Rspo2-LGR4 exacerbates hepatocellular carcinoma progression via activation of Wnt/b-catenin signaling pathway / Rspo2-LGR4 agrava la progresión del carcinoma hepatocelular al activar la vía de señalización Wnt/beta-catenina

Background: The leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) plays an important role in stem cell differentiation, organ development and cancer. Whether LGR4 affects the progression of hepatocellular carcinoma (HCC) remains unknown. This study aimed to reveal the role of LGR4 in HCC. Methods: Clinical samples of HCC were collected to assess the expression of LGR4 and its correlation with patients’ clinical characteristics. The expression level of LGR4 in HCC cells was altered by pharmacological and genetic methods, and the role of LGR4 in HCC progression was analyzed by in vivo and in vitro assays. HCC was induced by diethylnitrosamine (DEN) and carbon tetrachloride (CCl4) in wild-type and LGR4 deficient mice, the effect of LGR4 on HCC was examined by histopathological evaluation and biochemical assays. Results: LGR4 expression was up-regulated in HCC samples, and its expression level was positively correlated with tumor size, microvascular invasion (MVI), TNM stage and pathological differentiation grade of HCC patients. In the mouse HCC model induced by DEN+CCl4, knockdown of LGR4 effectively inhibited the progression of HCC. Silencing of LGR4 inhibited the proliferation, migration, invasion, stem cell-like properties and Warburg effect of HCC cells. These phenotypes were promoted by R-spondin2 (Rspo2), an endogenous ligand for LGR4. Rspo2 markedly increased the nuclear translocation of β-catenin, whereas IWR-1, an inhibitor of Wnt/β-catenin signaling, reversed its effect. Deficiency of LGR4 significantly reduced the nuclear translocation of β-catenin and the expression of its downstream target genes cyclinD1 and c-Myc. Conclusions: LGR4 promotes HCC progression via Wnt/β-catenin signaling pathway. (AU)

Antecedentes: El receptor de acoplamiento de proteínas G de secuencia repetida 4 (LGR4), rico en leucina, juega un papel importante en la diferenciación de células madre, el desarrollo de órganos y el cáncer. Se desconoce si LGR4 afecta la progresión del carcinoma hepatocelular (HCC). El objetivo de este estudio es revelar el papel de LGR4 en el HCC. Métodos: Se recolectaron muestras clínicas de HCC para evaluar la expresión de LGR4 y su correlación con los resultados clínicos de HCC. Alterar los niveles de expresión de LGR4 en las células de HCC mediante métodos farmacológicos y genéticos y analizar el papel de LGR4 en la progresión del cáncer de hígado mediante mediciones in vivo e in vitro. El HCC fue inducido en ratones de tipo salvaje y con defectos de LGR4 con Nitrosamina de dietilo (DEN) y cloruro de carbono (CCl4), y los efectos de LGR4 sobre el HCC fueron detectados por evaluación histopatológica y determinación bioquímica. Resultados: La expresión de LGR4 está regulada en HCC, y su nivel de expresión está positivamente relacionado con el tamaño tumoral, la infiltración microvascular (MVI), la etapa de TNM y el grado de diferenciación patológica en pacientes con HCC. En el modelo de HCC de ratón inducido por DEN+CCl4, golpear bajo LGR4 inhibió efectivamente la progresión del HCC. El silencio de LGR4 inhibe la proliferación, migración, invasión, propiedades similares a las células madre y el efecto Warburg de las células HCC. Estos fenotipos son promovidos por el ligando endógeno roof slab-specific sponge 2 (Rspo2)de LGR4. El Rspo2 aumentó significativamente la translocación nuclear de la proteína beta-catenina, mientras que el inhibidor de la señalización Wnt/beta-cateninaIWR-1 revirtió su acción... (AU)

Tratamiento de la pseudoartrosis de radio con injerto de cresta iliaca y células madre del aspirado de médula ósea. A propósito de un caso / Treatment of radial pseudarthrosis with iliac crest graft and bone marrow aspirate stem cells. A case report

La pseudoartrosis es una complicación caracterizada por la ausencia de consolidación del hueso a los 9 meses desde el inicio de la fractura, con falta de progresión radiológica los últimos 3 meses, siendo sus principales causas el exceso de movimiento en el foco de fractura y una insuficiente vascularización. A pesar de no tratarse de una complicación frecuente, los huesos del antebrazo ocupan el 4.º puesto en incidencia de presentación. El manejo anestésico de la patología quirúrgica del miembro superior se realiza generalmente en régimen ambulatorio con técnicas de anestesia regional guiadas por ecografía. Estas técnicas tienen una doble función: anestesia durante el propio acto quirúrgico con una mínima variabilidad sobre el estado basal del paciente y analgesia en el postoperatorio inmediato, permitiendo de esta manera el alta a domicilio de forma más segura y precoz. Presentamos el caso de un varón de 34 años, con desarrollo de pseudoartrosis atrófica tras fractura diafisaria de radio, en el que se realiza injerto óseo de cresta ilíaca y aspirado de células madre como estímulo de la osteogénesis.(AU)

Pseudarthrosis is a complication characterised by the absence of bone healing 9 months after the onset of the fracture, with a lack of radiological progressionin the last 3 months, and its main causes are excessive movement at the fracture site and insufficient vascularisation. Despite not being a frequent complica-tion, the bones of the forearm occupy the fourth place in incidence of presentation. The anaesthetic management of surgical pathology of the upper limb isgenerally performed on an outpatient basis with regional anaesthesia techniques guided by ultrasound. These techniques have a dual function: anaesthesiaduring the surgical act with minimal variability over the patient’s baseline condition and analgesia in the immediate postoperative period, thus allowing forsafer and earlier discharge home. We present the case of a 34-year-old male with diagnosis of atrophic pseudarthrosis following a diaphyseal fracture of theradius, in whom iliac crest bone grafting and stem cell aspiration were performed to stimulate osteogenesis.(AU)

El embrioide y sus leyes. Una breve aproximación al contexto internacional / L’embrioide i les seves lleis. Una breu aproximació al context internacional / The embryoid and its laws. A brief approach to the international context

En los últimos años el desarrollo de modelos in vitro con células madre humanas que simulan el desarrollo embrionario temprano ha vivido un gran progreso. Las dificultades para acceder a embriones humanos, la escasez de material embrionario y los desafíos técnicos, legales y éticos existentes sobre la investigación y experimentación con embriones humanos in vitro siguen siendo una barrera para avanzar en el conocimiento de la embriogénesis tras la gastrulación. Por ello distintos mecanismos celulares subyacentes a la formación de las líneas celulares en los seres humanos siguen siendo desconocidos. En el presente trabajo intentaremos reflejar varios de los aspectos que son motivo de incertidumbres jurídicas internacionales en relación con la investigación con embrioides como modelo experimental.(AU)

En els últims anys el desenvolupament de models in vitroamb cèl·lules mare humanes que simulen el desenvolupament embrionari primerenc ha viscut un gran progrés. Les dificultats per a accedir a embrions humans, l'escassetat de material embrionari i els desafiaments tècnics, legals i ètics existents sobre la recerca i experimentació amb embrions humans in vitrocontinuen sent una barrera per a avançar en el coneixement de la embriogènesi després de la gastrulació. Per això diferents mecanismes cel·lulars subjacents a la formació de les línies cel·lulars en els éssers humans continuen sent desconeguts. En el present treball intentaré reflectir diversos dels aspectes que són motiu d'incerteses jurídiques internacionals en relació amb la recerca amb embrioides com a model experimental.(AU)

In recent years, the development of in vitromodels with human stem cells that simulate early embryonic development has experienced great progress. The difficulties in accessing human embryos, the scarcity of embryonic material, and the existing technical, legal, and ethical challenges regarding research and experimentation with in vitrohuman embryos still represents a barrier to advancing in the knowledge of post-gastrulation embryogenesis. Therefore, different cellular mechanisms underlying the formation of cell lines in humans remain unknown. In the present work Iwill try to reflect several of the aspects that are the cause of internationallegal uncertainties in relation to research with embryoids as an experimental model.(AU)

Efficacy of stem cells in bone rehabilitation in patients with alveolar bone atrophy: a systematic review

Background: Biomedical engineering proposes the use of stem cells as a bone rehabilitation treatment in patients with alveolar bone defects. Many authors suggest that this innovative technique could represent the future of bone regeneration in dentistry. The present study systematically reviewed the efficacy of stem cells in bone regeneration in patients with alveolar bone atrophy. Material and methods: The study was developed following the criteria of the PRISMA guideline (2020). The literature review was conducted in Pubmed, Medline Complete, and Scopus. The search algorithms used the following key words: stem cells, bone regeneration, and alveolar ridge augmentation. To assess the risk of bias, the CASPe methodology was used. Results: Seven clinical trials in humans were included in this systematic review. In all the studies, the proposed objective of bone regeneration by using stem cells was achieved, although in a different way with different results. Although the authors of the analysed clinical trials achieved favourable results, they highlighted the presence of multiple limitations throughout bone regeneration treatments, such as scarce scientific literature on stem cells, a reduced number of follow-up studies, and a lack of a standardized international protocol. Conclusions: Based on the analysed studies, it is concluded that the therapy proposed by tissue engineering through the use of stem cells to rehabilitate patients with bone atrophies can be considered effective. In addition, the need for further studies and standardization of protocols is highlighted. (AU)

Las Células Madre Mesenquimales Endometriales modificadas genéticamente como Terapia Celular para la Endometriosis / Genetic modified endometrial mesenchymal stem cells as a cellular therapy for endometriosis

Se plantea una revisión e integración de tres conceptos fundamentales que son de alto interés científi-co y biomédico: la endometriosis, las Células Madre Mesenquimales Endometriales y su modificación genética para ser empleadas como terapia celular en esta patología. La endometriosis es una enferme-dad ginecológica, crónica y benigna caracterizada por la formación ectópica de estroma endometrial y glándulas principalmente en el peritoneo pélvico. Afecta entre el 6-15% de mujeres en edad reproducti-va con importantes consecuencias en su salud física, reproductiva y mental. Se ha planteado una importante participación de las células madre endometriales dentro de sus causas, por lo que se convierten en un objetivo terapéutico significativo. Se ha reportado múltiples características que establecen a las Células Madre Mesenquimales Endometriales como modelo idóneo para su utilización en terapia celular: caracterización definida, capacidad de migración y co-localización en la lesión, acción paracrina y juxtacrina, acción inmunomoduladora, acción moduladora de la angiogénesis y de la apop-tosis, baja tumorigenicidad, capacidad de transdiferenciación, entre otros, y se propone el uso concre-to de las células madre de sangre menstrual por su fácil obtención, caracterización y cultivo. (AU)

A review and integration of three fundamental concepts that are currently of high scientific and biome-dical interest is proposed: endometriosis, Endometrial Mesenchymal Stem Cells and their genetic modification to be used as cell therapy in this pathology. Endometriosis is a chronic, benign gynecolo-gical disease characterized by the ectopic formation of endometrial stroma and glands, mainly in the pelvic peritoneum. It affects between 6-15% of women in reproductive age with important consequen-ces on their physical, reproductive and mental health. It has been established an important participa-tion of endometrial stem cells within its causes, which is why they become a significant therapeutic target. It has been reported multiple characteristics that support Endometrial Mesenchymal Stem Cells as the optimal model for their use in cell therapy: defined characterization, migration capacity and co-location in the lesion, paracrine and juxtacrine action, immunomodulatory action, modulator action of angiogenesis and apoptosis, low tumorigenicity, transdifferentiation capacity, among others, and the specific use of menstrual blood stem cells is proposed because their easy obtaining, characteriza-tion and cell culture. It has been described that these stem cells have a dynamic and dual behavior, in the sense that they can contribute both to the formation of a lesion and to its modulation, for which, a better understanding of their activity is proposed before their direct use and, to mitigate this dynamic cellular behavior, it is proposed that cell therapy employ a strategy of genetic modification of cells for the expression of factors relevant to the treatment of endometriotic lesions and that have an antiproli-ferative, immunomodulatory action. (AU)

Eficacia y seguridad de la lipotransferencia asistida con células madre en el rejuvenecimiento facial / Efficacy and safety of stem cell-assisted fat transfer in facial rejuvenation

Introducción y objetivo: El lipoinjerto aporta un rejuvenecimiento volumétrico que mejora la calidad del tejido facial por un mecanismo que podría implicar participación de células madre. Realizamos la presente investigación con el objetivo de evaluar la eficacia y seguridad de la lipotransferencia por centrifugación asistida con células madre derivadas del tejido adiposo en el envejecimiento facial. Material y método: Ensayo clínico comparativo aleatorizado doble ciego (fase III) en 70 pacientes de los que 35 recibieron tratamiento de lipotransferencia facial asistido con células madre (grupo estudio) y 35 recibieron tratamiento con lipotransferencia con centrifugación convencional (grupo control). Resultados: El porcentaje de conservación del volumen del lipoinjerto disminuyó con el tiempo (grupo control, p < 0.001; grupo de estudio, p = 0.223); el tratamiento asistido con células madre redujo el riesgo de pérdida de conservación del lipoinjerto en un 86 %. Constatamos desaparición y disminución de arrugas; ningún paciente permaneció en la escala de envejecimiento más desfavorable. Los cambios ocurridos en los 2 grupos fueron significativos (p < 0.001) con mejores resultados en el grupo experimental. No recogimos complicaciones. El 100% de los pacientes expresó sentirse satisfecho. Referente a la evaluación del resultado final, en el 94.3% resultó buena en el grupo estudio y en igual proporción de los pacientes del grupo control fue regular. Conclusiones: En nuestro estudio la lipotransferencia por centrifugación asistida con células madre derivadas del tejido adiposo demostró ser eficaz y segura en rejuvenecimiento facial y superior a la lipotransferencia por centrifugación convencional. (AU)

Background and objective: Lipograft provides volumetric rejuvenation that improves the quality of facial tissue by a mechanism that could involve the participation of stem cells. Our objective is to evaluate the efficacy and safety of lipotransfer by assisted centrifugation with stem cells derived from adipose tissue in facial aging. Methods: Double-blind randomized comparative clinical trial (phase III) in 70 patients, 35 who received lipotransfer facial treatment assisted with stem cells (study group) and 35 who received conventional facial lipotransfer by centrifugation (control group). Results: Percentage of fat graft volume conservation required over time (control group, p < 0.001; study group, p = 0.223); stem cell-assisted treatment reduced the risk of loss of fat graft preservation by 86%. The patient's wrinkles disappeared and decreased, none remained on the most unfavorable aging scale. The changes occurred in the 2 groups were significant (p < 0.001) with better results in the experimental group. No complications were reported. All patients were satisfied. Regarding the evaluation of the final result, in 94.3% it was good in the study group and in the same proportion of the patients in the control group it was fair. Conclusions: In our study lipotransfer by centrifugation assisted with stem cells derived from adipose tissue stands out as being effective and safe in facial rejuvenation, superior to conventional lipotransfer by centrifugation. (AU)

The stem cell-specific long non-coding RNAs in leukemia

Leukemia is defined as a heterogeneous group of hematological cancers whose prevalence is on the rise worldwide. Despite the large body of studies, the etiology of leukemia has not been fully elucidated. Leukemia stem cells (LSCs) are a subpopulation of cancer cells that sustain the growth of the leukemic clone and are the main culprit for the maintenance of the neoplasm. In contrast to most leukemia cells, LSCs are resistant to chemo- and radiotherapy. Several recent studies demonstrated the altered expression profile of long non-coding RNAs (lncRNAs) in LSCs and shed light on the role of lncRNAs in the survival, proliferation, and differentiation of LSCs. LncRNAs are transcripts longer than 200 nucleotides that are implicated in several cellular and molecular processes such as gene expression, apoptosis, and carcinogenesis. Likewise, lncRNAs have shown a prognostic marker in leukemia patients and represent novel treatment options. Herein, we review the current knowledge concerning lncRNAs’ implication in the pathogenesis of LSCs and discuss their prognostic, diagnostic, and therapeutic potential (AU)

Use of stem cell-enriched fat grafts in facial reconstruction: have they demonstrated superiority over autologous fat grafting? / Uso de injertos de grasa enriquecidos con células madre en la reconstrución facial: ¿han demostrado superioridad sobre los injertos de grasa autólogos?

Since the 1990s, big strides have been made in plastic and reconstructive surgery, thanks to the implementation of new techniques and resources for its execution. In cases where restoration is sought for small defects, biosurgery, involving the utilization of stem cells, biomaterials, chemical engineering, and tissue engineering, can be employed. The potential to stimulate the innate regenerative capacity of tissue and generate a persistent response over time, minimizing surgical trauma and re-interventions, lies in stem cell transplantation. However, the evidence published on this matter is very scarce, leading to divergence of opinions, suggestions, and recommendations. The aim of this review is to analyze the most recent evidence concerning the outcomes of using fat grafts enriched with stem cells in facial reconstruction, as compared to autologous fat grafts. The review of the literature demonstrates a marked trend suggesting that fat grafts enriched with stem cells may be superior to autologous grafts in facial reconstruction, with potential benefits in medium-term volume retention and faster attainment of results. (AU)

La cirugía plástica y reconstructiva, ha dado pasos agigantados desde la década de los años 90, con la implementación de nuevas técnicas y recursos para su realización. En los casos de defectos pequeños donde se busca la restauración, se puede hacer uso de la biocirugía, que involucra el uso de células madre, biomateriales, ingeniería química y de tejidos. El trasplante de células madre, tiene el potencial de estimular la capacidad innata regenerativa del tejido, así como de generar una respuesta persistente a lo largo del tiempo, minimizando el trauma quirúrgico y la reintervención. A pesar, de la ventaja plausible, la calidad y cantidad de la evidencia publicada al respecto es muy escasa, ocasionando divergencia de opiniones, sugerencias y recomendaciones. Con base en lo anterior, el objetivo de esta revisión consiste en realizar un análisis de la evidencia más reciente sobre los resultados en el uso de injertos de grasa enriquecidos con células madre en la reconstrucción facial, comparado a los injertos de grasa autólogos. A través de la revisión de la literatura, se pudo evidenciar que, aunque la evidencia es heterogénea a la fecha, existe una tendencia marcada en que el uso de injertos de grasa enriquecidos con células madre, podría ser superior a los injertos autólogos en la reconstrucción facial, con potenciales beneficios en la retención de volumen a mediano plazo y mayor celeridad en la obtención de resultados. (AU)

Quimeras macaco-humanas: aspectos científicos y consideraciones bioéticas / Macaque-human chimeras: scientific aspects and bioethical reflections

Se describe la obtención de embriones quiméricos macaco-humanos por Izpisua y colaboradores (2021) mediante la inyección de células troncales pluripotentes humanas en blastocistos de macaco cultivados ex vivo y se analiza el destino de las líneas celulares humanas en su desarrollo posterior hasta el día 19 después de la fecundación. Se hace una reflexión bioética sobre la relación ciencia-ética en general y sobre dicha investigación en particular. (AU)

The obtention by Izpisua and collaborators (2021) of macaque-human chimeric embryos by microinjection of human pluripotent stem cells into early blastocysts of cynomolgus monkey is described. They studied the competency of human pluripotent stem cells in macaque embryos cultured ex vivo until 19 days post-fertilization. A reflection on these experiments is made from the bioethical point of view. (AU)

Metformin induces mitochondrial remodeling and differentiation of pancreatic progenitor cells into beta-cells by a potential mechanism including suppression of the T1R3, PLCB2, cytoplasmic Ca+2, and AKT

The main goal of this study was to investigate the molecular changes in pancreatic progenitor cells subject to high glucose, aspartame, and metformin in vitro. This scope of work glucose, aspartame, and metformin were exposed to pancreatic islet derived progenitor cells (PID-PCs) for 10 days. GLUT1’s role in beta-cell differentiation was examined by using GLUT1 inhibitor WZB117. Insulin+ cell ratio was measured by flow cytometry; the expression of beta-cell differentiation related genes was shown by RT-PCR; mitochondrial mass, mitochondrial ROS level, cytoplasmic Ca2+, glucose uptake, and metabolite analysis were made fluorometrically and spectrophotometrically; and proteins involved in related molecular pathways were determined by western blotting. Findings showed that glucose or aspartame exposed cells had similar metabolic and gene expression profile to control PID-PCs. Furthermore, relatively few insulin+ cells in aspartame treated cells were determined. Aspartame signal is transmitted through PLCβ2, CAMKK2 and LKB1 in PID-PCs. The most obvious finding of this study is that metformin significantly increased beta-cell differentiation. The mechanism involves suppression of the sweet taste signal’s molecules T1R3, PLCβ2, cytoplasmic Ca+2, and AKT in addition to the direct effect of metformin on mitochondria and AMPK, and the energy metabolism of PID-PCs is remodelled in the direction of oxidative phosphorylation. These findings are very important in terms of determining that metformin stimulates the mitochondrial remodeling and the differentiation of PID-PCs to beta-cells and thus it may contribute to the compensation step, which is the first stage of diabetes development. (AU)

Differential chemotherapeutic regimen cytotoxicity against pancreatic cancer stem cells: a preliminary in vitro study / Citotoxicidad diferencial según el régimen de quimioterapia contra las células madre del cáncer de páncreas: estudio preliminar in vitro

Introduction: Pancreatic cancer treatment in advanced stages is based on different chemotherapy regimens. Cancer stem cells are responsible for tumor chemoresistance and recurrence in adjuvant and metastatic settings. The objective of this article was to evaluate how these chemotherapeutic regimens affect the proportion of cancer stem cells and the expression of stemness markers. Method: We used the pancreatic adenocarcinoma cell line PANC-1 as a model to apply different chemotherapeutic protocols (monotherapy and combined therapy) using 5-Fluorouracil, Oxaliplatin, Irinotecan, Gemcitabine and Abraxane. Results: After analyzing different tumor stem cell markers (SOX2, OCT4, CD133, CD44 and CD24) in pancreatic cancer cells treated with different chemotherapeutic protocols by means of RT-qPCR, Oxaliplatin and Gemcitabine in monotherapy were the chemotherapies that selected the most cancer stem cells while the FOLFIRI protocol decreased them. Conclusions: Regarding the selection of markers, it has been much higher in the case of Gemcitabine alone. In conclusion, these findings could improve and personalize pancreatic cancer therapy. (AU)

Introducción: El tratamiento del cáncer de páncreas en estadios avanzados se basa en diferentes regímenes de quimioterapia. Las células madre cancerosas son responsables de la quimiorresistencia tumoral y la recurrencia tras tratamientos en etapa adyuvante y metastásica. El objetivo de este artículo fue evaluar cómo estos regímenes quimioterapéuticos afectan a la proporción de células madre cancerosas y la expresión de sus marcadores. Método: Utilizamos la línea celular de adenocarcinoma pancreático PANC-1 como modelo para aplicar diferentes protocolos quimioterapéuticos (monoterapia y terapia combinada) utilizando 5-Fluorouracilo, Oxaliplatino, Irinotecán, Gemcitabina y Abraxane. Resultados: Tras analizar mediante RT-qPCR diferentes marcadores de células madre tumorales (SOX2, OCT4, CD133, CD44 y CD24) en células de cáncer de páncreas tratadas con diferentes protocolos quimioterapéuticos, el Oxaliplatino y la Gemcitabina en monoterapia fueron los quimioterápicos que seleccionaron en mayor medida las células madre cancerosas mientras que el protocolo FOLFIRI las disminuyó. Conclusiones: En cuanto a la selección de marcadores, ha sido mucho mayor en el caso de Gemcitabina en monoterapia. En conclusión, estos hallazgos podrían mejorar y personalizar la terapia del cáncer de páncreas. (AU)

Isolated cancer stem cells from human liver cancer: morphological and functional characteristics in primary culture

BackgroundPrimary liver cancer cells (PLCs) could more directly simulate the human tumor microenvironment. Compared with liver cancer cell lines, PLCs could reflect the human situation. As in previous studies, tumor stem cells were a small number of cancer cells in the microenvironment and considered to be one of the origins of liver cancer. This study aimed to screen stem cells in PLCs, analyze their biological characteristics, propose the possibility that liver cancer originated from stem cells.MethodsLiver cancer tissues of 17 patients were taken from the Affiliated Hospital of Guangdong Medical College, and PLCs were isolated by tissue slice method. The proliferation, tumor formation in nude mice, stem protein expression of PLCs were observed. C-kit+ liver cancer cells were screened and their biological characteristics were analyzed.ResultsPLCs could be stably passaged. Transmission electron microscopy indicated that the nucleus was irregular, there were many mitochondria, and the endoplasmic reticulum was irregularly distributed. PLCs could express E-Cadherin, Oct-4, β-Catenin, Sox2, CD326, C-kit, GPC3, Nanog. The proliferation curve of PLCs and Hep3B cells were similar, and they all could form tumors in nude mice. Flow-sorted C-kit+ PLCs, as well as C-kit+ Hep3B cells could highly express Bmi1, Sox2, Oct4, Notch1, Nanog, C-kit, β-Catenin, Smo, Nestin, ABCG2, ABCB1. And they also could clone and form tumors in vivo. But C-kit+ PLCs were more sensitive to chemotherapy drugs than C-kit+ liver cancer cell lines.ConclusionC-kit+ PLCs had the characteristics of tumor stem cells and were more sensitive to chemotherapy drugs.

Tratamiento de la artrosis de rodilla con células mesenquimales estromales expandidas: revisión sistemática de la literatura / Expanded Mesenchymal Stromal Cells in Knee Osteoarthritis: A Systematic Literature Review

Objetivo: Analizar la eficacia y la seguridad de los implantes de células mesenquimales estromales expandidas (MSCs, por sus siglas en inglés) en la artrosis de rodilla. Métodos: Revisión sistemática de la literatura. Estrategia de búsqueda en Medline, Embase y Cochrane Library hasta febrero del 2018. Inclusión: artrosis de rodilla (II-IV Kellgren); inyección intra-articular de MSCs; ensayos controlados aleatorizados (ECA) y ensayos clínicos cuasi-experimentales (ECC)≥6 meses y N≥10. Resultados: De 252 artículos identificados, se incluyeron 3 ECA y 6 ECC (N=169). El 60% de los pacientes mejoraron clínicamente y el 50% estructuralmente hasta 2 años después del implante. El beneficio máximo se alcanzó al año con dosis ≥40×106 MSCs. La intervención fue bien tolerada, igualmente segura en implantes alogénicos y autólogos. Conclusiones: Las MSCs intra-articulares son seguras. La baja calidad de la evidencia analizada no admite conclusiones sobre la eficacia, pero prueba un fundamento para su uso en la artrosis como modificador de síntoma y de estructura que debe ser constatado en ensayos de alta calidad.(AU)

Objective: To analyse the efficacy and safety of intra-articular injection of expanded Mesenchymal Stromal Cells (MSCs) in knee osteoarthritis. Methods: Systematic Literature Review. A pre-defined search strategy was run in Medline, Embase and Cochrane Library until February 2018. Inclusion criteria: knee osteoarthritis (grades II-IV Kellgren-Lawrence); intra-articular injection of MSCs (without surgical co-treatments); Randomized Controlled Trials (RCTs) or Quasi-experimental Clinical Trials (QCTs) N≥10 and ≥6 months of follow-up were included. Evidence was assigned according to the Scottish Intercollegiate Guidelines Network (SIGN). Results: The search identified 252 articles. Nine proof-of-concept trials (3 RCTs, 6 QCTs) were included (N=169). Evidence showed clinical improvement in 60% of patients. Structural benefit was reported in half of patients. Clinical benefit was observed from the 3rd month and structural improvement from the 6th. All studies reported maximum clinical and structural benefit a year following the implant. This benefit was sustained for up to 24 months. Studies with doses ≥40×106 showed more consistent clinical and structural benefits than those with lower doses. No systemic adverse reactions were reported. The most common adverse effect was pain and/or inflammation in the puncture area (13-53%). The use of donor cells was as safe as autologous implants. Conclusions: Intra-articular implants of MSCs seem to be safe with no serious adverse effects. Low-quality evidence precludes conclusions regarding efficacy in this review. However, the clinical and structural benefits observed provide a rationale for using expanded MSCs implants in osteoarthritis patients. High-quality evidence trials are needed to further determine best protocols to maximize clinical and structural improvement.(AU)

El negocio de la medicina regenerativa y las células madre: confusión con implicaciones legales / The regenerative medicine and stem cell business: confusion with legal implications

El auge de la medicina regenerativa y el crecimiento de la oferta de terapias autólogas obtenidas a partir de sangre, células o tejidos de los propios pacientes se ha visto favorecido por la actual disponibilidad de diversos dispositivos comerciales de fácil manejo que permiten la elaboración de los productos y su aplicación dentro de un mismo procedimiento. Independientemente de las dudosas eficacia y seguridad de muchos de los tratamientos que se ofrecen bajo el reclamo de las células madre o la medicina regenerativa, la mayor parte de los centros y de los profesionales que ofrecen estos tratamientos desconocen los requisitos y las implicaciones legales de su uso. Una confusión frecuente consiste en no distinguir entre la autorización que requiere el propio dispositivo, considerado producto sanitario, y la autorización para el uso del producto obtenido, que en general se trata de un medicamento, ya sea de terapia avanzada o no, o de un trasplante. Por otra parte, es frecuente que estos tratamientos tengan un carácter experimental, por lo que su administración en ese caso, además de requerir la evaluación ética correspondiente y la autorización de diversos organismos reguladores, debe ofrecerse de forma gratuita y tras recabar el consentimiento informado del paciente y contratar una póliza de seguros específica. En este artículo se presentan, de forma resumida, los principales requisitos para la aplicación de estos productos biológicos autólogos, con el objetivo de que puedan servir de guía tanto para los profesionales que los prescriben como para aquellos que inspeccionan los centros donde se administran. Por último, se ofrecen algunas recomendaciones para los pacientes. (AU)

The rise of regenerative medicine and the growth of the offer of autologous therapies, obtained from blood, cells or tissues of the patients, have been favoured by the current availability of an increasing number of commercial devices. Most of these devices are easy to use, allowing the elaboration of products and its application within the same procedure. Regardless of the questionable efficacy and safety of many of the treatments offered under the claim of stem cells or regenerative medicine, most of the centres and professionals offering these treatments are unaware of the legal requirements and implications of their use. A common confusion consists in not distinguishing between the authorization required by the equipment itself, considered a medical device, and the authorization for the use of the product obtained, usually considered a medicinal product (whether advanced therapy or not) or a transplant. Moreover, these treatments frequently have an experimental nature. In that case, in addition to requiring the corresponding ethical evaluation and the authorization of various regulatory bodies, their administration must be offered free of charge, obtaining the patient's informed consent and after contracting a specific insurance policy. In this article we present a brief summary of the main requirements for the application of these autologous biological products with the aim of serving as a guide both for the professionals who prescribe them and for those who inspect the centres where the products are administered. Finally, we include some recommendations for patients. (AU)

Efectos biológicos de los cementos MTA Repair HP y ProRoot MTA tras ser expuestos a diferentes pH sobre células madre de ligamento periodontal / Biological effects of MTA Repair HP and ProRoot MTA exposed to different pH on human periodontal ligament stem cells

Objetivos. El propósito de este trabajo fue evaluar los efectos biológicos de MTA Repair HP y ProRoot MTA en células madre procedentes de ligamento periodontal (hPDLSCs) tras ser ex-puestos los cementos a ambiente ácido y neutro. Material y Métodos: Discos de cada material (n=30) fueron ex-puestos a un tampón fosfato-salino (pH 7.4) o a ácido butírico (pH 5.2) durante 7 días, posteriormente se realizaron pruebas biológicas in vitro usando hPDLSCs. A partir de eluatos de los diferentes materiales de obturación a retro, se analizaron pruebas de viabilidad celular y apoptosis. Para evaluar la adhesión celular, hPDLSCs se sembraron directamente sobre la superficie de los materiales y se observaron bajo microscopio electrónico de barrido. Para analizar estadísticamente los resultados se usaron los test ANOVA y Tukey test (p < 0.05). Resultados: Los cementos endodónticos expuestos a ambiente ácido mostraron un similar grado de adhesión celular, y sorprendentemente, MTA Repair HP a pH 5.2 exhibió una mayor viabilidad celular que ProRootMTA (p<0.05). A pH 7.4, ProRooT MTA obtuvo una tasa mayor de viabilidad celular que con MTA Repair HP. Conclusiones: Los materiales ProRoot MTA y MTA Repair HP presentaron adecuadas propiedades biológicas en los diferentes ambientes, en términos de viabilidad celular, apoptosis y adhesión

Objectives: The purpose of this work was to evaluate the biolo-gical effects of MTA Repair HP and ProRoot MTA on stem cells from periodontal ligament (hPDLSCs) after exposure to acidic and neutral environments. Material and Methods: Discs of each material (n=30) were ex-posed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05).Results: Endodontic cements exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than Pro-Root MTA, whereas ProRoot MTA 7.4 showed higher cell viability rates than MTA Repair HP. Conclusions: Adequate biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death and cell attach-ment were observed in both environments

Bisphosphonate-related osteonecrosis. Application of adipose-derived stem cells in an experimental murine model

Background: Bisphosphonate-related osteonecrosis of the jaw is a pathological condition without effective established treatment and preventive strategies. The aim of this study was to analyse the effect of adipose-derived stem cells (ASC) in an experimental murine model of osteonecrosis. Material and Methods: 38 Wistar rats were injected intraperitoneally with zoledronic acid. After treatment, upper jaw molars were extracted. The animals were randomly assigned to one of two groups. In the control group, saline solution was applied over the alveolar sockets after the tooth extractions. In the treatment group, ASCs were applied instead of saline solution. The control and treatment groups were subdivided based on the time of euthanasia. A clinical and histological analysis was performed. Results: The presence of osteonecrosis in alveolar bone was observed in a similar distribution in both groups. In the ASC-treated group, new bone formation was greater than in controls. Conclusions: In this study, application of ASCs showed greater new bone formation in an osteonecrosis-like murine model. Previous inhibited post-extraction bone remodelling could be reactivated, and these findings appeared to be secondary to implantation of ASCs

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Método de aislamiento, cultivo y precondicionamiento con hipoxia de células madre derivadas de tejido adiposo en ratas / Method of isolation, culture and hypoxia preconditioning of adipose tissue stem cells in rats

Introducción y objetivo. Las células madre son candidatas terapéuticas para una amplia gama de enfermedades. Resultan de gran interés en Cirugía Plástica para el tratamiento de heridas crónicas, transferencia de tejido adiposo y colgajos. El objetivo de este estudio es describir el método de aislamiento, cultivo y caracterización de células madre derivadas de tejido adiposo y cómo estas pueden precondicionarse con hipoxia y generar cambios in vitro en su capacidad proliferativa y migratoria. El trabajo es un complemento didáctico a otro publicado por nosotros en esta misma revista utilizando esta metodología en comparación al grupo de retardo de colgajo y grupo control en colgajos cutáneos aleatorizados en ratas. Métodos. Obtuvimos las células madre de tejido graso ínguino-abdominal de ratas adultas: 10 en el grupo de células madre derivadas de tejido adiposo y otras 10 en el grupo de células madre derivadas de tejido adiposo precondicionadas con hipoxia (2% O2 y 5% CO2). Realizamos análisis morfológico directo y con inmunofluorescencia con el marcador vimentina y CD90 y estudio de proliferación y migración celular in vitro. Resultados: Utilizamos en promedio 1.64 +/- 1.13 gr de tejido adiposo en el grupo sin precondicionamiento y 0.93 +/- 0.34 gr en el grupo con precondicionamiento con hipoxia para el aislamiento. Las células madre derivadas de tejido adiposo precondicionadas con hipoxia presentaron un aumento de la capacidad migratoria a las 24 horas de 2.44 +/- 0.85 mm frente a 2.24 +/- 0.82 mm (p ≤ 0.01) y proliferativa 5.42 x105 +/- 1.03 x105 céls/ml frente a 3.26 x105 +/- 8.61 x104 céls/ml) (p ≤ 0.001) de forma significativa en comparación a las sin precondicionamiento. Conclusiones. Describimos en detalle un método de precondicionamiento de células madre mediante hipoxia. Logramos potenciar el efecto de las células madre aumentando en forma significativa su capacidad migratoria y proliferativa de forma precoz

Background and objective. Stem cells are therapeutic candidates for a wide range of diseases. They are of great interest in Plastic Surgery for the management of chronic wounds, adipose tissue transfer and flaps. The objective of this study is to describe the method of isolation, culture and characterization of stem cells derived from adipose tissue and how these can be preconditioned with hypoxia and generate in vitro changes in their proliferative and migratory capacity. This study is a didactic supplement to a paper published by us in this same journal using this methodology in comparison to the group of flap delay and control group in skin flaps randomized in rats. Methods. Stem cells were obtained from inguinal-abdominal fatty tissue of adult rats: 10 in the group of stem cells derived from adipose tissue and another 10 in the group of stem cells derived from adipose tissue preconditioned with hypoxia (2% O2 and 5% CO2). Direct morphological analysis was carried out and with immunofluorescence (vimentin and CD90 marker). Study proliferation and in vitro cell migration was performed. Results. An average of 1.64 +/- 1.13 gr of adipose tissue of the inguinoabdominal area was used in the group of stem cells without preconditioning and 0.93 +/- 0.34 gr. in the group with hypoxic preconditioning for the isolation. Stem cells derived from adipose tissue preconditioned with hypoxia showed an increase in migratory capacity at 24 hours of 2.44 +/- 0.85 mm v/s 2.24 +/- 0.82 mm (p ≤ 0.01) and proliferative of 5.42 x105 +/- 1.03 x105 cells / ml v/s 3.26 x 105 +/- 8.61 x104 cells / ml) (p ≤0.001) significantly compared to those without preconditioning. Conclusions. A method of preconditioning stem cells by hypoxia is described in detail. It is possible to enhance the effect of the stem cells, significantly increasing their early migratory and proliferative capacity