RESUMEN
OBJECTIVES: To develop and evaluate a novel human stratum corneum (SC) mimetic phospholipid vesicle-based permeation assay (PVPASC) model for in vitro permeation studies. SIGNIFICANCE: Due to the increasing restrictions on the use of human and animal skins, artificial skin models have attracted substantial interest in pharmaceuticals and cosmetic industries. In this study, a modified PVPASC model containing both SC lipids and proteins was developed. METHODS: The PVPASC model was optimized by altering the lipid composition and adding keratin in the formulation of large liposomes. The barrier properties were monitored by measuring the electrical resistance (ER) and permeability of Rhodamine B (RB). The modified PVPASC model was characterized in terms of the surface topography, solvent influence and storage stability. The permeation studies of the active components in Compound Nanxing Zhitong Plaster (CNZP) were performed to examine the capability of PVPASC in the application of skin penetration. RESULTS: The ER and Papp values of RB obtained from the optimized PVPASC model indicated a similar barrier property to porcine ear skin. Scanning electron microscope analysis demonstrated a mimic 'brick-and-mortar' structure. The PVPASC model can be stored for three weeks at -20 °C, and withstand the presence of different receptor medium for 24 h. The permeation studies of the active components demonstrated a good correlation (r2 = 0.9136) of Papp values between the drugs' permeation through the PVPASC model and porcine ear skin. CONCLUSION: Keratin contained composite phospholipid vesicle-based permeation assay models have been proven to be potential skin tools in topical/transdermal permeation studies.
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Permeabilidad , Fosfolípidos , Absorción Cutánea , Humanos , Fosfolípidos/química , Absorción Cutánea/efectos de los fármacos , Absorción Cutánea/fisiología , Porcinos , Permeabilidad/efectos de los fármacos , Animales , Liposomas , Administración Cutánea , Epidermis/metabolismo , Epidermis/efectos de los fármacos , Piel/metabolismo , Piel/efectos de los fármacos , Piel Artificial , Rodaminas/farmacocinética , Rodaminas/química , Rodaminas/administración & dosificaciónRESUMEN
PURPOSE: Topical therapy of local disease (e.g. skin) is advantageous over oral therapy since there is less systemic drug distribution (so fewer side-effects), no first-pass effect, etc. However, patient compliance with topical therapy can be poor as it may require many applications a day and can last months. Here we propose a topical controlled release formulation with thermoresponsive gelation at body temperature and improved adhesiveness, making it easier to remain in contact with the body. METHODS: The formulation contains two excipients, poloxamer 407 (P407) and casein. Casein can modify the properties of the hydrogel through molecular entanglement. In addition, tissue reaction and drug release profile were evaluated. RESULTS: Changes in casein concentration affected adhesive strength, viscosity, mechanical properties and drug release, presumably by hydrophobic interactions between casein and P407. Two different concentrations of P407 were tested with two different concentrations of casein. Formulations containing 5% and 10% casein released 80% of model drug in 48 h, while formulations without casein released the same fraction in around 24 h hours. Formulations with 10% casein had almost twice the adhesive strength of those without casein. CONCLUSIONS: Addition of casein modified the mechanical properties and drug release rate of the hydrogel. There was no inflammation or injury after brief exposure in vivo.
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Bupivacaína/química , Caseínas/química , Portadores de Fármacos/química , Hidrogeles/química , Poloxámero/química , Rodaminas/química , Adhesividad , Administración Tópica , Animales , Bupivacaína/administración & dosificación , Bupivacaína/farmacocinética , Composición de Medicamentos , Liberación de Fármacos , Excipientes/química , Humanos , Masculino , Fenómenos Mecánicos , Ratas Sprague-Dawley , Reología , Rodaminas/administración & dosificación , Rodaminas/farmacocinética , Temperatura , ViscosidadRESUMEN
The aim of the present paper is to study the effect of common excipients on the permeability of atenolol (as drug absorbed mainly by passive diffusion) and rhodamine (as P-glycoprotein substrate). The apparent permeability was measured by an in situ perfusion method in Wistar rats using the closed loop Doluisio's method. Permeability values were characterized in the absence and presence of 18 commonly used excipients. Excipient concentrations were selected based on the amounts in oral immediate release dosage forms, which failed the test during the human bioequivalence studies. Atenolol was studied with and without excipients in the whole small intestine, whereas rhodamine was tested in three different intestinal segments to account for the differential expression of P-glycoprotein, and it was further on tested in the ileum, in the presence of excipients. Atenolol presented higher permeability values when it was administered with colloidal silica, croscarmellose, hydroxypropyl methylcellulose (HPMC), magnesium stearate, MgCO3, poly(ethylene glycol) 400, poly(vinylpyrrolidone), sorbitol, starch, and TiO2 rhodamine showed higher permeability values when it was administered with croscarmellose and HPMC. On the one hand, the mechanisms of action were not discernible with the proposed experiments. On the other hand, commercial formulations do not present a single excipient but several, which can counteract their effects. The in situ perfusion technique can be useful for a preliminary screening and risk analysis, while the in vivo pharmacokinetic results would be needed to define conclusive effects.
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Atenolol/farmacocinética , Composición de Medicamentos/métodos , Excipientes/farmacología , Íleon/metabolismo , Absorción Intestinal/efectos de los fármacos , Rodaminas/farmacocinética , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Administración Oral , Animales , Atenolol/administración & dosificación , Difusión/efectos de los fármacos , Masculino , Permeabilidad/efectos de los fármacos , Ratas , Ratas Wistar , Rodaminas/administración & dosificaciónRESUMEN
The blood-arachnoid barrier (BAB), which is formed by arachnoid epithelial cells linked by tight junctions, has generally been considered impermeable to water-soluble substances. However, we recently demonstrated that organic anion transporters 1 and 3 (Oat1 and Oat3) play roles in drug clearance at the BAB. Here, we examined whether an organic anion-transporting polypeptide (Oatp) also plays a role, using the fluorescent organic anion sulforhodamine-101 (SR-101) as a model substrate. SR-101 was injected into the cisterna magna of rats in order to minimize the contribution of choroid plexus transport. The in vivo cerebrospinal fluid (CSF) elimination clearance of SR-101 after intracisternal administration was ninefold greater than that of fluorescein-labeled inulin, a bulk flow marker. In the case of pre-administration of taurocholate, a broad-spectrum inhibitor of Oatps, or digoxin, a strong substrate/inhibitor for Oatp1a4 but not for Oatp1a1, Oat1, and Oat3, the CSF elimination of SR-101 was significantly reduced, becoming similar to that of inulin, and thus indicating complete inhibition of SR-101 clearance from the CSF. The distribution of SR-101 fluorescence was restricted to the arachnoid mater in the absence of inhibitor, whereas the fluorescence was increased in the parenchyma of the spinal cord after co-injection of taurocholate or digoxin. Immunostaining confirmed the localization of Oatp1a4 in the arachnoid mater. These results indicate that Oatp1a4 at the BAB acts as an avid clearance pathway of SR-101 in the CSF to the blood. Thus, Oatp1a4 appears to play a major role in CSF detoxification by limiting the distribution of organic anions to the brain and spinal cord.
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Aracnoides/metabolismo , Barrera Hematoencefálica/metabolismo , Líquido Cefalorraquídeo/metabolismo , Transportadores de Anión Orgánico/metabolismo , Rodaminas/farmacocinética , Animales , Encéfalo/metabolismo , Digoxina/farmacología , Colorantes Fluorescentes/farmacocinética , Masculino , Tasa de Depuración Metabólica , Transportadores de Anión Orgánico/antagonistas & inhibidores , Proteínas de Transporte de Catión Orgánico/metabolismo , Compuestos Orgánicos/farmacocinética , Ratas , Ratas Wistar , Rodaminas/administración & dosificación , Médula Espinal/metabolismo , Ácido Taurocólico/farmacología , Distribución TisularRESUMEN
Polymeric microneedles have attracted increasing attention as a minimally invasive platform for delivering drugs or vaccines in a more patient-friendly manner. However, traditional microfabrication techniques using negative molds with needle-shaped cavities usually require cumbersome centrifugation and vacuum degassing processes, which have restricted the scaled-up mass production of polymeric microneedles. Herein, a novel polydimethylsiloxane (PDMS)-based negative mold with cavities packed with silk fibroin scaffold is developed for rapid fabrication of polymeric microneedles, which comprise primarily the composition of poly(ethylene glycol) diacrylate (PEGDA) and sucrose as the needle matrix. Fibroin scaffolds can instantly adsorb prepolymer solution due to capillary force, and subsequently initiate the formation of microneedles via photoinduced polymerization. Based on three types of model drugs, including Rhodamine B (RhB), indocyanine green (ICG), and doxorubicin (DOX), the fabricated PEGDA/sucrose microneedles can realize effective transdermal delivery and controllable release of therapeutic molecules by regulating the sucrose content. The presented method provides a simple strategy for quick fabrication of polymeric microneedles toward transdermal drug delivery applications.
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Administración Cutánea , Sistemas de Liberación de Medicamentos , Fibroínas/química , Agujas , Polietilenglicoles/química , Sacarosa/química , Animales , Doxorrubicina/administración & dosificación , Células Endoteliales de la Vena Umbilical Humana , Humanos , Verde de Indocianina/administración & dosificación , Ratones , Porosidad , Rodaminas/administración & dosificaciónRESUMEN
Nanoparticles are a useful material in biomedicine given their unique properties and biocompatibility; however, there is increasing concern regarding the potential toxicity of nanoparticles with respect to cell metabolism. Some evidence suggests that nanoparticles can disrupt glucose and energy homeostasis. In this study, we investigated the metabolomic, transcriptomic, and integrated effects of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate dye [MNPs@SiO2(RITC)] on glucose metabolism in human embryonic kidney 293 (HEK293) cells. Using gas chromatography-tandem mass spectrometry, we analysed the metabolite profiles of 14 organic acids (OAs), 20 amino acids (AAs), and 13 fatty acids (FAs) after treatment with 0.1 or 1.0 µg/µl MNPs@SiO2(RITC) for 12 h. The metabolic changes were highly related to reactive oxygen species (ROS) generation and glucose metabolism. Additionally, effects on the combined metabolome and transcriptome or "metabotranscriptomic network" indicated a relationship between ROS generation and glucose metabolic dysfunction. In the experimental validation, MNPs@SiO2(RITC) treatment significantly decreased the amount of glucose in cells and was associated with a reduction in glucose uptake efficiency. Decreased glucose uptake efficiency was also related to ROS generation and impaired glucose metabolism in the metabotranscriptomic network. Our results suggest that exposure to high concentrations of MNPs@SiO2(RITC) produces maladaptive alterations in glucose metabolism and specifically glucose uptake as well as related metabolomic and transcriptomic disturbances via increased ROS generation. These findings further indicate that an integrated metabotranscriptomics approach provides useful and sensitive toxicological assessment for nanoparticles.
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Glucosa/metabolismo , Nanopartículas de Magnetita/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Dióxido de Silicio/química , Células HEK293 , Humanos , Nanopartículas de Magnetita/administración & dosificación , Metabolómica , Rodaminas/administración & dosificación , TranscriptomaRESUMEN
Background Direct quantitative measurement of GFR (mGFR) remains a specialized task primarily performed in research settings. Multiple formulas for estimating GFR have been developed that use the readily available endogenous biomarkers creatinine and/or cystatin C. However, eGFR formulas have limitations, and an accurate mGFR is necessary in some clinical situations and for certain patient populations. We conducted a prospective, open-label study to evaluate a novel rapid technique for determining plasma volume and mGFR.Methods We developed a new exogenous biomarker, visible fluorescent injectate (VFI), consisting of a large 150-kD rhodamine derivative and small 5-kD fluorescein carboxymethylated dextrans. After a single intravenous injection of VFI, plasma volume and mGFR can be determined on the basis of the plasma pharmacokinetics of the rhodamine derivative and fluorescein carboxymethylated dextrans, respectively. In this study involving 32 adults with normal kidney function (n=16), CKD stage 3 (n=8), or CKD stage 4 (n=8), we compared VFI-based mGFR values with values obtained by measuring iohexol plasma disappearance. VFI-based mGFR required three 0.5-ml blood draws over 3 hours; iohexol-based mGFR required five samples taken over 6 hours. Eight healthy participants received repeat VFI injections at 24 hours.Results VFI-based mGFR values showed close linear correlation with the iohexol-based mGFR values in all participants. Injections were well tolerated, including when given on consecutive days. No serious adverse events were reported. VFI-based mGFR was highly reproducible.Conclusions The VFI-based approach allows for the rapid determination of mGFR at the bedside while maintaining patient safety and measurement accuracy and reproducibility.
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Dextranos/farmacocinética , Fluoresceína/farmacocinética , Tasa de Filtración Glomerular , Volumen Plasmático , Sistemas de Atención de Punto , Insuficiencia Renal Crónica/fisiopatología , Rodaminas/farmacocinética , Adulto , Anciano , Estudios de Casos y Controles , Dextranos/administración & dosificación , Femenino , Fluoresceína/administración & dosificación , Colorantes Fluorescentes/administración & dosificación , Colorantes Fluorescentes/farmacocinética , Humanos , Inyecciones Intravenosas , Yohexol/farmacocinética , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Reproducibilidad de los Resultados , Rodaminas/administración & dosificación , Adulto JovenRESUMEN
Objective: To design and characterize aerosol microparticles (MP) to provide sustained release of the water-soluble compound sulforhodamine B (SRB) and achieve effective aerosol dispersion. Significance: Modulating the release of water-soluble compounds remains a challenge in pulmonary drug delivery. Methods: SRB and water made up an aqueous solution, while acetalated dextran (Ac-Dex) and isopropyl alcohol made up an organic solution. The two solutions were mixed together, and the solution was spray dried to produce MP. MP were characterized for morphology, size, release kinetics, aerosol dispersion, and cellular interactions. Results: Ac-Dex MP exhibited corrugated morphology and aerodynamic diameters from 2.06 to 2.86 µm. MP deposited in all stages of a Next Generation Impactor, with >90% fine particle fraction. MP exhibited encapsulation efficiencies >129% with SRB loading values up to 16.7 µg SRB/mg MP. MP exhibited sustained release of SRB at pH 7 and fast release at pH 5. In vitro experiments showed minimal cytotoxicity, successful uptake of MP in epithelial cells, and no disruption to the integrity of epithelial monolayers. Conclusions: Ac-Dex MP systems demonstrated the ability to provide sustained the release of a water-soluble therapeutic in addition to effective aerosol dispersion for pulmonary applications.
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Aerosoles/química , Preparaciones de Acción Retardada/química , Dextranos/química , Colorantes Fluorescentes/administración & dosificación , Rodaminas/administración & dosificación , Acetilación , Administración por Inhalación , Cristalización , Liberación de Fármacos , Inhaladores de Polvo Seco , Colorantes Fluorescentes/química , Polvos , Rodaminas/química , Agua/químicaRESUMEN
The aim of this work is to prepare ultraviolet (UV) triggered controlled release of compounds from microcapsule systems (MCs). Polyurethane (PU) and poly(methyl methacrylate) (PMMA) microcapsules were studied with/without chemical functionalization using photocatalytic TiO2 nanoparticles (NPs) on their surface. Once TiO2 nanoparticles are illuminated with UV light (λ = 370 nm), they initiate the rupture of the polymeric bonds of the microcapsule and subsequently initiate the encapsulated compound release, methotrexate (MTX) or rhodamine (Rh), in the present work. The size, polydispersity, charge, and yield of all MCs were measured, being the methotrexate drug release for all systems determined and compared with and without functionalization with TiO2 NPs, under dark, visible light and UV illumination in vitro. Finally, the Rh release was characterized using fluorescence microscopy. The TiO2 NPs size is around 10 nm, as determined by X-ray diffraction experiments. The PU MCs average size is around 60 µm, its electric charge +3.11 mV and yield around 85%. As for the PMMA MCs, the average size is around 280 µm, its electric charge -7.2 mV and yield around 25% and 30% for both MTX and Rh, respectively. In general, adding TiO2 NPs or the encapsulated products to the MCs does not affect the size but functionalization with TiO2 NPs lowers the electric charge. Microcapsules functionalized with TiO2 nanoparticles and irradiated with UV light presented the highest release of MTX and Rh. All other samples showed lower drug release levels when studied under the same conditions.
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Preparaciones de Acción Retardada/administración & dosificación , Composición de Medicamentos/métodos , Metotrexato/administración & dosificación , Cápsulas , Catálisis/efectos de la radiación , Liberación de Fármacos , Nanopartículas del Metal/química , Metotrexato/farmacocinética , Polimetil Metacrilato/química , Poliuretanos/química , Rodaminas/administración & dosificación , Rodaminas/farmacocinética , Titanio/química , Rayos UltravioletaRESUMEN
A mixed organosilane system for simultaneous formation of organosilica nanoparticles has been systematically studied for loading of various compounds with a wide range of log P values. The molecule-entrapping system was understood by investigating the effects of adjusting various experimental parameters on particle formation and molecule entrapment. Particularly, rhodamine 6 G (R6G) loaded colloidal particles were prepared and characterised in detail. The results show that whereas most molecules had entrapment efficiency (EE%) in the range of 20-80%, R6G exhibited near 100% efficiency. Moreover, the colloidal system can be tuned to incorporate R6G with the extent of entrapment spanning at least 2 orders of magnitude (i.e. from 0.04 to 4 mg) and a maximum EE% of 98%. In conclusion, the study demonstrates the promise of the proposed mixed organosilane system in forming colloidal particles containing multiple functional groups with selective loading of highly hydrophobic molecules.
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Antineoplásicos/administración & dosificación , Coloides/química , Portadores de Fármacos/química , Nanopartículas/química , Compuestos de Organosilicio/química , Rodaminas/administración & dosificación , Antineoplásicos/química , Línea Celular , Precipitación Química , Composición de Medicamentos/métodos , Liberación de Fármacos , Colorantes Fluorescentes/administración & dosificación , Colorantes Fluorescentes/química , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Nanopartículas/ultraestructura , Nanotecnología/métodos , Tamaño de la Partícula , Rodaminas/químicaRESUMEN
Intranasal delivery of mesenchymal stem cells (MSCs) to the olfactory bulb is a promising approach for treating olfactory injury. Additionally, using the homing phenomenon of MSCs may be clinically applicable for developing therapeutic cell carriers. Herein, using superparamagnetic iron oxide nanoparticles (SPIONs) and a permanent magnet, we demonstrated an enhanced homing effect in an olfactory model. Superparamagnetic iron oxide nanoparticles with rhodamine B (IRBs) had a diameter of 5.22 ± 0.9 nm and ζ-potential of +15.2 ± 0.3 mV. IRB concentration of 15 µg/mL was injected with SPIONs into MSCs, as cell viability significantly decreased when 20 μg/mL was used (p ≤ 0.005) compared to in controls. The cells exhibited magnetic attraction in vitro. SPIONs also stimulated CXCR4 (C-X-C chemokine receptor type 4) expression and CXCR4-SDF-1 (Stromal cell-derived factor 1) signaling in MSCs. After injecting magnetized MSCs, these cells were detected in the damaged olfactory bulb one week after injury on one side, and there was a significant increase compared to when non-magnetized MSCs were injected. Our results suggest that SPIONs-labeled MSCs migrated to injured olfactory tissue through guidance with a permanent magnet, resulting in better homing effects of MSCs in vivo, and that iron oxide nanoparticles can be used for internalization, various biological applications, and regenerative studies.
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Quimiocina CXCL12/inmunología , Nanopartículas de Magnetita/administración & dosificación , Bulbo Olfatorio/efectos de los fármacos , Receptores CXCR4/genética , Animales , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Compuestos Férricos/administración & dosificación , Compuestos Férricos/química , Imagen por Resonancia Magnética , Nanopartículas de Magnetita/química , Células Madre Mesenquimatosas/efectos de los fármacos , Ratones , Bulbo Olfatorio/lesiones , Bulbo Olfatorio/metabolismo , Rodaminas/administración & dosificación , Rodaminas/química , Transducción de Señal/efectos de los fármacosRESUMEN
Neonatal hypoxiaâ»ischemia is one of the main causes of mortality and disability of newborns. To study the mechanisms of neonatal brain cell damage, we used a model of neonatal hypoxiaâ»ischemia in seven-day-old rats, by annealing of the common carotid artery with subsequent hypoxia of 8% oxygen. We demonstrate that neonatal hypoxiaâ»ischemia causes mitochondrial dysfunction associated with high production of reactive oxygen species, which leads to oxidative stress. Targeted delivery of antioxidants to the mitochondria can be an effective therapeutic approach to treat the deleterious effects of brain hypoxiaâ»ischemia. We explored the neuroprotective properties of the mitochondria-targeted antioxidant SkQR1, which is the conjugate of a plant plastoquinone and a penetrating cation, rhodamine 19. Being introduced before or immediately after hypoxiaâ»ischemia, SkQR1 affords neuroprotection as judged by the diminished brain damage and recovery of long-term neurological functions. Using vital sections of the brain, SkQR1 has been shown to reduce the development of oxidative stress. Thus, the mitochondrial-targeted antioxidant derived from plant plastoquinone can effectively protect the brain of newborns both in pre-ischemic and post-stroke conditions, making it a promising candidate for further clinical studies.
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Hipoxia-Isquemia Encefálica/prevención & control , Fármacos Neuroprotectores/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Plastoquinona/análogos & derivados , Rodaminas/administración & dosificación , Animales , Animales Recién Nacidos , Modelos Animales de Enfermedad , Hipoxia-Isquemia Encefálica/metabolismo , Masculino , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/fisiología , Fármacos Neuroprotectores/farmacología , Plastoquinona/administración & dosificación , Plastoquinona/farmacología , Ratas , Especies Reactivas de Oxígeno/metabolismo , Recuperación de la Función/efectos de los fármacos , Rodaminas/farmacologíaRESUMEN
We report the control of guest release profiles by dialing-in desirable interactions between guest molecules and pores in metal-organic frameworks (MOFs). The interactions can be derived by the rate constants that were quantitatively correlated with the type of functional group and its proportion in the porous structure; thus the release of guest molecules can be predicted and programmed. Specifically, three probe molecules (ibuprofen, rhodamine B, and doxorubicin) were studied in a series of robust and mesoporous MOFs with multiple functional groups [MIL-101(Fe)-(NH2)x, MIL-101(Fe)-(C4H4)x, and MIL-101(Fe)-(C4H4)x(NH2)1-x]. The release rate can be adjusted by 32-fold [rhodamine from MIL-101(Fe)-(NH2)x], and the time of release peak can be shifted by up to 12 days over a 40-day release period [doxorubicin from MIL-101(Fe)-(C4H4)x(NH2)1-x], which was not obtained in the physical mixture of the single component MOF counterparts nor in other porous materials. The corelease of two pro-drug molecules (ibuprofen and doxorubicin) was also achieved.
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Complejos de Coordinación/química , Preparaciones de Acción Retardada/química , Compuestos de Hierro/química , Estructuras Metalorgánicas/química , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/química , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/química , Doxorrubicina/administración & dosificación , Doxorrubicina/química , Liberación de Fármacos , Ibuprofeno/administración & dosificación , Ibuprofeno/química , Modelos Moleculares , Porosidad , Rodaminas/administración & dosificación , Rodaminas/químicaRESUMEN
Mitochondrial gene therapy seems to be a valuable and promising strategy to treat mitochondrial disorders. The use of a therapeutic vector based on mitochondrial DNA, along with its affinity to the site of mitochondria, can be considered a powerful tool in the reestablishment of normal mitochondrial function. In line with this and for the first time, we successfully cloned the mitochondrial gene ND1 that was stably maintained in multicopy pCAG-GFP plasmid, which is used to transform E. coli. This mitochondrial-gene-based plasmid was encapsulated into nanoparticles. Furthermore, the functionalization of nanoparticles with polymers, such as cellulose or gelatin, enhances their overall properties and performance for gene therapy. The fluorescence arising from rhodamine nanoparticles in mitochondria and a fluorescence microscopy study show pCAG-GFP-ND1-based nanoparticles' cell internalization and mitochondria targeting. The quantification of GFP expression strongly supports this finding. This work highlights the viability of gene therapy based on mitochondrial DNA instigating further in vitro research and clinical translation.
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ADN Mitocondrial/genética , Genes Mitocondriales/genética , Mitocondrias/efectos de los fármacos , Mitocondrias/genética , Nanopartículas/administración & dosificación , Plásmidos/genética , Rodaminas/administración & dosificación , Animales , Clonación Molecular/métodos , Escherichia coli/genética , Fluorescencia , Gelatina/administración & dosificación , Terapia Genética/métodos , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes/genética , Humanos , Ratones , Polímeros/administración & dosificación , Transfección/métodosRESUMEN
One major challenge in the field of nanotherapeutics is to increase the selective delivery of cargo to targeted cells. Using polylactic-co-glycolic acid (PLGA), we recently highlighted the importance of polymer composition in the biological fate of the nanodrug delivery systems. However, the route of internalization of polymeric nanoparticles (NPs) is another key component to consider in the elaboration of modern and targeted devices. For that purpose, herein, we effectively synthesized and characterized glutathione-functionalized PLGA-nanoparticles (GSH-NPs) loaded with curcumin (GSH-NPs-Cur), using thiol-maleimide click reaction and determined their physicochemical properties. We found that GSH functionalization did not affect the drug loading efficiency (DLE), the size, the polydispersity index (PDI), the zeta potential, the release profile, and the stability of the formulation. While being nontoxic, the presence of GSH on the surface of the formulations exhibits a better neuroprotective property against acrolein. The neuronal internalization of GSH-NPs-Cur was higher than free curcumin. In order to track the intracellular localization of the formulations, we used a covalently attached rhodamine (PLGA-Rhod), into our GSH-functionalized matrix. We found that GSH-functionalized matrix could easily be taken up by neuronal cells. Furthermore, we found that GSH conjugation modifies the route of internalization enabling them to escape the uptake through macropinocytosis and therefore avoiding the lysosomal degradation. Taken together, GSH functionalization increases the uptake of formulations and modifies the route of internalization toward a safer pathway. This study shows that the choice of ideal ligand to develop NPs-targeting devices is a crucial step when designing innovative strategy for neuronal cells delivery.
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Curcumina/química , Curcumina/farmacología , Glutatión/metabolismo , Nanopartículas/química , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Poliésteres/química , Polietilenglicoles/química , Línea Celular Tumoral , Química Farmacéutica/métodos , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos/métodos , Estabilidad de Medicamentos , Humanos , Ligandos , Nanopartículas/administración & dosificación , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/farmacología , Tamaño de la Partícula , Poliésteres/administración & dosificación , Polietilenglicoles/administración & dosificación , Rodaminas/administración & dosificación , Rodaminas/químicaRESUMEN
The highly acidic gastric environment creates a physiological barrier for using therapeutic drugs in the stomach. While proton pump inhibitors have been widely used for blocking acid-producing enzymes, this approach can cause various adverse effects. Reported herein is a new microdevice, consisting of magnesium-based micromotors which can autonomously and temporally neutralize gastric acid through efficient chemical propulsion in the gastric fluid by rapidly depleting the localized protons. Coating these micromotors with a cargo-containing pH-responsive polymer layer leads to autonomous release of the encapsulated payload upon gastric-acid neutralization by the motors. Testing in a mouse model demonstrate that these motors can safely and rapidly neutralize gastric acid and simultaneously release payload without causing noticeable acute toxicity or affecting the stomach function, and the normal stomach pH is restored within 24â h post motor administration.
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Preparaciones de Acción Retardada/química , Ácido Gástrico/química , Magnesio/química , Polímeros/química , Animales , Liberación de Fármacos , Colorantes Fluorescentes/administración & dosificación , Oro/química , Concentración de Iones de Hidrógeno , Ratones , Ácidos Polimetacrílicos/química , Rodaminas/administración & dosificaciónRESUMEN
There is a great challenge in constructing pH-responsive drug delivery systems in biomedical application research. Many nanocomposites are intended to be pH-responsive as drug carriers because of a tumorous or intracellular mildly acidic environment. However, it is always difficult to find an appropriate system for quick response and release before the carrier is excreted from the living system. In this work, hyperbranched polymer, hyperbranched polyglycerol (hPG), and conjugated mesoporous silica nanoparticles (MSNs) were assembled as complexes to serve as drug carriers. Herein, the conjugated polymer-MSNs interacted through the Schiff base bond, which possessed a mildly acidic responsive property. Interestingly, the assembled system could rapidly respond and release guest molecules inside cancer cells. This would make the entrapped drug released before the carriers escape from the endosome counterpart. The results show that the assembled composite complexes can be considered to be a drug delivery system for cancer therapy.
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Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Nanopartículas/química , Células A549 , Reactivos de Enlaces Cruzados , Endocitosis , Colorantes Fluorescentes/administración & dosificación , Colorantes Fluorescentes/farmacocinética , Glicerol/química , Humanos , Concentración de Iones de Hidrógeno , Microscopía Confocal , Microscopía Electrónica de Transmisión , Nanopartículas/ultraestructura , Nanotecnología , Neoplasias/tratamiento farmacológico , Polímeros/química , Rodaminas/administración & dosificación , Rodaminas/farmacocinética , Bases de Schiff/química , Dióxido de Silicio/química , Microambiente TumoralRESUMEN
Mesoporous silica nanoparticles (MSNs) are highly attractive as supports in the design of controlled delivery systems that can act as containers for the encapsulation of therapeutic agents, overcoming common issues such as poor water solubility and poor stability of some drugs and also enhancing their bioavailability. In this context, we describe herein the development of polyglutamic acid (PGA)-capped MSNs that can selectively deliver rhodamine B and doxorubicin. PGA-capped MSNs remain closed in an aqueous environment, yet they are able to deliver the cargo in the presence of pronase because of the hydrolysis of the peptide bonds in PGA. The prepared solids released less than 20% of the cargo in 1 day in water, whereas they were able to reach 90% of the maximum release of the entrapped guest in ca. 5 h in the presence of pronase. Studies of the PGA-capped nanoparticles with SK-BR-3 breast cancer cells were also undertaken. Rhodamine-loaded nanoparticles were not toxic, whereas doxorubicin-loaded nanoparticles were able to efficiently kill more than 90% of the cancer cells at a concentration of 100 µg/mL.
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Sistemas de Liberación de Medicamentos , Nanopartículas/química , Ácido Poliglutámico/química , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/farmacocinética , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Doxorrubicina/administración & dosificación , Doxorrubicina/farmacocinética , Femenino , Humanos , Microscopía Confocal , Microscopía Electrónica de Transmisión , Nanopartículas/ultraestructura , Nanotecnología , Pronasa/administración & dosificación , Rodaminas/administración & dosificación , Rodaminas/farmacocinética , Dióxido de Silicio/químicaRESUMEN
PURPOSE: To assess the potential of triblock copolymers based on poly(ethylene glycol) (PEG) and functionalized poly(ε-caprolactone) as temperature/pH responsive gels for controlled delivery of macromolecules. METHODS: Poly(α-carboxylate-co-α-benzylcarboxylate-ε-caprolactone)-PEG-poly(α-carboxylate-co-α-benzylcarboxylate-ε-caprolactone) (PCBCL-PEG-PCBCL) was synthesized through ring opening polymerization of α-benzyl carboxylate-ε-caprolactone by PEG, followed by 30% debenzylation of the lateral blocks. The effect of Tris buffer and pH on the sol-gel transition temperature of PCBCL-PEG-PCBCL was assessed. The temperature/pH responsive release of tetramethylrhodamine-dextran (TMR-D) (10 and 40 kDa) from PCBCL-PEG-PCBCL was investigated. RESULTS: Replacement of water with Tris buffer reduced PCBCL-PEG-PCBCL sol-gel transition temperature. Thermo-reversible hydrogels were only formed at pHs ≥ 5.0, but PCBCL-PEG-PCBCL transition temperature was not affected by pH above pH 5.0. In contrast to Pluronic F127 that released 100% of TMR-D within 2 h, PCBCL-PEG-PCBCL hydrogel controlled TMR-D release efficiently at pH = 7.4 and 37°C (~27 and 11% TMRD 10 and 40 kDa release within 150 h, respectively). At 50°C or pH = 9.0, TMR-D release was increased slightly, while at room temperature or pH = 5.0, no control over TMR-D release was observed by PCBCL-PEG-PCBCL hydrogel. CONCLUSION: PCBCL-PEG-PCBCL hydrogel provides depot release of macromolecules at physiological conditions. This release can be triggered through changes in the temperature or pH.
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Preparaciones de Acción Retardada/química , Hidrogeles/química , Poliésteres/química , Polietilenglicoles/química , Dextranos/administración & dosificación , Colorantes Fluorescentes/administración & dosificación , Concentración de Iones de Hidrógeno , Rodaminas/administración & dosificación , TemperaturaRESUMEN
A series of tryptophan-based peptides W1a, b-W4a, b, with diverse architectures were designed and synthesized. These tryptophan containing peptides can self-assemble to spherical particle. This self-assembled system was demonstrated to encapsulate rhodamine B and penetrate the cell membrane.