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Este artículo se encuentra disponible en la siguiente URL: https://academic.oup.com/stcltm/article/11/12/1177/6786226 Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes progressive joint destruction. Despite the advances in the treatment of this condition there remains a clinical need for safe therapies leading to clinical remission. Mesenchymal stem/stromal cells (MSCs) play immunomodulatory and regenerative roles which can be partly mediated by their secretome. In recent years, the important contribution of extracellular vesicles (EVs) to MSC actions has received an increasing interest as a new therapeutic approach. We provide an extensive overview of the immunomodulatory properties of MSC EVs and their effects on articular cells such as fibroblast-like synoviocytes that play a central role in joint destruction. This review discusses the anti-arthritic effects of MSC EVs in vitro and in animal models of RA as well as their potential mechanisms. Recent preclinical data suggest that transfer of non-coding RNAs by MSC EVs regulates key signaling pathways involved in the pathogenesis of RA. We also examine a number of EV modifications for improving their anti-arthritic efficacy and carrier ability for drug delivery.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.sciencedirect.com/science/article/pii/S2214031X21000656?via%3Dihub Background: Osteoarthritis (OA) is a joint disease characterized by cartilage degradation, low-grade synovitis and subchondral bone alterations. In the damaged joint, there is a progressive increase of oxidative stress leading to disruption of chondrocyte homeostasis. The modulation of oxidative stress could control the expression of inflammatory and catabolic mediators involved in OA. We have previously demonstrated that extracellular vesicles (EVs) present in the secretome of human mesenchymal stem cells from adipose tissue (AD-MSCs) exert antiinflammatory and anti-catabolic effects in OA chondrocytes. In the current work, we have investigated whether AD-MSC EVs could regulate oxidative stress in OA chondrocytes as well as the possible contribution of peroxiredoxin 6 (Prdx6). Methods: Microvesicles (MV) and exosomes (EX) were isolated from AD-MSC conditioned medium by differential centrifugation with size filtration. The size and concentration of EVs were determined by resistive pulse sensing. OA chondrocytes were isolated from knee articular cartilage of advanced OA patients. 4-Hydroxynonenal adducts, IL-6 and MMP-13 were determined by enzyme-linked immunosorbent assay. Expression of Prdx6 and autophagic markers was assessed by immunofluorescence and Western blotting. Prdx6 was downregulated in AD-MSCs by transfection with a specific siRNA. Results: MV and to a lesser extent EX significantly reduced the production of oxidative stress in OA chondrocytes stimulated with IL-1β. Treatment with MV resulted in a dramatic upregulation of Prdx6. MV also enhanced the expression of autophagy marker LC3B. We downregulated Prdx6 in AD-MSCs by using a specific siRNA and then MV were isolated. These Prdx6-silenced MV failed to modify oxidative stress and the expression of autophagy markers. We also assessed the possible contribution of Prdx6 to the effects of MV on IL-6 and MMP-13 production. The reduction in the levels of both mediators induced by MV was partly reverted after Prdx6 silencing. Conclusion: Our results indicate that EVs from AD-MSCs regulate the production of oxidative stress in OA chondrocytes during inflammation. Prdx6 may mediate the antioxidant and protective effects of MV. The translational potential of this article: This study gives insight into the protective properties of EVs from AD-MSCs in OA chondrocytes. Our findings support the development of novel therapies based on EVs to prevent or treat cartilage degradation.

Este artículo se encuentra disponible en la siguiente URL: https://www.mdpi.com/1422-0067/23/9/4693 Este artículo de investigación pertenece al número especial "New Sources, Differentiation, and Therapeutic Uses of Mesenchymal Stem Cells 2.0". The use of mesenchymal stem cells constitutes a promising therapeutic approach, as it has shown beneficial effects in different pathologies. Numerous in vitro, pre-clinical, and, to a lesser extent, clinical trials have been published for osteoarthritis. Osteoarthritis is a type of arthritis that affects diarthritic joints in which the most common and studied effect is cartilage degradation. Nowadays, it is known that osteoarthritis is a disease with a very powerful inflammatory component that affects the subchondral bone and the rest of the tissues that make up the joint. This inflammatory component may induce the differentiation of osteoclasts, the bone-resorbing cells. Subchondral bone degradation has been suggested as a key process in the pathogenesis of osteoarthritis. However, very few published studies directly focus on the activity of mesenchymal stem cells on osteoclasts, contrary to what happens with other cell types of the joint, such as chondrocytes, synoviocytes, and osteoblasts. In this review, we try to gather the published bibliography in relation to the effects of mesenchymal stem cells on osteoclastogenesis. Although we find promising results, we point out the need for further studies that can support mesenchymal stem cells as a therapeutic tool for osteoclasts and their consequences on the osteoarthritic joint.

Este artículo se encuentra disponible en la siguiente URL: https://www.mdpi.com/2227-9059/9/11/1726 Este artículo pertenece al número especial "Adipose-Derived Mesenchymal Stem Cells, Cell-Based Therapies, and Biomaterials as New Regenerative Strategies". Osteoarthritis (OA) is the most common articular disease in adults and has a current prevalence of 12% in the population over 65 years old. This chronic disease causes damage to articular cartilage and synovial joints, causing pain and leading to a negative impact on patients’ function, decreasing quality of life. There are many limitations regarding OA conventional therapies—pharmacological therapy can cause gastrointestinal, renal, and cardiac adverse effects, and some of them could even be a threat to life. On the other hand, surgical options, such as microfracture, have been used for the last 20 years, but hyaline cartilage has a limited regeneration capacity. In recent years, the interest in new therapies, such as cell-based and cell-free therapies, has been considerably increasing. The purpose of this review is to describe and compare bioregenerative therapies’ efficacy for OA, with particular emphasis on the use of mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP). In OA, these therapies might be an alternative and less invasive treatment than surgery, and a more effective option than conventional therapies.

Este artículo se encuentra disponible en la siguiente URL: https://www.mdpi.com/1422-0067/22/3/1375 Este artículo pertenece al número especial "The Role of Mesenchymal Stem Cells on Inflammatory and Fibrotic Diseases". Adipose tissue represents an abundant source of mesenchymal stem cells (MSC) for therapeutic purposes. Previous studies have demonstrated the anti-inflammatory potential of adipose tissue-derived MSC (ASC). Extracellular vesicles (EV) present in the conditioned medium (CM) have been shown to mediate the cytoprotective effects of human ASC secretome. Nevertheless, the role of EV in the anti-inflammatory effects of mouse-derived ASC is not known. The current study has investigated the influence of mouse-derived ASC CM and its fractions on the response of mouse-derived peritoneal macrophages against lipopolysaccharide (LPS). CM and its soluble fraction reduced the release of pro-inflammatory cytokines, adenosine triphosphate and nitric oxide in stimulated cells. They also enhanced the migration of neutrophils or monocytes, in the absence or presence of LPS, respectively, which is likely related to the presence of chemokines, and reduced the phagocytic response. The anti-inflammatory effect of CM may be dependent on the regulation of toll-like receptor 4 expression and nuclear factor- B activation. Our results demonstrate the antiinflammatory effects of mouse-derived ASC secretome in mouse-derived peritoneal macrophages stimulated with LPS and show that they are not mediated by EV.

[EN] Mesenchymal stem/stromal cells (MSCs) represent a promising therapy for musculoskeletal diseases. There is compelling evidence indicating that MSC effects are mainly mediated by paracrine mechanisms and in particular by the secretion of extracellular vesicles (EVs). Many studies have thus suggested that EVs may be an alternative to cell therapy with MSCs in tissue repair. In this review, we summarize the current understanding of MSC EVs actions in preclinical studies of (1) immune regulation and rheumatoid arthritis, (2) bone repair and bone diseases, (3) cartilage repair and osteoarthritis, (4) intervertebral disk degeneration and (5) skeletal muscle and tendon repair. We also discuss the mechanisms underlying these actions and the perspectives of MSC EVs-based strategies for future treatments of musculoskeletal disorders., This work has been funded by grant SAF2017-85806-R (Ministerio de Ciencia, Innovación y Universidades, Spain, FEDER.

Este es el artículo que se ha publicado de forma definitiva en: https://www.mdpi.com/1996-1944/12/7/1015 Dentistry-applied bioceramic materials are ceramic materials that are categorized asbioinert, bioactive and biodegradable. They share a common characteristic of being specificallydesigned to fulfil their function; they are able to act as root canal sealers, cements, root repair or fillingmaterials. Bioactivity is only attributed to those materials which are capable of inducing a desiredtissue response from the host. The aim of this study is to present a systematic review of availableliterature investigating bioactivity of dentistry-applied bioceramic materials towards dental pulp stemcells, including a bibliometric analysis of such a group of studies and a presentation of the parametersused to assess bioactivity, materials studied and a summary of results. The research question, based onthe PICO model, aimed to assess the current knowledge on dentistry-based bioceramic materials byexploring to what extent they express bioactive properties inin vitroassays and animal studies whenexposed to dental pulp stem cells, as opposed to a control or compared to different bioceramic materialcompositions, for their use in the dentin-pulp complex therapy. A systematic search of the literaturewas performed in six databases, followed by article selection, data extraction, and quality assessment.Studies assessing bioactivity of one or more bioceramic materials (both commercially available ornovel/experimental) towards dental pulp stem cells (DPSCs) were included in our review. A total of37 articles were included in our qualitative review. Quantification of osteogenic, odontogenic andangiogenic markers using reverse transcriptase polymerase chain reaction (RT-PCR) is the prevailingmethod used to evaluate bioceramic material bioactivity towards DPSCs in the current investigativestate, followed by alkaline phosphatase (ALP) enzyme activity assays and Alizarin Red Staining(ARS) to assess mineralization potential. Mineral trioxide aggregate and Biodentine are the prevalentreference materials used to compare with newly introduced bioceramic materials. Available literaturecompares a wide range of bioceramic materials for bioactivity, consisting mostly ofin vitroassays.The desirability of this property added to the rapid introduction of new material compositions makesthis subject a clear candidate for future research.Keywords:bioactivity; bioceramic materials; dental pulp stem cells; systematic reviewMaterials2019,12, 1015; doi:10.3390/ma12071015www.mdpi.com/journal/materials Materials2019,12, 10152 of 301. IntroductionWithin the field of biomedical therapeutics, we can highlight the concept of tissue engineeringto refer to the development of procedures and biomaterials that aim to devise new tissues to replacethose damaged, following the principles of cellular and molecular biology and taking as a premise thesearch for “biological solutions for biological problems” [1].In 2007, the American Association of Endodontists adopted the term “regenerative endodontics”to refer to the concept of tissue engineering applied to the restoration of root canal health, in a waythat continuous development of the root and tissues surrounding it is promoted [2].The introduction of the so-called bioceramic materials meant a great advance for this newparadigm in endodontic therapy [3], given their biocompatible nature and excellent physicochemicalproperties [4]. Categorized as bioinert, bioactive and biodegradable [5], dentistry-applied bioceramicmaterials are ceramic materials which share a common characteristic of being specifically designed tofulfil their function; they are able to act as root canal sealers, cements, root repair or filling materials [4].Applied to vital pulp therapy, bioceramic materials can be used in cases of pulp exposition fromtrauma, caries or other mechanical causes, as direct pulp cappers [6].Properties like biocompatibility and bioactivity are to be expected in dentistry-applied bioceramicmaterials for their use in vital pulp therapy [7]. The first one refers to the “ability to perform as asubstrate that will support the appropriate cellular activity, including the facilitation of molecularand mechanical signaling systems, in order to optimize tissue regeneration, without eliciting anyundesirable local or systemic responses in the eventual host” [8], while bioactivity goes even further,and is only attributed to those materials which are capable of inducing a desired tissue responsefrom the host [9] by the use of biomimetic approaches [10]. The term differs depending on the fieldin which it is implemented, being related to the cellular effects induced by biologically active ionsand substances released from biomaterials in the field of tissue engineering, but referred to as thebiomaterial’s capability of forming hydroxyl apatite mineral on its surface bothin vitroandin vivointhe field of biomaterial science [11].Considering these desirable characteristics of bioceramic materials, it seems convenient to analyzethe interaction between human dental pulp stem cells (hDPSCs), which are post-natal stem cellswith mesenchymal stem cell (MSCs)-like characteristics, like auto-renewal ability and multilineagedifferentiation potential [12], and them; as their combined use could mean and advancement in thefield of regenerative endodontics.Cytotoxicity and biocompatibility of a wide range of bioceramic materials towards dental stemcells (DSCs) have been investigated in numerous studies [13–17]; among others. The well-knownPro-Root MTA (Dentsply Tulsa Dental Specialties, Tulsa, OK, USA) has been shown to increaseosteoblast, fibroblast, cementoblast, odontoblast and pulp cell differentiation, but its handling difficultyamong other limitations encourages for a search for alternative materials [13]. Materials like Biodentine(Septodont, Saint Maurdes-Fosses, France) and TheraCal LC (Bisco Inc., Schaumburg, IL, USA) areexamples of bioceramic materials introduced posteriorly in dentistry for their use in vital pulp therapyas blood clot protectors in pulpal revascularization procedures, standing out for their consistency,easier manipulation and tricalcium silicate composition [16].However, to the best of the authors’ knowledge, there has been no effort to sort and summarizestudies analyzing bioactivity of such materials into more homogenous subgroups that would allow foran easier analysis of the evidence.

Este es el artículo que se ha publicado de forma definitiva en: https://www.mdpi.com/1422-0067/20/12/3105 Chronic musculoskeletal (MSK) pain is one of the most common medical complaintsworldwide and musculoskeletal injuries have an enormous social and economical impact. Currentpharmacological and surgical treatments aim to relief pain and restore function; however,unsatiscactory outcomes are commonly reported. In order to find an accurate treatment to suchpathologies, over the last years, there has been a significantly increasing interest in cellular therapies,such as adipose-derived mesenchymal stem cells (AMSCs). These cells represent a relatively newstrategy in regenerative medicine, with many potential applications, especially regarding MSKdisorders, and preclinical and clinical studies have demonstrated their efficacy in muscle, tendon,bone and cartilage regeneration. Nevertheless, several worries about their safety and side effects atlong-term remain unsolved. This article aims to review the current state of AMSCs therapy in thetreatment of several MSK diseases and their clinical applications in veterinary and human medicine.

[EN] The inflammatory process is an essential phenomenon in the induction of immune responses. Monocytes are key effector cells during the inflammatory process. A wide range of evidence indicates that mesenchymal stem cells from adipose tissue (ASC) are endowed with immunomodulatory capacity. However, the interaction between ASC and monocytes in the innate immune response is not well understood. The aim of this work was to investigate the possible paracrine anti-inflammatory effects of ASC in human monocytes. Monocytes were isolated from buffy coats and ASC from fat of non-obese patients. Conditioned medium (CM) from ASC in primary culture was used. We have assessed the effects of CM on the production of inflammatory mediators, degranulation, migration, phagocytic activity, senescence, oxidative stress, mitochondrial membrane potential and macrophage polarization. We have shown that ASC exert paracrine anti-inflammatory actions on human monocytes. CM significantly reduced the production of TNF alpha, NO and PGE2 and the activation of NF-kappa B. In addition, we observed a significant reduction of degranulation, phagocytic activity and their migratory ability in the presence of the chemokine CCL2. The senescence process and the production of oxidative stress and mitochondrial dysfunction were inhibited by CM which also reduced the production of TNF alpha by M1 macrophages while enhanced TGF beta 1 and IL-10 release by M2 macrophages. This study have demonstrated relevant interactions of ASC with human monocytes and macrophages which are key players of the innate immune response. Our results indicate that ASC secretome mediates the anti-inflammatory actions of these cells. This paracrine mechanism would limit the duration and amplitude of the inflammatory response., This work has been funded by grants SAF2017-85806-R (MINECO and FEDER), PROMETEOII/2014/071 (Generalitat Valenciana) and PRCEU-UCH20/11.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.mdpi.com/2079-4983/9/1/10 The overall increase of chronic degenerative diseases associated with ageing makes wound care a tremendous socioeconomic burden. Thus, there is a growing need to develop novel wound healing therapies to improve cutaneous wound healing. The use of regenerative therapies is becoming increasingly popular due to the low-invasive procedures needed to apply them. Platelet-rich plasma (PRP) is gaining interest due to its potential to stimulate and accelerate the wound healing process. The cytokines and growth factors forming PRP play a crucial role in the healing process. This article reviews the emerging field of skin wound regenerative therapies with particular emphasis on PRP and the role of growth factors in the wound healing process.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://bmcvetres.biomedcentral.com/articles/10.1186/s12917-018-1577-y Background: The use of Plasma Rich in Growth Factors (PRGF) and Adipose Derived Mesenchymal Stem Cells (ASCs) are today extensively studied in the field of regenerative medicine. In recent years, human and veterinary medicine prefer to avoid using traumatic techniques and choose low or non-invasive procedures. The objective of this study was to evaluate the efficacy of PRGF, ASCs and the combination of both in wound healing of full-thickness skin defects in rabbits. With this purpose, a total of 144 rabbits were used for this study. The animals were divided in three study groups of 48 rabbits each depending on the administered treatment: PRGF, ASCs, and PGRF+ASCs. Two wounds of 8 mm of diameter and separated from each other by 20 mm were created on the back of each rabbit: the first was treated with saline solution, and the second with the treatment assigned for each group. Macroscopic and microscopic evolution of wounds was assessed at 1, 2, 3, 5, 7 and 10 days post-surgery. With this aim, 8 animals from each treatment group and at each study time were euthanized to collect wounds for histopathological study. Results: Wounds treated with PRGF, ASCs and PRGF+ASCs showed significant higher wound healing and epithelialization rates, more natural aesthetic appearance, significant lower inflammatory response, significant higher collagen deposition and angiogenesis compared with control wounds. The combined treatment PRGF +ASCs showed a significant faster cutaneous wound healing process. Conclusions: The combined treatment PRGF+ASCs showed the best results, suggesting this is the best choice to enhance wound healing and improve aesthetic results in acute wounds.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.karger.com/Article/FullText/489739 Background/Aims: Chronic inflammation contributes to cartilage degeneration during the progression of osteoarthritis (OA). Adipose tissue-derived mesenchymal stem cells (ADMSC) show great potential to treat inflammatory and degradative processes in OA and have demonstrated paracrine effects in chondrocytes. In the present work, we have isolated and characterized the extracellular vesicles from human AD-MSC to investigate their role in the chondroprotective actions of these cells. Methods: AD-MSC were isolated by collagenase treatment from adipose tissue from healthy individuals subjected to abdominal lipectomy surgery. Microvesicles and exosomes were obtained from conditioned medium by filtration and differential centrifugation. Chondrocytes from OA patients were used in primary culture and stimulated with 10 ng/ml interleukin(IL)-1β in the presence or absence of AD-MSC microvesicles, exosomes or conditioned medium. Protein expression was investigated by ELISA and immunofluorescence, transcription factor-DNA binding by ELISA, gene expression by real-time PCR, prostaglandin E2 (PGE2) by radioimmunoassay, and matrix metalloproteinase (MMP) activity and nitric oxide (NO) production by fluorometry. Results: In OA chondrocytes stimulated with IL-1β, microvesicles and exosomes reduced the production of inflammatory mediators tumor necrosis factor-α, IL-6, PGE2 and NO. The downregulation of cyclooxygenase-2 and microsomal prostaglandin E synthase-1 would lead to the decreased PGE2 production while the effect on NO could depend on the reduction of inducible nitric oxide synthase expression. Treatment of OA chondrocytes with extracellular vesicles also decreased the release of MMP activity and MMP-13 expression whereas the production of the anti-inflammatory cytokine IL-10 and the expression of collagen II were significantly enhanced. The reduction of inflammatory and catabolic mediators could be the consequence of a lower activation of nuclear factor-κB and activator protein-1. The upregulation of annexin A1 specially in MV may contribute to the anti-inflammatory and chondroprotective effects of AD-MSC. Conclusions: Our data support the interest of AD-MSC extracellular vesicles to develop new therapeutic approaches in joint conditions.

Se trata de un proyecto de investigación de translación en el área de Oftalmología, que plantea el desarrollo de un nuevo tratamiento, basado en las denominadas terapias avanzadas, para pacientes que sufren neuropatía óptica isquémica anterior no arterítica aguda (NOIA-NA). Esta enfermedad afecta a un número de pacientes significativo (2 a 10 personas por cada 100.000 habitantes/año), tiene un alto grado de recurrencia en el ojo contralateral (entre un 15 y un 25% a los 5 años) y no posee un tratamiento efectivo. Por tanto, el objetivo de este proyecto es evaluar la seguridad de la administración intravítrea de células madre mesenquimales (MSC), en concreto el producto en fase de investigación (PEI) nº 15-007 (Mesenchymal Stromal Cells from Valladolid-MSV), en pacientes con NOIA-NA., Instituto Universitario de Oftalmobiología Aplicada (IOBA), Doctorado en Ciencias de la Visión

Background Subjective pain assessment scales have been widely used for assessing lameness in response to pain, but the accuracy of these scales has been questioned. To assess scale accuracy, 10 lame, presa Canario dogs with osteoarthritis (OA) associated with bilateral hip dysplasia were first treated with mesenchymal stem cells. Then, potential lameness improvement was analyzed using two pain scales (Bioarth and visual analog scale). These data were compared with similar data collected using a force platform with the same animals during a period of 6 months after treatment. Results The F test for intraclass correlation showed that concordance in pain/lameness scores between the 2 measuring methodologies was not significant (P value ≥ 0.9213; 95 % confidence interval, –0.56, 0.11). Although subjective pain assessment showed improvement after 6 months, force platform data demonstrated those same animals had returned to the initial lameness state. Conclusion Use of pain assessment scales to measure lameness associated with OA did not have great accuracy and concordance when compared with quantitative force platform gait analysis. Electronic supplementary material The online version of this article (doi:10.1186/s12917-016-0852-z) contains supplementary material, which is available to authorized users.

Este artículo se encuentra disponible en la siguiente URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0170692 The aim of this study was to assess the static posturography in dogs as a useful tool for diagnosis of lameness by means of the use of a pressure platform. For this purpose, a series of different parameters (pressure distribution, area of support, mean pressure, maximum pressure and statokinesiograms) were obtained from five lame dogs with unilateral elbow osteoarthritis treated with plasma rich in growth factors. Data were obtained before and 3 months after treatment, and results were compared with a control group of sound dogs of similar conformation. Significant differences were found in the above mentioned parameters between sound and lame limbs. Improvement after 3 months of treatment was also detected, demonstrating that this multi-parametric technique is an effective and reliable method for the assessment of lameness in dogs.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://www.mdpi.com/1422-0067/19/7/1926 Osteoarthritis (OA) is a major cause of disability in elderly population around the world. More than one-third of people over 65 years old shows either clinical or radiological evidence of OA. There is no effective treatment for this degenerative disease, due to the limited capacity for spontaneous cartilage regeneration. Regarding the use of regenerative therapies, it has been reported that one option to restore degenerated cartilage are adipose-derived mesenchymal stem cells (ASCs). The purpose of this review is to describe and compare the efficacy of ASCs versus other therapies in OA. Methods: Recent studies have shown that ASCs exert paracrine effects protecting against degenerative changes in chondrocytes. According to the above, we have carried out a review of the literature using a combination of osteoarthritis, stem cells, and regenerative therapies as keywords. Results: Conventional pharmacological therapies for OA treatment are considered before the surgical option, however, they do not stop the progression of the disease. Moreover, total joint replacement is not recommended for patients under 55 years, and high tibia osteotomy (HTO) is a viable solution to address lower limb malalignment with concomitant OA, but some complications have been described. In recent years, the use of mesenchymal stem cells (MSCs) as a treatment strategy for OA is increasing considerably, thanks to their capacity to improve symptoms together with joint functionality and, therefore, the patients’ quality of life. Conclusions: ASC therapy has a positive effect on patients with OA, although there is limited evidence and little long-term follow-up.

Tesis - Universidad CEU Cardenal Herrera (Valencia), Facultad de Veterinaria, Departamento de Medicina y Cirugía Animal, leída el 09-02-2017.

Tesis - Universidad CEU Cardenal Herrera (Valencia), Facultad de Veterinaria, Departamento de Medicina y Cirugía Animal, leída el 16-04-2015. La Osteoartrosis (OA) es una de las enfermedades con más prevalencia y morbilidad en la sociedad actual, tanto en medicina humana como en medicina veterinaria, debido sobre todo al aumento de la esperanza de vida y a la prevalencia creciente de la obesidad. Además por su gravedad e irreversibilidad, cada vez existe un mayor interés y empeño en tratar de encontrar tratamientos que alivien los síntomas y retrasen la evolución de la patología. En los últimos años, se han buscado alternativas a los tratamientos convencionales ya que estos últimos únicamente palían la sintomatología de la enfermedad sin detener la progresión de la misma. Entre estas nuevas terapias englobadas dentro de la medicina regenerativa, encontramos el Plasma Rico en Factores de Crecimiento (PRGF) y las células mesenquimales, concretamente las obtenidas a partir del tejido adiposo (CMG). El objetivo del presente trabajo de Tesis Doctoral ha sido la evaluación del efecto de las CMG y del PRGF, así como de la combinación de ambos, en el tratamiento de la Enfermedad degenerativa articular (EDA), utilizando para ello un modelo clínico de la especie canina. Para llevar a cabo el estudio se seleccionaron 49 animales de la especie canina, con EDA en las articulaciones del codo, la cadera o la rodilla. A estos animales se les realizó previamente una exploración física completa, además de una minuciosa exploración traumatológica, y una analítica sanguínea para descartar otras enfermedades concomitantes. Una vez comprobado que todos los animales únicamente padecían EDA, se dividieron aleatoriamente en tres grupos de estudio, dependiendo del tratamiento aplicado (CMG, CMG+PRGF y PRGF), habiendo un total de 25 articulaciones en el grupo de CMG, 23 en el grupo de CMG+PRGF, y 47 en el grupo de PRGF. A continuación, tras la obtención del PRGF y de las CMG se realizó una infiltración intraarticular estándar a cada uno de los animales dependiendo del grupo asignado. Además se estudió la viabilidad de las células madre evaluando las características y naturaleza de estas células, y determinando los efectos de concentraciones crecientes de PRGF en la proliferación de las mismas. Posteriormente, a tiempo basal (antes de aplicar los tratamientos), uno, tres y seis meses tras a la aplicación de los mismos se les realizó a cada uno de los pacientes una evaluación de las variables del estudio entre las que se encuentran las variables de la escala bioarth (grado radiológico de artrosis, limitación funcional y movilidad articular), un cuestionario sobre la satisfacción ante el tratamiento, la valoración del dolor mediante la escala analógica visual (VAS) tanto por parte del propietario, como del veterinario y finalmente la extracción de suero para el posterior análisis de biomarcadores. Además, en los animales de las Palmas de Gran Canaria, se llevó a cabo en cada una de las revisiones un análisis cinético de las extremidades afectadas mediante la plataforma de fuerza. En el análisis de la viabilidad celular, además de obtener un crecimiento exponencial de las muestras con una curva proliferativa definida en un cultivo celular que contenía suero bovino fetal, observamos que los factores de crecimiento contenidos en PRGF pueden también ser suficientes para mejorar el rendimiento de las CMG en cultivos celulares. Tras el análisis estadístico de las variables contenidas en la escala bioarth, observamos que el grado de artrosis se mantuvo constante a lo largo del estudio en todos los grupos, sin existir diferencias significativas entre ellos. La limitación funcional mejoró de manera significativa en los tres grupos obteniendo mejores resultados a los 6 meses en los grupos tratados con CMG y con CMG+PRGF, lo mismo que ocurrió con la movilidad articular. En cuanto a la valoración del dolor mediante la VAS, tanto por parte del propietario como del veterinario, existió una disminución significativa del dolor que presentaban los pacientes desde el primer mes post tratamiento manteniéndose hasta los 6 meses posteriores a la aplicación de los mismos, obteniendo resultados significativamente más favorables en el grupo tratado con CMG+PRGF al finalizar el estudio. Respecto a las preguntas realizadas a los propietarios para determinar el grado de satisfacción que presentaban ante el tratamiento aplicado a sus mascotas, ya desde el primer mes post tratamiento se obtuvieron unos resultados muy satisfactorios, que se mantuvieron hasta el final del estudio en los 3 grupos de estudio. En el estudio de los biomarcadores analizados en el suero de los pacientes, observamos que tanto en el grupo de CMG como en el grupo de CMG+PRGF, existió un aumento progresivo y significativo del ácido hialurónico (HA) en el tiempo, no observándose estos cambios en el grupo tratado con PRGF. Resultados similares se obtuvieron con el C2C, en el cual también observamos tanto en el grupo de CMG como en el de CMG+PRGF una disminución progresiva de este biomarcador en el tiempo, sin existir cambios significativos en el grupo de PRGF. No existieron diferencias entre grupos en ninguno de los tiempos de estudio valorados. Por último, en el estudio de la plataforma de fuerza realizado en los animales de las Palmas de Gran Canaria se observó una mejora progresiva en las variables valoradas en el análisis cinético, alcanzando a los 6 meses de estudio los resultados significativamente más favorables en el grupo de estudio valorado. Tras los resultados logrados en el estudio, podemos decir que la terapia con CMG y PRGF, así como la combinación de ambos tratamientos, son una nueva vía de tratamiento que intenta conseguir una reparación lo más similar al cartílago nativo, sin necesidad de optar por técnicas más agresivas y que causen un daño mayor al organismo. Hay que destacar que en determinados parámetros existen diferencias entre el grupo tratado únicamente con PRGF y los tratados con CMG y la combinación de CMG con PRGF. Nuestra sospecha es que el uso de PRGF en inyección única produce inicialmente una mejora clara del paciente, cuyo efecto está mas limitado en el tiempo, por eso sería recomendable realizar una infiltración seriada de este tratamiento cuando queramos utilizarlo como terapia para la OA. Respecto a las diferencias entre el uso únicamente de CMG, o el uso de una combinación de CMG junto con PRGF, en esta tesis doctoral no hemos obtenido muchas diferencias significativas entre ambos grupos, aunque pensamos que esto es debido a la duración del estudio, ya que si el estudio se hubiese prolongado durante más tiempo, habríamos obtenido resultados con diferencias significativas obteniendo unos valores más favorables en el grupo tratado con CMG+PRGF debido al efecto sinérgico que tienes estos tratamientos. Osteoarthritis (OA) is one of the most prevalent diseases in today's society, both in human medicine and veterinary medicine, mainly due to increased life expectancy and the growing prevalence of obesity. Besides its severity and irreversibility, an increasing interest and commitment exists in trying to find treatments to alleviate symptoms and delay the evolution of the disease. In recent years, alternatives to conventional treatments have been sought after because the latter only palliates the symptoms of disease without halting its progression. Among these new therapies included within regenerative medicine, are Plasma Rich in Growth Factors (PRGF) and mesenchymal cells, particularly those derived from adipose tissue (ADMSC). The aim of this Thesis has been the evaluation of the effect of ADMSC and PRGF, as well as their combination in the treatment of degenerative joint disease (DJD), using a canine species clinical model. Forty nine animals were selected, with DJD in the elbow, hip or knee. A complete physical examination was performed on these animals previously, and a thorough orthopedic examination, and blood tests to rule out other comorbidities. Once verified that all animals were only suffering from DJD, they were randomly divided into three treatment groups, (ADMSC, ADMSC+PRGF and PRGF), having a total of 25 joints in the ADMSC group, 23 in the ADMSC+PRGF group and 47 in the PRGF group. Then, after obtaining the PRGF and ADMSC a standard intra-articular injection was performed. The viability of the stem cells was also studied, evaluating the characteristics and nature of these cells and determining the effects of the increasing concentrations of PRGF in proliferation. Patients were assessed, at baseline (before applying the treatments) and one, three and six months after the application studying variables including Bioath scale (degree of radiological osteoarthritis, functional limitation and joint mobility), questionnaire on satisfaction with the treatment, pain assessment by visual analogue scale (VAS) both by the owner and the Vet and finally serum extraction for subsequent analysis of biomarkers. Also in the animals at Palmas de Gran Canaria, a kinetic analysis of the affected limbs by force platform was performed at each follow up. In the cell viability analysis , in addition to obtaining an exponential growth in the samples with a proliferative curve defined in a cell culture containing fetal bovine serum, we observed that the growth factors contained in PRGF may also be sufficient to improve the performance of the AMSC cultures. After statistical analysis of the variables in the BIOARTH scale, we noted that the degree of osteoarthritis remained constant throughout the study in all groups, with no significant differences between them. Functional limitation improved significantly in all three groups with better results at 6 months in the groups treated with AMSC and ADMSC+PRGF, and the same happened with joint mobility. Regarding the questions owners were asked to determine their satisfaction with the treatment applied to their pets, from the first month post treatment very satisfactory results were observed, which remained constant throughout the study. As for the VAS assessment, by both the owner and vet, there was a significant decrease in pain in the patients from the first month after treatment which remained until 6 months after the application thereof, obtaining significantly more favorable results in the group treated with AMDSC+PRGF group up to the end of the study. With reference to the study of the markers in the serum, we observed that in both the ADMSC and ADMSC+PRGF, there was a progressive and significant increase of hyaluronic acid (HA) at the time and these changes were not observed in the group treated with PRGF alone. Similar results were obtained with the C2C, which we also observed in both the ADMSC group and the ADMSC+PRGF group showing a progressive decrease of this biomarker over time and existed without significant changes in the PRGF group. There were no differences between groups at any of the study times. Finally, the force platform evaluation conducted in animals at Las Palmas de Gran Canaria, progressive improvement was observed in the variables assessed in the kinetic analysis, reaching significantly more favorable results in the study group evaluated at 6 months after treatment. Following the results achieved in the study, we can say that therapy with ADMSC and PRGF, and the combination of both treatments, are a novel treatment attempting to achieve repair as similar to native cartilage without opting for more aggressive techniques and cause further damage to the body. It is important to note that there are differences between certain parameters in the group treated with PRGF alone and that treated with ADMSC and the combination of ADMSC with PRGF. We suspect that using a single PRGF injection initially produces a clear improvement for the patient, the effect is more limited in time, so it would be advisable to perform a serie of injections with this treatment when used as a therapy for OA. Regarding the differences between the use of only ADMSC, or using a combination of ADMSC with PRGF, in this thesis we have not observed any significant differences between the groups, although we think that this is due to the duration of the study and believe that had the studied continued for longer, significant differences would have been obtained with more favorable values in the group treated with ADMSC+PRGF due to the synergistic effect of these treatments.

Tesis - Universidad CEU Cardenal Herrera (Valencia), Facultad de Veterinaria, Departamento de Medicina y Cirugía Animal, leída el 15-04-2015. La incidencia anual de lesiones en la médula espinal en los países desarrollados es de entre un 11,5 y un 53,4 por millón de habitantes, encontrándose entre las principales causas los accidentes de trabajo, los deportes y actividades de aventura, accidentes de trabajo, caídas domésticas y violencia [1]. El paciente presenta parálisis motora y pérdida de la sensibilidad de los segmentos corporales por debajo de la zona de la lesión que se cronifica en el tiempo. La lesión medular provoca la muerte de neuronas, oligodendrocitos, astrocitos y precursores neurales y provoca también, la formación de cavidades que interrumpen los tractos axonales. Todo ello acompañado de mecanismos secundarios que culminan en la formación de una cicatriz glial impenetrable para los axones. A pesar de ello, existe una recuperación espontánea tras la lesión medular debida a la plasticidad de la médula espinal acompañada de la proliferación de nuevos precursores neurales en la médula espinal procedente del canal ependimario, con potencial reparador. Sin embargo, la reparación espontánea es incompleta y se requiere intervención terapéutica. Estudios previos demuestran que la activación de dichas células ependimarias tras una lesión medular (epSPCi) hace que proliferen 10 veces mas rápido in vitro y su trasplante en lesión medular aguda es capaz de obtener una recuperación funcional importante. En el modelo de lesión medular crónico, la aplicación de una terapia celular de administración única, por inyección de epSPCi vía intramedular ha revelado una mejoría funcional significativa tras 4 meses de estudio en lesiones severas. Este rescate de la función locomotriz está acompañado de una mejor distribución en el espesor medular en el epicentro de la lesión con abundantes oligodendrocitos activamente produciendo envolturas nuevas de mielina y una mayor reorganización de la misma en comparación con aquellos animales no trasplantados. El trasplante de células ependimarias ha migrado y se ha mantenido residente en la zona de la lesión, incluida la cicatriz glial, favoreciendo un mayor número de fibras nerviosas dentro de la misma. La terapia celular por lo tanto, resulta efectiva en el rescate de la función neuronal en una lesión crónica, con una cicatriz glial ya establecida. Sin embargo, y a pesar de que el rescate de la función es muy rápido tras el trasplante, la regeneración no es completa, por lo que intervenciones múltiples, con trasplantes seriados y/o combinación de terapias serán necesarias para mejorar las expectativas de éxito. Spinal Cord injuries shows an anual incidence between 11.5 and 53.4 million people. The main causes are sports and adventure activities, work accidents, domestic incidents and falls [5]. The patient develops a motor paralysis and loss of sensation of body segments below the site of injury that becomes chronic in time. Spinal cord injury provokes the death of neurons, oligodendrocytes, astrocytes and neural precursors and also causes the formation of cavities which disrupt axonal tracts. All accompanied by secondary mechanisms that culminate in the formation of a glial scar impenetrable for axons. However, there is a spontaneous recovery after spinal cord injury due to spinal cord plasticity accompanied by the proliferation of new neural precursors from the ependymal canal. However, the spontaneous repair is incomplete and additional therapeutic intervention are required. Previous studies demonstrate that activation of these ependymal cells after spinal cord injury (epSPCi) inducing 10 times faster proliferative activity, moreover, epSPCi transplantation in acute spinal cord injury rescue lost locomotion activity. In the model of chronic spinal cord injury, the application of a single cell therapy administration of epSPCis via intramedullary injection in severe injury revealed significant functional improvement after four months of study. This rescue of locomotor function is accompanied by a better distribution in the thickness of the epicenter of lesion with abundant oligodendrocytes actively producing new myelin sheaths and a major reorganization. In addition, transplantation of ependymal cells migrated and remained resident in the area of the injury, including the glial scar, favoring a greater number of nerve fibers within it. Therefore, this cell therapy is effective in the rescue of neuronal function in a chronic injury with a glial scar already established. However, despite the recovery of the function is very fast after transplantation, regeneration is not complete, so multiple interventions, with serial transplants and / or combination of therapies are needed to improve the success.

Tesis - Universidad CEU Cardenal Herrera (Valencia), Facultad de Ciencias de la Salud, Departamento de Ciencias Biomédicas, leída el 19-05-2015.

Este artículo se encuentra disponible en la siguiente URL: https://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-9-131 Background: Adipose-derived mesenchymal stem cell (ADMSC) therapy in regenerative medicine is a rapidly growing area of research and is currently also being used to treat osteoarthritis (OA). Force platform analysis has been consistently used to verify the efficacy of different therapeutic strategies for the treatment of OA in dogs, but never with AD-MSC. The aim of this study was to use a force platform to measure the efficacy of intraarticular ADMSC administration for limb function improvement in dogs with severe OA. Results: Eight lame dogs with severe hip OA and a control group of 5 sound dogs were used for this study. Results were statistically analyzed to detect a significant increase in peak vertical force (PVF) and vertical impulse (VI) in treated dogs. Mean values of PVF and VI were significantly improved after treatment of the OA groups, reaching 53.02% and 14.84% of body weight, respectively, at day 180, compared with only 43.56% and 12.16% at day 0. Conclusion: This study objectively demonstrated that intraarticular ADMSC therapy resulted in reduced lameness due to OA.

Este artículo se encuentra disponible en la siguiente URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0149472 The aim of this study was to evaluate the use of serum type II collagen cleavage epitope and serum hyaluronic acid as biomarkers for treatment monitoring in osteoarthritic dogs. For this purpose, a treatment model based on mesenchymal stem cells derived from adipose tissue combined with plasma rich in growth factors was used. This clinical study included 10 dogs with hip osteoarthritis. Both analytes were measured in serum at baseline, just before applying the treatment, and 1, 3, and 6 months after treatment. These results were compared with those obtained from force plate analysis using the same animals during the same study period. Levels of type II collagen cleavage epitope decreased and those of hyaluronic acid increased with clinical improvement objectively verified via force plate analysis, suggesting these two biomarkers could be effective as indicators of clinical development of joint disease in dogs.

Este artículo se encuentra disponible en la siguiente URL: https://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-10-143 Background: Regenerative medicine using Mesenchymal Stem Cells (MSC) alone or combined with Plasma Rich in Growth Factors (PRGF) is a rapidly growing area of clinical research and is currently also being used to treat osteoarthritis (OA). Force platform analysis has been consistently used to verify and quantify the efficacy of different therapeutic strategies for the treatment of OA in dogs including MSC associated to PRGF, but never with AD-MSC alone. The aim of this study was to use a force platform to measure the efficacy of intraarticular ADMSC administration for limb function improvement in dogs with severe OA. Results: Ten lame dogs with severe hip OA and a control group of 5 sound dogs were used for this study. Results were statistically analyzed to detect a significant increase in peak vertical force (PVF) and vertical impulse (VI) in treated dogs. Mean values of PVF and VI were significantly improved within the first three months post-treatment in the OA group, increasing 9% and 2.5% body weight, respectively, at day 30. After this, the effect seems to decrease reaching initial values. Conclusion: Intraarticular ADMSC therapy objectively improved limb function in dogs with hip OA. The duration of maximal effect was less than 3 months.