[Supply of pharmaceutical care services: The key to a new model of health services]. / Oferta de servicios de atención farmacéutica: clave para un nuevo modelo de servicios de salud.

Interdisciplinary collaboration between health professionals is essential to improve health outcomes. The competences of pharmacists make them professionals capable of contributing to the comprehensive management of pharmacotherapy in collaboration with other healthcare professionals. The Council of Europe adopted resolution CM/Res(2020)3 on implementation of pharmaceutical care for the benefit of patients and health services in order to promote the appropriate and safe use of medicines. One of the activities to be carried out through the pharmaceutical care process is the detection of drug related problems, such as contraindications, duplications, prescription errors, interactions, etc. In many cases, it involves regular patient follow-up and needs an evaluation of the interventions performed, requiring the establishment of an appropriate interprofessional collaboration framework. This article discusses the issues to be addressed to face the change of model towards a care-based pharmacy.

Nucleic Acid Delivery by Solid Lipid Nanoparticles Containing Switchable Lipids: Plasmid DNA vs. Messenger RNA.

The development of safe and effective nucleic acid delivery systems remains a challenge, with solid lipid nanoparticle (SLN)-based vectors as one of the most studied systems. In this work, different SLNs were developed, by combination of cationic and ionizable lipids, for delivery of mRNA and pDNA. The influence of formulation factors on transfection efficacy, protein expression and intracellular disposition of the nucleic acid was evaluated in human retinal pigment epithelial cells (ARPE-19) and human embryonic kidney cells (HEK-293). A long-term stability study of the vectors was also performed. The mRNA formulations induced a higher percentage of transfected cells than those containing pDNA, mainly in ARPE-19 cells; however, the pDNA formulations induced a greater protein production per cell in this cell line. Protein production was conditioned by energy-dependent or independent entry mechanisms, depending on the cell line, SLN composition and kind of nucleic acid delivered. Vectors containing 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) as unique cationic lipid showed better stability after seven months, which improved with the addition of a polysaccharide to the vectors. Transfection efficacy and long-term stability of mRNA vectors were more influenced by formulation-related factors than those containing pDNA; in particular, the SLNs containing only DOTAP were the most promising formulations for nucleic acid delivery.

Role of endocytic uptake in transfection efficiency of solid lipid nanoparticles-based nonviral vectors.

BACKGROUND: As has been shown for different vector systems, the entry pathway(s) impacts upon the transfection efficiency. The present study aimed to explore the cellular uptake mechanisms of three different vectors based on solid lipid nanoparticles (SLN) in HeLa cells. The use of endocytosis inhibitors that affect specific internalization pathways provides a tool for the study of these routes. METHODS: We prepared three vectors based on solid lipid nanoparticles: without protamine, with protamine, and with protamine and dextran. Uptake, percentage of transfected HeLa cells and enhanced green fluorescent protein (EGFP) production were all analyzed in the presence or absence of different endocytosis inhibitors. In addition, co-localization studies using lysosomal markers were carried out to determine the influence of the trafficking to late endosomal compartments on the transfection capacity of the vectors. RESULTS: Uptake and transfection of each vector was affected differently by each endocytosis inhibitor. Ethylisopropylamiloride (EIPA) did not affect uptake of the DNA-SLN vector, whereas all of the inhibitors affected transfection. In the case of protamine-DNA-SLN and dextran-protamine-DNA-SLN vectors, EIPA affected uptake and dynasore did not decrease transfection. CONCLUSIONS: DNA-SLN vector appear to enter productively by multiple pathways in HeLa cells. By contrast, dynamin does not appear to be essential in the productive entry of protamine-containing vectors. In addition, enhancement of the macropinocytic route increases EGFP production when dextran is added to the vector.

α-Galactosidase A Augmentation by Non-Viral Gene Therapy: Evaluation in Fabry Disease Mice.

Fabry disease (FD) is a monogenic X-linked lysosomal storage disorder caused by a deficiency in the lysosomal enzyme α-Galactosidase A (α-Gal A). It is a good candidate to be treated with gene therapy, in which moderately low levels of enzyme activity should be sufficient for clinical efficacy. In the present work we have evaluated the efficacy of a non-viral vector based on solid lipid nanoparticles (SLN) to increase α-Gal A activity in an FD mouse model after intravenous administration. The SLN-based vector incremented α-Gal A activity to about 10%, 15%, 20% and 14% of the levels of the wild-type in liver, spleen, heart and kidney, respectively. In addition, the SLN-based vector significantly increased α-Gal A activity with respect to the naked pDNA used as a control in plasma, heart and kidney. The administration of a dose per week for three weeks was more effective than a single-dose administration. Administration of the SLN-based vector did not increase liver transaminases, indicative of a lack of toxicity. Additional studies are necessary to optimize the efficacy of the system; however, these results reinforce the potential of lipid-based nanocarriers to treat FD by gene therapy.

Topical Administration of SLN-Based Gene Therapy for the Treatment of Corneal Inflammation by De Novo IL-10 Production.

One of the main challenges in gene therapy is the issue of delivery, and it is especially relevant for the success of gene therapy in the cornea. In the present work, eye drops containing biocompatible non-viral vectors based on solid lipid nanoparticles (SLNs) as gene delivery systems to induce the expression of interleukin 10 (IL-10) were designed to address the treatment of corneal inflammation. Two kinds of SLNs combined with different ligands (protamine, dextran, or hyaluronic acid (HA)) and formulated with polyvinyl alcohol (PVA) were prepared. SLN-based vectors were characterized in terms of size, adhesiveness, viscosity, and pH, before topical administration to wild type and IL-10 knock out (KO) mice. The formulations showed a homogenous particle size below 400 nm and a positive surface charge to favor bioadhesion; the incorporation of PVA improved the corneal penetration. After three days of treatment by topical instillation, SLN-based vectors mainly transfected corneal epithelial cells, HA-formulations being the most effective ones. IL-10 was capable of reaching even the endothelial layer. Corneal sections showed no histological change and formulations seemed to be well tolerated after repeated topical administration. These promising results highlight the possible contribution of non-viral gene augmentation therapy to the future clinical approach of corneal gene therapy.

Oferta de servicios de atención farmacéutica: clave para un nuevo modelo de servicios de salud / Supply of pharmaceutical care services: The key to a new model of health services

La colaboración interdisciplinar entre profesionales sanitarios es esencial para mejorar los resultados en salud de la población. Las capacidades que poseen los farmacéuticos les convierten en profesionales que pueden contribuir al control integral de la farmacoterapia en coordinación con otros profesionales de la salud. El Consejo de Europa aprobó la resolución CM/Res(2020)3 sobre implementación de la atención farmacéutica en el sistema sanitario para promover el uso apropiado y seguro de los medicamentos. Entre las actividades a realizar mediante el proceso de atención farmacéutica destaca la detección de problemas relacionados con la farmacoterapia como son las contraindicaciones, duplicidades, errores de prescripción, interacciones, etc. La intervención farmacéutica en ese ámbito requiere establecer un marco de colaboración interprofesional adecuado. En el presente artículo se comentan los aspectos a abordar para afrontar el cambio de modelo hacia una farmacia asistencial, con una mayor integración e implicación en el sistema, todo ello bajo el amparo del Consejo de Europa.

Interdisciplinary collaboration between health professionals is essential to improve health outcomes. The competences of pharmacists make them professionals capable of contributing to the comprehensive management of pharmacotherapy in collaboration with other healthcare professionals. The Council of Europe adopted resolution CM/Res(2020)3 on implementation of pharmaceutical care for the benefit of patients and health services in order to promote the appropriate and safe use of medicines. One of the activities to be carried out through the pharmaceutical care process is the detection of drug related problems, such as contraindications, duplications, prescription errors, interactions, etc. In many cases, it involves regular patient follow-up and needs an evaluation of the interventions performed, requiring the establishment of an appropriate interprofessional collaboration framework. This article discusses the issues to be addressed to face the change of model towards a care-based pharmacy.

Nanopartículas sólidas lipídicas para terapia génica / Solid lipid nanoparticles for gene therapy

La terapia génica consiste en la administración de ácidos nucleicos con el fin de modular la expresión de proteínas específicas y revertir así una enfermedad. Es una nueva área de la medicina con gran potencial para el tratamiento de enfermedades tanto hereditarias como adquiridas. Hasta ahora, los sistemas virales de administración de ácidos nucleicos han resultado eficaces, pero presentan importantes problemas de seguridad. Los vectores no virales, en cambio, son más seguros pero menos eficaces, aunque su eficacia ha aumentado significativamente en los últimos años. Esta revisión recoge la contribución de nuestro grupo de investigación al diseño de vectores no virales basados en nanopartículas sólidas lipídicas (SLNs) para terapia génica. Hemos estudiado la relación entre factores de la formulación con los procesos de internalización y disposición intracelular del material genético, que condicionan la eficacia de transfección, y por primera vez demostramos la capacidad de las SLNs para inducir la síntesis de una proteína tras su administración endovenosa a ratones. Esta revisión también recoge nuestros trabajos sobre la aplicación de las SLNs en el tratamiento de enfermedades infecciosas y enfermedades raras, como la retinosquisis juvenil ligada al cromosoma X, enfermedad en la que la retina está desestructurada debido a la deficiencia de la proteína retinosquisina. La administración de SLNs con el gen que codifica esta proteína en un modelo animal de esta enfermedad indujo la recuperación estructural de la retina. Los trabajos aquí recogidos muestran el gran potencial de las SLNs como sistemas de administración de ácidos nucleicos (AU)

Gene therapy is a rapidly advancing field with great potential for the treatment of genetic and acquired systemic diseases. This therapy requires the introduction of foreign genetic material in the target cells to modify a genetic sequence. Viral vectors are the most effective, but their application is limited by their immunogenicity, oncogenicity and the small size of the DNA they can transport. Non-viral vectors, however, are safer, lowered cost, and more reproducible and do not present DNA size limit. The main problem of nonviral systems is their low transfection efficiency, although it has improved during the last years. This review presents the contribution of our research group to the design and evaluation of SLNs based non-viral vectors for gene transfer. We report our studies about the relationship between formulation factors and cell uptake and intracellular trafficking of the genetic material, very important for transfection. We have shown, for the first time, the ability to induce transgene protein expression of the SLNs after endovenous administration to mice. This revision also reports our work about the potential application of SLNs to the treatment of infectious and rare diseases, as the X-linked juvenile retinoschisis, a retinal disorder due to a deficiency in the protein retinoschisin, and characterized by poor eyesight and degeneration of the retina. The intraocular injection of SLNs bearing the gene that encodes retinoschisin to diseased mice led to the partial recovery of the retina. All together, our results show the potential of SLNs as a system for gene delivery (AU)