPs for drug delivery applications are summarized. Recent advances in controlled energy release for thermal therapy and nanoscale energy therapy are addressed as well. Additionally, studies that utilize MNP-based thermal therapy in combination with other treatments such as chemotherapy or radiation to enhance the efficacy of the conventional treatment are discussed.
Assuntos
Sistemas de Liberação de Medicamentos , Nanocompostos/administração & dosagem , Nanopartículas/administração & dosagem , Robótica , Animais , Terapia Combinada , Tratamento Farmacológico , Humanos , Fenômenos Magnéticos , Nanocompostos/uso terapêutico , Nanopartículas/uso terapêuticoRESUMO
The surfaces of iron oxide nanoparticles are capable of catalytically generating reactive oxygen species (ROS) through the Fenton and Haber-Weiss reactions. Fenton chemistry has been shown to be temperature dependent with an increase in activity up to 40 °C and then a decrease above this temperature as the hydrogen peroxide degrades into oxygen and water which limits the reaction. When exposed to an alternating magnetic field (AMF), iron oxide nanoparticles absorb the energy from the magnetic field and convert it into heat. In this study, we observed an increase in the degradation of methylene blue when a suspension of magnetite nanoparticles (Fe3O4) was exposed to an AMF indicating there was an increase in the ROS generation in response to the AMF. The increase in ROS generation compared to the Arrhenius prediction was both time and concentration dependent; in which we observed a decrease in ROS enhancement with increased time of exposure and concentration. We postulate that the decrease is due to agglomeration in the presence of the field. As the nanoparticles agglomerate, there is a decrease in surface area per mass limiting the reaction rate.
RESUMO
Monosaccharide coated iron oxide nanoparticles were developed to selectively target colon cancer cell lines for magnetically mediated energy delivery therapy. The nanoparticles were prepared using a coupling reaction to attach the glucose functional group to the iron oxide core, and functionality was confirmed with physicochemical characterization techniques. The targeted nanoparticles were internalized into CT26 cells at a greater extent than non-targeted nanoparticles, and the nanoparticles were shown to be localized within lysosomes. Cells with internalized nanoparticles were exposed to an AMF to determine the potential to delivery therapy. Cellular ROS generation and apoptotic cell death was enhanced with field exposure. The nanoparticle coatings inhibit the Fenton-like surface generation of ROS suggesting a thermal or mechanical effect is more likely the source of the intracellular effect, unless the nanoparticle coating is unstable in the cellular environment. STATEMENT OF SIGNIFICANCE: This is the first study to assess glucose coated MNPs for the delivery of MagMED therapy. With exposure of an AMF, the glucose-coated nanoparticles displayed a significant increase in cellular ROS and apoptotic cell death with no measurable increase in media temperature. To determine the mechanism of toxicity, we investigated the surface generation of ROS through Fenton-like chemistry. The coated systems displayed negligible ROS generation compared to uncoated nanoparticles. These observations suggest the cellular ROS measured is attributed to a thermal or mechanical effect of the internalized nanoparticles. In summary, this manuscript reports on some new insights as to the mechanism of MagMED therapies, which are of high interest to the biomaterials and cancer nanomedicine fields.
Assuntos
Campos Magnéticos , Nanopartículas de Magnetita/química , Espécies Reativas de Oxigênio/metabolismo , Animais , Caspase 3/metabolismo , Caspase 7/metabolismo , Morte Celular , Linhagem Celular Tumoral , Ácido Cítrico/química , Difusão Dinâmica da Luz , Endocitose , Glucose/química , Peróxido de Hidrogênio/química , Ferro/química , Ferro/metabolismo , Camundongos , Monossacarídeos/química , Espectroscopia de Infravermelho com Transformada de Fourier , Eletricidade EstáticaRESUMO
One of the current challenges in the systemic delivery of nanoparticles in cancer therapy applications is the lack of effective tumor localization. Iron oxide nanoparticles (IONPs) coated with crosslinked dextran were functionalized with the tumor-homing peptide CREKA, which binds to fibrinogen complexes in the extracellular matrix of tumors. This allows for the homing of these nanoparticles to tumor tissue. The IONP core allows for particle heating upon exposure to an alternating magnetic field (AMF), while the dextran coating stabilizes the particles in suspension and decreases the cytotoxicity of the system. Magnetically mediated hyperthermia (MMH) allows for the heating of tumor tissue to increase the efficacy of traditional cancer treatments using IONPs. While MMH provides the opportunity for localized heating, this method is currently limited by the lack of particle penetration into tumor tissue, even after effective targeted delivery to the tumor site. The CREKA-conjugated nanoparticles presented were characterized for their size, stability, heating capabilities and biocompatibility. The particles had a hydrated diameter of 52nm, were stable in phosphate buffered saline solution and media with 10% v/v fetal bovine serum over at least 12h, and generated enough heat to raise solution temperatures well into the hyperthermia range (41-45°C) when exposed to an AMF, owing to an average specific absorption rate of 83.5Wg(-1). Cytotoxicity studies demonstrated that the particles have low cytotoxicity over long exposure times at low concentrations. A fibrinogen clotting assay was used to determine the binding affinity of CREKA-conjugated particles, which was significantly greater than the binding affinity of dextran, only coated IONPs demonstrating the potential for this particle system to effectively home to a variety of tumor locations. Finally, it was shown that in vitro MMH increased the effects of cisplatin compared with cisplatin or MMH treatments alone.
Assuntos
Compostos Férricos/química , Hipertermia Induzida/métodos , Nanopartículas Metálicas , Oligopeptídeos/administração & dosagem , Microscopia Eletrônica de Transmissão , Oligopeptídeos/síntese química , Espectrofotometria Ultravioleta , TermogravimetriaRESUMO
In this study, core-shell nanoparticles were developed to achieve thermal therapy that can ablate cancer cells in a remotely controlled manner. The core-shell nanoparticles were prepared using atomic transfer radical polymerization (ATRP) to coat iron oxide (Fe3O4) nanoparticles with a poly(ethylene glycol) (PEG) based polymer shell. The iron oxide core allows for the remote heating of the particles in an alternating magnetic field (AMF). The coating of iron oxide with PEG was verified through Fourier transform infrared spectroscopy and thermal gravimetric analysis. A thermoablation (55°C) study was performed on A549 lung carcinoma cells exposed to nanoparticles and over a 10 min AMF exposure. The successful thermoablation of A549 demonstrates the potential use of polymer coated particles for thermal therapy.
Assuntos
Materiais Biocompatíveis/síntese química , Óxido Ferroso-Férrico/química , Nanopartículas de Magnetita/química , Polietilenoglicóis/química , Polímeros/química , Animais , Materiais Biocompatíveis/uso terapêutico , Materiais Biocompatíveis/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Hipertermia Induzida , Neoplasias Pulmonares/terapia , Campos Magnéticos , Nanopartículas de Magnetita/toxicidade , Camundongos , Células NIH 3T3 , Polímeros/síntese química , TemperaturaRESUMO
Pulmonary inhalation chemotherapeutic drug delivery offers many advantages for lung cancer patients in comparison to conventional systemic chemotherapy. Inhalable particles are advantageous in their ability to deliver drug deep in the lung by utilizing optimally sized particles and higher local drug dose delivery. In this work, spray-dried and co-spray dried inhalable lung surfactant-mimic PEGylated lipopolymers as microparticulate/nanoparticulate dry powders containing paclitaxel were rationally designed via organic solution advanced spray drying (no water) in closed-mode from dilute concentration feed solution. Dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylethanolamine poly(ethylene glycol) (DPPE-PEG) with varying PEG chain length were mixed with varying amounts of paclitaxel in methanol to produce co-spray dried microparticles and nanoparticles. Scanning electron microscopy showed the spherical particle morphology of the inhalable particles. Thermal analysis and X-ray powder diffraction confirmed the retention of the phospholipid bilayer structure in the solid-state following spray drying, the degree of solid-state molecular order, and solid-state phase transition behavior. The residual water content of the particles was very low as quantified analytically Karl Fisher titration. The amount of paclitaxel loaded into the particles was quantified which indicated high encapsulation efficiencies (43-99%). Dry powder aerosol dispersion performance was measured in vitro using the Next Generation Impactor (NGI) coupled with the Handihaler dry powder inhaler device and showed mass median aerodynamic diameters in the range of 3.4-7 µm. These results demonstrate that this novel microparticulate/nanoparticulate chemotherapeutic PEGylated phospholipid dry powder inhalation aerosol platform has great potential in lung cancer drug delivery.
Assuntos
1,2-Dipalmitoilfosfatidilcolina/química , Antineoplásicos Fitogênicos/química , Inaladores de Pó Seco , Paclitaxel/química , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Tensoativos/química , Administração por Inalação , Aerossóis , Antineoplásicos Fitogênicos/administração & dosagem , Dessecação/métodos , Neoplasias Pulmonares/tratamento farmacológico , Paclitaxel/administração & dosagem , Tamanho da Partícula , Tecnologia FarmacêuticaRESUMO
Hyperthermia, the heating of tissue from 41 to 45 °C, has been shown to improve the efficacy of cancer therapy when used in conjunction with irradiation and/or chemotherapy. In this work, hydrogel nanocomposites have been developed that can control the delivery of both heat and a chemotherapeutic agent (e.g. paclitaxel). The nanocomposites studied involve a stealth, poly(ethylene glycol) (PEG)-based system comprised of PEG (n = 1000) methyl ether methacrylate and PEG (n = 400) dimethacrylate with iron oxide nanoparticles physically entrapped within the hydrogel matrices. The capability of the hydrogel nanocomposites to be heated in an alternating magnetic field was demonstrated. The heating of the hydrogel systems was dependent on the crosslinking of the hydrogel network where hydrogels with lower swelling ratios were found to heat to a greater extent than those with higher ratios. In addition, paclitaxel was shown to exhibit non-Fickian release from the hydrogel systems, with the amount of drug released dependent on the hydrogel network structure. Three cell lines: M059K (glioblastoma), MDA MB 231 (breast carcinoma), and A549 (lung adenocarcinoma) were exposed to paclitaxel only, hyperthermia only, and both paclitaxel and hyperthermia to determine if a synergistic cytotoxic effect was possible for these cell lines. The efficacy of paclitaxel was greater with hyperthermia for the A549 cells; however, the M059K and MDA MB 231 did not show the same response.
Assuntos
Portadores de Fármacos , Compostos Férricos/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hipertermia Induzida/métodos , Nanopartículas/uso terapêutico , Paclitaxel/administração & dosagem , Polietilenoglicóis/química , Antineoplásicos/administração & dosagem , Linhagem Celular Tumoral , Portadores de Fármacos/síntese química , Portadores de Fármacos/química , Temperatura Alta/uso terapêutico , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/síntese química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacocinética , Hipertermia Induzida/instrumentação , Nanopartículas/administração & dosagem , Nanopartículas/química , Paclitaxel/farmacocinéticaRESUMO
The biomedical use of superparamagnetic iron oxide nanoparticles has been of continued interest in the literature and clinic. Their ability to be used as contrast agents for imaging and/or responsive agents for remote actuation makes them exciting materials for a wide range of clinical applications. Recently, however, concern has arisen regarding the potential health effects of these particles. Iron oxide toxicity has been demonstrated in in vivo and in vitro models, with oxidative stress being implicated as playing a key role in this pathology. One of the key cell types implicated in this injury is the vascular endothelial cells. Here, we report on the development of a targeted polymeric antioxidant, poly(trolox ester), nanoparticle that can suppress oxidative damage. As the polymer undergoes enzymatic hydrolysis, active trolox is locally released, providing a long term protection against pro-oxidant agents. In this work, poly(trolox) nanoparticles are targeted to platelet endothelial cell adhesion molecules (PECAM-1), which are able to bind to and internalize in endothelial cells and provide localized protection against the cytotoxicity caused by iron oxide nanoparticles. These results indicate the potential of using poly(trolox ester) as a means of mitigating iron oxide toxicity, potentially expanding the clinical use and relevance of these exciting systems.
Assuntos
Antioxidantes/uso terapêutico , Cromanos/uso terapêutico , Compostos Férricos/toxicidade , Nanopartículas/toxicidade , Nanopartículas/uso terapêutico , Polímeros/uso terapêutico , Antioxidantes/administração & dosagem , Antioxidantes/química , Cromanos/administração & dosagem , Cromanos/química , Compostos Férricos/química , Células Endoteliais da Veia Umbilical Humana , Humanos , Nanopartículas/administração & dosagem , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Polímeros/administração & dosagem , Polímeros/química , Espécies Reativas de Oxigênio/metabolismoRESUMO
Novel advanced spray-dried and co-spray-dried inhalable lung surfactant-mimic phospholipid and poly(ethylene glycol) (PEG)ylated lipopolymers as microparticulate/nanoparticulate dry powders of biodegradable biocompatible lipopolymers were rationally formulated via an organic solution advanced spray-drying process in closed mode using various phospholipid formulations and rationally chosen spray-drying pump rates. Ratios of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylethanolamine PEG (DPPE-PEG) with varying PEG lengths were mixed in a dilute methanol solution. Scanning electron microscopy images showed the smooth, spherical particle morphology of the inhalable particles. The size of the particles was statistically analyzed using the scanning electron micrographs and SigmaScan® software and were determined to be 600 nm to 1.2 µm in diameter, which is optimal for deep-lung alveolar penetration. Differential scanning calorimetry (DSC) and powder X-ray diffraction (PXRD) were performed to analyze solid-state transitions and long-range molecular order, respectively, and allowed for the confirmation of the presence of phospholipid bilayers in the solid state of the particles. The residual water content of the particles was very low, as quantified analytically via Karl Fischer titration. The composition of the particles was confirmed using attenuated total-reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy and confocal Raman microscopy (CRM), and chemical imaging confirmed the chemical homogeneity of the particles. The dry powder aerosol dispersion properties were evaluated using the Next Generation Impactor™ (NGI™) coupled with the HandiHaler® dry powder inhaler device, where the mass median aerodynamic diameter from 2.6 to 4.3 µm with excellent aerosol dispersion performance, as exemplified by high values of emitted dose, fine particle fraction, and respirable fraction. Overall, it was determined that the pump rates defined in the spray-drying process had a significant effect on the solid-state particle properties and that a higher pump rate produced the most optimal system. Advanced dry powder inhalers of inhalable lipopolymers for targeted dry powder inhalation delivery were successfully achieved.
Assuntos
1,2-Dipalmitoilfosfatidilcolina/química , Nanopartículas/química , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Administração por Inalação , Aerossóis/química , Varredura Diferencial de Calorimetria , Inaladores de Pó Seco , Microscopia , Nanomedicina , Tamanho da Partícula , Pós/química , Surfactantes Pulmonares/química , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Água/análise , Difração de Raios XRESUMO
Poly(ß-amino ester) (PBAE) biodegradable hydrogels were investigated for potential combined chemotherapeutic and heat delivery in the synergistic treatment of cancer. Hyperthermia, the heating of cancerous tissue from 41 to 45 °C, increases the efficacy of conventional cancer therapies such as irradiation and chemotherapy. The hydrogel nanocomposites in this work provide a drug delivery vehicle (via the biodegradable PBAE polymer network) and the ability to be heated remotely upon exposure to an alternating magnetic field (via iron oxide nanoparticles incorporated into the hydrogel matrix). PBAE macromers composed of poly(ethylene glycol) (N=400) diacrylate (PEG400DA) or diethylene glycol diacrylate (DEGDA) with isobutylamine (IBA) were synthesized. Hydrogel nanocomposites were fabricated via free-radical polymerization to form a bulk hydrogel matrix entrapping both iron oxide nanoparticles and paclitaxel. The 2EG-IBA hydrogel exhibited complete degradation after approximately 7 weeks whereas the 9EG-IBA hydrogel degraded completely in 11h. The hydrogels heated upon exposure to an alternating magnetic field throughout the degradation process. Additionally, the cytotoxicity of the degradation products was evaluated. Paclitaxel release was controlled via bulk degradation of the hydrogels. The tailorability of these nanocomposites makes them solid candidates for the synergistic treatment of cancer.