RESUMEN
Drug carriers typically require both stealth and targeting properties to minimize nonspecific interactions with healthy cells and increase specific interaction with diseased cells. Herein, the assembly of targeted poly(ethylene glycol) (PEG) particles functionalized with cyclic peptides containing Arg-Gly-Asp (RGD) (ligand) using a mesoporous silica templating method is reported. The influence of PEG molecular weight, ligand-to-PEG molecule ratio, and particle size on cancer cell targeting to balance stealth and targeting of the engineered PEG particles is investigated. RGD-functionalized PEG particles (PEG-RGD particles) efficiently target U-87 MG cancer cells under static and flow conditions in vitro, whereas PEG and cyclic peptides containing Arg-Asp-Gly (RDG)-functionalized PEG (PEG-RDG) particles display negligible interaction with the same cells. Increasing the ligand-to-PEG molecule ratio improves cell targeting. In addition, the targeted PEG-RGD particles improve cell uptake via receptor-mediated endocytosis, which is desirable for intracellular drug delivery. The PEG-RGD particles show improved tumor targeting (14% ID g-1) when compared with the PEG (3% ID g-1) and PEG-RDG (7% ID g-1) particles in vivo, although the PEG-RGD particles show comparatively higher spleen and liver accumulation. The targeted PEG particles represent a platform for developing particles aimed at balancing nonspecific and specific interactions in biological systems.
Asunto(s)
Sistemas de Liberación de Medicamentos , Neoplasias/tratamiento farmacológico , Oligopéptidos/farmacología , Polietilenglicoles/farmacología , Animales , Línea Celular Tumoral , Citoplasma/efectos de los fármacos , Endocitosis/efectos de los fármacos , Humanos , Ligandos , Oligopéptidos/química , Polietilenglicoles/química , Transducción de Señal/efectos de los fármacos , Dióxido de Silicio/química , Dióxido de Silicio/farmacología , Propiedades de SuperficieRESUMEN
In vitro experiments provide a solid basis for understanding the interactions between particles and biological systems. An important confounding variable for these studies is the difference between the amount of particles administered and that which reaches the surface of cells. Here, we engineer a hydrogel-based nanoparticle system and combine in situ characterization techniques, 3D-printed cell cultures, and computational modeling to evaluate and study particle-cell interactions of advanced particle systems. The framework presented demonstrates how sedimentation and diffusion can explain differences in particle-cell association, and provides a means to account for these effects. Finally, using in silico modeling, we predict the proportion of particles that reaches the cell surface using common experimental conditions for a wide range of inorganic and organic micro- and nanoparticles. This work can assist in the understanding and control of sedimentation and diffusion when investigating cellular interactions of engineered particles.
Asunto(s)
Simulación por Computador , Hidrogeles , Nanopartículas , Nanotecnología/métodos , Comunicación Celular , Difusión , Citometría de Flujo , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Microscopía Electrónica de Transmisión , Modelos Estadísticos , Tamaño de la Partícula , Impresión Tridimensional , Dióxido de Silicio , Propiedades de SuperficieRESUMEN
The quantification of nano- and microparticles is critical for diverse applications relying on the exact knowledge of the particle concentration. Although many techniques are available for counting particles, there are some limitations in regards to counting with low-scattering materials and facile counting in harsh organic solvents. Herein, we introduce an easy and rapid particle counting technique, termed "immobilized particle imaging" (IPI), to quantify fluorescent particles with different compositions (i.e., inorganic or organic), structures (i.e., solid, porous, or hollow), and sizes (50-1000 nm) dispersed in either aqueous or organic solutions. IPI is achieved by immobilizing particles of interest in a cell matrix-like scaffold (e.g., agarose) and imaging using standard microscopy techniques. Imaging a defined volume of the immobilized particles allows for the particle concentration to be calculated from the count numbers in a fixed volume. IPI provides a general and facile approach to quantify advanced nano- and microparticles, which may be helpful to researchers to obtain new insights for different applications (e.g., nanomedicine).
RESUMEN
Smart poly(2-oxazoline) (POx)-based multifunctional polymer capsules that specifically target glycoprotein (GP) IIb/IIIa on the surface of activated platelets are degraded by the serine protease thrombin and release the urokinase plasminogen activator loaded into the polymer capsules, only in the area of acute thrombosis.
Asunto(s)
Plaquetas/efectos de los fármacos , Plaquetas/fisiología , Portadores de Fármacos/química , Oxazoles/química , Activación Plaquetaria/efectos de los fármacos , Trombina/metabolismo , Secuencia de Aminoácidos , Cápsulas , Humanos , Oligopéptidos/química , Trombosis/fisiopatología , Activador de Plasminógeno de Tipo Uroquinasa/química , Activador de Plasminógeno de Tipo Uroquinasa/metabolismo , Activador de Plasminógeno de Tipo Uroquinasa/farmacologíaRESUMEN
Over the last ten years, there has been considerable research interest in the development of polymeric carriers for biomedicine. Such delivery systems have the potential to significantly reduce side effects and increase the bioavailability of poorly soluble therapeutics. The design of carriers has relied on harnessing specific variations in biological conditions, such as pH or redox potential, and more recently, by incorporating specific peptide cleavage sites for enzymatic hydrolysis. Although much progress has been made in this field, the specificity of polymeric carriers is still limited when compared with their biological counterparts. To synthesize the next generation of carriers, it is important to consider the biological rationale for materials design. This requires a detailed understanding of the cellular microenvironments and how these can be harnessed for specific applications. In this review, several important physiological cues in the cellular microenvironments are outlined, with a focus on changes in pH, redox potential, and the types of enzymes present in specific regions. Furthermore, recent studies that use such biologically inspired triggers to design polymeric carriers are highlighted, focusing on applications in the field of therapeutic delivery.
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Biología/métodos , Portadores de Fármacos , Diseño de Fármacos , Animales , Portadores de Fármacos/síntesis química , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Enzimas/metabolismo , Humanos , Oxidación-Reducción , Polímeros/química , Polímeros/metabolismoRESUMEN
Polymer microcapsules are of particular interest for applications including self-healing coatings, catalysis, bioreactions, sensing, and drug delivery. The primary way that polymer capsules can exhibit functionality relevant to these diverse fields is through the incorporation of functional cargo in the capsule cavity or wall. Diverse functional and therapeutic cargo can be loaded into polymer capsules with ease using polymer-stabilized calcium carbonate (CaCO3) particles. A variety of examples are demonstrated, including 15 types of cargo, yielding a toolbox with effectively 500+ variations. This process uses no harsh reagents and can take less than 30 min to prepare, load, coat, and form the hollow capsules. For these reasons, it is expected that the technique will play a crucial role across scientific studies in numerous fields.
RESUMEN
We report the preparation of polymer-peptide blend replica particles via the mesoporous silica (MS) templated assembly of poly(ethylene glycol)-block-poly(2-diisopropylaminoethyl methacrylate-co-2-(2-(2-(prop-2-ynyloxy)ethoxy)ethoxy)ethyl methacrylate) (PEG45-b-P(DPA55-co-PgTEGMA4)) and poly(l-histidine) (PHis). PEG45-b-P(DPA55-co-PgTEGMA4) was synthesized by atom transfer radical polymerization (ATRP), and was coinfiltrated with PHis into poly(methacrylic acid) (PMA)-coated MS particles assembled from different peptide-to-polymer ratios (1:1, 1:5, 1:10, or 1:15). Subsequent removal of the sacrificial templates and PMA resulted in monodisperse, colloidally stable, noncovalently cross-linked polymer-peptide blend replica particles that were stabilized by a combination of hydrophobic interactions between the PDPA and the PHis, hydrogen bonding between the PEG and PHis backbone, and π-π stacking of the imidazole rings of PHis side chains at physiological pH (pH â¼ 7.4). The synergistic charge-switchable properties of PDPA and PHis, and the enzymatic degradability of PHis, make these particles responsive to pH and enzymes. In vitro studies, in simulated endosomal conditions and inside cells, demonstrated that particle degradation kinetics could be engineered (from 2 to 8 h inside dendritic cells) based on simple adjustment of the peptide-to-polymer ratio used.
Asunto(s)
Polímeros/química , Animales , Línea Celular , Células Dendríticas/química , Histidina/química , Concentración de Iones de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Metacrilatos/química , Ratones , Tamaño de la Partícula , Polietilenglicoles/química , Dióxido de Silicio/síntesis químicaRESUMEN
We report the preparation of polymer particles via convective polymer assembly (CPA). Convection is used to move polymer solutions and cargo through an agarose gel that contains immobilized template particles. This method both coats and washes the particles in a process that is amenable to automation, and does not depend on passive diffusion or electrical currents, thus facilitating incorporation of fragile and nanoscale objects, such as liposomes and gold nanoparticles, into the thin polymer films. Template dissolution leads to the formation of stable polymer particles and capsules.
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Polymer microcapsules can be used as bioreactors and artificial cells; however, preparation methods for cell-like microcapsules are typically time-consuming, low yielding, and/or involve custom microfluidics. Here, we introduce a rapid (â¼30 min per batch, eight layers), scalable (up to 500 mg of templates), and efficient (98% yield) microcapsule preparation technique utilizing a fluidized bed for the layer-by-layer (LbL) assembly of polymers, and we investigate the parameters that govern the formation of robust capsules. Fluidization in water was possible for particles of comparable diameter to mammalian cells (>5 µm), with the experimental flow rates necessary for fluidization matching well with the theoretical values. Important variables for polymer film deposition and capsule formation were the concentration of polymer solution and the molecular weight of the polymer, while the volume of the polymer solution had a negligible impact. In combination, increasing the polymer molecular weight and polymer solution concentration resulted in improved film deposition and the formation of robust microcapsules. The resultant polymer microcapsules had a thickness of â¼5.5 nm per bilayer, which is in close agreement with conventionally prepared (quiescent (nonflow) adsorption/centrifugation/wash) LbL capsules. The technique reported herein provides a new way to rapidly generate microcapsules (approximately 8 times quicker than the conventional means), while being also amenable to scale-up and mass production.
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Cápsulas/química , Polímeros/química , AdsorciónRESUMEN
The engineering of layer-by-layer (LbL) hybrid click capsules that are responsive to biological stimuli is reported. The capsules comprise a pH-sheddable, non cross-linked outer coating that protects enzyme-cleavable inner layers. Upon cellular uptake, the outer coating is released and the capsules are enzymatically degraded. In vitro cell degradation results in rapid capsule degradation (10 min) upon cellular internalization.
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Cápsulas , Enzimas/metabolismo , Concentración de Iones de Hidrógeno , Química Clic , Microscopía Electrónica de Transmisión , Polímeros/químicaRESUMEN
We report a templating approach for the preparation of functional polymer replica particles via surface-initiated polymerization in mesoporous silica templates. Subsequent removal of the template resulted in discrete polymer particles. Furthermore, redox-responsive replica particles could be engineered to disassemble in a reducing environment. Particles, made of poly(methacryloyloxyethyl phosphorylcholine) (PMPC) or poly[oligo(ethylene glycol) methyl ether methacrylate] (POEGMA), exhibited very low association to human cancer cells (below 5%), which renders the reported charge-neutral polymer particles a modular and versatile class of highly functional carriers with potential applications in drug delivery.
Asunto(s)
Sistemas de Liberación de Medicamentos , Polímeros/química , Dióxido de Silicio/química , Diseño de Fármacos , Células HeLa , Humanos , Espectroscopía de Resonancia Magnética , Ensayo de Materiales , Metacrilatos/química , Microscopía Electrónica de Transmisión , Oxidación-Reducción , Tamaño de la Partícula , Fosforilcolina/análogos & derivados , Fosforilcolina/química , Polietilenglicoles/química , Ácidos Polimetacrílicos/química , Porosidad , Propiedades de Superficie , Agua/químicaRESUMEN
A new class of polymer capsules with an in-built endocytic pH-coupled fluorescence switch is reported. These capsules display reversible "on/off" fluorescence in response to cellular pH variations. Using this system, the high-throughput quantification between surface-bound and internalized capsules is demonstrated. This system allows a fundamental study of the interaction between nanoengineered materials and biological systems at a cellular level.
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Portadores de Fármacos/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Endocitosis/fisiología , Colorantes Fluorescentes/administración & dosificación , Ácidos Polimetacrílicos/administración & dosificación , Animales , Células de la Médula Ósea/química , Células de la Médula Ósea/metabolismo , Cápsulas/administración & dosificación , Cápsulas/química , Cápsulas/farmacocinética , Línea Celular , Células Dendríticas/química , Células Dendríticas/metabolismo , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Sistemas de Liberación de Medicamentos/instrumentación , Colorantes Fluorescentes/química , Colorantes Fluorescentes/farmacocinética , Concentración de Iones de Hidrógeno , Ratones , Microscopía Fluorescente , Modelos Moleculares , Ácidos Polimetacrílicos/química , Ácidos Polimetacrílicos/farmacocinéticaRESUMEN
Click chemistry has had a significant impact in the field of materials science over the last 10 years, as it has enabled the design of new hybrid building blocks, leading to multifunctional and responsive materials. One key application for such materials is in the biomedical field, such as gene or drug delivery. However, to meet the functional requirements of such applications, tailored degradability of these materials under biological conditions is critical. There has been an increasing interest in combining click chemistry techniques with a range of degradable or responsive building blocks as well as investigating new or milder chemistries to design click delivery systems that are capable of physiologically relevant degradation. This Feature Article will cover some of the different approaches to synthesize degradable click delivery systems and their investigation for therapeutic release.
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Materiales Biocompatibles/química , Sistemas de Liberación de Medicamentos/instrumentación , Técnicas de Transferencia de Gen/instrumentación , Polímeros/química , Materiales Biocompatibles/síntesis química , Química Clic , Portadores de Fármacos/síntesis química , Portadores de Fármacos/química , Polímeros/síntesis químicaRESUMEN
Hollow polymer capsules were prepared from linear as well as brushlike poly(2-oxazoline)s (POxs). Linear POxs containing alkene functionalities were obtained by cationic ring-opening polymerization (CROP), whereas the brush POxs bearing alkyne moieties were synthesized by a combination of CROP and reversible addition-fragmentation chain transfer (RAFT) polymerization. Multilayers consisting of POx/poly(methacrylic acid) (PMA) were sequentially deposited onto silica particle templates, and the films were stabilized either by thiol-ene (TE) chemistry or copper-catalyzed azide-alkyne cycloaddition (CuAAc). Stable, monodisperse capsules were formed after removal of the silica particles with hydrofluoric acid and were observed using fluorescence and atomic force microscopy (AFM). Both architectures exhibited low-fouling behavior, an essential criteria for therapeutic carriers to be utilized in bioapplications. In particular, the brush-like POx capsules show potential as a viable alternative material for the fabrication of low-fouling capsules.