RESUMEN
Surfactant protein A (SP-A), a lung anti-infective protein, is a lectin with affinity for sugars found on fungal and micrococcal surfaces such as mannose. We synthesized a mannosylated poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) copolymer and used it to produce nanoparticles with a polyester (PLGA/PLA) core and a PEG shell decorated with mannose residues, designed to be strongly associated with SP-A for an increased uptake by alveolar macrophages. Nanoparticles made of the copolymers were obtained by nanoprecipitation and displayed a size of around 140 nm. The presence of mannose on the surface was demonstrated by zeta potential changes according to pH and by a strong aggregation in the presence of concanavalin A. Mannosylated nanoparticles bound to SP-A as demonstrated by dynamic light scattering and transmission electron microscopy. The association with SP-A increased nanoparticle uptake by THP-1 macrophages in vitro. In vivo experiments demonstrated that after intratracheal administration of nanoparticles with or without SP-A, SP-A-coated mannosylated nanoparticles were internalized by alveolar macrophages in greater proportion than SP-A-coated nonmannosylated nanoparticles. The data demonstrate for the first time that the pool of nanoparticles available to lung cells can be changed after surface modification, using a biomimetic approach.
Asunto(s)
Macrófagos Alveolares/metabolismo , Nanopartículas/química , Polímeros/química , Proteína A Asociada a Surfactante Pulmonar/metabolismo , Animales , Células Cultivadas , Femenino , Humanos , Macrófagos Alveolares/citología , Ratones , Ratones Endogámicos BALB C , Nanopartículas/administración & dosificación , Polímeros/administración & dosificación , Propiedades de SuperficieRESUMEN
Most nanoparticles produced for drug delivery purposes are spherical. However, the literature suggests that elongated particles are advantageous, notably in terms of cellular uptake. Thus, we synthesized biocompatible polylactide-b-poly(ethylene glycol) (PLA-PEG) polymers bearing carboxylate moieties, and used them to formulate worm-like nanoparticles by a simple emulsion-evaporation process. Worm-like nanoparticles with variable aspect ratio were obtained by simply adjusting the molar mass of the PLA block: the shorter the molar mass of the PLA block, the more elongated the particles. As PLA molar mass decreased from 80,000 g/mol to 13,000 g/mol, the proportion of worm-like nanoparticles increased from 0 to 46%, in contradiction with the usual behavior of block polymers based on their packing parameter. To explain this unusual phenomenon, we hypothesized the shape arises from a combination of steric and electrostatic repulsions between PEG chains bearing a carboxylate moiety present at the dichloromethane-water interface during the evaporation process. Worm-like particles turned out to be unstable when incubated at 37 °C, above polymer glass transition temperature. Indeed, above Tg, a Plateau-Rayleigh instability occurs, leading to the division of the worm-like particles into spheres. However, this instability was slow enough to assess worm-like particles uptake by murine macrophages. A slight but significant increase of internalization was observed for worm-like particles, compared to their spherical counterparts, confirming the interest of developing biocompatible anisotropic nanoparticles for pharmaceutical applications such as drug delivery.
Asunto(s)
Nanopartículas , Polímeros , Ratones , Animales , Polietilenglicoles , Poliésteres , Sistemas de Liberación de Medicamentos , Tamaño de la PartículaRESUMEN
The toxicity of polymeric biodegradable nanoparticles was evaluated on a co-culture made from direct contact of human lung alveolar epithelial cells (A459) and macrophages (differentiated THP-1 monocytes). The co-culture was characterized by its phenotype and by confocal laser scanning microscopy. Cytokine secretion induced by lipopolysaccharide was synergistically increased in the co-culture confirming cell-cell interactions. Poly(lactide-co-glycolide) (PLGA)-based nanoparticles of 200 nm were prepared in presence of hydrophilic polymers commonly used as stabilizers [poly(vinyl alcohol), chitosan and poloxamer 188] through their interaction with particle surface. Stabilizer-free PLGA nanoparticles and stabilizers alone were also evaluated as controls. Selective uptake kinetics of PLGA nanoparticles by cell subpopulations, as well as apoptosis/necrosis detection, was achieved using a specific label for each cell type, while cytokine secretions were quantified in culture supernatants. Both cell subpopulations took up PLGA nanoparticles with similar profiles, and induced only little cytotoxicity (mostly necrosis). A mild inflammatory response to stabilized nanoparticles was detected (compared to well-known inflammatory compounds), slightly higher than the one observed for stabilizer-free PLGA nanoparticles or stabilizing agents taken individually. These results demonstrate that although biodegradable nanoparticles can be considered as safe, they can internalize compounds such as the stabilizing agents which enhance their toxicity.
Asunto(s)
Células Epiteliales/inmunología , Inflamación/inducido químicamente , Inflamación/inmunología , Ácido Láctico/toxicidad , Macrófagos/inmunología , Nanocápsulas/toxicidad , Ácido Poliglicólico/toxicidad , Implantes Absorbibles , Línea Celular , Técnicas de Cocultivo , Citocinas/inmunología , Células Epiteliales/efectos de los fármacos , Humanos , Inflamación/patología , Activación de Macrófagos/efectos de los fármacos , Activación de Macrófagos/inmunología , Macrófagos/efectos de los fármacos , Nanocápsulas/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Propiedades de SuperficieRESUMEN
To design nanoparticle (NP)-based drug delivery systems for pulmonary administration, biodegradable materials are considered safe, but their potential toxicity is poorly explored. We here explore the lung toxicity in mice of biodegradable nanoparticles (NPs) and compare it to the toxicity of non-biodegradable ones. NP formulations of poly(d,l-lactide-co-glycolide) (PLGA) coated with chitosan (CS), poloxamer 188 (PF68) or poly(vinyl alcohol) (PVA), which renders 200 nm NPs of positive, negative or neutral surface charge respectively, were analyzed for their biodistribution by in vivo fluorescence imaging and their inflammatory potential after single lung nebulization in mice. After exposure, analysis of bronchoalveolar lavage (BAL) cell population, protein secretion and cytokine release as well as lung histology were carried out. The inflammatory response was compared to the one induced by non-biodegradable counterparts, namely, TiO2 of rutile and anatase crystal form and polystyrene (PS). PLGA NPs were mostly present in mice lungs, with little passage to other organs. An increase in neutrophil recruitment was observed in mice exposed to PS NPs 24 h after nebulization, which declined at 48 h. This result was supported by an increase in interleukin (IL)-6 and tumor necrosis factor α (TNFα) in BAL supernatant at 24 h. TiO2 anatase NPs were still present in lung cells 48 h after nebulization and induced the expression of pro-inflammatory cytokines and the recruitment of polymorphonuclear cells to BAL. In contrast, regardless of their surface charge, PLGA NPs did not induce significant changes in the inflammation markers analyzed. In conclusion, these results point out to a safe use of PLGA NPs regardless of their surface coating compared to non-biodegradable ones.
Asunto(s)
Quitosano/toxicidad , Ácido Láctico/toxicidad , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Nanopartículas/toxicidad , Poloxámero/toxicidad , Ácido Poliglicólico/toxicidad , Alcohol Polivinílico/toxicidad , Administración por Inhalación , Aerosoles/administración & dosificación , Aerosoles/farmacocinética , Aerosoles/toxicidad , Animales , Lavado Broncoalveolar , Quitosano/química , Quitosano/farmacocinética , Mediadores de Inflamación/metabolismo , Ácido Láctico/química , Ácido Láctico/farmacocinética , Pulmón/patología , Ratones , Nanopartículas/administración & dosificación , Nanopartículas/química , Poloxámero/química , Poloxámero/farmacocinética , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacocinética , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Poliestirenos/química , Poliestirenos/farmacocinética , Poliestirenos/toxicidad , Alcohol Polivinílico/química , Alcohol Polivinílico/farmacocinética , Distribución Tisular , Titanio/química , Titanio/toxicidadRESUMEN
The purpose of this study was to investigate the toxicity of a series of poly(lactide-co-glycolic) (PLGA) nanoparticles on human-like THP-1 macrophages. Positively-, negatively-charged and neutral nanoparticles (200 nm) were prepared using chitosan (CS), poloxamer 188 (PF68) and poly(vinyl alcohol) (PVA) as stabilizer. Stabilizer-free PLGA nanoparticles were obtained as well. When used at therapeutically relevant concentrations (up to 0.1 mg/mL in vitro), all tested nanoparticles showed no or scarce signs of toxicity, as assessed by cell mitochondrial activity, induction of apoptosis and necrosis, production of intracellular reactive oxygen species (ROS) and secretion of pro-inflammatory cytokines. At high concentrations (above 1mg/mL), cytotoxicity was found to be induced by the presence of stabilizers, whatever the toxicological pattern of the stabilizer itself. While stabilizer-free PLGA nanoparticles exerted no cytotoxicity, the slightly cytotoxic CS polymer conferred PLGA nanoparticles significant cytotoxicity when used as nanoparticle stabilizer; more surprisingly, the otherwise innocuous PVA and PF68 polymers also conferred a significant cytotoxicity to PLGA nanoparticles. These results unveiled the critical toxicological contribution played by stabilizers used for the formulation of PLGA nanoparticles when used at high concentrations, which may have implications for local toxicities of PLGA-based nanomedicine, and provided additional insight in cytotoxic effects of internalized nanoparticles.
Asunto(s)
Excipientes/toxicidad , Ácido Láctico/toxicidad , Macrófagos/efectos de los fármacos , Nanopartículas/química , Nanopartículas/toxicidad , Ácido Poliglicólico/toxicidad , Supervivencia Celular/efectos de los fármacos , Quitosano/química , Quitosano/toxicidad , Excipientes/química , Humanos , Ácido Láctico/química , Poloxámero/química , Poloxámero/toxicidad , Ácido Poliglicólico/química , Alcohol Polivinílico/química , Alcohol Polivinílico/toxicidadRESUMEN
Biodegradable nanoparticles exhibit high potentialities for local or systemic drug delivery through lung administration making them attractive as nanomedicine carriers. However, since particulate matter or some inorganic manufactured nanoparticles exposed to lung cells have provoked cytotoxic effects, inflammatory and oxidative stress responses, it becomes important to investigate nanomedicine toxicity towards the lungs. This is the reason why, in the present review, the behavior of biodegradable nanoparticles towards the different parts of the respiratory tract as well as the toxicological consequences, measured on several models in vitro, ex vivo or in vivo, are described. Taken all together, the different studies carried out so far conclude on no or slight toxicity of biodegradable nanoparticles.
Asunto(s)
Materiales Biocompatibles/toxicidad , Pulmón/efectos de los fármacos , Pulmón/patología , Nanopartículas/toxicidad , Animales , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Humanos , Pruebas de ToxicidadRESUMEN
In vitro cytotoxicity and inflammatory response following exposure to nanoparticles (NPs) made of poly(lactide-co-glycolide) (PLGA) have been investigated on A549 human lung epithelial cells. Three different PLGA NPs (230 nm) were obtained using different stabilizers (polyvinyl alcohol, chitosan, or Pluronic(®) F68) to form respectively neutral, positively or negatively charged NPs. Polystyrene NPs were used as polymeric but non-biodegradable NPs, and titanium dioxide (anatase and rutile) as inorganic NPs, for comparison. Cytotoxicity was evaluated through mitochondrial activity as well as membrane integrity (lactate dehydrogenase release, trypan blue exclusion, propidium iodide staining). The cytotoxicity of PLGA-based and polystyrene NPs was lower or equivalent to the one observed after exposure to titanium dioxide NPs. The inflammatory response, evaluated through the release of the IL-6, IL-8, MCP-1, TNF-α cytokines, was low for all NPs. However, some differences were observed, especially for negative PLGA NPs that led to a higher inflammatory response, which can be correlated to a higher uptake of these NPs. Taken together, these results show that both coating of PLGA NPs and the nature of the core play a key role in cell response.