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Vitamin D plays a major role in bone health and probably also in multiple extraskeletal acute and chronic diseases. Although supplementation with calcifediol, a vitamin D metabolite, has demonstrated efficacy and safety in short-term clinical trials, its effects after long-term monthly administration have been studied less extensively. This report describes the results of a 1-year, phase III-IV, double-blind, randomized, controlled, parallel, multicenter superiority clinical trial to assess the efficacy and safety of monthly calcifediol 0.266 mg versus cholecalciferol 25,000 IU (0.625 mg) in postmenopausal women with vitamin D deficiency (25(OH)D

Vitamin D has shown to play a role in multiple diseases due to its skeletal and extraskeletal actions. Furthermore, vitamin D deficiency has become a worldwide health issue. Few supplementation guidelines mention calcifediol treatment, despite being the direct precursor of calcitriol and the biomarker of vitamin D status. This 1-year, phase III-IV, double-blind, randomized, controlled, multicenter clinical trial assessed the efficacy and safety of calcifediol 0.266 mg soft capsules in vitamin D-deficient postmenopausal women, compared to cholecalciferol. Results reported here are from a prespecified interim analysis, for the evaluation of the study's primary endpoint: the percentage of patients with serum 25-hydroxyvitamin D (25(OH)D) levels above 30 ng/ml after 4 months. A total of 303 patients were enrolled, of whom 298 were included in the intention-to-treat (ITT) population. Patients with baseline levels of serum 25(OH)D

BACKGROUND Mesenchymal stem cells (MSC) of bone marrow are the progenitor of osteoblasts and adipocytes. MSC tend to differentiate into adipocytes, instead of osteoblasts, with aging. This favors the loss of bone mass and development of osteoporosis. Hypoxia induces hypoxia inducible factor 1α gene encoding transcription factor, which regulates the expression of genes related to energy metabolism and angiogenesis. That allows a better adaptation to low O2 conditions. Sustained hypoxia has negative effects on bone metabolism, favoring bone resorption. Yet, surprisingly, cyclic hypoxia (CH), short times of hypoxia followed by long times in normoxia, can modulate MSC differentiation and improve bone health in aging. AIM To evaluate the CH effect on MSC differentiation, and whether it improves bone mineral density in elderly. METHODS MSC cultures were induced to differentiate into osteoblasts or adipocytes, in CH (3% O2 for 1, 2 or 4 h, 4 d a week). Extracellular-matrix mineralization and lipid-droplet formation were studied in MSC induced to differentiate into osteoblast or adipocytes, respectively. In addition, gene expression of marker genes, for osteogenesis or adipogenesis, have been quantified by quantitative real time polymerase chain reaction. The in vivo studies with elderly (> 75 years old; n = 10) were carried out in a hypoxia chamber, simulating an altitude of 2500 m above sea level, or in normoxia, for 18 wk (36 CH sessions of 16 min each). Percentages of fat mass and bone mineral density from whole body, trunk and right proximal femur (femoral, femoral neck and trochanter) were assessed, using dual-energy X-ray absorptiometry. RESULTS CH (4 h of hypoxic exposure) inhibited extracellular matrix mineralization and lipid-droplet formation in MSC induced to differentiate into osteoblasts or adipocytes, respectively. However, both parameters were not significantly affected by the other shorter hypoxia times assessed. The longest periods of hypoxia downregulated the expression of genes related to extracellular matrix formation, in MSC induced to differentiate into osteoblasts. Interestingly, osteocalcin (associated to energy metabolism) was upregulated. Vascular endothelial growth factor an expression and low-density lipoprotein receptor related protein 5/6/dickkopf Wnt signaling pathway inhibitor 1 (associated to Wnt/β-catenin pathway activation) increased in osteoblasts. Yet, they decreased in adipocytes after CH treatments, mainly with the longest hypoxia times. However, the same CH treatments increased the osteoprotegerin/receptor activator for nuclear factor kappa B ligand ratio in both cell types. An increase in total bone mineral density was observed in elderly people exposed to CH, but not in specific regions. The percentage of fat did not vary between groups. CONCLUSION CH may have positive effects on bone health in the elderly, due to its possible inhibitory effect on bone resorption, by increasing the osteoprotegerin / receptor activator for nuclear factor kappa B ligand ratio.

Dipeptidyl peptidase IV (DPP4) is a ubiquitous protease that can be found in membrane-anchored or soluble form. Incretins are one of the main DPP4 substrates. These hormones regulate glucose levels, by stimulating insulin secretion and decreasing glucagon production. Because DPP4 levels are high in diabetes, DPP4 inhibitor (DPP4i) drugs derived from gliptin are widespread used as hypoglycemic agents for its treatment. However, as DPP4 recognizes other substrates such as chemokines, growth factors and neuropeptides, pleiotropic effects have been observed in patients treated with DPP4i. Several of these substrates are part of the stem-cell niche. Thus, they may affect different physiological aspects of mesenchymal stem-cells (MSC). They include viability, differentiation, mobilization and immune response. MSC are involved in tissue homeostasis and regeneration under both physiological and pathological conditions. Therefore, such cells and their secretomes have a high clinical potential in regenerative medicine. In this context, DPP4 activity may modulate different aspects of MSC regenerative capacity. Therefore, the aim of this review is to analyze the effect of different DPP4 substrates on MSC. Likewise, how the regulation of DPP4 activity by DPP4i can be applied in regenerative medicine. That includes treatment of cardiovascular and bone pathologies, cutaneous ulcers, organ transplantation and pancreatic beta-cell regeneration, among others. Thus, DPP4i has an important clinical potential as a complement to therapeutic strategies in regenerative medicine. They involve enhancing the differentiation, immunomodulation and mobilization capacity of MSC for regenerative purposes.

Extra virgin olive oil is characterized by its high content of unsaturated fatty acid residues in triglycerides, mainly oleic acid, and the presence of bioactive and antioxidant compounds. Its consumption is associated with lower risk of suffering chronic diseases and unwanted processes linked to aging, due to the antioxidant capacity and capability of its components to modulate cellular signaling pathways. Consumption of olive oil can alter the physiology of mesenchymal stem cells (MSCs). This may explain part of the healthy effects of olive oil consumption, such as prevention of unwanted aging processes. To date, there are no specific studies on the action of olive oil on MSCs, but effects of many components of such food on cell viability and differentiation have been evaluated. The objective of this article is to review existing literature on how different compounds of extra virgin olive oil, including residues of fatty acids, vitamins, squalene, triterpenes, pigments and phenols, affect MSC maintenance and differentiation, in order to provide a better understanding of the healthy effects of this food. Interestingly, most studies have shown a positive effect of these compounds on MSCs. The collective findings support the hypothesis that at least part of the beneficial effects of extra virgin olive oil consumption on health may be mediated by its effects on MSCs.

Preclinical data strongly suggest that the vitamin D endocrine system (VDES) may have extraskeletal effects. Cells of the immune and cardiovascular systems and lungs can express the vitamin D receptor, and overall these cells respond in a coherent fashion when exposed to 1,25-dihydroxyvitamin D, the main metabolite of the VDES. Supplementation of vitamin D-deficient subjects may decrease the risk of upper respiratory infections. The VDES also has broad anti-inflammatory and anti-thrombotic effects, and other mechanisms argue for a potential beneficial effect of a good vitamin D status on acute respiratory distress syndrome, a major complication of this SARS-2/COVID-19 infection. Activation of the VDES may thus have beneficial effects on the severity of COVID-19. Meta-analysis of observational data show that a better vitamin D status decreased the requirement of intensive care treatment or decreased mortality. A pilot study in Cordoba indicated that admission to intensive care was drastically reduced by administration of a high dose of calcifediol early after hospital admission for COVID-19. A large observational study in Barcelona confirmed that such therapy significantly decreased the odds ratio (OR) of mortality (OR = 0.52). This was also the conclusion of a retrospective study in five hospitals of Southern Spain. A retrospective study on all Andalusian patients hospitalized because of COVID-19, based on real-world data from the health care system, concluded that prescription of calcifediol (hazard ratio [HR] = 0.67) or vitamin D (HR = 0.75), 15 days before hospital admission decreased mortality within the first month. In conclusion, a good vitamin D status may have beneficial effects on the course of COVID-19. This needs to be confirmed by large, randomized trials, but in the meantime, we recommend (rapid) correction of 25 hydroxyvitamin D (25OHD) deficiency in subjects exposed to this coronavirus. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. ispartof: JBMR PLUS vol:5 issue:12 ispartof: location:England status: published

Context COVID-19 is a major health problem because of saturation of intensive care units (ICU) and mortality. Vitamin D has emerged as a potential treatment able to reduce the disease severity. Objective This work aims to elucidate the effect of 25(OH)D3 (calcifediol) treatment on COVID-19–related outcomes. Methods This observational cohort study was conducted from March to May 2020, among patients admitted to COVID-19 wards of Hospital del Mar, Barcelona, Spain. A total of 930 patients with COVID-19 were included; 92 were excluded because of previous calcifediol intake. Of the remaining 838, a total of 447 received calcifediol (532 μg on day 1 plus 266 μg on days 3, 7, 15, and 30), whereas 391 were not treated at the time of hospital admission (intention-to-treat). Of the latter, 53 patients were treated later during ICU admission and were allocated in the treated group in a second analysis. In healthy individuals, calcifediol is about 3.2-fold more potent on a weight basis than cholecalciferol. Main outcome measures were ICU admission and mortality. Results ICU assistance was required by 102 (12.2%) participants. Out of 447 patients treated with calcifediol at admission, 20 (4.5%) required the ICU, compared to 82 (21%) out of 391 nontreated (P Conclusion In patients hospitalized with COVID-19, calcifediol treatment significantly reduced ICU admission and mortality.

Dear Preprints with the Lancet Readers, We removed this preprint due to concerns about the description of the research in this paper. This led us to initiate an investigation into this study. Following the conclusion of an investigation into this study by the National Centre of Biomedical Research on Frailty and Healthy Aging (CIBERFES), Spain, the report concluded that: 1) In the elaboration of the manuscript and the correspondence of the authors with the Journal, there was a series of mistakes made by the authors in the qualification of the study and its description; and 2) At all times good practices for clinical research were carefully followed and in no case was the health of the patients put at any risk. The comments that have been posted on this preprint will remain available on this page. Please note that this comment thread is now closed to further posts.

Highlights • The vitamin D endocrine system have a variety of actions on cells and tissues involved in COVID-19 progression. • Early calcifediol (25-hydroxyvitamin D) treatment to hospitalized COVID-19 patients significantly reduced intensive care unit admissions-Calcifediol seems to be able to reduce severity of the COVID-19. • Calcifediol seems to be able to reduce severity of the disease., Objective The vitamin D endocrine system may have a variety of actions on cells and tissues involved in COVID-19 progression especially by decreasing the Acute Respiratory Distress Syndrome. Calcifediol can rapidly increase serum 25OHD concentration. We therefore evaluated the effect of calcifediol treatment, on Intensive Care Unit Admission and Mortality rate among Spanish patients hospitalized for COVID-19. Design Parallel pilot randomized open label, double-masked clinical trial. Setting University hospital setting (Reina Sofia University Hospital, Córdoba Spain.) Participants 76 consecutive patients hospitalized with COVID-19 infection, clinical picture of acute respiratory infection, confirmed by a radiographic pattern of viral pneumonia and by a positive SARS-CoV-2 PCR with CURB65 severity scale (recommending hospital admission in case of total score > 1). Procedures All hospitalized patients received as best available therapy the same standard care, (per hospital protocol), of a combination of hydroxychloroquine (400 mg every 12 h on the first day, and 200 mg every 12 h for the following 5 days), azithromycin (500 mg orally for 5 days. Eligible patients were allocated at a 2 calcifediol:1 no calcifediol ratio through electronic randomization on the day of admission to take oral calcifediol (0.532 mg), or not. Patients in the calcifediol treatment group continued with oral calcifediol (0.266 mg) on day 3 and 7, and then weekly until discharge or ICU admission. Outcomes of effectiveness included rate of ICU admission and deaths. Results Of 50 patients treated with calcifediol, one required admission to the ICU (2%), while of 26 untreated patients, 13 required admission (50 %) p value X2 Fischer test p < 0.001. Univariate Risk Estimate Odds Ratio for ICU in patients with Calcifediol treatment versus without Calcifediol treatment: 0.02 (95 %CI 0.002−0.17). Multivariate Risk Estimate Odds Ratio for ICU in patients with Calcifediol treatment vs Without Calcifediol treatment ICU (adjusting by Hypertension and T2DM): 0.03 (95 %CI: 0.003-0.25). Of the patients treated with calcifediol, none died, and all were discharged, without complications. The 13 patients not treated with calcifediol, who were not admitted to the ICU, were discharged. Of the 13 patients admitted to the ICU, two died and the remaining 11 were discharged. Conclusion Our pilot study demonstrated that administration of a high dose of Calcifediol or 25-hydroxyvitamin D, a main metabolite of vitamin D endocrine system, significantly reduced the need for ICU treatment of patients requiring hospitalization due to proven COVID-19. Calcifediol seems to be able to reduce severity of the disease, but larger trials with groups properly matched will be required to show a definitive answer.

The cells secrete extracellular vesicles (EV) that may have an endosomal origin, or from evaginations of the plasma membrane. The former are usually called exosomes, with sizes ranging from 50 to 100 nm. These EV contain a lipid bilayer associated to membrane proteins. Molecules such as nucleic acids (DNA, mRNA, miRNA, lncRNA, etc.) and proteins may be stored inside. The EV composition depends on the producer cell type and its physiological conditions. Through them, the cells modify their microenvironment and the behavior of neighboring cells. That is accomplished by transferring factors that modulate different metabolic and signaling pathways. Due to their properties, EV can be applied as a diagnostic and therapeutic tool in medicine. The mesenchymal stromal cells (MSC) have immunomodulatory properties and a high regenerative capacity. These features are linked to their paracrine activity and EV secretion. Therefore, research on exosomes produced by MSC has been intensified for use in cell-free regenerative medicine. In this area, the use of EV for the treatment of chronic skin ulcers (CSU) has been proposed. Such sores occur when normal healing does not resolve properly. That is usually due to excessive prolongation of the inflammatory phase. These ulcers are associated with aging and diseases, such as diabetes, so their prevalence is increasing with the one of such latter disease, mainly in developed countries. This has very important socio-economic repercussions. In this review, we show that the application of MSC-derived EV for the treatment of CSU has positive effects, including accelerating healing and decreasing scar formation. This is because the EV have immunosuppressive and immunomodulatory properties. Likewise, they have the ability to activate the angiogenesis, proliferation, migration, and differentiation of the main cell types involved in skin regeneration. They include endothelial cells, fibroblasts, and keratinocytes. Most of the studies carried out so far are preclinical. Therefore, there is a need to advance more in the knowledge about the conditions of production, isolation, and action mechanisms of EV. Interestingly, their potential application in the treatment of CSU opens the door for the design of new highly effective therapeutic strategies.

Resumen Antecedentes y objetivos El deficit de 25(OH)D se ha relacionado con un riesgo cardiovascular aumentado, aunque los estudios de intervencion son contradictorios. El objetivo principal fue evaluar el efecto del tratamiento con calcifediol (25(OH)D3) sobre el sistema cardiovascular en pacientes con sindrome coronario agudo sin elevacion de segmento ST. Pacientes y metodo Estudio prospectivo que incluyo a 41 pacientes (70,6 ±6,3 anos) ≥ 60 anos con sindrome coronario agudo sin elevacion de segmento ST y enfermedad coronaria revascularizada percutaneamente. Se aleatorizaron a recibir calcifediol + tratamiento habitual o tratamiento habitual exclusivo, con evaluacion de major adverse cardiovascular events (MACE, «episodios cardiovasculares mayores adversos») a los 3 meses. Se estudio la 25(OH)D en relacion con otras variables analiticas y con la extension de la enfermedad coronaria. Resultados Niveles basales de 25(OH)D ≤ 50 nmol/l se asociaron a enfermedad coronaria multivaso (RR: 2,6 [IC 95%: 1,1-7,1], p = 0,027) y 25(OH)D ≤ 50nmol/l + paratohormona ≥ 65pg/ml identificaron a pacientes con mayor riesgo de MACE (RR: 4 [IC 95%: 1,1-21,8], p = 0,04). Se registro un MACE en el grupo de pacientes suplementados y 5 en el de tratamiento convencional (p = 0,66). Entre los pacientes con niveles sericos de 25(OH)D ≤ 50 nmol/l al final del estudio el 28,6% presentaron MACE frente al 0% si los niveles eran > 50 nmol/l (RR: 1,4; p = 0,037). Conclusiones El deficit de vitamina D que implica un hiperparatiroidismo secundario puede ser un buen predictor de MACE. En pacientes suplementados con calcifediol se observo una tendencia a la disminucion de MACE en el seguimiento. Niveles finales de 25(OH)D ≤ 50 nmol/l se asociaron significativamente a un mayor numero de MACE, por lo que la normalizacion de 25(OH)D, ademas de mejorar la salud osea, puede mejorar la salud cardiovascular.

Atypical femoral fractures (AFFs) are a rare but potentially devastating event, often but not always linked to bisphosphonate (BP) therapy. The pathogenic mechanisms underlying AFFs remain obscure, and there are no tests available that might assist in identifying those at high risk of AFF. We previously used exome sequencing to explore the genetic background of three sisters with AFFs and three additional unrelated AFF cases, all previously treated with BPs. We detected 37 rare mutations (in 34 genes) shared by the three sisters. Notably, we found a p.Asp188Tyr mutation in the enzyme geranylgeranyl pyrophosphate synthase, a component of the mevalonate pathway, which is critical to osteoclast function and is inhibited by N-BPs. In addition, the CYP1A1 gene, responsible for the hydroxylation of 17β-estradiol, estrone, and vitamin D, was also mutated in all three sisters and one unrelated patient. Here we present a detailed list of the variants found and report functional analyses of the GGPS1 p.Asp188Tyr mutation, which showed a severe reduction in enzyme activity together with oligomerization defects. Unlike BP treatment, this genetic mutation will affect all cells in the carriers. RNAi knockdown of GGPS1 in osteoblasts produced a strong mineralization reduction and a reduced expression of osteocalcin, osterix, and RANKL, whereas in osteoclasts, it led to a lower resorption activity. Taken together, the impact of the mutated GGPPS and the relevance of the downstream effects in bone cells make it a strong candidate for AFF susceptibility. We speculate that other genes such as CYP1A1 might be involved in AFF pathogenesis, which remains to be functionally proved. The identification of the genetic background for AFFs provides new insights for future development of novel risk assessment tools. © 2018 American Society for Bone and Mineral Research.

Modest and even severe vitamin D deficiency is widely prevalent around the world. There is consensus that a good vitamin D status is necessary for bone and general health. Similarly, a better vitamin D status is essential for optimal efficacy of antiresorptive treatments. Supplementation of food with vitamin D or using vitamin D supplements is the most widely used strategy to improve the vitamin status. Cholecalciferol (vitamin D3) and ergocalciferol (vitamin D2) are the most widely used compounds and the relative use of both products depends on historical or practical reasons. Oral intake of calcifediol (25OHD3) rather than vitamin D itself should also be considered for oral supplementation. We reviewed all publications dealing with a comparison of oral cholecalciferol with oral calcifediol as to define the relative efficacy of both compounds for improving the vitamin D status. First, oral calcifediol results in a more rapid increase in serum 25OHD compared to oral cholecalciferol. Second, oral calcifediol is more potent than cholecalciferol, so that lower dosages are needed. Based on the results of nine RCTs comparing physiologic doses of oral cholecalciferol with oral calcifediol, calcifediol was 3.2-fold more potent than oral cholecalciferol. Indeed, when using dosages ≤ 25 μg/day, serum 25OHD increased by 1.5 ± 0.9 nmol/l for each 1 μg cholecalciferol, whereas this was 4.8 ± 1.2 nmol/l for oral calcifediol. Third, oral calcifediol has a higher rate of intestinal absorption and this may have important advantages in case of decreased intestinal absorption capacity due to a variety of diseases. A potential additional advantage of oral calcifediol is a linear dose-response curve, irrespective of baseline serum 25OHD, whereas the rise in serum 25OHD is lower after oral cholecalciferol, when baseline serum 25OHD is higher. Finally, intermittent intake of calcifediol results in fairly stable serum 25OHD compared with greater fluctuations after intermittent oral cholecalciferol.

Phloretin (a flavonoid abundant in apple), has antioxidant, anti-inflammatory, and glucose-transporter inhibitory properties. Thus, it has interesting pharmacological and nutraceutical potential. Bone-marrow mesenchymal stem cells (MSC) have high differentiation capacity, being essential for maintaining homeostasis and regenerative capacity in the organism. Yet, they preferentially differentiate into adipocytes instead of osteoblasts with aging. This has a negative impact on bone turnover, remodeling, and formation. We have evaluated the effects of phloretin on human adipogenesis, analyzing MSC induced to differentiate into adipocytes. Expression of adipogenic genes, as well as genes encoding OPG and RANKL (involved in osteoclastogenesis), protein synthesis, lipid-droplets formation, and apoptosis, were studied. Results showed that 10 and 20 µM phloretin inhibited adipogenesis. This effect was mediated by increasing beta-catenin, as well as increasing apoptosis in adipocytes, at late stages of differentiation. In addition, this chemical increased OPG gene expression and OPG/RANKL ratio in adipocytes. These results suggest that this flavonoid (including phloretin-rich foods) has interesting potential for clinical and regenerative-medicine applications. Thus, such chemicals could be used to counteract obesity and prevent bone-marrow adiposity. That is particularly useful to protect bone mass and treat diseases like osteoporosis, which is an epidemic worldwide.

40 Páginas; 3 Figuras; 3 Tablas, Aging may enhance both oxidative stress and bone-marrow mesenchymal stem-cell (MSC) differentiation into adipocytes. That reduces osteoblastogenesis, thus favoring bone-mass loss and fracture, representing an important worldwide health-issue, mainly in countries with aging populations. Intake of antioxidant products may help to retain bone-mass density. Interestingly, a novel olive-pomace physical treatment to generate olive oil also yields by-products rich in functional antioxidants. Thus, diet of postmenopausal women was supplemented for two months with one of such by-products (distillate 6; D6), being rich in squalene. After treatment, serum from such women showed reduced both lipidic peroxidation and oxidized low-density lipoprotein (LDL). Besides, vitamin E and coenzyme Q10 levels increased. Furthermore, culture medium containing 10% of such serum both increased osteoblastogenesis and reduced adipogenesis in human MSC from bone marrow. Therefore, highly antioxidant by-products like D6 may represent a relevant source for development of functional products, for both prevention and treatment of degenerative pathologies associated with aging, like osteoporosis., Supported by grants AGL2013-45110-R of “Ministerio de Ciencia e Innovación” (MICINN); PI081692, PI15/01857. “Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad” (RETICEF) and “CIBER de Fragilidad y Envejecimiento Saludable” (CIBERFES), with support from “Instituto de Salud Carlos III” (ISCIII), “Ministerio de Economía y Competitividad” (MINECO) and European Union (EU); “Ayudas de Intensificación de la Investigación” and “Grupo CTS413” of “Junta de Andalucía”, Spain.

IntroductionPain is an under-diagnosed problem in elderly people, especially in those with cognitive impairment who are unable to verbalise their pain. Although the Pain assessment in advanced dementia scale (PAINAD) scale is a tool recognised for its clinical interest in this type of patients, its correlation with the saliva biomarkers reinforced its utility. The aim of this research will be to correlate the scores of this scale with the levels of biomarkers of pain found in saliva samples of patients with cognitive impairment and inability to communicate.Methods and analysisThis is an observational study. The level of pain will be evaluated using the PAINAD scale. Moreover, pain biomarkers, in particular secretory IgA and soluble tumour necrosis factor receptor type II, will be determined in saliva. Both assessments will be conducted in 75 patients aged over 65 years with advanced cognitive impairment and inability to communicate. The PAINAD scores will be correlated with the levels of these biomarkers of pain. A control group consisting of 75 healthy subjects aged over 65 years will be included in the study. Moreover, sociodemographic variables and variables related to pain, dementia and other clinical conditions will be recorded. The analysis will be performed with the statistical package SPSS V.22 and the software R.Ethics and disseminationThe study has been reviewed and approved by the Andalusian Human Research Ethics Committee. In addition, this study has been financed by the Junta de Andalucía through a regional health research fund (Research code: PI-0357–2017). The results will be actively disseminated trough a high-impact journal in our study area, conference presentations and social media.

Extra virgin olive oil has positive effects on health. Oleuropein is a polyphenolic compound present in olive-tree leaves, fruits (olives) and olive oil. It is responsible for the relevant organoleptic and biological properties of olive oil, including antiadipogenic properties. Thus, the effects of oleuropein on the adipogenesis of human bone-marrow mesenchymal stem cells were studied by transcriptomics and differential gene-expression analyses. Oleuropein could upregulate expression of 60% of adipogenesis-repressed genes. Besides, it could activate signaling pathways such as Rho and β-catenin, maintaining cells at an undifferentiated stage. Our data suggest that mitochondrial activity is reduced by oleuropein, mostly during adipogenic differentiation. These results shed light on oleuropein activity on cells, with potential application as a "nutraceutical" for the prevention and treatment of diseases such as obesity and osteoporosis.

Despite the discussion on the optimal threshold of 25-hydroxyvitamin D serum level continues, there is now consensus on the fact that post-menopausal and elderly populations have inadequate Vitamin D serum levels worldwide. The adjustment of these levels is necessary to improve both bone and general health, as it is to optimize bone response to antiresortive treatments. It is recommended, as endorsed by international clinical guides, to use Vitamin D3, the physiological form of Vitamin D, in a dose range between 600-2000IU. It should be administered on a daily basis or on its weekly or monthly equivalents. In Spain, the use of calcidiol (25(OH)D3) at the same dose than Vitamin D3 is the most extended prescription, notwithstanding the available evidence stating that they are not equipotent. This may lead to over-dosage. In order to provide evidence on this circumstance, a convenience study was performed. Four groups of ten post-menopausal osteoporotic women each (average age 67), deficient in Vitamin D ((25(OH)D 37.5±10 nmol/L)) were enrolled. Each group followed a different treatment regimen: (G1) vitamin D3 20μg/day [800IU/day]; (G2) 25 (OH)D3 20μg/day; (G3) 25(OH)D3 266μg/week and (G4) 25(OH)D3 0.266mg every two weeks. 25(OH)D levels were measured for each group at 0, 6 and 12 months, with the following results: G1 (40.5±4.7;80.0±2; 86.2±23.7), G2 (37,2±4.2; 161±21.7;188.0±24.0), G3 (38±3.7;213.5±80.0; 233.0±81.2), G4 (39.5±4;164.5±41,7;210.5±22.2). These data reveal that both metabolites are not equipotent. Calcidiol is faster and 3-6 times more potent to obtain serum levels of 25(OH)D in the medium to long term. This circumstance must be assessed and included in the therapeutic prescription guides for Osteoporosis, since it should be of concern when planning and prescribing treatments to normalize serum levels of 25(OH)D3 and avoid potential adverse impacts.

Adipogenesis is a physiological process required for fat-tissue development, mainly involved in regulating the organism energetic-state. Abnormal distribution-changes and dysfunctions in such tissue are associated to different pathologies. Adipocytes are generated from progenitor cells, via a complex differentiating process not yet well understood. Therefore, we investigated differential mRNA and miRNA expression patterns of human mesenchymal stromal-cells (MSC) induced and not induced to differentiate into adipocytes by next (second)-generation sequencing. A total of 2,866 differentially expressed genes (101 encoding miRNA) were identified, with 705 (46 encoding miRNA) being upregulated in adipogenesis. They were related to different pathways, including PPARG, lipid, carbohydrate and energy metabolism, redox, membrane-organelle biosynthesis, and endocrine system. Downregulated genes were related to extracellular matrix and cell migration, proliferation, and differentiation. Analyses of mRNA-miRNA interaction showed that repressed miRNA-encoding genes can act downregulating PPARG-related genes; mostly the PPARG activator (PPARGC1A). Induced miRNA-encoding genes regulate downregulated genes related to TGFB1. These results shed new light to understand adipose-tissue differentiation and physiology, increasing our knowledge about pathologies like obesity, type-2 diabetes and osteoporosis. J. Cell. Physiol. 232: 771-784, 2017. © 2016 Wiley Periodicals, Inc.

Natural phenols may have beneficial properties against oxidative stress, which is associated with aging and major chronic aging-related diseases, such as loss of bone mineral mass (osteoporosis) and diabetes. The main aim of this study was to analyze the effect of quercetin, a major nutraceutical compound present in the "Mediterranean diet", on mesenchymal stem-cell (MSC) differentiation. Such cells were induced to differentiate into osteoblasts or adipocytes in the presence of two quercetin concentrations (0.1 and 10μM). Several physiological parameters and the expression of osteoblastogenesis and adipogenesis marker genes were monitored. Quercetin (10μM) inhibited cell proliferation, alkaline phosphatase (ALPL) activity and mineralization, down-regulating the expression of ALPL, collagen type I alpha 1 (COL1A1) and osteocalcin [bone gamma-carboxyglutamate protein (BGLAP)] osteoblastogenesis-related genes in MSC differentiating into osteoblasts. Moreover, in these cultures, CCAAT/enhancer-binding protein alpha (CEBPA) and peroxisome proliferator-activated receptor gamma 2 (PPARG2) adipogenic genes were induced, and cells differentiated into adipocytes were observed. Quercetin did not affect proliferation, but increased adipogenesis, mainly at 10-μM concentration in MSC induced to differentiate to adipocytes. β- and γ-catenin (plakoglobin) nuclear levels were reduced and increased, respectively, in quercetin-treated cultures. This suggests that the effect of high concentration of quercetin on MSC osteoblastic and adipogenic differentiation is mediated via Wnt/β-catenin inhibition. In conclusion, quercetin supplementation inhibited osteoblastic differentiation and promoted adipogenesis at the highest tested concentration. Such possible adverse effects of high quercetin concentrations should be taken into account in nutraceutical or pharmaceutical strategies using such flavonol.

Highlights • Corona virus infections can cause Acute Respiratory Distress Syndrome (ARDS), mediated by a variety of mechanisms, including cytokine storm, dysregulation of the renin-angiotensin system, neutrophil activation, and the coagulation cascade. • Vitamin D receptor activation act on many cellular and biochemical mechanisms responsible for ARDS. Most data suggest that VDR activation may taper down ARDS. • Vitamin D supplementation, either as vitamin D or calcifediol (25OHD), is being tested in several RCTs as to demonstrate its potential effects on the course of corona virus infections. • A pilot study in Cordoba suggested that calcifediol treatment may reduce the need for ICU treatment of patients hospitalized for COVID19 infections., Coronavirus infection is a serious health problem awaiting an effective vaccine and/or antiviral treatment. The major complication of coronavirus disease 2019 (COVID-19), the Acute Respiratory Distress syndrome (ARDS), is due to a variety of mechanisms including cytokine storm, dysregulation of the renin-angiotensin system, neutrophil activation and increased (micro)coagulation. Based on many preclinical studies and observational data in humans, ARDS may be aggravated by vitamin D deficiency and tapered down by activation of the vitamin D receptor. Several randomized clinical trials using either oral vitamin D or oral Calcifediol (25OHD) are ongoing. Based on a pilot study, oral calcifediol may be the most promising approach. These studies are expected to provide guidelines within a few months.

Osteoporosis long-term treatment with nitrogen-containing bisphosphonates, has been associated with uncommon adverse effects, as atypical femoral fractures (AFF). Thus, treatment with teriparatide (TPTD; fragment of human parathyroid hormone; PTH1−34) has been proposed for such patients. Besides its anabolizing effect on bone, TPTD may affect stem-cell mobilization and expansion. Bone marrow mononuclear cells (BMMNC) were isolated from five women that had suffered AFF associated to bisphosphonate treatment, before and after 6 months of TPTD therapy. The presence of mesenchymal stromal cells (CD73, CD90 and CD105 positive cells), gene expression of NANOG, SOX2 and OCT4, proliferation, senescence and capacity to differentiate into osteoblasts and adipocytes were analyzed. After TPTD treatment, BMMNC positive cells for CD73, CD90 and CD105 increased from 6.5 to 37.5% (p, Supported by Grants AGL2013-45110-R of “Ministerio de Ciencia e Innovación” (MICINN); PI081692 and PI15/01857 “Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad” (RETICEF) and CIBER “Fragilidad y Envejecimiento Saludable” (CIBERFES) of “Instituto de Salud Carlos III” (ISCIII), “Ministerio de Economía y Competitividad” (MINECO) and European Union (EU); and “Ayudas de Intensificación de la Investigación” and “Grupo CTS413” of “Junta de Andalucía”, Spain.

The nitrogen-containing or nitrogenous bisphosphonates (N-BF) are currently the main class of drugs used for the treatment of diseases characterized by an increased bone resorption. Preliminary data suggest that N-BF have also direct or indirect anti-tumoral effects, and recent evidence suggests that part of the anti-tumoral activity of N-BF may be attributed to their anti-angiogenic capacity when they are used at high concentrations. On the other hand, an optimal vitamin-D status seems to be necessary to maximize the bone response to N-BF. Our aim has been to evaluate the effect of risedronate, alone or in combination with either 1,25(OH)2D3 or 24,25(OH)2D3 (two main vitamin-D metabolites) on parameters related to the angiogenic capacity of human umbilical-vein endothelial cells (HUVEC). The studies of tube formation through in-vitro angiogenesis assays with Matrigel, chemotaxis and migration in a scratch assay showed that low concentrations of risedronate (0.01 to 1 μM) stimulated angiogenesis and cellular migration in vitro. The presence of 1,25(OH)2D3 in the medium inhibited tubular-structure formation and cellular migration. In addition, the presence of 1,25 or 24,25(OH)2D3 in the culture medium also decreased the pro-angiogenic effects of low-concentrations of risedronate. These data show the differential effects of different concentrations of vitamin-D metabolites and risedronate on angiogenesis, thus stressing the importance of an adequate vitamin D status during medical treatment with risedronate. This article is part of a Special Issue entitled ‘17th Vitamin D Workshop’.

There is controversial information about the impact of vitamin A on bone. Some epidemiological studies show that excessive intake of vitamin A, or an excess of serum vitamin A, has related with adverse impact on bone mass; however, other studies did not find these links, and some authors have proposed that this vitamin might promote a better bone health. The present work aims to contribute to clarify the real role of vitamin A in bone tissue. For this purpose, a cross-sectional study of 154 osteoporotic non-treated postmenopausal women (> 65 years old) was carried out. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. We assessed concentrations of serum retinol, osteocalcin, parathyroid hormone, alkaline phosphatase, calcium, and phosphorus. We also studied demographic and anthropometric parameters. Spearman’s correlations between retinol levels and other variables found negative correlations with BMD in both lumbar spine (R = − 0.162, P

Background and objectives Vitamin D deficiency has been consistently linked with cardiovascular diseases. However, results of intervention studies are contradictory. The aim of this study was to evaluate the effect of treatment with calcifediol (25(OH)D3) on the cardiovascular system of patients with non-ST-elevation acute coronary syndrome after percutaneous coronary intervention. Patients and methods A prospective study assessing ≥60-year-old patients with non-ST-elevation acute coronary syndrome, coronary artery disease and percutaneous revascularisation. We randomly assigned 41 patients (70.6 ±6.3 years) into 2 groups: Standard treatment + 25(OH)D3 supplementation or standard treatment alone. Major adverse cardiovascular events (MACE) were evaluated at the conclusion of the 3-month follow-up period. 25(OH)D levels were analyzed with regard to other relevant analytical variables and coronary disease extent. Results Basal levels of 25(OH)D ≤ 50 nmol/L were associated with multivessel coronary artery disease (RR: 2.6 [CI 95%:1.1–7.1], p =0.027) and 25(OH)D ≤ 50 nmol/L + parathormone ≥ 65 pg/mL levels correlated with increased risk for MACE (RR: 4 [CI 95%: 1.1–21.8], p = 0.04]. One MACE was detected in the supplemented group versus five in the control group (p = 0.66). Among patients with 25(OH)D levels ≤50 nmol/L at the end of the study, 28.6% had MACE versus 0% among patients with 25(OH)D > 50 nmol/L (RR: 1,4; p = 0.037). Conclusions Vitamin D deficiency plus secondary hyperparathyroidism may be an effective predictor of MACE. A trend throughout the follow up period towards a reduction in MACE among patients supplemented with 25(OH)D3 was detected. 25(OH)D levels ≤ 50 nmol/L at the end of the intervention period were significantly associated with an increased number of MACE, hence, 25(OH)D level normalization could improve cardiovascular health in addition to bone health.

cope β-Cryptoxanthin is an abundant carotenoid in fruits and vegetables that can be quantified in human blood-serum. Yet, contrary to other carotenoids, its effects on endothelial cells and angiogenesis remain unknown. Methods and results Human umbilical-vein endothelial cells (HUVEC) were treated with 0.01, 0.1 or 1 μM of β-cryptoxanthin. Antioxidant activity was determined by its free radical scavenging and oxygen-radical absorbance capacity (ABTS and ORAC assays). Effect on migration and formation of tubular structures was studied. Additionally, effect on angiogenesis was also analyzed using an in vivo model. β-Cryptoxanthin exhibited scavenging ability, having antioxidant effect on HUVEC. Interestingly, β-cryptoxanthin reduced their migration and angiogenesis, even in presence of vascular endothelial growth-factor (VEGF). Additionally, such carotenoid inhibited in vivo angiogenesis induced by VEGF. Besides, treatment of HUVEC with LE540 [retinoic-acid receptor (RAR) panantagonist] inhibited β-cryptoxanthin antiangiogenic effect on HUVEC. Conclusion β-Cryptoxanthin inhibits angiogenesis through RAR. Thus, this carotenoid and food containing it may be useful for prevention and treatment of angiogenic pathologies. That includes tumoral growth and wet macular-degeneration associated to aging. To the best of our knowledge, this is the first report of the antioxidant effect and antiangiogenic activity of such carotenoid on HUVEC, both in vitro and in vivo. This article is protected by copyright. All rights reserved

Background and Aims Bisphosphonates are widely used for the treatment of bone pathologies, mainly due to their ability to inhibit osteoclastic activity and thus bone resorption. Yet, their potential effect on bone formation is unclear. Our aim was to determine whether risedronate can affect osteoblastic differentiation of mesenchymal stromal cells (MSC). Methods We evaluated the effect of the risedronate, which is a bisphosphonate widely used in clinical settings, on the differentiation of precursor cells of osteoblasts and adipocytes. Thus, MSC from human bone marrow were induced to differentiate into osteoblasts or adipocytes in the presence or absence of two risedronate concentrations (10 −8 M and 10 −9 M). Results Risedronate increased the levels of osteogenic markers including the extracellular matrix mineralization on cells induced to osteoblasts. On the other hand, such bisphosphonates did not have significant effects on the pparγ2 and lpl adipogenic genes or the fat vesicle accumulation on the cells induced into adipocytes. In addition, it increased the expression of early osteogenic differentiation genes like runx2 on MSC not induced to differentiate. Conclusions Risedronate concentrations used favor the osteogenic differentiation and the MSC commitment towards osteoblasts. This suggests the possibility of using this bisphosphonate in cyclic bone treatments with anabolic drugs, enhancing the effect of the latter via a previous MSC commitment to preosteoblasts.

The aim of this study was to evaluate the relationship between vitamin E status and osteoporosis in early postmenopausal women. Anthropometric data, osteoporosis risk factors, vitamin E serum levels, bone mineral density (BMD) and other serum parameters which may influence bone mineral density in postmenopausal women were analyzed in a cross-sectional study. The association between osteoporosis and age, age of menopause, body mass index, osteocalcin, calcium, vitamin D, vitamin E (measured as 25 hydroxyvitamin D and as α-tocopherol:lipids ratio, respectively), bone alkaline phosphatase, smoking status, leisure physical activity and alcohol intake were modeled by a multivariate logistic regression and multi-linear regression analysis in 232 early postmenopausal women. A lower vitamin E:lipid ratio was associated with osteoporosis in multivariate logistic regression. In a multivariate linear model with BMD of the lumbar spine as a dependent variable, the vitamin E:lipid ratio was clearly related with BMD of the lumbar spine (F ratio = 6.30, p = 0.002). BMD of the lumbar spine was significantly higher in the highest tertile of the vitamin E:lipid ratio than in the lowest tertile. The mean vitamin E:lipid ratio was significantly lower in osteoporotic postmenopausal women (T score ≤−2.5) (3.0 ± 0.6 μmol/mmol) than normal (neither osteoporotic nor osteopenic) postmenopausal women (T score >−1) (3.5 ± 0.7 μmol/mmol) using multivariable-adjusted BMD. These findings highlight that vitamin E may increase BMD in healthy postmenopausal women.

Se ha desarrollado el programa iErrors-F, disponible a modo de juego para aprender los errores comunes del uso del inglés (F) en . Dicho proyecto (2015-2-2009) complementa otros nueve (07NA2037, 08A4087, 092016, 102008, 112007, 122022, 2013-12-2009, 2014-12-2013 y 2016-1-2009), que permitieron desarrollar, respectivamente, los programas iFalseFriends, iPartialFalseFriends, iAnglicisms, iErrors-A, iErrors-B, iErrors-C, iErrors-D, iErrors-E and iErrors-G, disponibles a modo de juego para aprender los “falsos amigos” o cognados entre inglés y español , los “amigos inconsistentes” o cognados parciales entre dichos idiomas , los anglicismos , los errores comunes en inglés (A) , los errores comunes en inglés (B) , los errores comunes en inglés (C) , los errores comunes en inglés (D) los errores comunes en inglés (E) y los errores comunes en inglés (G) . Conviene resaltar que estos desarrollos se basan en el aprendizaje de forma lúdica, mediante un juego educativo, lo cual potencia su uso por todos en general y por las nuevas generaciones en particular. Esto diferencia a estos desarrollos de otras alternativas, como son las listas de textos escritos o diccionarios existentes. Dicho con otras palabras, el objetivo es innovar desde el diccionario clásico al juego educativo. Consideramos que ello es un elemento clave para incentivar y motivar al posible usuario. Dichos juegos educativos están diseñados para una visualización óptima desde el navegador web Apple Safari con iPhone e iPod touch (también se pueden usar en el iPad, así como con Mac o Windows). Es indudable que el inglés es el idioma universal y la lingua franca de nuestros días. Sin embargo, el nivel general de inglés en la Universidad de Córdoba puede ser significativamente mejorado, incluyendo Personal Docente e Investigador (PDI), alumnos y Personal de Administración y Servicios (PAS). Esta realidad puede extrapolarse a Andalucía y a España entera. Se trata de un hecho de notable importancia, sobre todo si se tiene en cuenta que la tendencia progresiva es hacia la impartición de clases en inglés. Por tanto, la situación actual nos pone en desventaja frente al proceso de Convergencia Europea (ECTS/EEES), la competitividad empresarial y el acceso al mercado laboral de nuestros egresados, por citar sólo algunos ejemplos. Esta realidad cobra especial relevancia en un mundo cada vez más globalizado. Faltan, por otra parte, herramientas específicas de enseñanza, aprendizaje, uso y perfeccionamiento del inglés (con aplicación académica general, docente, investigadora, asistencial y de gestión) que incidan en las peculiaridades de los hispanoparlantes en general y de los universitarios en particular. El proyecto realizado ha usado las nuevas Tecnologías de la Información y la Comunicación (TIC) para el desarrollo y aplicación de herramientas multimedia y multiplataforma que ayuden a mejorar dichas carencias. El programa contiene una base de datos de errores comunes en el uso del inglés (F). El funcionamiento del recurso es tan intuitivo que no requiere instrucciones especiales: basta navegar hasta iErrors-F y seguir las instrucciones del juego educativo que aparecen en pantalla. La metodología utilizada es doble. Por una parte, se ha desarrollado dicho programa a modo de juego de aprendizaje para su acceso universal vía Internet. Por otra parte, los alumnos han jugado con el mismo de forma presencial y no presencial, analizándose después en clase los avances alcanzados en el aprendizaje del inglés, mediante las correspondientes evaluaciones de progreso. Con ello se ha conseguido una significativa mejora en el aprendizaje de inglés de los alumnos, en un campo en el que los propios angloparlantes cometen errores. El uso de esta herramienta permite una mejora de la docencia, aprendizaje y uso de dicho idioma, mejorando también con ello la calidad de las diferentes actividades desarrolladas en la universidad: académica general, docente, investigadora, asistencial y de gestión. De este modo se contribuye a aumentar la competitividad de la universidad, preparando a la misma para una mejor adaptación al Sistema de Convergencia Europea, actualizando la docencia en las nuevas tecnologías y mejorando la formación pedagógica del profesorado. Asimismo, todo ello genera un perfil más internacional y abierto de nuestra universidad, incrementando su calidad, y con ello el número de alumnos y profesores que la elijan. The application iErrors-F has been developed, being available as a game to learn common errors in English usage (F) in . Such project (2015-2-2009) complement other nine (07NA2037, 08A4087, 092016, 102008, 112007, 122022, 2013-12-2009, 2014-12-2013 and 2016-1-2009), which allowed to develop, respectively, the applications iFalseFriends, iPartialFalseFriends, iAnglicisms, iErrors-A, iErrors-B, iErrors-C, iErrors-D, iErrors-E and iErrors-G, available as games to learn “false friends” or cognates between English and Spanish , “inconsistent friends” or partial cognates between such languages , anglicisms , common errors in English (A) , common errors in English (B) , l common errors in English (C) , common errors in English (D) , common errors in English (E) and common errors in English (G) . It is important to remark that these developments are based on ludic learning, by means of educational games (edutainment), which enhance their use by all in general, and the new generations in particular. That sets apart these developments from other alternatives, like lists of available written texts or dictionaries. In other words, the objective is to innovate from classic dictionaries to educational games. We consider that this is a key element to encourage and motivate the potential user. Such educational games are designed for an optimal visualization from Apple Safari web browser, with iPhone and iPod touch (they can be also used with iPad, as well as Mac o Windows). It is obvious that English is the universal langue and lingua franca of our days. Yet, the general English level at the University of Cordoba (including faculty, students and administrative staff) can be significantly improved. This situation can be extrapolated to Andalusia and the whole Spain. That is a critical fact, mainly when taking into account that there is a progressive trend towards lecturing in English language. Therefore, the current situation represents a disadvantage for us in the context of European Convergence [European Credit Transfer and Accumulation System (ECTS)/European Higher Education Area (EHEA)], business competitiveness and access to labor market of graduates, among others. This situation takes special relevance in a world that is becoming globalized. On the other hand, there is a lack of specific tools for teaching, learning, use and improvement of English language (with general academic application, education, research, assistance and management), taking into account the Spanish-speaking people in general, and university students in particular. The project carried out has used the new Information and Communication Technologies (ICT) for development and deployment of multimedia and cross-platform tools to help improve such shortcomings. The application contains a database of common errors of English usage (F). Operation of the resource is so intuitive that it does not require special instructions: just navigate to iErrors-F and follow the simple directions of the edutainment game, showing on the screen. The methodology used is twofold. On one side, such application has been developed as a learning game for universal access via Internet. On other side, students have played with it in classrooms and outside locations (distance-learning), analyzing later on in classrooms the goals reached in English learning, by means of corresponding progress assessments. That has allowed to accomplish a significant improvement in English learning of students, in an area in which even English-speaking people make mistakes. Such tool allows an improvement of lecturing, learning and usage of such language, further improving that way the quality of different activities carried out at the university: academic in general, education, investigation, assistance and management. This is, therefore, a contribution to increase the university competitiveness, training for a better adaptation to the European Convergence System, updating teaching in new technologies and improving pedagogical level of teachers. Additionally, all that generates a more international and open profile of our university, increasing its quality, and thus the number of students and teachers applying for it.

Adipogenesis is a physiological process required for fat-tissue development, mainly involved in regulating the organism energetic-state. Abnormal distribution-changes and dysfunctions in such tissue are associated to different pathologies. Adipocytes are generated from progenitor cells, via a complex differentiating process not yet well understood. Therefore, we investigated differential mRNA and miRNA expression patterns of human mesenchymal stromal-cells (MSC) induced and not induced to differentiate into adipocytes by next (second)-generation sequencing. A total of 2,866 differentially expressed genes (101 encoding miRNA) were identified, with 705 (46 encoding miRNA) being upregulated in adipogenesis. They were related to different pathways, including PPARG, lipid, carbohydrate and energy metabolism, redox, membrane-organelle biosynthesis, and endocrine system. Downregulated genes were related to extracellular matrix and cell migration, proliferation, and differentiation. Analyses of mRNA-miRNA interaction showed that repressed miRNA-encoding genes can act downregulating PPARG-related genes; mostly the PPARG activator (PPARGC1A). Induced miRNA-encoding genes regulate downregulated genes related to TGFB1. These results shed new light to understand adipose-tissue differentiation and physiology, increasing our knowledge about pathologies like obesity, type-2 diabetes and osteoporosis. J. Cell. Physiol. 232: 771-784, 2017. © 2016 Wiley Periodicals, Inc.

Background and Aims Natural phenolic compounds are known for their antioxidant capacity, showing biological activity in numerous physiological processes. Such chemicals have been proposed for prevention or treatment of pathologies like osteoporosis and diabetes. One of these is hydroxytyrosol (HT), which may be involved in the differentiation of human mesenchymal stromal/stem cells (MSCs), which are precursors of osteoblasts and adipocytes. Yet, little information is available. Therefore, our objective was to study the possible effect of HT on MSC differentiation. Methods Differentiation markers were analyzed while human bone marrow MSCs were differentiated into osteoblasts or adipocytes in the presence of 1 or 100 μmol HT. Results High HT concentrations repressed the expression of osteoblastic markers in MSCs differentiating into osteoblasts, whereas they increased the expression of adipogenic genes and the formation of fat vesicles in MSCs differentiating into adipocytes. Conclusions High HT concentrations may inhibit osteoblastogenesis and promote adipogenesis, which can lead to bone loss. Therefore, the possible pharmacological use of extracts rich in HT should take into account this undesirable effect.

Atypical femoral fractures (AFF) associated with long-term bisphosphonates (LTB) are a growing concern. Their etiology is unknown, but bone material properties might be deteriorated. In an AFF series, we analyzed the bone material properties by microindentation. Four groups of patients were included: 6 AFF, 38 typical osteoporotic fractures, 6 LTB, and 20 controls without fracture. Neither typical osteoporotic fractures nor controls have received any antiosteoporotic medication. A general laboratory workup, bone densitometry by dual-energy X-ray absorptiometry (DXA), and microindentation testing at the tibia were done in all patients. Total indentation distance (Total ID), indentation distance increase (IDI), and creep indentation distance (Creep ID) were measured (microns). Age-adjusted analysis of covariance (ANCOVA) was used for comparisons. Controls were significantly younger than fracture groups. Bisphosphonate exposure was on average 5.5 years (range 5 to 12 years) for the AFF and 5.4 years (range 5 to 8 years) for the LTB groups. Total ID (microns) showed better material properties (lower Total ID) for controls 36 (± 6; mean ± SD) than for AFF 46 (± 4) and for typical femoral fractures 47 (± 13), respectively. Patients on LTB showed values between controls and fractures, 38 (± 4), although not significantly different from any of the other three groups. IDI values showed a similar pattern 13 (± 2), 16 (± 6), 19 (± 3), and 18 (± 5). After adjusting by age, significant differences were seen between controls and typical (p

Arachidonic fatty acid (AA) induces adipogenesis in human mesenchymal stem cells cultures, and high concentrations inhibit osteoblastogenesis; whereas eicosapentaenoic and docosahexaenoic fatty acids do not induce adipogenesis and do not inhibit osteoblastogenesis. In mesenchymal stem cells, omega-6 arachidonic polyunsaturated fatty acid promotes the differentiation of adipocytes and inhibits the osteoblast differentiation. While omega-3 fatty acids do not affect the adipogenic differentiation their effects on osteoblastogenesis are less relevant. An increased ratio of omega-3/omega-6 fatty acid consumption can prevent bone mass loss. Consumption of omega-3 may protect against osteoporosis since they may inhibit osteoclastogenesis. However, with aging, MSC in bone marrow are increasingly differentiated into adipocytes, reducing the number of osteoblasts. Products derived from omega-6 and omega-3 metabolism may affect MSC differentiation into osteoblasts and adipocytes. Human MSC have been differentiated into osteoblasts or adipocytes in the presence of omega-6 (AA), or omega-3 (DHA and EPA), and osteoblastic and adipocytic markers have been analyzed. AA decreases the expression of osteogenic markers and the osteoprotegerin/receptor activator of nuclear factor kappa β ligand gene expression ratio (opg/rankl). High concentrations of AA inhibit the mineralization and cause the appearance of adipocytes in MSC differentiating into osteoblasts to a higher extent than DHA or EPA. In MSC differentiated into adipocytes, AA increases adipogenesis, while DHA and EPA do not affect it. AA caused the appearance of adipocytes in undifferentiated MSC. The lipoxygenase gene (alox15b) is induced by omega-3 in MSC induced to osteoblasts, and by omega-6 in MSC induced to adipocytes. An increase in the intake of omega-3 respect to omega-6 may provide protection against the loss of bone mass, since omega-6 favors the osteoclastic activity by diminishing the opg/rankl gene expression in osteoblasts and promotes MSC differentiation into adipocytes, thus diminishing the production of osteoblasts.

The combination of calcium with vitamin D (vitamin D(3) [colecalciferol]) forms the basis of preventive and therapeutic regimens for osteoporosis. A number of studies have suggested that the combination of calcium and vitamin D is effective when administered at respective dosages of at least 1200 mg and 800 IU per day, although efficacy is, as expected, affected by patient compliance. Overall, treatment with this combination appears to be effective in reducing the incidence of non-vertebral and hip fractures. Also, in all drug studies (of antiresorptive and anabolic agents and strontium ranelate) that demonstrated a reduction in risk of osteoporotic fractures, patients also took calcium and vitamin D supplements. An important finding in this regard is that vitamin D levels have been demonstrated to be inadequate in more than half of women treated for osteoporosis in the US and Europe. The capacity of the small intestine to absorb calcium salts depends on the solubility and ionization of the salts. These properties vary for different salts, with fasting calcium citrate absorption being greater than that of calcium lactogluconate and calcium carbonate. Calcium citrate formulations taken between meals may help to prevent abdominal distension and flatulence, as well as minimize the risk of renal calculus formation, thus helping to optimize patient compliance. Therefore, calcium citrate combined with vitamin D is the combination of choice for the prevention or treatment of osteoporosis.

Objectives Association between vitamin D deficiency and excess of vitamin A as a potential risk factor of osteoporosis and fracture has been evaluated. Design and methods 232 healthy postmenopausal women were studied. Serum parameters were analyzed by standard methods and fat-soluble vitamins by an own HPLC method. QUS measurement of the calcaneal bone was carried out by Sahara. Results 124 patients were considered non-osteoporotic and 101 (44.9%) were osteoporotic. The prevalence of high serum levels of retinol was 36.4% and vitamin D deficiency was 70.1%. 60.4% of women with vitamin D deficiency have high serum levels of retinol. In the whole population, the increased risk of osteoporosis was up to three times higher for the highest retinol quintile, as compared with the lowest retinol quintile. Whereas in women with vitamin D deficiency the risk of osteoporosis increased was up 5 times higher than women in the lowest quintile of retinol. Conclusions Our results show that high retinol levels together with vitamin D deficiency are hitherto an overlooked risk factor for osteoporosis.

The effects of oleuropein on the processes of osteoblastogenesis and adipogenesis in mesenchymal stem cells (MSCs) from human bone marrow have been studied. We report that oleuropein, a polyphenol abundant in olive tree products, reduces the expression of peroxisome proliferator-activated receptor gamma (PPARγ), inhibits adipocyte differentiation, and enhances differentiation into osteoblast.Age-related bone loss is associated with osteoblast insufficiency during continuous bone remodeling. It has been suggested that the formation of osteoblasts in bone marrow is closely associated with adipogenesis, and age-related changes in this relationship could be responsible for the progressive adiposity of bone marrow which occurs with osteoporosis. In addition, the consumption of oleuropein, a major polyphenol in olive leaves and olive oil, has been associated with a reduction in bone loss.We have analyzed the effects of oleuropein-at concentrations between 10(-6) and 10(-4) M-on the processes of osteoblastogenesis and adipogenesis in MSCs from human bone marrow.The results show an increase in osteoblast differentiation and a decrease in adipocyte differentiation when there is oleuropein in the culture media. The gene expression of osteoblastogenesis markers, RUNXII, osterix, collagen type I, osteocalcin, or alkaline phosphatase (ALP), was higher in osteoblast-induced oleuropein-treated cells. Also, the ALP activity and extracellular matrix mineralization were higher when oleuropein was present in the media. Oleuropein in MSCs induced adipocytes to produce a decrease in the expression of the genes involved in adipogenesis, the PPARγ, lipoprotein lipase, or fatty acid-binding protein 4, and minor fat accumulation.Our data suggest that oleuropein, highly abundant in olive tree products included in the traditional Mediterranean diet, could prevent age-related bone loss and osteoporosis.