Aplicación clínica de las terapias con células, genes y tejidos en España / Clinical application of cell, gene and tissue therapies in Spain

Los avances científicos y técnicos en el área biomédica y la medicina regenerativa han permitido el desarrollo de nuevos tratamientos, denominados «terapias avanzadas», que engloban la terapia celular, la génica y la ingeniería tisular. Los productos biológicos que pueden fabricarse a partir de estos elementos se clasifican desde el punto de vista de la Agencia Española del Medicamento y Productos Sanitarios en medicamentos de terapias avanzadas, productos derivados de la sangre y trasplantes. Esta revisión pretende aportar información científica y administrativa, de utilidad para el clínico, sobre el uso de estos recursos biológicos

Scientific and technical advances in the areas of biomedicine and regenerative medicine have enabled the development of new treatments known as "advanced therapies", which encompass cell therapy, genetics and tissue engineering. The biologic products that can be manufactured from these elements are classified from the standpoint of the Spanish Agency of Medication and Health Products in advanced drug therapies, blood products and transplants. This review seeks to provide scientific and administrative information for clinicians on the use of these biologic resources

Clinical application of cell, gene and tissue therapies in Spain. / Aplicación clínica de las terapias con células, genes y tejidos en España.

Scientific and technical advances in the areas of biomedicine and regenerative medicine have enabled the development of new treatments known as "advanced therapies", which encompass cell therapy, genetics and tissue engineering. The biologic products that can be manufactured from these elements are classified from the standpoint of the Spanish Agency of Medication and Health Products in advanced drug therapies, blood products and transplants. This review seeks to provide scientific and administrative information for clinicians on the use of these biologic resources.

Nanoemulsions for dermal controlled release of oleanolic and ursolic acids: In vitro, ex vivo and in vivo characterization.

The aim of the present study was to design and optimize a nanoemulsion for dermal administration of mixtures of natural or synthetic pentacyclic triterpenes with recognized anti-inflammatory activity. The composition of the developed nanoemulsions was obtained from pseudo-ternary phase diagrams, composed of castor oil as the oil phase, labrasol as the surfactant, transcutol-P as co-surfactant and propylene glycol as the aqueous phase. Different ratios of surfactant/co-surfactant mixture (Smix) (4:1, 3:1, 2:1, 1:1, 1:2 and 1:4) were produced, and Smix 4:1 was chosen based on the greater area of optimal nanoemulsion conditions. Two different nanoemulsions of mean droplet size below 600 nm were produced, loading mixtures of natural or synthetic pentacyclic triterpenes, respectively. The viscosity of nanoemulsion containing natural pentacyclic triterpenes was 51.97±4.57 mPas and that loaded with synthetic mixtures was 55.33±0.28 mPas. The studies of release and skin permeation were performed using Franz diffusion cells, adjusting the release kinetics of both formulations to Korsmeyer-Peppas model. No significant differences in permeation parameters between the two nanoemulsions were observed. The amount of drug retained in the skin was higher than the amount of drug that has permeated, favoring a local action. The results of the in vivo tests demonstrated that the developed formulations were not toxic and not irritant to the skin. The formulation loading a mixture of natural triterpenes showed greater ability to inhibit inflammation than that loading the synthetic mixture. The findings clearly corroborate the added value of o/w nanoemulsions for dermal delivery of pentacyclic triterpenes.

Design, development and characterization of buccal bioadhesive films of Doxepin for treatment of odontalgia.

Tricyclic antidepressants, as doxepin hydrochloride (DH), may have analgesic local effect due to its biochemical mechanism of action. Delivery of DH directly to the oral cavity could be an interesting alternative for toothache due to its analgesic local effect. One problem associated with the mucosal administration routes is the short residence time of the dosage form on the mucosal membranes. In this sense, we have developed new doxepin mucoadhesive films able of reducing pain and increasing the effectiveness of treatment. For this purpose, we tested three different polymers: chitosan, sodium hydroxypropylmethylcellulose (HPMC) and sodium carboxymethylcellulose (SCMC) in film elaboration. The results obtained show that all films are hydrophilic matrices that absorb water when placed in an aqueous media. All the films hydrated very quickly, reaching high percentage of swelling after just few minutes (5 min for SCMC, 2 min for HPMC and 30 min for chitosan). Moreover, the SCMC and HPMC films were dissolved whereas chitosan was not dissolved. Dissolution also leads to viscous liquids with a higher retention time over mucosal surfaces what may lead to adhesive interactions. In vitro permeation studies showed that for all the formulations studied, SCMC (19.91%), HPMC (69.5%) and chitosan (24.17%), the percentage of drug permeated increased compared to the drug solution (8.26%). Specifically the HPMC film presents greater amounts of doxepin permeated (49.27 ± 4.47 µg/cm(2)).

Permeation studies through porcine small intestine of furosemide solutions for personalised paediatric administration.

Personalized medicine is a challenging research area in paediatric drug design since no suitable pharmaceutical forms are currently available. Furosemide is an anthranilic acid derivative used in paediatric practice to treat cardiac and pulmonary disorders in premature infants and neonates. However, it is not commercialized in suitable dosage forms for paediatrics. Elaborating new paediatric formulations when no commercial forms are available is a common practice in pharmacy laboratories; amongst these, oral liquid formulations are the most common. We developed two extemporaneous paediatric oral solutions of furosemide (pure powder). The characterization and stability study were also performed. Parameters such as organoleptic characteristics, rheology, pH, content of active substance, and microbial stability were evaluated at three temperatures for two months. Evaluation of all these parameters showed that both solutions were stable for 60 days at 4 and 25 °C. Moreover, ex vivo studies were performed to evaluate the permeation behaviour of developed solutions through porcine small intestine to evaluate the potential paediatric biological parameters influencing the bioavailability and efficacy. A validated spectrofluorometric method was also used for this purpose. Our results guarantee a correct dosification, administration and potential efficacy of furosemide when is formulated in liquid oral forms for the treatment of cardiac and pulmonary disorders in children.

Obtaining fast dissolving disintegrating tablets with different doses of melatonin.

Fast dissolving disintegrating tablets (FDDTs) containing different dosages of melatonin have been manufactured for administration to a specific target population: pediatric patients, having potential difficulties taking other oral forms. The lower dosages (3 and 5mg) are intended for epileptic children, migraine prevention, neurodevelopmental disability, sleep disorders and blindness. Dosages of 10 and 60 mg are intended for Duchenne muscular dystrophy. Two FDDT groups have been designed, one which has excipients for direct compression and others having direct compression and effervescent excipients. Tablets have been produced having disintegration times of less than 25s and with friability and hardness values that require no special storage or packaging conditions.

Design and physicochemical stability studies of paediatric oral formulations of sildenafil.

Personalized medicine is a challenging research area in paediatric treatments. Elaborating new paediatric formulations when no commercial forms are available is a common practice in pharmacy laboratories; among these, oral liquid formulations are the most common. But due to the lack of specialized equipment, frequently studies to assure the efficiency and safety of the final medicine cannot be carried out. Thus the purpose of this work was the development, characterization and stability evaluation of two oral formulations of sildenafil for the treatment of neonatal persistent pulmonary hypertension. After the establishment of a standard operating procedure (SOP) and elaboration, the physicochemical stability parameters appearance, pH, particle size, rheological behaviour and drug content of formulations were evaluated at three different temperatures for 90 days. Equally, prediction of long term stability, as well as, microbiological stability was performed. Formulations resulted in a suspension and a solution slightly coloured exhibiting fruity odour. Formulation I (suspension) exhibited the best physicochemical properties including Newtonian behaviour and uniformity of API content above 90% to assure an exact dosification process.

Drug delivery to inflammation based on nanoparticles surface decorated with biomolecules.

Anti-inflammatory molecules often display little affinity for inflamed tissues, leading to low accumulation into this site of action (and inefficiency), and high incidence of severe side effects. To face the problem, numerous strategies have been proposed, i.e., chemical modifications to the drug molecule, and engineering of drug nanocarriers. The later approach to the problem can result in optimized drug biodistribution and concentration into the target region, thus enhancing the anti-inflammatory effect while reducing the associated drug toxicity. Such nanoparticulate systems offer remarkable possibilities when they are made of biodegradable polymers, lipid-based structures, and/or inorganic particles. Recent advances in the field have been devoted to the optimization of the in vivo fate and effectiveness of these drug nanocarriers, e.g., passive targeting strategies based on the functionalization of nanoparticle surface with special biomolecules. In this contribution, we analyze the possibilities and future perspectives of nanoparticle therapy in inflammatory processes.

Elaboration, characterization, and stability study of a sunscreen emulsion for use as a towelette application in pediatric photoprotection.

The World Health Organization (WHO) estimates that as much as 80% of the solar radiation that an adult receives throughout his/her life is received during the first 18 years (1). Skin protection against harmful solar radiation during this early stage of life is therefore a highly important factor in the prevention of future skin-related diseases. In this respect, recent developments in pediatric dermatology and cosmetic technology have led to remarkable improvements in child skin protection products. However, in spite of these scientific breakthroughs, many currently available commercial sunscreen formulations have not been well received by the general public, due to inadequate sensory properties, chemical instability, undesirable side effects, and low effectiveness. These disadvantages are not only attributable to the formulations themselves, active principle, and excipients, but also, to a large extent, galenic aspects. The objective of this work was to develop and characterize a sunscreen emulsion for pediatric use, using a towelette as vehicle, to overcome problems of ineffectiveness and formulation instability, and to improve skin-sensory properties. The composition of the towelette, the emulsion, and the presentation format were selected on the basis of the differences between children's and adult skin. In order to evaluate the chemical stability of the formulation, a study of the organoleptic, physicochemical, microbiological, and rheological characteristics was carried out at 4°, 25°, and 40°C over a period of 30 days. Tests were performed on both the sunscreen emulsion only and the same formulation impregnated within a towel, to test the influence the towel may have on the stability of the emulsion.

Lipid-based drug delivery systems for cancer treatment.

It is a fact that chemotherapy agents have little specificity for cancer cells, this leading to low concentrations into the tumor interstititum and severe side effects on healthy tissues. The formulation of lipid-based nanomedicines against cancer has been hypothesized to improve drug localization into the tumor tissue and to increase the anticancer efficacy of concentional drugs, while minimizing their systemic adverse effects. In this review, special attention is devoted to the analysis of the state-of-the-art in the development of lipid-based drug carriers against cancer. Specifically, the most significant in vitro and in vivo results on the use of niosomes, liposomes, and solid lipid nanoparticles are revised. It is concluded that biodistribution profiles of chemotherapy agents can be controlled by their loading to such nanoplatforms. Lipid-based nanomedicines offer an interesting approach to the delivery of anticancer drugs to brain tumors, and to reverse multi-drug resistance of cancer cells. Finally, a deep evaluation of the applicability of drug delivery strategies in the formulation of lipid-based nanoplatforms is carried out. They involve active drug targeting (including ligand-mediated delivery, and stimuli-sensitive carriers), and passive drug targeting (through the enhanced permeability and retention effect) to tumors.

Structural characterization and stability of dimethylaminoethanol and dimethylaminoethanol bitartrate for possible use in cosmetic firming.

2-Dimethylaminoethanol (DMAE) (also known as deanol) has been used as an ingredient in skin care, and in cognitive function- and mood-enhancing products. It is marketed as a free base or salt, and in theory, the two forms should be equally effective and able to substitute for each other in pharmaceutical formulations. Detecting possible alterations in the active principle is a basic part of preformulation studies. Accordingly, this study compared DMAE and DMAE bitartrate to identify potential alterations or differences between the free base and the salt that might compromise the long-term stability of cosmetic preparations at different temperatures, and also compared the behavior of the base substance and derivative alone and in solution. Samples were analyzed with different physicochemical methods such as differential scanning calorimetry, ultraviolet and infrared spectroscopy, and nuclear magnetic resonance spectroscopy.

Diagnóstico y terapia del cáncer mediante magnetosomas / Cancer therapy and diagnosis by magnetosomes

Los magnetosomas (magnetoliposomas) han permitido en los últimos años un gran avance en la terapia del cáncer, gracias a su capacidad para transportar de forma eficiente diferentes agentes quimioterápicos hasta la masa tumoral. Estos sistemas no sólo se usan en el transporte de fármacos sino que también presentan importantes posibilidades en el diagnóstico de esta severa enfermedad, en terapia génica (transfección magnética o magnetofección) y en hipertermia. Sin embargo, su destino biológico, una vez que son administrados, se encuentra fuertemente condicionado por su geometría y propiedades fisicoquímicas. El presente trabajo se centra en la investigación de los principales aspectos relacionados con el diseño y utilización clínica de los magnetosomas. En concreto, estrategias de formulación, biocompatibilidad, toxicidad y situación actual a nivel preclínico(AU)

During the last years, magnetosomes (magnetoliposomes) have contributed to a significant improvement in cancer therapy, thanks to their capability to transport very efficiently the antitumor drug dose to the malignant cells. Interestingly, these systems also offer very promising possibilities in cancer diagnosis, gene therapy (magnetic transfection or magnetofection), and hyperthermia. However, the biological fate of magnetosomes, upon administration, is markedly conditioned by their geometry and physicochemical properties. This work is focussed on the investigation of the main aspects related to the engineering and clinical use of liposomes. Concretely, formulation strategies, biocompatibility, toxicity, and current state of the art at the preclinical level(AU)

Motivos de elección de una asignatura por parte de los alumnos / Some reasons which could induce to the students to choose an academic subject

El motivo de elección de una determinada asignatura por parte de los alumnos, ha sidofrecuentemente interpretado bajo diferentes puntos de vista. En esta comunicación nuestro objetivo hasido analizar los motivos de elección en distintos tipos de asignaturas con características peculiarescada una de ellas, concretamente dos de ellas son de libre configuración, siendo una de impartición enel campus andaluz virtual –Fotoprotección- y la otra en el campus virtual de la Universidad deGranada –Aplicación de la Tecnología Farmacéutica en el tratamiento del cáncer y del dolor-. Latercera asignatura seleccionada es una optativa -Farmacia Práctica- que se imparte actualmente en laLicenciatura en Farmacia. Se ha realizado un análisis de los diferentes motivos que impulsan a unalumno a la elección de una disciplina frente a otra, para ello se ha utilizado una de las herramientas demayor uso, la encuesta anónima a todos aquellos que estaban matriculados. Las conclusiones de esteestudio son diversas pero ante todo cabe señalar en el caso de las virtuales la facilidad de acceso yrealización de las asignaturas virtuales, y destacar también que un título atractivo es fundamental en suelección. En el caso de la optativa el motivo fundamental ha sido aprender algo nuevo relacionado consu licenciatura y seguido muy de cerca por la facilidad de horario. Por tanto como conclusióndefinitiva es interesante destacar que tanto en un caso como en otro el alumno desea aprender algoatractivo y que se acomode fácilmente a su horario de estudio(AU)

The reason why a specific subject is chosen by the students has been frequently interpretedfrom several points of view. The aim of this study is to analyze the reasons why different types ofsubjects with particular characteristics are chosen. Specifically, two of these subjects are freeelectives: one of them –Fotoprotección- is imparted through the Andalusian Virtual Campus, and theother one –Aplicación de la Tecnología Farmacéutica en el tratamiento del cáncer y del dolorthroughthe virtual campus of the University of Granada. The third subject chosen is optional -Farmacia Práctica- and it is at present imparted within the Degree in Pharmacy. The different reasonswhy a student chooses one subject against other have been analyzed using one of the most used tools:an anonymous survey to all the students enrolled. The conclusions of this study are diverse, but it ismainly to be remarked in the case of the virtual subjects their accessibility and easy implementation aswell as the importance of an attractive title for the selection of a subject. The main reason in the caseof the optional subject is to learn something new in relation to their Degree, followed very closely byits timetable. Therefore, as final conclusion, it is interesting to remark that the students want in bothcases to learn something attractive that can be easily adapted to their study schedule(AU)

Study and description of hydrogels and organogels as vehicles for cosmetic active ingredients.

Cellulite, a clinical syndrome mainly affecting women, involves specific changes in conjunctive dermic and subcutaneous tissue, leading to vascular and hypertrophic alterations in adipose tissues and the consequent alteration of tissue structure. This paper describes the design of hydrogels and pluronic-lecithin organogels elaborated as vehicles of Aloe vera (Aloe vera linné) and Hydrocotyle asiatica (Centella asiatica) for the treatment of cellulite. The objective of this work was to carry out a complete evaluation of the proposed formulae through the study of the organoleptic and rheological properties of the formulae. Our work revealed that, in appearance, hydrogels show better organoleptic characteristics than organogels. On the other hand, from a rheological point of view, both hydrogels and organogels display a plastic behavior. However, the main difference between the two is that the more complex internal structure of the organogel bestows it with more viscosity. Finally, in vitro tests with Franz-type diffusion cells revealed that the release of cosmetic active principle from the tested excipients was appropriate, both in terms of magnitude and velocity.

Evaluation of the anti-wrinkle efficacy of cosmetic formulations with an anti-aging peptide (Argireline®)

The purpose of this research was to evaluate, by means of in vivo studies, the efficacy of new cosmetic active ingredients which effect of botox, called Argireline®), so that width and depth of wrinkles could be established. For this, it is prepared two formulations: an emulsion with an external aqueous phase for normal to dry skin, and a gel for oily skin. We likewise study the water content of the skin after the application of both formulas, as this must be one of the priority functions of facial treatments in general, as well as the level of satisfaction from the subjective point of view, fundamental for patients and their continuation of the treatment.After the designed tests, it is possible to verify that there is a remarkable diminution of the wrinkles size tested in each patient during the month of treatment. Besides, it is possible to review how the moisturizing capacity has been increased in all cases.At the end of the visual test, all the volunteers experienced a reduction in the depth of wrinkles, and from the subjective point of view, the appearance and elasticity of the skin were improved. Finally it is possible to conclude that Argireline® (acetyl hexapeptide-8 ) shows a great antiaging capacity in all the cases that have been studied and the tried compounds have increased moisturizing power(AU)

Preparation, characterization, and in vitro release of new transdermal methimazole as alternative to oral delivery.

Our objective has been the development and study of the stability of transdermal methimazole formulae as alternative to oral administration. Preparation of F-127 and PLO Pluronic gel (Pluronic lecitin organogel) are described, as well as their characteristics from transmission electron microscopy. The possible structural and rheological changes to both transdermal forms were studied in terms of composition, temperature and time. The trial period was from 24 hr to 3 months after preparation. Furthermore, identical tests were carried out on formulae conserved for 1 year after production to check their integrity. Studies of release in vitro were carried out showing that the selected excipients do not pose an obstacle to the cession of methimazole, even though the PLO samples were made more slowly.

Multilamellar liposomes of triamcinolone acetonide: preparation, stability, and characterization.

The aim of this study was to assess and characterize the stability of multilamellar liposomes as a delivery vehicle for triamcinolone acetonide. A standardized preparation method for a liposomal delivery vehicle was developed, after varying composition and storage conditions, and assessing encapsulation efficiency and loss of active principle. The assessment of temperature as a factor in formula stability during storage showed that stability improved under refrigeration (4-6 degrees C) (less early diffusion of active principle through the liposomal wall), in comparison with samples stored at room temperature. To improve stability, cholesterol was added to some formulae, which although resulting in a decrease in average encapsulation efficiency, mitigated subsequent losses of retained active principle (formulae 4, 5, and 6), in comparison with those without cholesterol (formulae 1, 2, and 3). This was evident both under refrigerated and room-temperature conditions. Finally, after testing the effects of adding an antioxidant and/or preservative to the formulae, a liposomal design was achieved with acceptable stability, vesicle dimensions, and encapsulation efficiency.

Preparation, rheological study, and characterization of an organogel as a system for transdermal release of active principles.

The use of formulations containing Pluronic gel as a vehicle and permeabilizing agent for transdermal preparations has increased in recent years. We prepared and compared two transdermal formulations for drug administration as an alternative to oral or parenteral administration. In formulations containing Pluronic F127 gel or pluronic lecithin organogel (PLO), rheological, structural (transmission electron microscopy) and physicochemical characteristics were studied under different conditions of composition, temperature, and time from 24 hr to 3 months after preparation. Rheological studies at 20-25 degrees C and at 4 degrees C to study the influence of refrigeration on viscosity and pH showed that both formulas were thermoreversible. Unilamellar vesicles smaller than 1 microm in diameter were seen in the PLO formulation on TEM observation. The characteristics of these excipients may facilitate the application and may avoid the gastrointestinal tract and the first-pass effect.

Preparation and stability of cosmetic formulations with an anti-aging peptide.

Wrinkling of the skin is the most obvious sign of deterioration of the human body with age. This process involves a number of genetic, constitutional, hormonal, nutritional, and environmental factors, in addition to the influence of frequently repeated facial movements during laughing, smoking, etc. This article reviews the physiological basis and mechanism of action of the active cosmetic ingredient acetyl hexapeptide-8 (Argireline). We prepared two formulations: an emulsion with an external aqueous phase for normal to dry skin, and a gel for oily skin. Laboratory analyses, rheology tests and in vitro release assays were used to evaluate the stability of these formulations for cosmetic treatment.

Liposomas de acetónido de triamcinolona: influencia de la composición en el rendimiento y estabilidad / Liposomal triamcinolone acetonide: influence of composition in performance and stability

El acetónido de triamcinolona, antiinfl amatorio esteroideo, presenta por vía tópica los inconvenientes de la corticoterapia. No obstante, su incorporación a un sistema transportador, como los liposomas permitiría prolongar la dosis efectiva en el lugar de acción (dermis y epidermis), reduciendo los efectos secundarios. Así pues, se ha normalizado el método de elaboración de liposomas multilaminares portadores de acetónido de triamcinolona y evaluado el grado de captación del fármaco seleccionado, en función de la composición de los liposomas, determinando los componentes más idóneos para su obtención y los rendimientos proporcionados al variar las concentraciones de los mismos. La adición de colesterol para mejorar su estabilidad, provoca una reducción media en la captación; no obstante la encapsulación sigue siendo bastante elevada. La evaluación de la estabilidad muestra la infl uencia de la temperatura de conservación, de tal manera que los liposomas mantenidos a temperatura de refrigeración (4-6ºC) poseen una estabilidad mayor que las muestras a temperatura ambiente

In topical presentation, Triamcinolone acetonide, a steroidal anti-infl ammatory preparation, presents all the disadvantages of corticotherapy. However, on incorporation into a liposomal drug delivery system, the effectiveness of each dosage within the area of its activity (dermis and epidermis) is prolonged, serving to reduce secondary side effects. For this reason, an attempt has been made to standardize a method for the preparation of a multilaminar liposomal delivery system of triamcinolone acetonide and to assess how much of the drug could be encapsulated by the varying liposomal formulations tested and consequently, which of these would prove to be the most suitable. The addition of cholesterol to such formulations was found to improve stability. However, although such an addition was found to reduce levels of encapsulation of the drug, these still remained suffi ciently high. In the assessment of stability, storage temperature was found to bear an infl uence. Liposomes kept under cold storage (4-6ºC) presented higher stability than samples stored at room temperature