RESUMEN
Over the last years, there has been an increasing number of proposals for the use of nanomaterials in medicine. The safety of novel technologies must be verified, prior to their clinical application. Pathology has much to contribute towards this end. In this study, we compared the in vivo toxicity effects of poly- (lactic-co-glycolic acid) nanoparticles with and without chitosan shell. Both nanoparticle types were loaded with curcumin. The nanoparticles were assessed in vitro for potential cytotoxicity with cell viability studies. For the in vivo test, 36 adult Wistar rats were used, four of which were the control group. The remaining 32 were divided into 2 groups, each of which was administered differentially coated drug carriers: (A) nanoparticles without chitosan coating and (B) nanoparticles with chitosan coating. For both groups, the subcutaneous route was used for administration. Each group was further divided into 2 sub-groups of 8 animals each. The animals of the first sub-groups were sacrificed 24 h after the injection and those of the second on the 7th day. The control group was also divided into 2 subgroups of 2 animals each. At the appointed post-administrative date, the rats were sacrificed, and specimens from the brain, liver, kidneys, heart, stomach, lungs, and from the skin at the injection site were collected and studied histopathologically. The evaluation of both in vitro and in vivo testing shows that nanoparticles with chitosan have significantly less, if any, toxic effects compared to those without chitosan.
Asunto(s)
Quitosano , Nanopartículas , Ratas , Humanos , Animales , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Quitosano/toxicidad , Ácido Poliglicólico/toxicidad , Ácido Láctico , Nanomedicina , Ratas Wistar , Nanopartículas/toxicidadRESUMEN
Among the different synthetic polymers developed for biomedical applications, poly(lactic-co-glycolic acid) (PLGA) has attracted considerable attention because of its excellent biocompatibility and biodegradability. Nanocomposites based on PLGA and metal-based nanostructures (MNSs) have been employed extensively as an efficient strategy to improve the structural and functional properties of PLGA polymer. The MNSs have been used to impart new properties to PLGA, such as antimicrobial properties and labeling. In the present review, the different strategies available for the fabrication of MNS/PLGA nanocomposites and their applications in the biomedical field will be discussed, beginning with a description of the preparation routes, antimicrobial activity, and cytotoxicity concerns of MNS/PLGA nanocomposites. The biomedical applications of these nanocomposites, such as carriers and scaffolds in tissue regeneration and other therapies are subsequently reviewed. In addition, the potential advantages of using MNS/PLGA nanocomposites in treatment illnesses are analyzed based on in vitro and in vivo studies, to support the potential of these nanocomposites in future research in the biomedical field.
Asunto(s)
Antiinfecciosos/química , Antiinfecciosos/farmacología , Metales/química , Nanocompuestos/química , Ácido Poliglicólico/química , Animales , Humanos , Metales/farmacología , Nanocompuestos/toxicidad , Ácido Poliglicólico/toxicidadRESUMEN
The use of herbicides plays a vital role in controlling weeds and conserving crops; however, its usage generates both environmental and economic problems. For example, herbicides pose a financial issue as farmers must apply large quantities to protect crops due to absorption rates of less than 0.1%. Therefore, there is a great need for the development of new methods to mitigate these issues. Here, we report for the first time the synthesis of poly(lactic- co-glycolic-acid) (PLGA) nanoherbicides loaded with atrazine as an active ingredient. We used potato plants as a biological model to assess the herbicidal activity of the engineered PLGA nanoherbicides. Our method produced nanoherbicides with an average size of 110 ± 10 nm prior to lyophilization. Fifty percent of the loaded atrazine in the PLGA matrix is released in 72 h. Furthermore, we performed Monte Carlo simulations to determine the chemical interaction among atrazine, PLGA, and the solvent system. One of the most significant outcomes of these simulations was to find the formation of a hydrogen bond of 1.9 Å between PLGA and atrazine, which makes this interaction very stable. Our in vitro findings showed that as atrazine concentration is increased in PLGA nanoparticles, potato plants undergo a significant decrease in stem length, root length, fresh weight, dry weight, and the number of leaves, with root length being the most affected. These experimental results suggest the herbicidal effectiveness of atrazine-loaded PLGA nanoherbicides in inhibiting the growth of the potato plant. Hence, we present the proof-of-concept for using PLGA nanoherbicides as an alternative method for inhibiting weed growth. Future studies will involve a deep understanding of the mechanism of plant-nanoherbicide interaction as well as the role of PLGA as a growth potentiator.
Asunto(s)
Atrazina/química , Portadores de Fármacos/química , Herbicidas/química , Ácido Láctico/química , Nanopartículas/química , Ácido Poliglicólico/química , Portadores de Fármacos/toxicidad , Composición de Medicamentos , Herbicidas/toxicidad , Ácido Láctico/toxicidad , Nanopartículas/toxicidad , Tamaño de la Partícula , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Solanum tuberosum/efectos de los fármacos , Solanum tuberosum/crecimiento & desarrolloRESUMEN
Plasmid DNA (pDNA) vaccines have the potential for protection against a wide range of diseases including rabies but are rapid in degradation and poor in uptake by antigen-presenting cells. To overcome the limitations, we fabricated a pDNA nanoparticulate vaccine. The negatively charged pDNA was adsorbed onto the surface of cationic PLGA (poly (d, l-lactide-co-glycolide))-chitosan nanoparticles and were used as a delivery vehicle. To create a hydrogel for sustainable vaccine release, we dispersed the pDNA nanoparticles in poloxamer 407 gel which is liquid at 4⯰C and turns into soft gels at 37⯰C, providing ease of administration and preventing burst release of pDNA. Complete immobilization of pDNA to cationic nanoparticles was achieved at a pDNA to nanoparticles ratio (P/N) of 1/50. Cellular uptake of nanoparticles was both time and concentration dependent and followed a saturation kinetics with Vmax of 11.389⯵g/mLâ¯h and Km of 139.48⯵g/mL. The in vitro release studies showed the nanoparticulate vaccine has a sustained release for up to 24â¯days. In summary, pDNA PLGA-chitosan nanoparticles were non-cytotoxic, their buffering capacity and cell uptake were enhanced, and sustained the release of pDNA. We expect our pDNA vaccine's potency will be greatly improved in the animal studies.
Asunto(s)
Quitosano/química , Portadores de Fármacos , Ácido Láctico/química , Nanopartículas , Poloxámero/química , Ácido Poliglicólico/química , Vacunas Antirrábicas/química , Animales , Línea Celular , Quitosano/toxicidad , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Composición de Medicamentos , Liberación de Fármacos , Estabilidad de Medicamentos , Ensayo de Cambio de Movilidad Electroforética , Hidrogeles , Cinética , Ácido Láctico/toxicidad , Ratones Endogámicos C57BL , Nanotecnología , Poloxámero/toxicidad , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Vacunas Antirrábicas/administración & dosificación , Vacunas Antirrábicas/inmunología , Vacunas Antirrábicas/metabolismo , Solubilidad , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Tecnología Farmacéutica/métodos , Vacunas de ADN/química , Vacunas de ADN/inmunologíaRESUMEN
Magnetic nanoparticles (MNPs) are used as contrast agents and targeted drug delivery systems (TDDS) due to their favorable size, surface charge, and magnetic properties. Unfortunately, the toxicity associated with MNPs limits their biological applications. Surface functionalization of MNPs with selective polymers alters the surface chemistry to impart better biocompatibility. We report the preparation of surface functionalized MNPs using iron oxide NPs (MNPs), poly (lactic-co-glycolic acid) (PLGA), and sodium alginate via co-precipitation, emulsification, and electro-spraying, respectively. The NPs are in the nanosize range and negatively charged. Morphological and structural analyses affirm the surface functionalized nanostructure of the NPs. The surface functionalized MNPs are biocompatible, and demonstrate enhanced intracellular delivery under an applied magnetic field (H), which evinces the targeting ability of MNPs. After NP treatment, the physico-mechanical properties of fibroblasts are decided by the selective MNP uptake under "on" or "off" magnetic field conditions. We envision potential use of biocompatible surface functionalized MNP for intracellular-, targeted-DDS, imaging, and for investigating cellular mechanics.
Asunto(s)
Alginatos/química , Materiales Biocompatibles/química , Reactivos de Enlaces Cruzados/química , Ácido Láctico/química , Campos Magnéticos , Magnetismo/métodos , Nanopartículas de Magnetita/química , Nanomedicina/métodos , Ácido Poliglicólico/química , Alginatos/metabolismo , Alginatos/toxicidad , Animales , Materiales Biocompatibles/metabolismo , Materiales Biocompatibles/toxicidad , Transporte Biológico , Supervivencia Celular/efectos de los fármacos , Reactivos de Enlaces Cruzados/metabolismo , Reactivos de Enlaces Cruzados/toxicidad , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/patología , Ácido Glucurónico/química , Ácido Glucurónico/metabolismo , Ácido Glucurónico/toxicidad , Ácidos Hexurónicos/química , Ácidos Hexurónicos/metabolismo , Ácidos Hexurónicos/toxicidad , Ácido Láctico/metabolismo , Ácido Láctico/toxicidad , Nanopartículas de Magnetita/toxicidad , Ratones , Células 3T3 NIH , Tamaño de la Partícula , Ácido Poliglicólico/metabolismo , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Propiedades de SuperficieRESUMEN
Frequent intravitreal injections are currently used to overcome the ocular barriers and provide sufficient drug to the posterior eye segment. However, intravitreal injections have been associated with a number of complications and high treatment costs. To overcome these limitations, peptide-loaded poly(d,l-lactic-co-glycolic acid) nanoparticles (PLGA NPs) were developed using the nanoprecipitation technique and were optimized via Box-Behnken Design (BBD) and Response Surface Methodology (RSM). Developed NPs were evaluated for potential toxicity and cell apoptosis using the zebrafish embryo toxicity (ZET) model with titanium dioxide NPs and ethanol (1% v/v) serving as positive controls. Developed NPs had a size of 75.6-153.8 nm, a polydispersity index between 0.11 and 0.25 and a zeta potential of -9.4 to -46.0 mV. Loaded peptide was found to be stable under various experimental conditions tested. BBD and RSM were validated through the characterization of optimized formulations. Survival and hatching rates of NP-treated zebrafish 0-144 h post-fertilization were found to be normal with no significant malformations. Cellular apoptosis studies also endorsed the non-cytotoxic nature of the NPs. The overall results indicate that optimized PLGA nanoparticles could be a promising platform for efficient peptide delivery to the posterior segment of the eye.
Asunto(s)
Portadores de Fármacos/toxicidad , Ácido Láctico/toxicidad , Nanopartículas/toxicidad , Péptidos/administración & dosificación , Ácido Poliglicólico/toxicidad , Animales , Apoptosis/efectos de los fármacos , Portadores de Fármacos/administración & dosificación , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos/efectos adversos , Embrión no Mamífero/citología , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/fisiología , Inyecciones Intravítreas , Ácido Láctico/administración & dosificación , Ácido Láctico/química , Nanopartículas/administración & dosificación , Nanopartículas/química , Péptidos/química , Ácido Poliglicólico/administración & dosificación , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Pruebas de Toxicidad , Pez Cebra/embriologíaRESUMEN
In response to the demand for new implant materials characterized by high biocompatibility and bioresorption, two prototypes of fibrous nanocomposite implants for osseous tissue regeneration made of a newly developed blend of poly(l-lactide-co-glycolide) (PLGA) and syntheticpoly([R,S]-3-hydroxybutyrate), PLGA/PHB, have been developed and fabricated. Afibre-forming copolymer of glycolide and l-lactide (PLGA) was obtained by a unique method of synthesis carried out in blocksusing Zr(AcAc)4 as an initiator. The prototypes of the implants are composed of three layers of PLGA or PLGA/PHB, nonwoven fabrics with a pore structure designed to provide the best conditions for the cell proliferation. The bioactivity of the proposed implants has been imparted by introducing a hydroxyapatite material and IGF1, a growth factor. The developed prototypes of implants have been subjected to a set of in vitro and in vivobiocompatibility tests: in vitro cytotoxic effect, in vitro genotoxicity and systemic toxicity. Rabbitsshowed no signs of negative reactionafter implantation of the experimental implant prototypes.
Asunto(s)
Implantes Absorbibles , Regeneración Ósea , Hidroxibutiratos , Ácido Láctico/química , Ácido Láctico/farmacología , Nanocompuestos , Poliésteres , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacología , Andamios del Tejido , Animales , Biomarcadores , Línea Celular , Supervivencia Celular , Humanos , Hidroxibutiratos/química , Ácido Láctico/toxicidad , Ratones , Nanocompuestos/química , Poliésteres/química , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Porosidad , Prohibitinas , Conejos , Ingeniería de TejidosRESUMEN
Purpose: Poly(lactic-co-glycolic) acid (PLGA) inserts have been successfully developed for the treatment of posterior eye disease as a means of reducing injection frequency of intravitreally administered therapeutics. PLGA microspheres are also of interest for the delivery of intravitreal drugs, since they offer the advantage of being easily injected without surgical procedures or large injectors. Methods: In the current study, the toxicity of PLGA microspheres and rods was investigated in nonhuman primates (NHPs) and rabbits. An in vitro assessment of cytokine responses to PLGA in peripheral blood mononuclear cells (PBMCs) and macrophages was also performed. Results: Intravitreal administration of 3, 10, or 12.5 mg/eye of PLGA microspheres in NHPs resulted in a severe immune response characterized by a foreign body response. Follow-up studies in the rabbit confirmed this finding for PLGA microspheres ranging in size from 20 to 100 µm. In contrast, administration of PLGA rod implants with a similar PLGA mass did not elicit a significant immune response. In vitro assays in PBMCs and macrophages confirmed proinflammatory cytokine release upon treatment with PLGA microspheres but not PLGA rods. Conclusions: These data demonstrate a lack of tolerability of PLGA microspheres upon intravitreal injection, and suggest that the size, shape, and/or surface area of PLGA depots are critical attributes in determining ocular toxicity.
Asunto(s)
Materiales Biocompatibles/toxicidad , Sistemas de Liberación de Medicamentos/efectos adversos , Ácido Láctico/toxicidad , Microesferas , Ácido Poliglicólico/toxicidad , Animales , Materiales Biocompatibles/administración & dosificación , Citocinas/metabolismo , Modelos Animales de Enfermedad , Reacción a Cuerpo Extraño/metabolismo , Reacción a Cuerpo Extraño/patología , Inyecciones Intravítreas , Ácido Láctico/administración & dosificación , Macrófagos/metabolismo , Ácido Poliglicólico/administración & dosificación , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Conejos , Retina/efectos de los fármacos , Retina/patología , Cuerpo Vítreo/efectos de los fármacos , Cuerpo Vítreo/patologíaRESUMEN
This study aimed to evaluate the teratogenic and hepatotoxic potential of the usnic acid encapsulated into PLGA-microspheres. In total, 12 female Wistar rats in pregnancy were randomly distributed in the control group (n= 6) that received 1.0 mL of physiological solution and treatment group (n= 6) that received 25 mg/kg of encapsulated usnic acid by oral administration. All females were euthanized at day 20 of pregnancy and their fetuses were removed and analyzed. During the pregnancy was observed a reduction in weight gain. There was no difference in serum transaminases levels analyzed as well as any difference in liver weight in both groups. The histomorphometric analysis of the liver from the treatment group revealed an increase in number of hepatocytes and a decrease in nuclear area of these cells. Moreover, no alteration was observed in cell area of hepatocytes or number of Kupffer cells. The fetuses had an increase in total number of hepatocytes and a reduction in the amount of megakaryocytes. These results show the hepatotoxic potential of usnic acid during pregnancy. However, its toxicity can be minimized by encapsulation in microspheres.
Asunto(s)
Ascomicetos/química , Benzofuranos/toxicidad , Feto/efectos de los fármacos , Ácido Láctico/toxicidad , Líquenes/química , Hígado/efectos de los fármacos , Ácido Poliglicólico/toxicidad , Anomalías Inducidas por Medicamentos , Animales , Benzofuranos/química , Femenino , Peso Fetal/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Ácido Láctico/química , Hígado/patología , Exposición Materna , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Embarazo , Distribución Aleatoria , Ratas Wistar , Valores de ReferenciaRESUMEN
ABSTRACT This study aimed to evaluate the teratogenic and hepatotoxic potential of the usnic acid encapsulated into PLGA-microspheres. In total, 12 female Wistar rats in pregnancy were randomly distributed in the control group (n= 6) that received 1.0 mL of physiological solution and treatment group (n= 6) that received 25 mg/kg of encapsulated usnic acid by oral administration. All females were euthanized at day 20 of pregnancy and their fetuses were removed and analyzed. During the pregnancy was observed a reduction in weight gain. There was no difference in serum transaminases levels analyzed as well as any difference in liver weight in both groups. The histomorphometric analysis of the liver from the treatment group revealed an increase in number of hepatocytes and a decrease in nuclear area of these cells. Moreover, no alteration was observed in cell area of hepatocytes or number of Kupffer cells. The fetuses had an increase in total number of hepatocytes and a reduction in the amount of megakaryocytes. These results show the hepatotoxic potential of usnic acid during pregnancy. However, its toxicity can be minimized by encapsulation in microspheres.
Asunto(s)
Animales , Femenino , Embarazo , Ácido Poliglicólico/toxicidad , Ascomicetos/química , Benzofuranos/toxicidad , Ácido Láctico/toxicidad , Feto/efectos de los fármacos , Líquenes/química , Hígado/efectos de los fármacos , Ácido Poliglicólico/química , Valores de Referencia , Anomalías Inducidas por Medicamentos , Benzofuranos/química , Distribución Aleatoria , Ratas Wistar , Exposición Materna , Ácido Láctico/química , Peso Fetal/efectos de los fármacos , Hepatocitos/efectos de los fármacos , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Hígado/patologíaRESUMEN
Paclitaxel (PTX) incorporation in poly(lactic-co-glycolic acid) (PLGA) matrices produce films with high tensile rigidity and slow release that fail to deliver the required release rate for most biomedical applications such as in drug eluting stents and cancer treatments. To modify and improve this behavior, a set of poly(diol sebacate)s were synthesized and fully characterized as possible additives. The tensile properties of PLGA blends were evaluated as these materials could be used as coatings in drug eluting stent applications. A significant improvement in mechanical flexibility was observed with 20% additive content, as it reduced the Young's modulus value and increased the maximum deformation at break. PTX release was studied and correlated with the release of additive from PLGA films. An increase in the initial burst release phase was observed on all blends when compared to the control films of PLGA. Modulation of PTX release was achieved by altering the hydrophilicity degree of the additive or its percentage content on the blend. This supports the possibility that PTX was partitioned into the additive phase. Cytotoxicity analyses of novel additives were performed on mouse embryonic fibroblasts NIH/3T3.
Asunto(s)
Antineoplásicos Fitogénicos/administración & dosificación , Materiales Biocompatibles/química , Ácidos Decanoicos/química , Ácidos Dicarboxílicos/química , Portadores de Fármacos/química , Ácido Láctico/química , Paclitaxel/administración & dosificación , Ácido Poliglicólico/química , Polímeros/química , Animales , Materiales Biocompatibles/toxicidad , Ácidos Decanoicos/toxicidad , Ácidos Dicarboxílicos/toxicidad , Portadores de Fármacos/toxicidad , Módulo de Elasticidad , Ácido Láctico/toxicidad , Ratones , Células 3T3 NIH , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Polímeros/toxicidadRESUMEN
Amphotericin B (AMB) is used most commonly in severe systemic life-threatening fungal infections. There is currently an unmet need for an efficacious (AMB) formulation amenable to oral administration with better bioavailability and lower nephrotoxicity. Novel PEGylated polylactic-polyglycolic acid copolymer (PLGA-PEG) nanoparticles (NPs) formulations of AMB were therefore studied for their ability to kill Candida albicans (C. albicans). The antifungal activity of AMB formulations was assessed in C. albicans. Its bioavalability was investigated in nine groups of rats (n = 6). Toxicity was examined by an in vitro blood hemolysis assay, and in vivo nephrotoxicity after single and multiple dosing for a week by blood urea nitrogen (BUN) and plasma creatinine (PCr) measurements. The MIC of AMB loaded to PLGA-PEG NPs against C. albicans was reduced two to threefold compared with free AMB. Novel oral AMB delivery loaded to PLGA-PEG NPs was markedly systemically available compared to Fungizone® in rats. The addition of 2% of GA to the AMB formulation significantly (p < 0.05) improved the bioavailability from 1.5 to 10.5% and the relative bioavailability was > 790% that of Fungizone®. The novel AMB formulations showed minimal toxicity and better efficacy compared to Fungizone®. No nephrotoxicity in rats was detected after a week of multiple dosing of AMB NPs based on BUN and PCr, which remained at normal levels. An oral delivery system of AMB-loaded to PLGA-PEG NPs with better efficacy and minimal toxicity was formulated. The addition of glycyrrhizic acid (GA) to AMB NPs formulation resulted in a significant oral absorption and improved bioavailability in rats.
Asunto(s)
Anfotericina B/administración & dosificación , Antifúngicos/administración & dosificación , Candida albicans/efectos de los fármacos , Portadores de Fármacos , Ácido Láctico/química , Nanopartículas , Polietilenglicoles/química , Ácido Poliglicólico/química , Administración Oral , Anfotericina B/química , Anfotericina B/farmacocinética , Anfotericina B/toxicidad , Animales , Antifúngicos/química , Antifúngicos/farmacocinética , Antifúngicos/toxicidad , Disponibilidad Biológica , Biomarcadores/sangre , Nitrógeno de la Urea Sanguínea , Candida albicans/crecimiento & desarrollo , Creatinina/sangre , Composición de Medicamentos , Ácido Glicirrínico/administración & dosificación , Ácido Glicirrínico/química , Hemólisis/efectos de los fármacos , Enfermedades Renales/sangre , Enfermedades Renales/inducido químicamente , Ácido Láctico/toxicidad , Pruebas de Sensibilidad Microbiana , Polietilenglicoles/toxicidad , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas Sprague-Dawley , Tecnología Farmacéutica/métodosRESUMEN
Endothelial dysfunction is initial and critical step of atherosclerosis. Impaired bioavailability of endothelial nitric oxide synthase (eNOS) is one of the main reasons of endothelial dysfunction. Improving bioavailability of eNOS by increasing its expression or activity using statins is an effective therapeutic strategy in restoring endothelial dysfunction. In this study, simvastatin (SIM) as a poorly water-soluble drug was loaded in poly (lactic-co-glycolic acid) (PLGA) nanoparticles (SIM-PLGA-NPs). NPs were then conjugated with mZD7349 peptide (mZD7349-SIM-PLGA-NPs) and directed against vascular cell adhesion molecule 1 (VCAM-1). In vitro evaluation of the NPs for targeted delivery of SIM was performed on activated Human Umbilical Cord Vascular Endothelial Cells (HUVECs) by tumor necrosis factor alpha (TNF-α). Effect of mZD7349-SIM-PLGA-NPs and SIM-PLGA-NPs was compared on eNOS phosphorylation (ser-1177). Results of western blot showed SIM post-treatment increased significantly phosphor-eNOS (Ser1177) expression but no total eNOS expression. The study showed that mZD7349-SIM-PLGA-NPs have particle size, zeta potential value, polydispersity index (PDI) and encapsulation efficacy % of 233±18nm, -9.6±1.1mV, 0.59±0.066 and 69±17.3%, respectively. Also phosphor-eNOS (Ser1177) expression in activated HUVECs treated with mZD7349-SIM-PLGA-NPs was significantly (p<0.05) better than treated cells with SIM-PLGA-NPs. The results suggest that mZD7349-SIM-PLGA-NPs may be usable as an appropriate drug carrier for restoring endothelial dysfunction.
Asunto(s)
Antiinflamatorios/farmacología , Portadores de Fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Inflamación/prevención & control , Ácido Láctico/química , Nanopartículas , Péptidos Cíclicos/metabolismo , Ácido Poliglicólico/química , Simvastatina/farmacología , Molécula 1 de Adhesión Celular Vascular/metabolismo , Antiinflamatorios/química , Antiinflamatorios/metabolismo , Antiinflamatorios/toxicidad , Células Cultivadas , Composición de Medicamentos , Liberación de Fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/química , Inhibidores de Hidroximetilglutaril-CoA Reductasas/metabolismo , Inhibidores de Hidroximetilglutaril-CoA Reductasas/toxicidad , Inflamación/metabolismo , Inflamación/patología , Ácido Láctico/toxicidad , Óxido Nítrico Sintasa de Tipo III/metabolismo , Péptidos Cíclicos/química , Péptidos Cíclicos/toxicidad , Fosforilación , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Serina , Simvastatina/química , Simvastatina/metabolismo , Simvastatina/toxicidad , Solubilidad , Factores de Tiempo , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Biocompatible and biodegradable polymers like PLGA have revolutionized the drug delivery approaches. However, poor drug loading and substantially high lipophilicity, pave a path for further tailing of this promising agent. In this regard, PLGA was feathered with biocompatible phospholipid and polymeric micelles were developed for delivery of Methotrexate (MTX) to cancer cells. The nanocarriers (114.6nm±5.5nm) enhanced the cytotoxicity of MTX by 2.13 folds on MDA-MB-231 cells. Confocal laser scanning microscopy confirmed the increased intracellular delivery. The carrier decreased the protein binding potential and enhanced the bioavailable fraction of MTX. Pharmacokinetic studies vouched substantial enhancement in AUC and bioresidence time, promising an ideal carrier to effectively deliver the drug to the site of action. The developed nanocarriers offer potential to deliver the drug in the interiors of cancer cells in an effective manner for improved therapeutic action.
Asunto(s)
Glycine max/química , Ácido Láctico/química , Ácido Láctico/metabolismo , Lecitinas/química , Metotrexato/química , Micelas , Ácido Poliglicólico/química , Ácido Poliglicólico/metabolismo , Animales , Transporte Biológico , Bovinos , Línea Celular Tumoral , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/toxicidad , Liberación de Fármacos , Humanos , Ácido Láctico/farmacocinética , Ácido Láctico/toxicidad , Ácido Poliglicólico/farmacocinética , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas , Albúmina Sérica Bovina/metabolismoRESUMEN
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
UNLABELLED: : The regeneration of tissue-engineered cartilage in an immunocompetent environment usually fails due to severe inflammation induced by the scaffold and their degradation products. In the present study, we compared the tissue remodeling and the inflammatory responses of engineered cartilage constructed with bone marrow mesenchymal stem cells (BMSCs), chondrocytes, or both and scaffold group in pigs. The cartilage-forming capacity of the constructs in vitro and in vivo was evaluated by histological, biochemical, and biomechanical analyses, and the inflammatory response was investigated by quantitative analysis of foreign body giant cells and macrophages. Our data revealed that BMSC-based engineered cartilage suppressed in vivo inflammation through the alteration of macrophage phenotype, resulting in better tissue survival compared with those regenerated with chondrocytes alone or in combination with BMSCs. To further confirm the macrophage phenotype, an in vitro coculture system established by engineered cartilage and macrophages was studied using immunofluorescence, enzyme-linked immunosorbent assay, and gene expression analysis. The results demonstrated that BMSC-based engineered cartilage promoted M2 polarization of macrophages with anti-inflammatory phenotypes including the upregulation of CD206, increased IL-10 synthesis, decreased IL-1ß secretion, and alterations in gene expression indicative of M1 to M2 transition. It was suggested that BMSC-seeded constructs have the potential to ameliorate scaffold-induced inflammation and improve cartilaginous tissue regeneration through M2 polarization of macrophages. SIGNIFICANCE: Finding a strategy that can prevent scaffold-induced inflammation is of utmost importance for the regeneration of tissue-engineered cartilage in an immunocompetent environment. This study demonstrated that bone marrow mesenchymal stem cell (BMSC)-based engineered cartilage could suppress inflammation by increasing M2 polarization of macrophages, resulting in better tissue survival in a pig model. Additionally, the effect of BMSC-based cartilage on the phenotype conversion of macrophages was further studied through an in vitro coculture system. This study could provide further support for the regeneration of cartilage engineering in immunocompetent animal models and provide new insight into the interaction of tissue-engineered cartilage and macrophages.
Asunto(s)
Células de la Médula Ósea , Trasplante de Médula Ósea , Cartílago/trasplante , Condrocitos/trasplante , Condrogénesis , Reacción a Cuerpo Extraño/prevención & control , Ácido Láctico/química , Macrófagos/metabolismo , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Ácido Poliglicólico/química , Ingeniería de Tejidos/métodos , Andamios del Tejido , Animales , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/metabolismo , Trasplante de Médula Ósea/efectos adversos , Cartílago/inmunología , Cartílago/metabolismo , Células Cultivadas , Condrocitos/inmunología , Condrocitos/metabolismo , Técnicas de Cocultivo , Colágeno/metabolismo , Módulo de Elasticidad , Reacción a Cuerpo Extraño/inducido químicamente , Reacción a Cuerpo Extraño/genética , Reacción a Cuerpo Extraño/inmunología , Regulación de la Expresión Génica , Glicosaminoglicanos/metabolismo , Supervivencia de Injerto , Ácido Láctico/toxicidad , Macrófagos/inmunología , Trasplante de Células Madre Mesenquimatosas/efectos adversos , Células Madre Mesenquimatosas/inmunología , Células Madre Mesenquimatosas/metabolismo , Fenotipo , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Porcinos , Factores de TiempoRESUMEN
AIM: Quantify the biodistribution and assess the toxicity of PLGA (poly-lactic-co-glycolic acid) and surface-modified PLGA chitosan (PLGA/Chi) nanoparticles (NPs) orally administered for 7, 14 and 21 days to F344 rats. MATERIALS & METHODS: Fluorescent NPs were tracked in F344 rat tissues, and toxicity was evaluated by alkaline phosphatase and alanine transaminase levels, and by histologic examination of tissue samples. RESULTS: Biodistribution of PLGA and PLGA/Chi were similar, with highest amounts found in the intestine and liver. Alkaline phosphatase increased significantly in treated rats. Mild histological differences were detected in the intestine and liver. CONCLUSION: PLGA and PLGA/Chi NPs behaved similarly presenting minimal toxicity in the liver and intestine, but not in kidney, lung and brain.
Asunto(s)
Ácido Láctico/química , Ácido Láctico/farmacocinética , Nanopartículas/química , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacocinética , Administración Oral , Alanina Transaminasa/sangre , Fosfatasa Alcalina/sangre , Animales , Quitosano/química , Portadores de Fármacos , Humanos , Ácido Láctico/toxicidad , Nanopartículas/toxicidad , Tamaño de la Partícula , Ácido Poliglicólico/toxicidad , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas Endogámicas F344 , Propiedades de Superficie , Distribución Tisular , Pruebas de Toxicidad SubagudaRESUMEN
Multiple synthetic polymer nanoparticles (NPs) have been widely used as drug delivery systems. However, their toxicity to the retinal pigment epithelium and retinal endothelium remains unclear. In this study, we analyze the cytotoxic effects of three different kinds of NPs, made of poly lactic-co-glycolic acid (PLGA), polycaprolactone (PCL), and PEGylated PLGA (PEG-PLGA), in a retinal pigment epithelium cell line (ARPE-19) and in primary human retinal vascular endothelial cells (RVEC). PEG-PLGA NPs presented the lowest cytotoxicity on ARPE-19 cells and RVEC as assessed by MTT viability assay. While PLGA and PCL exhibited variable amounts of toxicity, no significant toxicity was observed when incubating cells with high PEG-PLGA concentrations (100 µg/ml), for up to 6 days. On both transmission electron microscopy and confocal microscopy, Rhodamine 6G-loaded PEG-PLGA NPs were observed intracellularly in multiple subcellular organelles. PEG-PLGA NPs are a potentially viable option for the treatment of eye diseases.
Asunto(s)
Sistemas de Liberación de Medicamentos , Células Endoteliales/efectos de los fármacos , Ácido Láctico/toxicidad , Poliésteres/toxicidad , Polietilenglicoles/toxicidad , Poliglactina 910/toxicidad , Ácido Poliglicólico/toxicidad , Epitelio Pigmentado de la Retina/efectos de los fármacos , Vasos Retinianos/citología , Línea Celular , Humanos , Nanopartículas , Copolímero de Ácido Poliláctico-Ácido PoliglicólicoRESUMEN
With the ongoing commercialization of nanotechnology products, human exposure to nanoparticles (NPs) is set to increase dramatically and an evaluation of their potential adverse effects is essential. Surface charge, among other physico-chemicals parameters, is a key criterion that should be considered when using a definition for nanomaterials in a regulatory context. It has recently been recognized as an important factor in determining the toxicity of NPs; however, a complete understanding of the mechanisms involved is still lacking. In this context, the aim of the present study was to investigate the influence of the surface charge modification of NPs on in vitro toxicity assays. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles bearing different surface charges, positive(+), neutral(n) or negative(-), were synthesized. In vitro genotoxicity assays (micronucleus and comet assays) coupled with an assessment of cytotoxicity, were performed in different cell lines (L5178Y mouse lymphoma cells, TK6 human B-lymphoblastoid cells and 16HBE14o- human bronchial epithelial cells). Reactive oxygen species (ROS) production and endocytosis studies were also performed. Our results showed that PLGA(+) NPs were cytotoxic. They are endocytosed by the clathrin pathway and induced ROS in the three cell lines. They led to chromosomal aberrations without primary DNA damage in 16HBE14o- cells, suggesting that aneuploidy may be considered as an important biomarker when assessing the genotoxic potential of NPs. Moreover, 16HBE14o- cells seem to be more suitable for the in vitro screening of inhaled NPs than the regulatory L5178Y and TK6 cells.
Asunto(s)
Daño del ADN , Endocitosis , Ácido Láctico/toxicidad , Micronúcleos con Defecto Cromosómico/inducido químicamente , Nanopartículas/toxicidad , Estrés Oxidativo/efectos de los fármacos , Ácido Poliglicólico/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Aneuploidia , Animales , Bronquios/efectos de los fármacos , Bronquios/metabolismo , Bronquios/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Clatrina/metabolismo , Ensayo Cometa , Relación Dosis-Respuesta a Droga , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Células Epiteliales/patología , Humanos , Ácido Láctico/química , Ácido Láctico/metabolismo , Linfoma/genética , Linfoma/metabolismo , Linfoma/patología , Ratones , Pruebas de Micronúcleos , Nanopartículas/química , Nanopartículas/metabolismo , Ácido Poliglicólico/química , Ácido Poliglicólico/metabolismo , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Medición de Riesgo , 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.