Mesenchymal stem cells and cell-free preparations for treating atopic dermatitis

Atopic dermatitis (AD) is a chronic cutaneous inflammatory disease caused by an interaction between genetic, immune and epidermal barrier factors. Several treatments can be used to treat this disease but there are patients that do not respond to actual drugs. So, there is a need to develop effective therapies for AD. Mesenchymal stem cells (MSCs) are non-hematopoietic multipotent adult progenitor cells with immunomodulatory power and self-regenerating capacity to repair tissue damage, so they could be a potential effective treatment for AD. MSCs-Conditioned Medium (CM) and MSCs-exosomes are cell-free preparation with molecules secreted by stem cells that could be also beneficial for AD. This viewpoint reviews the actual development of MSCs, MSCs-CM and MSCs-exosomes for treating patients with AD

Current Advanced Therapies Based on Human Mesenchymal Stem Cells for Skin Diseases

Skin disease may be related with immunological disorders, external aggressions, or genetic conditions. Injuries or cutaneous diseases such as wounds, burns, psoriasis, and scleroderma among others are common pathologies in dermatology, and in some cases, conventional treatments are ineffective. In recent years, advanced therapies using human mesenchymal stem cells (hMSCs) from different sources has emerged as a promising strategy for the treatment of many pathologies. Due to their properties; regenerative, immunomodulatory and differentiation capacities, they could be applied for the treatment of cutaneous diseases. In this review, a total of thirteen types of hMSCs used as advanced therapy have been analyzed, considering the last 5 years (2015–2020). The most investigated types were those isolated from umbilical cord blood (hUCB-MSCs), adipose tissue (hAT-MSCs) and bone marrow (hBM-MSCs). The most studied diseases were wounds and ulcers, burns and psoriasis. At preclinical level, in vivo studies with mice and rats were the main animal models used, and a wide range of types of hMSCs were used. Clinical studies analyzed revealed that cell therapy by intravenous administration was the advanced therapy preferred except in the case of wounds and burns where tissue engineering was also reported. Although in most of the clinical trials reviewed results have not been posted yet, safety was high and only local slight adverse events (mild nausea or abdominal pain) were reported. In terms of effectiveness, it was difficult to compare the results due to the different doses administered and variables measured, but in general, percentage of wound’s size reduction was higher than 80% in wounds, Psoriasis Area and Severity Index and Severity Scoring for Atopic Dermatitis were significantly reduced, for scleroderma, parameters such as Modified Rodnan skin score (MRSC) or European Scleroderma Study Group activity index reported an improvement of the disease and for hypertrophic scars, Vancouver Scar Scale (VSS) score was decreased after applying these therapies. On balance, hMSCs used for the treatment of cutaneous diseases is a promising strategy, however, the different experimental designs and endpoints stablished in each study, makes necessary more research to find the best way to treat each patient and disease.Instituto de Salud Carlos III (European Regional Development Fund "A way to make Europe") PI13/02576 PI17/02083Andalusian Regional Government SAS PI-0458-2016 PIGE-0242-2019Instituto de Salud Carlos III (European Social Fund "Investing in your future") FI18/0026

Real world effectiveness and tolerability of candesartan in the treatment of migraine: a retrospective cohort study

To date, two randomized, controlled studies support the use of candesartan for migraine prophylaxis but with limited external validity. We aim to evaluate the effectiveness and tolerability of candesartan in clinical practice and to explore predictors of patient response. Retrospective cohort study including all patients with migraine who received candesartan between April 2008-February 2019. The primary endpoint was the number of monthly headache days during weeks 8–12 of treatment compared to baseline. Additionally, we evaluated the frequency during weeks 20–24. We analysed the percentage of patients with 50% and 75% response rates and the retention rates after three and 6 months of treatment. 120/4121 patients were eligible, aged 45.9 [11.5]; 100 (83.3%) female. Eighty-four patients (70%) had chronic migraine and 53 (42.7%) had medication-overuse headache. The median number of prior prophylactics was 3 (Inter-quartile range 2–5). At baseline, patients had 20.5 ± 8.5 headache days per month, decreasing 4.3 ± 8.4 days by 3 months (weeks 12–16) and by 4.7 ± 8.7 days by 6 months (paired Student’s t-test, p < 0.001). The percentage of patients with a 50% response was 32.5% at 3 months and 31.7% at 6 months, while the retention rate was 85.0% and 58.3%. The number of prior treatments (Odds ratio 0.79, 95% CI 0.64–0.97) and the presence of daily headache (Odds ratio 0.39, 95% CI 0.16–0.97) were associated with a lower probability of response. Candesartan showed beneficial effects in the preventive treatment of migraine in clinical practice, including patients with chronic migraine, medication-overuse headache and resistance to prior prophylactics

Técnicas de digitalización para el levantamiento gráfico y de diagnóstico mediante pruebas no destructivas para el estudio de lesiones en el patrimonio construido

[EN]Historic buildings and heritage elements located in urban or rural environments are studied for the detection of the degradation of their materials and the characterization of their geometry.Due to the complex structures and constructions that are part of the Cultural Heritage, currently, without the help of advanced instruments and techniques in the collection of digital data, the representation of this falls into an idealized simplification, often ignoring geometry, hidden spaces and specific details.There are numerous tests that facilitate the examination and analysis of Building of Cultural Interest and systems without damaging them or affecting their integrity. They are called non-destructive tests, such as short-range photogrammetry, terrestrial 3D scanner and / or structured light, among others.The technique used should be chosen according to the object or building to be studied, that is, the morphological characteristics, workspace, the expected level of detail, the execution time and the metric quality of the final result, among others, are fundamental parameters that determine the most appropriate working technique.Therefore, this article analyzes and compares several of the digitization techniques, and the data obtained in each of them, on the Heritage applied to Building of Cultural Interest such as "La Puerta de la Colada" of the walls of Ciudad Rodrigo (Salamanca, Spain), the Castle of Sagunto (Valencia, Valencia, Spain) and "Salón de Reinos" of Prado Museum (Madrid, Spain). All the examples presented have been the subject of an exhaustive analysis and subsequent intervention for its consolidation and restoration, so the techniques used have been chosen depending on the subsequent intervention to be carried out.Sánchez Corrochano, Á.; Martínez Sierra, E.; Greco, A. (2024). Técnicas de digitalización para el levantamiento gráfico y de diagnóstico mediante pruebas no destructivas para el estudio de lesiones en el patrimonio construido. Editorial Universitat Politècnica de València. https://doi.org/10.4995/FORTMED2024.2024.1809

Digital technicals for the study of the Defensive Heritage: “Puerta de Almenara” and south walls of “Palacio del Gobernador” and “Plaza de Armas” of the Castle of Sagunto (Valencia)

[EN] The use of digital documentation and registration techniques in Cultural Heritage is becoming more common every day, thanks to its ability to capture a large amount of data in a fast and efficient process. Its high geometric precision, thoroughness, performance retrieved and especially the generation of high fidelity and precision of architectural good assets make these tools optimal for the planimetric surveys. The work of intervention or conservation of cultural heritage requires a previous graphic registration using different techniques available. This article presents a combined method of implementation of various digital techniques that allow to achieve the most accurate graphic documentation possible. The different results obtained from the use of photogrammetry by drone or by manual camera are discussed. It is intended to seek the standardization and optimization of the process of documentation and value of the Cultural Heritage by combining these techniques. These techniques have been used in a real case: the three-dimensional modeling of various parts of the defensive set of the Castle of Sagunto (Valencia), called the “Puerta de Almenara”, which gives access to the square of the same name, on the eastern side and some walls of the fortification. The Castle of Sagunto is a mosaic of the different cultures who occupied it (Iberians, Romans, Goths, Arabs...). The fortification is located on top of a hill of the Sierra Calderona, controlling even the Mediterranean coastal road and the communication route with Aragon. During the last years, the castle has been immersed, for almost 20 years, in various works of consolidation and restoration to initiatives of the Institute of Cultural Heritage of Spain.Sánchez Corrochano, Á.; Martínez Sierra, E.; Greco, A.; Besana, D. (2020). Técnicas digitales para el estudio del Patrimonio Defensivo: “Puerta de Almenara” y lienzos sur del Palacio del Gobernador y Plaza de Armas del Castillo de Sagunto (Valencia). Editorial Universitat Politècnica de València. 455-462. https://doi.org/10.4995/FORTMED2020.2020.11387OCS45546

The Role of Exosomes Derived From Mesenchymal Stromal Cells in Dermatology

This study has been funded by the Carlos III Health Institute of Spain through the PI13/02576 and PI17/02083 projects [cofunded by European Regional Development Fund "A way to make Europe" and Andalusian Regional Government Finance (SAS PI-0458-2016)]. The work of MQ-V was supported by a predoctoral fellowship (BOE 22/10/2019) from the Spanish Ministry of Science, Innovation and Universities. This study is part of her doctoral research in the Biomedicine program at the University of Granada.The skin is the largest organ of the human body and its main functions include providing protection from external harmful agents, regulating body temperature, and homeostatic maintenance. Skin injuries can damage this important barrier and its functions so research focuses on approaches to accelerate wound healing and treat inflammatory skin diseases. Due to their regenerative and immunomodulatory properties, mesenchymal stromal cells (MSCs) have been reported to play a significant role in skin repair and regeneration. However, it seems that the secretome of these cells and exosomes in particular may be responsible for their functions in skin regeneration and the immunomodulation field. The present review aims to gather the available information about the role of MSC-derived exosomes for both in vitro and in vivo models of different skin conditions and to highlight the need for further research in order to overcome any limitations for clinical translation.Carlos III Health Institute of Spain [European Regional Development Fund "A way to make Europe"] PI13/02576 PI17/02083Carlos III Health Institute of Spain [Andalusian Regional Government Finance] PI13/02576 PI17/02083 SAS PI-0458-2016Spanish Ministry of Science, Innovation and Universities BOE 22/10/201

Erythema Increase Predicts Psoriasis Improvement after Phototherapy

Psoriasis is a major global health problem. There is a need to develop techniques to help physicians select the most appropriate cost-effective therapy for each patient. The main objectives of this study are (1) to evaluate changes in epidermal barrier function and skin homeostasis after phototherapy and (2) to explore potentially predictive values in epidermal barrier function and skin homeostasis to assess clinical improvement after fifteen sessions of phototherapy. A total of 76 subjects, 38 patients with plaque-type psoriasis and 38 gender- and age-matched healthy volunteers, were included in the study. Erythema, transepidermal water loss (TEWL), temperature, stratum corneum hydration (SCH), pH, sebum, and antioxidant capacity were measured before and after the first and fifteenth phototherapy session. Erythema (401.09 vs. 291.12 vs. 284.52 AU, p < 0.001) and TEWL (18.23 vs. 11.44 vs. 11.41 g·m−2 ·h −1 , p < 0.001) were significantly higher at psoriatic plaques than in uninvolved psoriatic skin and healthy volunteers, respectively, while SCH was lower (9.71 vs. 44.64 vs. 40.00 AU, p < 0.001). After fifteen phototherapy sessions, TEWL (–5.19 g·m−2 ·h −1 , p = 0.016) decreased while SCH (+7.01 AU, p = 0.013) and erythema (+30.82 AU, p = 0.083) increased at psoriatic plaques. An erythema increase exceeding 53.23 AU after the first phototherapy session, with a sensitivity of 71.4% and specificity of 84.2%, indicates that a patient may improve Psoriasis Area and Severity Index (PASI) by ≥3 points after fifteen phototherapy sessions. In conclusion, phototherapy improves epidermal barrier function in psoriatic patients and the erythema increase after one phototherapy session could help doctors select psoriasis patients who are more likely to respond to phototherapy

Autoimmune inflammation triggers aberrant astrocytic calcium signaling to impair synaptic plasticity

Cortical pathology involving inflammatory and neurodegenerative mechanisms is a hallmark of multiple sclerosis and a correlate of disease progression and cognitive decline. Astrocytes play a pivotal role in multiple sclerosis initiation and progression but astrocyte-neuronal network alterations contributing to gray matter pathology remain undefined. Here we unveil deregulation of astrocytic calcium signaling and astrocyte-to-neuron communication as key pathophysiological mechanisms of cortical dysfunction in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis. Using two-photon imaging ex vivo and fiber photometry in freely behaving mice, we found that acute EAE was associated with the emergence of spontaneously hyperactive cortical astrocytes exhibiting dysfunctional responses to cannabinoid, glutamate and purinoreceptor agonists. Abnormal astrocyte signaling by Gi and Gq protein coupled receptors was observed in the inflamed cortex. This was mirrored by treatments with pro-inflammatory factors both in vitro and ex vivo, suggesting cell-autonomous effects of the cortical neuroinflammatory environment. Finally, deregulated astrocyte calcium activity was associated with an enhancement of glutamatergic gliotransmission and a shift of astrocyte-mediated short-term and long-term plasticity mechanisms towards synaptic potentiation. Overall, our data identify astrocyte-neuronal network dysfunctions as key pathological features of gray matter inflammation in multiple sclerosis and potentially additional neuroimmunological disorders.This work was funded by the Instituto de Salud Carlos III (PI21/00629, to S.M. and A.R.-A; CB06/05/00, to C.M.) and cofounded by the European Union, Basque Government (PIBA_2023_1_0046; 2023111031; IT1473-22, to S.M.; IT1203-19, to C.M.), ARSEP Foundation (ARSEP - 1310 to S.M. and G.M.), INSERM (to G.M.), the European Research Council (MiCaBra, ERC-2017-AdG-786467, to G.M.), Fondation pour la Recherche Medicale (FRM, DRM20101220445 to G.M.), Region Aquitaine (CanBrain, AAP2022A-2021-16763610 and -17219710 to G.M.); French State/Agence Nationale de la Recherche (HippObese, ANR-23-ce14-0004-03; ERA-Net Neuron CanShank, ANR-21-NEU2-0001-04, to G.M), La Caixa Research Health 2023 (PsychoCannabis, HR23-00793, to G.M.), Spanish Ministry of Science and Innovation (PID2019-109724RB-100 to C.M.; PGC2018-093990-A-I00, to E.S.-G.), National Institutes of Health-MH (MH, R01MH119355; NINDS, R01NS097312; NIDA, R01DA048822, to A.A.), Postdoctoral and Predoctoral Programs of the Basque Government (to A.M.B., T.C. A.M.-G., and C.U), Predoctoral Program of the UPV/EHU (to E.S.)

Pilot study of cutaneous tolerability of fibrin-agarose substitutes in healthy volunteers

Objetivos: En el presente estudio se persigue comprobar posibles reacciones adversas, derivadas del uso tópico de láminas de fibrina-agarosa en el antebrazo de voluntarios sanos. Metodología: Se llevó a cabo un estudio experimental en siete voluntarios sanos, cinco varones y dos mujeres, que no presentaban ningún tipo de lesión cutánea visible. En el antebrazo de cada voluntario se colocaron dos láminas de fibrina-agarosa de 4 cm2 . Cada lámina se cubrió con un apósito impregnado y sobre una de las láminas se aplicó pomada antibiótica con mupirocina. Ambas láminas se cubrieron finalmente con un apósito protector y se mantuvieron en contacto directo sobre la piel durante 48 horas. Resultados: Los resultados determinaron que no se detectaron reacciones adversas después de 48 horas de evolución ni en los siguientes 7 días en ningún voluntario. Se observaron diferencias entre las dos láminas implantadas en cada voluntario, ya que al retirar el apósito cubierto con pomada antibiótica, la lámina presentaba un aspecto más hidratado que la que no llevaba pomada antibiótica. Conclusiones: El uso tópico de las láminas de fibrina-agarosa en voluntarios sanos no presenta reacciones adversas del tipo irritación o alergia al aplicarse directamente por vía tópica. Aunque el tamaño muestral del estudio es limitado, sugiere que la combinación de fibrina-agarosa se presenta como el biomaterial idóneo para el desarrollo de un modelo de piel artificial humana.Purpose: This study aims to analyse possible adverse reactions resulting from the topical use of fibrin-agarose substitutes in the forearm of healthy volunteers. Methods: An experimental study was carried out in seven healthy volunteers, five males and two females, who did not have any cutaneous lesion. Two fibrin-agarose substitutes of 4 cm2 were placed in the forearm of each volunteer. Each substitute was covered with an impregnated dressing and one of the substitutes was covered with antibiotic ointment (mupirocin). Both substitutes were finally covered with a protective dressing. The substitutes were maintained for 48 hours. Results: The results determined that no adverse reactions were detected in any volunteer after 48 hours and a week of evolution. Differences were observed between the two substitutes implanted in each volunteer, since when removing the covered dressing with antibiotic ointment, the substitute presented a more hydrated appearance than the one without antibiotic cream. Conclusions: The implant of fibrin-agarose substitutes in healthy volunteers does not present irritation or allergic type adverse reactions when they applied directly topically on the skin. Although the sample size is low, the fibrin-agarose combination is presented as the biomaterial suitable for the development of an artificial human skin model

Epithelial in vitro differentiation of human mesenchymal stem cells (hMSCs) from adipose tissue (AT) and bone marrow (BM): cellular characterization and study of HLA I and II expression

AGRADECIMIENTOS Laboratorio de Citogenética del servicio de Análisis Clínicos del Hospital Universitario Virgen de las Nieves. Servicio de Análisis Clínicos (Sección de Citometría/Biopatología tumoral) del Hos- pital Universitario Virgen de las Nieves.Introducción: Las células troncales mesenquimales derivadas de tejido adiposo o médula ósea constituyen uno de los tratamientos de terapia celular más utilizados en los ensayos clínicos actuales por su capacidad inmunomoduladora. Además, por su potencial de diferenciación a células epiteliales pueden ser utilizadas en ingeniería tisular incorporadas a tejidos artificiales como la piel o córnea, sustituyendo a las células epiteliales autólogas de estos tejidos. Es necesario realizar una correcta caracterización de estas células diferenciadas y estudiar el efecto de la diferenciación en la expresión del HLA de clase I y II. Objetivos: Caracterizar y realizar los controles de calidad GMP en dos líneas de células mesenquimales troncales humanas de distintos orígenes (tejido adiposo y médula ósea) tras diferenciarlas a células epiteliales in vitro, y analizar si se modifica la expresión de los marcadores HLA I y II antes y después del proceso diferenciador. Metodología: Se ha realizado el aislamiento y expansión de las dos líneas celulares de células mesenquimales troncales a partir del tejido fuente y se ha procedido a su diferenciación in vitro a células epiteliales mediante medios de cultivos suplementados con factores de crecimiento específico. Se han realizado controles de calidad siguiendo los requerimientos de las normas de correcta fabricación y se ha estudiado por citometría de flujo la expresión de HLA tipo I y II antes y después del proceso diferenciador. Finalmente se ha comprobado mediante estudio histológico e inmunohistoquímico las características de las células diferenciadas. Resultados: Se han aislado dos líneas de células mesenquimales troncales de tejido adiposo y médula ósea que cumplen los controles de calidad propuestos. Tras el proceso diferenciador in vitro, las células mesenquimales troncales humanas no expresan marcadores HLA (I y II) importantes en la respuesta inmune, pero sí expresan débilmente proteínas relacionadas con los principales estratos epiteliales (CK5, CK6 y CK14). Conclusión: La ausencia de expresión de marcadores de HLA I y II por citometría de flujo en las células diferenciadas favorecería su uso con carácter alogénico en la construcción de piel y córneas humanas por ingeniería de tejidos, sin embargo, son necesarios más estudios que confirmen estos resultados preliminares y protocolos que optimicen el proceso diferenciador in vitro de las células mesenquimales troncales.Background: Human mesenchymal stem cells derived from adipose tissue and bone marrow are one of the most common cell therapy procedures used in recent clinical trials due to their immunomodulation capacity. Furthermore, for their epithelial differentiation potential can be used in tissue engineering, incorporated in artificial tissues such as skin and cornea, replacing autologous epithelial cells. It is necessary to make a correct cellular characterization of differentiated cells and to study the effect in HLA I and II expression. Objetives: Characterization and quality controls under GMP conditions of in vitro differentiated human mesenchymal stem cells from different sources (adipose tissue and bone marrow) to epithelial lineage, and study of HLA I and II expression before and after differentiation. Methods: Isolation and expansion of two human mesenchymal stem cells lines from their tissues of origin and in vitro differentiation to epithelial cells using culture mediums supplemented with specific growth factors. Quality controls according Good Manufacturing Practices have been made and HLA I and II expression before and after differentiation have been studied. Finally, characteristics of differentiated cells have been demonstrated by histological and immunohistochemical analysis. Results: Two human mesenchymal stem cells lines from adipose tissue and bone marrow have been isolated complying with the proposed quality controls. After in vitro differentiation, human mesenchymal stem cells do not express HLA (I and II) markers, which are important in immune response, but weakly express proteins related to main epithelial layers of human skin (CK5, CK6 and CK14). Conclusion: The absence of expression of HLA I and II by flow cytometry in differentiated cells would promote the use of them with allogenic character to construct human skin and cornea by tissue engineering, however, more studies and protocols are required to confirm these preliminary results and to optimize in vitro differentiation of human mesenchymal stem cells.FIS ISC-III and FEDER PI13/0257