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[Abstract] The increase in life expectancy associated with developments in the field of medicine has prompted the prevalence of degenerative diseases related to aging. Specifically, articular cartilage is a tissue with a very limited ability to self-repair upon injury due to its avascular and aneural nature. Hence, osteoarthritis (OA) represents the most common cause of long-term pain and physical disability in developed countries. However, none of the current therapeutic options has been able to completely restore the function of hyaline cartilage, generally leading to the formation of fibrocartilage. In this context, gene therapy has emerged as a promising alternative to treat articular cartilage injuries by transferring therapeutic genes into the lesion site. Non-viral vectors represent the safest tools to accomplish this aim as they avoid the main drawbacks of viral carriers, including the risk of eliciting insertional mutagenesis or immune responses in the host. Nonetheless, the existence of several extracellular and intracellular barriers considerably reduced their efficiency compared to their viral counterparts. Noteworthy, the design of gene-activated matrices (GAMs) may help to overcome these issues by promoting a controlled delivery of the candidate genes into the cellular microenvironment. This dissertation focuses on the production of a gene-activated cryogel (G-HACG) based on a combination of non-viral vectors, a shape memory hyaluronic acid based cryogel (HACG) and a source of primary mesenchymal stem cells (MSCs). Developed cryogels systems showed a macroporous structure mimicking the composition of the cartilage extracellular matrix with great biocompatibility and promoting cell proliferation. Various non-viral gene delivery systems based on niosomes were produced and their composition was optimized to obtain high levels of transfection in MSCs with a reduced cytotoxicity. Best niosome formulations were tested as carriers of a plasmid encoding for th, [Resumen] El aumento de la esperanza de vida asociado a los avances médicos ha propiciado la prevalencia de enfermedades degenerativas relacionadas con el envejecimiento. En concreto, el cartílago articular es un tejido con una capacidad muy limitada de autorreparación debido a su naturaleza avascular y aneural. Por lo tanto, la osteoartritis (OA) representa la causa más frecuente de dolor a largo plazo y discapacidad física en los países desarrollados. Además, ninguna de las opciones terapéuticas actuales ha sido capaz de restaurarlo por completo, dando lugar generalmente a la formación de fibrocartílago. En este contexto, la terapia génica ha surgido como una alternativa prometedora para tratar las lesiones del cartílago articular mediante la transferencia de genes terapéuticos en el lugar de la lesión. Los vectores no virales son las herramientas más seguras para este fin, evitando los principales inconvenientes de los transportadores virales, como el riesgo de inducir mutagénesis por inserción o de desencadenar una respuesta inmune en el paciente. No obstante, la existencia de barreras extracelulares e intracelulares reduce considerablemente su eficiencia en comparación con sus homólogos virales. El diseño de matrices activadas por genes (GAMs) constituye una alternativa para soslayar dichas barreras a través de la liberación controlada de los genes candidatos en el microambiente celular. Esta tesis abordó la producción de un criogel activado por genes (G-HACG) mediante la combinación de una fuente de vectores no virales, un criogel de ácido hialurónico (HACG) con memoria de forma y una población de células madre mesenquimales (MSCs). Los criogeles desarrollados mostraron una estructura macroporosa que mimetiza la composición de la matriz del cartílago, y una elevada biocompatibilidad capaz de promover la proliferación celular. Se produjeron varios sistemas no virales de liberación de genes basados en niosomas y se optimizó su composición para obtener niveles el, [Resumo] O aumento da esperanza de vida asociado aos avances no campo da medicina provocou a prevalencia de enfermidades dexenerativas relacionadas co envellecemento. En concreto, a cartilaxe articular é un tecido cunha capacidade limitada de autorrepararse debido a súa natureza avascular e aneural. Por isto, a osteoartrite (OA) é a causa máis común de dor a longo prazo e discapacidade física nos países desenvolvidos. Non obstante, ningunha das opcións terapéuticas actuais é capaz de restaurar a cartilaxe hialina, derivando xeralmente na formación de fibrocartilaxe. Neste contexto, a terapia xénica xurdiu como unha alternativa prometedora para tratar as lesións da cartilaxe articular mediante a transferencia de xenes terapéuticos no lugar da lesión. Os vectores non virais son as ferramentas máis seguras para cumprir esta función, xa que evitan os principais inconvenientes dos transportadores virais, incluíndo o risco de inducir mutaxénese por inserción ou de desencadear unha resposta inmune no paciente. Non obstante, a existencia de barreiras extracelulares e intracelulares reduce a súa eficiencia en comparación cos seus homólogos virais. Cabe destacar que o deseño de matrices activadas por xenes (GAMs) pode axudar a superar estes inconvenientes proporcionando unha liberación controlada dos xenes candidatos no microambiente celular. Esta tese centrouse na produción dun crioxel activado por xenes (G-HACG ) baseada na combinación de vectores non virais, un crioxel de ácido hialurónico (HACG) con memoria de forma e unha fonte de células nai mesenquimais (MSCs). Os crioxeles desenvolvidos mostraron unha estrutura macroporosa que mimetizou a composición da matriz extracelular da cartilaxe, e unha alta biocompatibilidade que promoveu a proliferación celular. Producíronse varios sistemas non virais de liberación de xenes baseados en niosomas e optimizouse a súa composición para obter niveis de transfección elevados en MSCs, cunha reducida citotoxicidade. As mellores formul

[Abstract] Recovery from a traumatic spinal cord injury (TSCI) is challenging due to the limited regenerative capacity of the central nervous system to restore cells, myelin, and neural connections. Cell therapy, particularly with mesenchymal stem cells (MSCs), holds significant promise for TSCI treatment. This systematic review aims to analyze the efficacy, safety, and therapeutic potential of MSC-based cell therapies in TSCI. A comprehensive search of PUBMED and COCHRANE databases until February 2023 was conducted, combining terms such as "spinal cord injury," "stem cells," "stem cell therapy," "mesenchymal stem cells," and "traumatic spinal cord injury". Among the 53 studies initially identified, 22 (21 clinical trials and 1 case series) were included. Findings from these studies consistently demonstrate improvements in AIS (ASIA Impairment Scale) grades, sensory scores, and, to a lesser extent, motor scores. Meta-analyses further support these positive outcomes. MSC-based therapies have shown short- and medium-term safety, as indicated by the absence of significant adverse events within the studied timeframe. However, caution is required when drawing generalized recommendations due to the limited scientific evidence available. Further research is needed to elucidate the long-term safety and clinical implications of these advancements. Although significant progress has been made, particularly with MSC-based therapies, additional studies exploring other potential future therapies such as gene therapies, neurostimulation techniques, and tissue engineering approaches are essential for a comprehensive understanding of the evolving TSCI treatment landscape.

[Abstract] Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a biodegradable and biocompatible biopolymer that has gained popularity in the field of biomedicine. This review provides an overview of recent advances and potential applications of PHBV, with special emphasis on drug encapsulation and scaffold construction. PHBV has shown to be a versatile platform for drug delivery, offering controlled release, enhanced therapeutic efficacy, and reduced side effects. The encapsulation of various drugs, such as anticancer agents, antibiotics, and anti-inflammatory drugs, in PHBV nanoparticles or microspheres has been extensively investigated, demonstrating enhanced drug stability, prolonged release kinetics, and increased bioavailability. Additionally, PHBV has been used as a scaffold material for tissue engineering applications, such as bone, cartilage, and skin regeneration. The incorporation of PHBV into scaffolds has been shown to improve mechanical properties, biocompatibility, and cellular interactions, making them suitable for tissue engineering constructs. This review highlights the potential of PHBV in drug encapsulation and scaffold fabrication, showing its promising role in advancing biomedical applications.

[Abstract] Gene transfer to mesenchymal stem cells constitutes a powerful approach to promote their differentiation into the appropriate cartilage phenotype. Although viral vectors represent gold standard vehicles, because of their high efficiency, their use is precluded by important concerns including an elevated immunogenicity and the possibility of insertional mutagenesis. Therefore, the development of new and efficient non-viral vectors is under active investigation. In the present study, we developed new non-viral carriers based on niosomes to promote the effective chondrogenesis of human MSCs. Two different niosome formulations were prepared by varying their composition on non-ionic surfactant, polysorbate 80 solely (P80), or combined with poloxamer 407 (P80PX). The best niosome formulation was proven to transfer a plasmid, encoding for the potent chondrogenic transcription factor SOX9 in hMSC aggregate cultures. Transfection of hMSC aggregates via nioplexes resulted in an increased chondrogenic differentiation with reduced hypertrophy. These results highlight the potential of niosome formulations for gene therapy approaches focused on cartilage repair.

[Resumo] A queratoplastia ou transplante de córnea é o único tratamento dispoñible para moitas enfermidades corneais. Sen embargo, o acceso ás queratoplastias vese limitado pola dispoñibilidade das córneas doadas para transplante. Para incrementar o número de tecidos corneais, nesta tese investigáronse métodos de preservación de córneas alternativos e outras fontes de tecido corneal para queratoplastias. O primeiro obxectivo consistiu en analizar o efecto, en córneas humanas, de protocolos de criopreservación convencional con dimetil sulfóxido (DMSO) con (P1) e sen albúmina (P2), e de protocolos de vitrificación sen (P3) e con formamida (P4). O segundo obxectivo consistiu no desenvolvemento dun enxerto mediante enxeñaría de tecidos empregando membranas de Descemet como andamios e células endoteliais (CEs) para construír o endotelio. Tanto células como andamios procederon de córneas descartadas para o seu uso en clínica. Os resultados obtidos para o primeiro obxectivo mostran que só o P1 proporcionou córneas criopreservadas con CEs viables e unha estrutura xeral deformada. Sen embargo, a variabilidade dos resultados impide aplicar este protocolo na clínica. Os resultados do segundo obxectivo mostraron que é posible crear un enxerto a partir de córneas descartadas. Non obstante, a duración do estudio impediu que se crease un endotelio completamente funcional. Estas dúas prometedoras metodoloxías representan alternativas coas que, nun futuro, e tras máis estudos para a súa mellora, se podería reciclar e incrementar o número de tecidos corneais transplantables e mellorar o acceso ás queratoplastias., [Resumen] La queratoplastia o trasplante de córnea es el único tratamiento disponible para muchas enfermedades corneales. Sin embargo, el acceso a las queratoplastias se ve limitado por la disponibilidad de las córneas donadas para trasplante. Para incrementar el número de tejidos corneales, en esta tesis se investigaron métodos de preservación de córneas alternativos y otras fuentes de tejido corneal para queratoplastias. El primer objetivo consistió en analizar el efecto, en córneas humanas, de protocolos de criopreservación convencional con dimetil sulfóxido con (P1) y sin albúmina (P2), y de protocolos de vitrificación sin (P3) y con formamida (P4). El segundo objetivo consistió en el desarrollo un injerto mediante ingeniería de tejidos empleando membranas de Descemet como andamios y células endoteliales (CEs) para construir el endotelio. Tanto células como andamios procedieron de córneas descartadas para su uso en clínica. Los resultados obtenidos para el primer objetivo muestran que sólo el P1 proporcionó córneas criopreservadas con CEs viables y una estructura general deformada. Sin embargo, la variabilidad de los resultados impide aplicar dicho protocolo en la clínica. Los resultados del segundo objetivo mostraron que es posible crear un injerto a partir de córneas descartadas. No obstante, la duración del estudio impidió que se crease un endotelio completamente funcional. Estas dos prometedoras metodologías representan alternativas con las que, en un futuro, y tras más estudios para su mejora, se podría reciclar e incrementar el número de tejidos corneales trasplantables y mejorar el acceso a las queratoplastias., [Abstract] Many corneal diseases can only be treated through keratoplasty, a corneal transplantation whose access is dependent on the limited availability of transplantable donor corneas. To increase the number of corneal tissues, this thesis investigated alternative preservation methods and alternative sources of transplantable corneal tissues. The first objective was to investigate the effect, on human corneas, of cryopreservation protocols using dimethyl sulfoxide (DMSO) with (P1) and without albumin (P2), and of vitrification protocols without (P3) and with formamide (P4). The second objective was to engineer new transplantable tissues using Descemet’s membranes (DMs) and human endothelial cells (hECs) isolated from discarded corneas which were unfit for clinical use. Results showed that only P1 provided corneas with viable ECs and a slightly distorted general structure, but variability among C1-cryorpeserved corneas did not allow to use this protocol in the clinical practice. A corneal graft was successfully engineered using discarded corneas with an intact decellularized DM seeded with primary hECs. However, a fully functional endothelium was not observed within the timeframe used in this study. Consequently, additional research will be needed to improve both approaches for clinical applications. In the future and after further studies, these approaches would represent promising alternatives to improve the availability of transplantable corneal tissues and the access to keratoplasty.

[Resumen] INTRODUCCIÓN Y OBJETIVOS: La pandemia por el nuevo coronavirus (COVID-19) desde que fuera identificado en diciembre de 2019 ha traído consigo múltiples medidas restrictivas para la población mundial. En España hemos vivido diferentes etapas de confinamiento y normas de distanciamiento social que han tenido repercusión no solo en la actividad diaria de la población, sino también, en el funcionamiento de los hospitales de todo el territorio español. En el campo de la Cirugía Oral y Maxilofacial, los traumatismos faciales son la urgencia quirúrgica más frecuente. Dado que la incidencia de los traumatismos guarda relación directa la forma de vida y las costumbres de la población es lógico especular que las medidas adoptadas para frenar la pandemia han ocasionado un cambio en la incidencia y los patrones de las fracturas maxilofaciales. El objetivo principal del estudio es evaluar el impacto de la pandemia por coronavirus sobre la incidencia y las características de las fracturas maxilofaciales. MATERIAL Y MÉTODOS: Se ha diseñado un protocolo de investigación de un estudio observacional de cohorte retrospectiva. Se incluirán en el estudio los pacientes intervenidos por fracturas faciales por los servicios de Cirugía Oral y Maxilofacial de los hospitales Virgen del Rocío y Virgen Macarena de Sevilla. Se compararán tres grupos de pacientes según el año de intervención quirúrgica. El grupo 1, formado por los pacientes intervenidos el año previo a la pandemia; grupo 2, los pacientes intervenidos el primer año de la pandemia; y, grupo 3, los pacientes operados el segundo año de la pandemia. Se estudiarán cambios en la incidencia, epidemiología, etiología, patrones y tratamientos de las fracturas faciales utilizando técnicas de análisis estadístico descriptivo e inferencial., [Resumo] INTRODUCIÓN E OBXECTIVOS. A pandemia debida ao novo coronavirus (COVID-19) desde que se identificou en decembro de 2019 trouxo consigo múltiples medidas restritivas para a poboación mundial. En España vivimos diferentes etapas de confinamento e normas de distanciamento social. Estas medidas repercutiron non só na actividade diaria da poboación, senón tamén no funcionamento dos hospitais de todo o territorio español. No campo da Cirurxía Oral e Maxilofacial, os traumatismos faciais son a urxencia cirúrxica máis frecuente. Dado que a incidencia dos traumatismos garda relación directa a forma de vida e as costumes da poboación é lóxico especular as medidas adoptadas para frear a pandemia ocasionaron un cambio na incidencia e os patróns das fracturas maxilofaciais. O obxectivo principal do estudo é avaliar o impacto que tivo a pandemia de coronavirus na incidencia e características das fracturas maxilofaciais. MATERIAIS E MÉTODOS. Deseñouse un protocolo de investigación para un estudo de cohortes observacional retrospectivo. Incluiranse no estudo os pacientes intervidos por fracturas faciais polos servizos de Cirurxía Oral e Maxilofacial dos hospitais Virxe do Rocío e Virxe Macarena de Sevilla. O grupo 1, formado polos pacientes intervidos o ano previo á pandemia; grupo 2, os pacientes intervidos o primeiro ano de pandemia; e, grupo 3, os pacientes operados o segundo ano de pandemia Estudaranse os cambios de incidencia, epidemioloxía, etioloxía, patróns e tratamentos das fracturas faciais empregando técnicas de análise estatística descritiva e inferencial., [Abstract] INTRODUCTION AND OBJECTIVES. The coronavirus pandemic (COVID-19) since it was identified in December 2019 has brought with it multiple restrictive measures for the world population. In Spain we have lived through different stages of confinement and social distancing rules. These measures have had an impact not only on the daily activity of the population, but also on the functioning of Spanish Hospitals. As for the field of Oral and Maxillofacial Surgery, facial trauma is the most frequent surgical emergency. Since the incidence of trauma is directly related to the lifestyle and customs of the population, it is logical to speculate that the measures taken to curb the pandemic have caused a change in the incidence and patterns of maxillofacial fractures. The main objective of the study is to evaluate the impact of the coronavirus pandemic on the incidence and characteristics of maxillofacial fractures. MATERIAL AND METHODS. A research protocol of a retrospective cohort observational study has been designed. We will include patients operated for facial fractures by the Oral and Maxillofacial Surgery Departments of the Virgen del Rocío and Virgen Macarena Hospitals in Seville. Three groups of patients will be compared according to the year of surgical intervention. Group 1, patients operated on in the year prior to the pandemic; group 2, patients operated on in the first year of the pandemic; and group 3, patients operated on in the second year of the pandemic. Changes in the incidence, epidemiology, etiology, patterns and treatments of facial fractures will be studied using descriptive and inferential statistical analysis techniques.

[Resumen] Introducción. El dolor lumbar es la principal causa mundial de años de vida perdidos por discapacidad y supone un importante problema de salud con una carga personal y económica significativa. El 10% de los casos se identifican de forma crónica y ha sido reconocido como una afección de origen multifactorial, reconociéndose principalmente desacondicionamiento muscular y alteraciones en el control motor funcional. Es por ello que el entrenamiento de fuerza de levantamiento ha sido considerado como una modalidad de ejercicio efectiva para disminuir el dolor lumbar crónico. Objetivo. Realizar un proyecto de investigación para comparar la eficacia del ejercicio de Peso Muerto y el ejercicio de Sentadilla, en términos de sus efectos en la disminución de dolor lumbar crónico y variables asociadas. Material y métodos. Se expone el protocolo de un ensayo clínico controlado aleatorizado, de grupos paralelos y a simple ciego. La muestra está formada por 60 participantes con dolor lumbar crónico, que se dividirán a través de una aleatorización simple en tres grupos. Uno realizará el ejercicio de Peso Muerto, otro realizará el ejercicio de Sentadilla y un tercero combinará ambos ejercicios en un grupo mixto. Las mediciones se llevarán a cabo inicialmente, al final de la intervención y en un seguimiento a los 12 y 24 meses. La duración del programa será de 12 semanas, con una frecuencia de 2 días a la semana. Las variables medidas serán: intensidad de dolor, discapacidad, rango de movimiento de la columna lumbar, resistencia de los músculos lumbares y máxima fuerza isométrica de levantamiento., [Resumo] Introdución. A dor lumbar é a principal causa mundial de anos de vida perdidos por mor da discapacidade e é un importante problema de saúde cunha importante carga persoal e económica. O 10% dos casos identifícanse de forma crónica e recoñeceuse como unha condición de orixe multifactorial, recoñecendo fundamentalmente o desacondicionamento muscular e alteracións no control motor funcional. É por iso que o adestramento de forza de levantamento considerouse unha modalidade de exercicio eficaz para reducir a dor lumbar crónica. Obxectivo. Realizar un proxecto de investigación para comparar a efectividade do exercicio de Peso Morto e do exercicio de Agachamento, en canto aos seus efectos na redución da lumbalxia crónica e variables asociadas. Material e métodos. Preséntase o protocolo dun ensaio clínico controlado aleatorizado, de grupos paralelos e a simple cego. A mostra está formada por 60 participantes con dor lumbar crónica, que se dividirán mediante aleatorización simple en tres grupos. Un realizará o exercicio de Peso Morto outro realizará o exercicio de Agachamento e un terceiro combinará ambos exercicios nun grupo mixto. As medicións realizaranse inicialmente, ao final da intervención e nun seguimento aos 12 e 24 meses. A duración do programa será de 12 semanas, cunha frecuencia de 2 días á semana. As variables medidas serán: intensidade da dor, discapacidade, amplitude de movemento da columna lumbar, resistencia dos músculos lumbares e forza isométrica máxima de elevación., [Abstract] Introduction. Low back pain is the world's leading cause of years of life lost due to disability and is a major health problem with a significant personal and economic burden. 10% of cases are identified chronically and it has been recognized as a condition of multifactorial origin, mainly recognizing muscular deconditioning and alterations in functional motor control. That is why lifting strength training has been considered an effective exercise modality to reduce chronic low back pain. Objective. Carry out a research project to compare the effectiveness of the Deadlift exercise and the Squat exercise, in terms of their effects on the reduction of chronic low back pain and associated variables. Methods. The protocol of a single-blind, parallel-group, randomized controlled clinical trial is presented. The sample consists of 60 participants with chronic low back pain, who will be divided through simple randomization into three groups. One will perform the Deadlift exercise, another will perform the Squat exercise and a third will combine both exercises in a mixed group. The measurements will be carried out initially, at the end of the intervention and in a follow-up at 12 and 24 months. The duration of the program will be 12 weeks, with a frequency of 2 days a week. The variables measured will be: intensity of pain, disability, range of motion of the lumbar spine, resistance of the lumbar muscles and maximum lifting isometric force.

[Resumen] Introducción. La patología inguinal o pubalgia es una afección muy común en los deportes y, más aún, en aquellos que implican cambios de dirección. Para su tratamiento, lo que más eficacia ha demostrado, es el fortalecimiento de la musculatura de la cadera y de la musculatura abdominal. Pese a ello, los estudios disponibles son de baja calidad, no resultan fáciles de reproducir y no especifican un protocolo de abordaje concreto. Objetivos. Comparar la eficacia de un nuevo protocolo de abordaje de las pubalgias con la propuesta evidenciada en la literatura existente (en concreto con la de Weir et al.) tanto a corto como a largo plazo. Material y métodos. Se plantea un ensayo clínico aleatorio, tomando una muestra de 60 sujetos con edades de entre 18 y 30 años con patología inguinal que practiquen fútbol, baloncesto o atletismo de forma no profesional en el algún equipo de la ciudad de A Coruña. Se dividen en dos grupos, A y B, de 30 sujetos cada uno. El primero recibe el tratamiento evidenciado por el estudio de Weir et al., mientras que el segundo recibe un nuevo tratamiento experimental que incluye fortalecimiento de la musculatura abdominal, de la musculatura de la cadera y de miembros inferiores en general, así como un programa de regreso a la carrera. Todos los sujetos son valorados al inicio del tratamiento, al final, y a los 6 y 12 meses, tomando como ítems principales la reducción del dolor, el tiempo de recuperación y las recaídas existentes., [Abstract] Background. Inguinal pathology or pubalgia is a very common condition in sports and, even more so, in those that involve changes of direction. For its treatment, what has been shown to be most effective is the strengthening of the hip and abdominal muscles. Despite this, the available studies are of low quality, are not easy to reproduce and do not specify a specific approach protocol. Objective. To compare the efficacy of a new protocol for dealing with pubalgia with the proposal evidenced in the existing literature (specifically with that of Weir et al.) both in the short and long term. Methods. A randomized clinical trial is proposed, taking a sample of 60 subjects aged between 18 and 30 years with inguinal pathology who practice football, basketball or athletics non-professionally in a team in the city of A Coruña. They are divided into two groups, A and B, of 30 subjects each. The first receives the treatment evidenced by the study by Weir et al., while the second receives a new experimental treatment that includes strengthening of the abdominal muscles, the hip muscles and the lower limbs in general, as well as a program of back to the race. All subjects are assessed at the beginning of treatment, at the end, and at 6 and 12 months, taking as main items pain reduction, recovery time and existing relapses., [Resumo] Introdución A patoloxía inguinal ou pubalxia é unha afección moi común nos deportes e, máis aínda, naqueles que conlevan cambios de dirección. Para o seu tratamento o que más eficacia demostrou, é o fortalecemento da musculatura da cadeira e da musculatura abdominal. A pesar disto, os estudos dispoñibles son de baixa calidade, non resultan doados de reproducir ou non especifican un protocolo de intervención concreto. Obxectivo Comparar a eficacia dun novo protocolo de tratamento de pubalxias coa proposta evidenciada na literatura existente (en concreto coa de Weir et al.) tanto a curto como a longo prazo. Material e método Plantéxase un ensaio clínico aleatorio, tomando unha mostra de 60 suxeitos con idades de entre 18 e 30 anos con patoloxía inguinal que practiquen fútbol, baloncesto ou atletismo de forma non profesional nalgún equipo da cidade de A Coruña. Divídense en dous grupos, A e B, de 30 integrantes cada un. O primeiro recibe o tratamento evidenciado polo estudo de Weir et al., mentres que o segundo recibe o novo tratamento experimental que inclúe fortalecemento da musculatura abdominal, da musculatura da cadeira e dos membros inferiores en xeral, así como un programa de regreso á carreira. Todos os suxeitos son valorados ao comezo e ao final do tratamento, así como aos 6 e aos 12 meses, tomando como ítems principais a redución da dor, o tempo de recuperación e as recaídas existentes.

[Abstract] Background: Peri-implantitis (PIT) is highly prevalent in patients with dental implants and is a challenging condition to treat due to the limited outcomes reported for non-surgical and surgical therapies. Therefore, epigenetic therapeutics might be of key importance to treat PIT. However, developing epigenetic therapeutics is based on understanding the relationship between epigenetics and disease. To date, there is still scarce knowledge about the relationship between epigenetic modifications and PIT, which warrants further investigations. Aim: The purpose of this study was to evaluate the level of global DNA methylation associated with implant failure (IF) due to PIT compared to periodontally healthy (PH) patients. Material and Methods: A total of 20 participants were initially enrolled in this pilot, exploratory, single-blinded, cross-sectional clinical human study in two groups: 10 in the PH group and 10 in the IF group. In the participants who have completed the study, gingival tissue and bone samples were harvested from each participant and were used to perform global DNA methylation analysis. The percentage of global DNA methylation (5-mC%) was compared (1) between groups (PH and IF); (2) between the subgroups of gingival tissue and bone separately; (3) in the whole sample, comparing gingival tissue and bone; (4) within groups, comparing gingival tissue and bone. Demographic, periodontal, and peri-implant measurements as well as periodontal staging, were also recorded. All statistical comparisons were made at the 0.05 significance level. Results: Out of the initially enrolled 20 patients, only 19 completed the study and, thus, were included in the final analysis; 10 patients in the PH group and 9 patients in the IF group, contributing to a total of 38 samples. One patient from the IF group was excluded from the study due to systemic disease. The mean implant survival time was 10.8 years (2.17–15.25 years). Intergroup comparison, stratified by group

[Abstract] Reprogramming somatic cells toward pluripotency became possible over a decade ago. Since then, induced pluripotent stem cells (iPSCs) have served as a versatile and powerful tool not only for basic research but also with the long-term goal of using them in human cell transplantation after differentiation. Nonetheless, downstream applications are frequently blurred by the difficulties that researchers have to face when working with iPSCs, such as trouble with clonal selection, in vitro culture and cryopreservation, adaptation to feeder-free conditions, or expansion of the cells. Therefore, in this article we aim to provide other researchers with practical and detailed information to successfully culture and adapt iPSCs. Specifically, we (1) describe the most common problems when in-vitro culturing iPSCs onto feeder cells as well as its possible troubleshooting, and (2) compare different matrices and culture media for adapting the iPSCs to feeder-free conditions. We believe that the troubleshooting and recommendations provided in this article can be of use to other researchers working with iPSCs and who may be experiencing similar issues, hopefully enhancing the appeal of this promising cell source to be used for biomedical investigations, such as tissue engineering or regenerative medicine applications.

[Abstract] Background: Mesenchymal stromal cells (MSCs) have the capacity for self-renewal and multi-differentiation, and for this reason they are considered a potential cellular source in regenerative medicine of cartilage and bone. However, research on this field is impaired by the predisposition of primary MSCs to senescence during culture expansion. Therefore, the aim of this study was to generate and characterize immortalized MSC (iMSC) lines from aged donors. Methods: Primary MSCs were immortalized by transduction of simian virus 40 large T antigen (SV40LT) and human telomerase reverse transcriptase (hTERT). Proliferation, senescence, phenotype and multi-differentiation potential of the resulting iMSC lines were analyzed. Results: MSCs proliferate faster than primary MSCs, overcome senescence and are phenotypically similar to primary MSCs. Nevertheless, their multi-differentiation potential is unbalanced towards the osteogenic lineage. There are no clear differences between osteoarthritis (OA) and non-OA iMSCs in terms of proliferation, senescence, phenotype or differentiation potential. Conclusions: Primary MSCs obtained from elderly patients can be immortalized by transduction of SV40LT and hTERT. The high osteogenic potential of iMSCs converts them into an excellent cellular source to take part in in vitro models to study bone tissue engineering.

[Abstract] Corneal cryopreservation can partially solve the worldwide concern regarding donor cornea shortage for keratoplasties. In this study, human corneas were cryopreserved using two standard cryopreservation protocols that are employed in the Tissue Bank of the Teresa Herrera Hospital (Spain) to store corneas for tectonic keratoplasties (TK protocol) and aortic valves (AV protocol), and two vitrification protocols, VS55 and DP6. Endothelial viability and general corneal state were evaluated to determine the protocol that provides the best results. The potential corneal cryopreservation protocol was studied in detail taking into consideration some cryopreservation-related variables and the endothelial integrity and stroma arrangement of the resulting cryopreserved corneas. TK corneas showed mostly viable endothelial cells, while the others showed few (AV) or none (DP6 and VS55). The corneal structure was well maintained in TK and AV corneas. TK corneas showed endothelial acellular areas surrounded by injured cells and a normal-like stromal fiber arrangement. Cryoprotectant solutions of the TK protocol presented an increasing osmolality and a physiological pH value. Cooling temperature rate of TK protocol was of 1 °C/min to −40 °C and 3 °C/min to −120 °C, and almost all of dimethyl sulfoxide left the tissue after washing. Future studies should be done changing cryopreservation-related variables of the TK protocol to store corneas of optical grade.

[Abstract] Hydrogel-based nonviral gene delivery constitutes a powerful strategy in various regenerative medicine scenarios, as those concerning the treatment of musculoskeletal, cardiovascular, or neural tissues disorders as well as wound healing. By a minimally invasive administration, these systems can provide a spatially and temporarily defined supply of specific gene sequences into the target tissue cells that are overexpressing or silencing the original gene, which can promote natural repairing mechanisms to achieve the desired effect. In the present work, we provide an overview of the most avant-garde approaches using various hydrogels systems for controlled delivery of therapeutic nucleic acid molecules in different regenerative medicine approaches.

[Abstract] Induced pluripotent stem cells (iPSCs) represent an unlimited source of pluripotent cells capable of di erentiating into any cell type of the body. Several studies have demonstrated the valuable use of iPSCs as a tool for studying the molecular and cellular mechanisms underlying disorders a ecting bone, cartilage and muscle, as well as their potential for tissue repair. Musculoskeletal diseases are one of the major causes of disability worldwide and impose an important socio-economic burden. To date there is neither cure nor proven approach for e ectively treating most of these conditions and therefore new strategies involving the use of cells have been increasingly investigated in the recent years. Nevertheless, some limitations related to the safety and di erentiation protocols among others remain, which humpers the translational application of these strategies. Nonetheless, the potential is indisputable and iPSCs are likely to be a source of di erent types of cells useful in the musculoskeletal field, for either disease modeling or regenerative medicine. In this review, we aim to illustrate the great potential of iPSCs by summarizing and discussing the in vitro tissue regeneration preclinical studies that have been carried out in the musculoskeletal field by using iPSCs.

[Abstract] Human bone marrow-derived mesenchymal stromal cells (MSCs) obtained from aged patients are prone to senesce and diminish their differentiation potential, therefore limiting their usefulness for osteochondral regenerative medicine approaches or to study age-related diseases, such as osteoarthiritis (OA). MSCs can be transduced with immortalizing genes to overcome this limitation, but transduction of primary slow-dividing cells has proven to be challenging. Methods for enhancing transduction efficiency (such as spinoculation, chemical adjuvants, or transgene expression inductors) can be used, but several parameters must be adapted for each transduction system. In order to develop a transduction method suitable for the immortalization of MSCs from aged donors, we used a spinoculation method. Incubation parameters of packaging cells, speed and time of centrifugation, and valproic acid concentration to induce transgene expression have been adjusted. In this way, four immortalized MSC lines (iMSC#6, iMSC#8, iMSC#9, and iMSC#10) were generated. These immortalized MSCs (iMSCs) were capable of bypassing senescence and proliferating at a higher rate than primary MSCs. Characterization of iMSCs showed that these cells kept the expression of mesenchymal surface markers and were able to differentiate towards osteoblasts, adipocytes, and chondrocytes. Nevertheless, alterations in the CD105 expression and a switch of cell fate-commitment towards the osteogenic lineage have been noticed. In conclusion, the developed transduction method is suitable for the immortalization of MSCs derived from aged donors. The generated iMSC lines maintain essential mesenchymal features and are expected to be useful tools for the bone and cartilage regenerative medicine research.

[Abstract] Here, we report the establishment of the human iPS cell line N1-FiPS4F#7 generated from skin cells of a patient with no rheumatic diseases, thus obtaining an appropriate control iPS cell line for researchers working in the field of rheumatic diseases. The reprogramming factors Oct4, Sox2, Klf4 and c-Myc were introduced using a non-integrating reprogramming strategy involving Sendai Virus.

[Abstract] Knowledge and research results about hand osteoarthritis (hOA) are limited due to the lack of samples and animal models of the disease. Here, we report the generation of two induced pluripotent stem cell (iPSC)-lines from patients with radiographic hOA. Furthermore, we wondered whether these iPSC-lines carried single nucleotide polymorphisms (SNPs) within genes that have been associated with hOA. Finally, we performed chondrogenic differentiation of the iPSCs in order to prove their usefulness as cellular models of the disease. We performed a non-integrative reprogramming of dermal fibroblasts obtained from two patients with radiographic rhizarthrosis and non-erosive hOA by introducing the transcriptional factors Oct4, Sox2, Klf4 and c-Myc using Sendai virus. After reprogramming, embryonic stem cell-like colonies emerged in culture, which fulfilled all the criteria to be considered iPSCs. Both iPSC-lines carried variants associated with hOA in the four studied genes and showed differences in their chondrogenic capacity when compared with a healthy control iPSC-line. To our knowledge this is the first time that the generation of iPSC-lines from patients with rhizarthrosis and non-erosive hOA is reported. The obtained iPSC-lines might enable us to model the disease in vitro, and to deeper study both the molecular and cellular mechanisms underlying hOA.