Nanoparticles as carrier for improve therapeutic efficacy of pioglitazone in ocular inflammatory disorders: development and validation of a high throughput LC-MS/MS method for quantitation in ocular tissues

Abstract: Pioglitazone is an oral anti-hyperglycemic agent and it is used for the treatment of diabetes mellitus type 2. The anti-inflammatory activity has also been demonstrated in the literature. Pioglitazone belongs to Class II of Biopharmaceutical Classification System, i.e., slightly soluble and highly permeable. Polymeric nanoparticle formulations play an important role in the improvement of the efficacy of ocular therapies. These systems are non-toxic and biodegradable, show appropriate physicochemical characteristics as well as prolonged release profile suitable for ocular delivery. An accurate, sensitive, selective, reproducible and high throughput HPLC-MS/MS method was validated to quantitate pioglitazone in ocular tissues (cornea, sclera, lens, aqueous humour and vitreous humour). The chromatographic separation was achieved in 10 min on a Kinetex C18 column. Linear response of pioglitazone was observed over the range of 5-100 ng/ml. The limit of quantitation was 10 ng/ml (in extract). The recovery of pioglitazone or pioglitazone encapsulated in nanoparticles of polylactic-co-glycolic acid-polyethylene glycol (PLGA-PEG) was in the range 85-115 % in all tissues and levels tested. The intra-day and inter-day precision were <5 % and <10 % respectively. The obtained extracts demonstrated to be stable under various experimental conditions in all the studied matrices. This method can be applied to in vivo and ex vivo biodistribution studies related to the ocular administration of pioglitazone nanoparticles

Validated spectrofluorometric method for determination of gemfibrozil in self nanoemulsifying drug delivery systems (SNEDDS)

A spectrofluorometric method has been developed and validated for the determination of gemfibrozil. The method is based on the excitation and emission capacities of gemfibrozil with excitation and emission wavelengths of 276 and 304 nm respectively. This method allows de determination of the drug in a self-nanoemulsifying drug delivery system (SNEDDS) for improve its intestinal absorption. Results obtained showed linear relationships with good correlation coefficients (r(2)>0.999) and low limits of detection and quantification (LOD of 0.075 μg mL(-1) and LOQ of 0.226 μg mL(-1)) in the range of 0.2-5 μg mL(-1), equally this method showed a good robustness and stability. Thus the amounts of gemfibrozil released from SNEDDS contained in gastro resistant hard gelatine capsules were analysed, and release studies could be performed satisfactorily

Percepción de los nuevos graduados de Farmacia sobre la adquisición de competencias frente a los egresados del ciclo formativo

Podeu consultar la Vuitena trobada de professorat de Ciències de la Salut completa a: http://hdl.handle.net/2445/66524Tras la implantación en 2009-10 del nuevo plan de estudios de Farmacia-UB, conviene explorar desde el punto de vista de los nuevos graduados (GF) el papel de las metodologías docentes y de la evaluación en el desarrollo de sus competencias. Para ello, se ha pasado un cuestionario cerrado a la promoción del 2013 y, paralelamente, a egresados del Ciclo formativo (FP) Fabricación de productos farmacéuticos i afines, con el fin de identificar y contrastar qué competencias les parecen haber desarrollado y saber qué factores atribuyen a dicho desarrollo así como posibles carencias. Según las respuestas recibidas (15 FP y 58 GF), ambos grupos coinciden en que los centros contribuyen mucho a adquirir las competencias específicas del área y poco la capacidad comunicativa y ética-sostenibilidad. En las demás, los FP se sienten claramente mejor preparados en competencias que los GF, en particular para el trabajo en equipo. En Farmacia, el método expositivo/lección magistral es el más usado, pero valoran más positivamente el aprendizaje basado en experimentos, casos o problemas. En el Ciclo formativo, lo son el aprendizaje colaborativo y experimentos, igualmente importantes que usados. Entre las actividades realizadas, a ambos grupos les parecen más útiles las prácticas de laboratorio y las externas que el Trabajo de Fin de Grado o el de Síntesis. Test, informe/memoria y trabajo/proyecto son las estrategias evaluativas más empleadas y, además, el examen de desarrollo en FP. Destacan proyectos y prácticas para la adquisición de varias competencias y exposición oral para la capacidad comunicativa y trabajo en equipo..

Study of the stability of packaging and storage conditions of human mesenchymal stem cell for intra-arterial clinical application in patient with critical limb ischemia

Critical limb ischemia (CLI) is associated with significant morbidity and mortality. In this study, we developed and characterized an intra-arterial cell suspension containing human mesenchymal stem cells (hMSCs) for the treatment of CLI. Equally, the stability of cells was studied in order to evaluate the optimal conditions of storage that guarantee the viability from cell processing to the administration phase. Effects of various factors, including excipients, storage temperature and time were evaluated to analyze the survival of hMSCs in the finished medicinal product. The viability of hMSCs in different packaging media was studied for 60 h at 4 °C. The best medium to maintain hMSCs viability was then selected to test storage conditions (4, 8, 25 and 37 °C; 60 h). The results showed that at 4 °C the viability was maintained above 80% for 48 h, at 8 °C decreased slightly, whereas at room temperature and 37 °C decreased drastically. Its biocompatibility was assessed by cell morphology and cell viability assays. During stability study, the stored cells did not show any change in their phenotypic or genotypic characteristics and physicochemical properties remained constant, the ability to differentiate into adipocytes and osteocytes and sterility requirements were also unaltered. Finally, our paper proposes a packing media composed of albumin 20%, glucose 5% and Ringer's lactate at a concentration of 1 × 106 cells/mL, which must be stored at 4 °C as the most suitable to maintain cell viability (>80%) and without altering their characteristics for more than 48 h

Development, physical-chemical stability and release studies of four alcohol-free spironolactone suspensions for use in pediatrics

Dissolution studies have become of great significance because, in most cases, drug dissolution is the rate-limiting step in the absorption process. As occurs with solid oral dosage forms, heterogeneous disperse systems (suspensions) could also have some problems with their in vitro dissolution. The objective of this study was to evaluate influence of the excipients on the release of spironolactone from four alcohol free suspensions (pharmaceutical compounding) of spironolactone 5 mg/mL suitable for pediatric use. Also the comparison of the physical and chemical stability of the suspensions stored at 4, 25 and 40 ºC over a 60- day period has been studied. Rheological behavior, particle size, a prediction of long-term physical stability, pH and assay of spironolactone by HPLC were assessed at prefixed times. The dissolution profile of each suspension was determined and compared with that of the commercial tablets. A microbiological study of the best formula was also performed. Chemically, the four spironolactone suspensions were stable for 60 days stored at three temperatures; Suspension IV had optimum pH values and the highest recovery percentage. In terms of physical stability, sedimentation occurred in Suspension IV and flotation of spironolactone in Suspensions I, II and III. Suspension III had the highest viscosity and the slowest drug release. Suspension IV was also microbiologically stable for 60 days. In conclusion, Suspension IV had the best properties and the least suitable form was Suspension III, as its high viscosity made it difficult to achieve homogeneous redispersion, and it had the slowest dissolution profile

Ética en la docencia. El profesor universitario como modelo de actuación personal y profesional

Podeu consultar la Vuitena trobada de professorat de Ciències de la Salut completa a: http://hdl.handle.net/2445/66524El profesor universitario debe ser un modelo de actuación personal y profesional para sus alumnos; un ejemplo que estimule a los estudiantes tanto en el ejercicio de la profesión, como con el respeto por su entorno. De tal modo que el conocimiento que adquieran los alumnos puedan aplicarlo a la vida cotidiana. Esta práctica se ha llevado a cabo en la asignatura de Trabajo Dirigido. El profesor debe manifestar una coherencia entre acción y el pensamiento, debe ser justo y equitativo en la evaluación y tener en cuenta las necesidades particulares de los alumnos; también debe conseguir motivarlos, de forma amigable, con el fin que ellos se esfuercen para lograr las metas académicas. El objetivo es fomentar el logro de competencias transversales tales como responsabilidad, igualdad y capacidad de trabajo en equipo; mediante la realización de sesiones de debate entre el alumno, el tutor, y/o otros investigadores. Por otra parte, también se pretende un análisis reflexivo de nuestra propia actividad docente con el fin de tener en cuenta el potencial y necesidades particulares de cada alumno para lograr una mejora continua. Para alcanzar estos objetivos, el método consiste en la colaboración directa del alumno con los investigadores de la Unidad en proyectos reales y, sesiones semanales de discusión de los resultados que se van obteniendo. De forma que los alumnos desarrollan capacidad de trabajar en equipo, responsabilidad, respeto entre géneros, pensamiento crítico y una verdadera integración. Finalmente, creemos que la ética en la docencia también se pone de manifiesto a la hora de evaluar el aprendizaje de los alumnos. Por este motivo, es de vital importancia que el profesor universitario fomente un clima de integración, de comunicación y de trabajo en equipo en las sesiones docentes. El análisis reflexivo que ha realizado el grupo docente ha permitido examinar las debilidades, las fortalezas y oportunidades acaecidas de la actividad docente para potenciar mejoras en el nuevo curso

Study of Melatonin as Preventive Agent of Gastrointestinal Damage Induced by Sodium Diclofenac

: Safety profile of nonsteroidal anti‐inflammatory drugs (NSAIDs) has been widely studied and both therapeutic and side effects at the gastric and cardiovascular level have been generally associated with the inhibitory effect of isoform 1 (COX‐1) and 2 (COX‐2) cyclooxygenase enzymes. Now there are evidences of the involvement of multiple cellular pathways in the NSAIDs‐mediated‐gastrointestinal (GI) damage related to enterocyte redox state. In a previous review we summarized the key role of melatonin (MLT), as an antioxidant, in the inhibition of inflammation pathways mediated by oxidative stress in several diseases, which makes us wonder if MLT could minimize GI NSAIDs side effects. So, the aim of this work is to study the effect of MLT as preventive agent of GI injury caused by NSAIDs. With this objective sodium diclofenac (SD) was administered alone and together with MLT in two experimental models, ex vivo studies in pig intestine, using Franz cells, and in vivo studies in mice where stomach and intestine were studied. The histological evaluation of pig intestine samples showed that SD induced the villi alteration, which was prevented by MLT. In vivo experiments showed that SD altered the mice stomach mucosa and induced tissue damage that was prevented by MLT. The evaluation by quantitative reverse transcription PCR (RT‐qPCR) of two biochemical markers, COX‐2 and iNOS, showed an increase of both molecules in less injured tissues, suggesting that MLT promotes tissue healing by improving redox state and by increasing iNOS/NO that under non‐oxidative condition is responsible for the maintenance of GI‐epithelium integrity, increasing blood flow and promoting angiogenesis and that in presence of MLT, COX‐2 may be responsible for wound healing in enterocyte. Therefore, we found that MLT may be a preventive agent of GI damages induced by NSAIDs. Keywords: melatonin; NSAIDs; gastric injuries; antioxidan

Carprofen Permeation Test through Porcine Ex Vivo Mucous Membranes and Ophthalmic Tissues for Tolerability Assessments: Validation and Histological Study

Carprofen (CP), a non-steroidal anti-inflammatory drug (NSAID), is profusely used in veterinary medicine for its analgesic and anti-inflammatory activity. Some undesirable effects are associated with its systemic administration. Alternative local routes are especially useful to facilitate its administration in animals. The main aim of this paper is to validate the suitability of ex vivo permeation experiments of CP with porcine mucous membranes (buccal, sublingual and vaginal) and ophthalmic tissues (cornea, sclera and conjunctiva) intended to be representative of naïve in vivo conditions. Chromatographic analysis of CP in membrane-permeated samples and drug-retained have been validated following standard bioanalytical guidelines. Then, recovery levels of drugs in tissue samples were assessed with aqueous phosphate buffered saline (PBS) buffer to preserve the histological integrity. Finally, as a proof of concept, a series of CP permeation tests in vertical Franz diffusion cells has been performed to evaluate permeation flux and permeability constants in all tissues, followed by a histological study for critical evaluation. Furthermore, synthetic tissue retention-like samples were prepared to verify the value of this experimental study. Results show linear relationships with good determination coefficient (R2 > 0.998 and R2 > 0.999) in the range of 0.78 to 6.25 mg/mL and 3.125 mg/mL to 100 mg/mL, respectively. Low limits of quantification around 0.40 µg/mL were allowed to follow permeation levels until a minimum of 0.40% of the locally-applied dose. This method showed a good accuracy and precision with values lower than 2%. After the recovery technique, reproducible values below 30% were achieved in all tissues, suggesting it is a non-damaging method with low efficiency that requires the use of further solvents to enhance the extraction percentages. After permeation and histology tests, no relevant peak interferences were detected, and no cell or tissue damage was found in any tissue. In conclusion, results demonstrate the suitability of this test to quantify the distribution of CP with good histological tolerability

Novel Polymeric Formulation for Removal of Gastrointestinal Polyps by Digestive Endoscopy

Álvaro Gimeno Sandig from Animal House Bellvitge University of Barcelona is acknowledged for his technical support.Endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD) are two techniques used in the resection of gastrointestinal mucosal polyps. The aim of this work is the development and evaluation of an innovative polymeric solution containing sodium carboxymethylcellulose and hyaluronic acid. For this purpose, several mixtures of these two main components, as well as other components such as fructose, citric acid, and zinc, are evaluated in terms of physicochemical and microbiological properties, rheological behavior, extensibility, syringeability, and stability at different storage conditions. Furthermore, the potential production of mucosal elevation and duration is also studied by an ex vivo model using porcine stomach and colon. Results show that the developed polymeric solutions possess optimal values of pH, from 4.58 to 6.63, for their use in the gastrointestinal tract. The formulations exhibit both Newtonian and pseudoplastic behaviors with different viscosity values as a function of their composition. All formulations exhibit high stability properties and no bacterial or fungal growth is detected. MCS01 and MCS05 are the polymeric solutions with the best syringeability results. In this line, MCS05 is the formulation that provides the highest, 2.20 ± 0.18 cm and 1.40 ± 0.11 cm, and longest-lasting, for more than 120 min, elevation effect on porcine submucosal stomach and colon tissues, respectively. Thus, it can be concluded that polymeric solution MCS05 might be considered as a promising tool for use in human EMR and ESD.This research was funded by the local Regional Government of Andalusia (Junta de Andalucía), grant number PIN-0479-2016

Influence of freeze-drying and γ-irradiation in preclinical studies of flurbiprofen polymeric nanoparticles for ocular delivery using d-(+)-trehalose and polyethylene glycol.

This study investigated the suspension of poly(ε-caprolactone) nanoparticles as an ocular delivery system for flurbiprofen (FB-PεCL-NPs) in order to overcome the associated problems, such as stability, sterility, tolerance, and efficacy, with two different FB-PεCL-NP formulations. The formulations were stabilized with poloxamer 188 (1.66% and 3.5%) and submitted individually for freeze-drying and γ-irradiation with polyethylene glycol 3350 (PEG3350) and d-(+)-trehalose (TRE). Both formulations satisfied criteria according to all physicochemical parameters required for ocular pharmaceuticals. The FB-PεCL-NP formulations showed non-Newtonian behavior and sustained drug release. Ex vivo permeation analysis using isolated ocular pig tissues suggested that the presence of PEG3350 results in a reduction of FB transcorneal permeation. Moreover, TRE improved the penetration of FB across the cornea, especially after γ-irradiation. In addition, both formulations did not show a significant affinity in increasing FB transscleral permeation. Both formulations were classified as nonirritating, safe products for ophthalmic administration according to hen's egg test-chorioallantoic membrane and Draize eye test. Furthermore, an in vivo anti-inflammatory efficacy test showed that irradiated FB-PεCL-NPs prepared with PEG3350 (IR-NPsPEG) have longer anti-inflammatory effects than those presented with irradiated FB-PεCL-NPs prepared with TRE (IR-NPsTRE). IR-NPsPEG showed a suitable physical stability after an aqueous reconstitution over .30 days. This study concludes that both formulations meet the Goldman's criteria and demonstrate how irradiated nanoparticles, with innovative permeation characteristics, could be used as a feasible alternative to a flurbiprofen solution for ocular application in clinical trials