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Neuroblastoma (NB) is a heterogeneous developmental tumor occurring in childhood, which arises from the embryonic sympathoadrenal cells of the neural crest. Although the recent progress that has been done on this tumor, the mechanisms involved in NB are still partially unknown. Despite some genetic aberrations having been identified, the sporadic cases represent the majority. Due to its wide heterogeneity in clinical behavior and etiology, NB represents a challenge in terms of prevention and treatment. Since a definitive therapy is lacking so far, there is an urgent necessity to unveil the molecular mechanisms behind NB onset and progression to develop new therapeutic approaches. Long non-coding RNAs (lncRNAs) are a group of RNAs longer than 200 nucleotides. Whether lncRNAs are destined to become a protein or not, they exert multiple biological functions such as regulating gene expression and functions. In recent decades, different research has highlighted the possible role of lncRNAs in the pathogenesis of many diseases, including cancer. Moreover, lncRNAs may represent potential markers or targets for diagnosis and treatment of diseases. This mini-review aimed to briefly summarize the most recent findings on the involvement of some lncRNAs in NB disease by focusing on their mechanisms of action and possible role in unveiling NB onset and progression.

Background and aims Several chronic multifactorial diseases originate from energy unbalance between food intake and body energy expenditure, including non-alcoholic fatty liver disease (NAFLD), diabetes, and cardiovascular disorders. Vascular endothelium plays a central role in body homeostasis, and NAFLD is often associated with endothelial dysfunction (ED), the first step in atherosclerosis. Both sugars and fatty acids (FAs) are fuel sources for energy production, but their excess leads to liver steatosis which may trigger ED through a network of mechanisms which need to be clarified. Here, we investigated the crosstalk pathways between in vitro cultured steatotic hepatocytes (FaO) and endothelial cells (HECV) being mediated by soluble factors. Methods and results We employed the conditioned medium approach to test how different extent and features of hepatic steatosis distinctively affect endothelium leading to ED. The steatogenic media collected from steatotic hepatocytes were characterized by high triglyceride content and led to lipid accumulation and fat-dependent dysfunction in HECV cells. We found a parallelism between (i) extent of hepatocyte steatosis and level of lipid accumulation in HECV cells; (ii) type of hepatocyte steatosis (with macro- or microvesicular LDs) and extent of oxidative stress, lipid peroxidation, nitric oxide release and expression of ED markers in HECV cells. Conclusions The present findings seem to suggest that, in addition to triglycerides, other soluble mediators should be released by steatotic hepatocytes and may influence lipid accumulation and function of HECV cells. Further studies need to depict the exact profile of soluble factors involved in steatotic hepatocyte-endothelium crosstalk.

The solutions provided through natural evolution of living creatures serve as an ingenious source of inspiration for many technological and applicative fields. Along these lines, bone-inspired concepts lead to fascinating advances in product design, architecture and garments, thanks to the bone’s exceptional combination of strength, toughness and lightness. Structural applications are inspired by the bone’s ability to resist fracture under a large spectrum of forces, while the high surface area and pore connectivity of bone architecture present exciting opportunities from an aesthetic point of view. Behind these inspirations, a disruptive common belief emerges: “down to the bone”, a journey in search of equality, universality and substantiality. Herein, we explore the current state of the art in bone-inspired applications in these fields, considering the two major categories of structural and aesthetic inspirations and discussing further technological developments.

The comprehension of trabecular bone damage processes could be a crucial hint for understanding how bone damage starts and propagates. Currently, different approaches to bone damage identification could be followed. Clinical approaches start from dual X-ray absorptiometry (DXA) technique that can evaluate bone mineral density (BMD), an indirect indicator of fracture risk. DXA is, in fact, a two-dimensional technology, and BMD alone is not able to predict the effective risk of fractures. First attempts in overcoming this issue have been performed with finite element (FE) methods, combined with the use of three-dimensional high-resolution micro-computed tomographic images. The purpose of this work is to evaluate damage initiation and propagation in trabecular vertebral porcine samples using 2D linear-elastic FE models from DXA images and 3D linear FE models from micro-CT images. Results show that computed values of strains with 2D and 3D approaches (e.g., the minimum principal strain) are of the same order of magnitude. 2D DXA-based models still remain a powerful tool for a preliminary screening of trabecular regions that are prone to fracture, while from 3D micro-CT-based models, it is possible to reach details that permit the localization of the most strained trabecula. Graphical abstract

The complexity of bone tissue, its hierarchical multi-scale structure and the delicate mechanobiological equilibrium that regulates its architecture make the analysis of bone local mechanical characteristics highly challenging. The strong dependence between bone’s mechanical properties and meso- and micro-architectural arrangement is particularly evident in the human femur, where loads are transmitted from the pelvis along the stress trajectories. An alteration in loads distribution generates a consequent deterioration in the structural pattern of trabecular groups, that worsen in presence of pathologies such as osteoporosis. Considering this complexity, it is crucial to map femur’s local strength values, in order to capture their variability and to evaluate the possible causes of fragility and fracture. For this purpose, a portable micro-compression device compatible with a synchrotron is designed and realized: it allows to test millimetric samples cut along the stress trajectories from three healthy and three osteoporotic femoral heads and to real-time monitor the apparent stress-apparent strain curves. Intra-patients variability shows increased resistance to compression in the proximal sub-region, anatomically close to the acetabulum, where the loads are transmitted from the pelvis to the femur. Inter-patients variability show an interesting trend: non-osteoporotic femoral heads are characterized by a more uniform distribution in terms of Young moduli in the whole head, while non-uniform values are present in osteoporotic subjects.

Health locus of control (HLOC) may influence people’s behavior regarding their health as well as their desires to be involved in the medical decision-making. Our study aimed to examine HLOC’s relations with people’s control preferences about the medical decision-making. A total of153 people filled out the self-administered version of the Control Preference Scale and the Multidimensional Health Locus of Control Scale–form C. The most preferred role is the collaborative one. However, HLOC explained heterogeneity in people’s control preferences: lower scores in external HLOC were related to a greater preference for the active and the collaborative role. From the personalized medicine perspective, an accurate evaluation of the patient’s HLOC could help tailoring the decision-making process within the clinical context.

Aims Adipocyte hypertrophy is the main cause of obesity. A deeper understanding of the molecular mechanisms regulating adipocyte dysfunction may help to plan strategies to treat/prevent obesity and its metabolic complications. Here, we investigated in vitro the molecular alterations associated with early adipocyte hypertrophy, focusing on mitochondrial dysfunction. Main methods As model of adipocyte hypertrophy, we employed 3T3-L1 preadipocytes firstly differentiated into mature adipocytes, then cultured with long-chain fatty acids. As a function of differentiation and hypertrophy, we assessed triglyceride content, lipid droplet size, radical homeostasis by spectrophotometry and microscopy, as well as the expression of PPARγ, adiponectin and metallothioneins. Mitochondrial status was investigated by electron microscopy, oxygraph 2 k (O2K) high-resolution respirometry, fluorimetry and western blot. Key findings Compared to mature adipocytes, hypertrophic adipocytes showed increased triglyceride accumulation and lipid peroxidation, larger or unique lipid droplet, up-regulated expression of PPARγ, adiponectin and metallothioneins. At mitochondrial level, early-hypertrophic adipocytes exhibited: (i) impaired mitochondrial oxygen consumption with parallel reduction in the mitochondrial complexes; (ii) no changes in citrate synthase and HSP60 expression, and in the inner mitochondrial membrane polarization; (iii) no stimulation of mitochondrial fatty acid oxidation. Our findings indicate that the content, integrity, and catabolic activity of mitochondria were rather unchanged in early hypertrophic adipocytes, while oxygen consumption and oxidant production were altered. Significance In the model of early adipocyte hypertrophy exacerbated oxidative stress and impaired mitochondrial respiration were observed, likely depending on reduction in the mitochondrial complexes, without changes in mitochondrial mass and integrity.

The investigation of bone damage processes is a crucial point to understand the mechanisms of age-related bone fractures. In order to reduce their impact, early diagnosis is key. The intricate architecture of bone and the complexity of multiscale damage processes make fracture prediction an ambitious goal. This review, supported by a detailed analysis of bone damage physical principles, aims at presenting a critical overview of how multiscale imaging techniques could be used to implement reliable and validated numerical tools for the study and prediction of bone fractures. While macro- and meso-scale imaging find applications in clinical practice, micro- and nano-scale imaging are commonly used only for research purposes, with the objective to extract fragility indexes. Those images are used as a source for multiscale computational damage models. As an example, micro-computed tomography (micro-CT) images in combination with micro-finite element models could shed some light on the comprehension of the interaction between micro-cracks and micro-scale bone features. As future insights, the actual state of technology suggests that these models could be a potential substitute for invasive clinical practice for the prediction of age-related bone fractures. However, the translation to clinical practice requires experimental validation, which is still in progress.

Disease prevention is a multifaceted construct that has been widely studied. Nevertheless, in spite of its importance, it is still not sufficiently considered by the general population. Since the reasons for this lack of consideration are not yet fully understood, we created an Online Prevention Survey (OPS) to investigate the role of both sociodemographic and psychological factors in predicting individuals&rsquo, spontaneous participation in secondary prevention programs. The results revealed that younger people, men, manual workers, unemployed people, and those who do not regularly practise physical activity were less likely to spontaneously participate in such programs. Furthermore, an analysis of the psychological determinants of the willingness to participate in secondary prevention programs showed that depressive symptoms negatively predict it, while an individual&rsquo, s perception of receiving high social support acts as a positive predictor. Based on these results, we suggest the need for implementing new tailored approaches to promote prevention initiatives to those segments of the population which are more reluctant to spontaneously undertake prevention paths.

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Refinery equipment subjected to high pressure is commonly made of Vanadium high strength steels (2¼Cr1Mo¼V), characterized by high allowable stress and low toughness in the as welded condition, leading to potential wall cracking before the application of thermal treatments. Therefore, the decision to perform specific thermal treatments after welding is of paramount importance. These thermal treatments, which are quite expensive and time demanding for the manufacturer, are still under discussion and not supported by evident scientific findings. The paper presents a numerical and experimental study on a plate-to-plate weld and on a nozzle-to-plate weld, created as ad-hoc mock-ups. Experimental residual stresses are collected by an X-ray diffractometer in the as welded configurations. These values are used to validate a complex 3D numerical model, implemented with the finite element software Abaqus and its AWI plugin. Finally, this validated model allows for the identification of joint criticality through two parameters: the volume of plasticized material per unit of welded length and the strain-based assessment according with ASME code. Their application as tools to compare the criticality of different welded geometries and the effect of thermal treatments on the residual stress field are discussed.

Hepatic steatosis is the hallmark of non-alcoholic fatty liver disease (NAFLD), the hepatic manifestation of the metabolic syndrome and insulin resistance with potential evolution towards non-alcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma. Key roles of autophagy and oxidative stress in hepatic lipid accumulation and NAFLD progression are recognized. Here, we employed a rat hepatoma cell model of NAFLD progression made of FaO cells exposed to oleate/palmitate followed or not by TNFα treatment to investigate the molecular mechanisms through which silybin, a lipid-lowering nutraceutical, may improve hepatic lipid dyshomeostasis. The beneficial effect of silybin was found to involve amelioration of the fatty acids profile of lipid droplets, stimulation of the mitochondrial oxidation and upregulation of a microRNA of pivotal relevance in hepatic fat metabolism, miR-122. Silybin was also found to restore the levels of Aquaporin-9 (AQP9) and glycerol permeability while reducing the activation of the oxidative stress-dependent transcription factor NF-κB, and autophagy turnover. In conclusion, silybin was shown to have molecular effects on signaling pathways that were previously unknown and potentially protect the hepatocyte. These actions intersect TG metabolism, fat-induced autophagy and AQP9-mediated glycerol transport in hepatocytes.

In the Additive Manufacturing (AM) field, Selective Laser Melting (SLM) process is one of the most promising manufacturing technologies for the production of 3D complex parts. High-tech sectors, like aerospace, show growing interest for the use of AM, because it can offer unique opportunities in terms of freedom in the design and integration of some specific functionalities in lightweight structures. Together with lightness of materials, another important requirement is the capacity of vibrations damping, relevant e.g. for the vibro-acoustic environment during launch in the aerospace field. This work focuses on the correlation between the damping behavior of AlSi10Mg specimens and residual stresses induced during SLM, i.e. in as-built condition, or after different post-additive heat treatments, specifically studied for this Al alloy. Indeed, residual stresses have a well-known influence on the static and fatigue mechanical behavior, but a few literature works are present relating residual stresses with damping capacity. Results of the performed experimental tests showed that the damping capacity decreases with the increase of the heat treatment temperatures, i.e. damping can be modulated by customized thermal treatments.

Background: Patient preferences regarding their involvement in shared treatments decisions is fundamental in clinical practice. Previous evidences demonstrated a large heterogeneity in these preferences. However, only few studies have analysed the influence of patients’ individual differences, contextual and situational qualities, and their complex interaction in explaining this variability. Methods: We assessed the role of the interaction of patient’s sociodemographic and psychological factors with a physician’s gender. Specifically, we focused on patient gender and attitudes toward male or female physicians. One hundred fifty-three people participated in this randomised controlled study and were randomly assigned to one of two experimental conditions in which they were asked to imagine discussing their treatment with a male and a female doctor. Results: Analyses showed an interplay between attitude towards women and the gender of patients and doctors, explaining interindividual variability in patient preferences. Conclusions: In conclusion, patients’ attitudes toward the physicians’ gender constitutes a relevant characteristic that may influence the degree of control patients want to have and the overall patient-physician relationship.

Integrating the psychosocial perspective in oncology is warranted. Here, we introduce a structured psychological intervention, the clinical-care focused psychological interview (CLiC), to address patients' needs in the relationship with health professionals, clinical pathway and decision-making process. The perceived utility and feasibility of the CLiC were evaluated in a preliminary sample of 30 patients who were candidates to undergo a radical cystectomy at the European Institute of Oncology, Milan, Italy. Patients reported a very high satisfaction with the interview, because it prepared them for their upcoming surgery by gathering more information about their clinical pathway and allowed them to discover the information they still needed. 30% stated that CLiC helped them to reorganise their thoughts and 36.7% understood the role of psychological intervention in the clinical pathway. Only 20% considered the CLiC useful in helping to build their relationship with the clinical staff. Before an invasive surgery such as radical cystectomy, patients' need for information regarding the upcoming surgery seems to prevail. Knowing the future consequences and adjusting toward the disease could be paramount for patients in facing uncertainty and they might feel that creating a relationship with physicians could be a secondary issue. However, our data show that a structured psychological intervention such as the CLiC interview can collect important information for patients and healthcare professionals to develop real patient-centred care.

Combining thermography and Digital Image Correlation (DIC) measurements, this paper aims to experimentally capture the fatigue damage phenomenon in steels, with a methodology allowing to rapidly estimate the fatigue limit. Surface temperatures and displacement fields are collected during fatigue tests performed with blocks of progressively increased stress amplitude, at different stress ratios. Results show that both thermography and DIC-based parameters have bilinear trends as a function of the stress amplitude. These changes in the thermal and mechanical behaviours, highly sensitive to the selected stress ratio, are related to damage initiation due to microplasticity and give a precise and rapid estimation of the fatigue limit.

The response of an AlSi10Mg alloy produced by selective laser melting (SLM) to specifically designed heat treatments is explored. Thermal phenomena of the rapidly solidified material were studied by differential scanning calorimetry (DSC), and the results have been used to define specific post-processing thermal treatment temperatures. In detail, three heat treatments were studied: two innovative ones, able to selectively trigger (1) Mg2Si precipitation and (2) the rupture and spheroidization of the silicon network, and (3) a T5-like aging were applied for different times and their effects on mechanical properties were studied. The first and the second heat treatments were selected by combining DSC analysis and aging curves for the optimization of the temperature and duration. And these were more deeply investigated in terms of microstructural evolution and of residual stresses. The optimized thermal treatments for SLM-built AlSi10Mg are recommended as valid alternatives to the usually applied T6 or high-temperature stress release treatments.

S-adenosylmethionine (SAMe) is an endogenous methyl donor derived from ATP and methionine that has pleiotropic functions. Most SAMe is synthetized and consumed in the liver, where it acts as the main methylating agent and in protection against the free radical toxicity. Previous studies have shown that the administration of SAMe as a supernutrient exerted many beneficial effects in various tissues, mainly in the liver. In the present study, we aimed to clarify the direct effects of SAMe on fatty acid-induced steatosis and oxidative stress in hepatic and endothelial cells. Hepatoma FaO cells and endothelial HECV cells exposed to a mixture of oleate/palmitate are reliable models for hepatic steatosis and endothelium dysfunction, respectively. Our findings indicate that SAMe was able to significantly ameliorate lipid accumulation and oxidative stress in hepatic cells, mainly through promoting mitochondrial fatty acid entry for &beta, oxidation and external triglyceride release. SAMe also reverted both lipid accumulation and oxidant production (i.e., ROS and NO) in endothelial cells. In conclusion, these outcomes suggest promising beneficial applications of SAMe as a nutraceutical for metabolic disorders occurring in fatty liver and endothelium dysfunction.

Metal-polymer-metal sandwiches can find promising applications in the automotive field thanks to their lightweight and formability . The paper focuses on the effect of low velocity impacts on the residual mechanical behavior. Experimental stepwise tests are run on undamaged and impacted specimens with different combinations of thickness and grade for the outer steel skins and the inner polymeric core. Surface temperature evolution is thermally monitored during the tests with the aim to characterize the induced damage and to identify a parameter able to quantify the residual strength of the panel. Several approaches have been considered. The analysis of the thermal amplitude trend with the lock-in thermography evidences a variation in the thermal behavior of the specimens, defining a corresponding damage stress σD. We found a 20% σD difference between undamaged and damaged specimens. Moreover, impacted specimens experience a temperature and stress concentration at the impact area dependent on the indentation . Based on these results, we evidence the possibility to relate impact indentation with the damage stress estimated by thermography and with the stress concentration factor induced by the impact. Therefore, thermography is a useful and valid tool for post-impact damage detection, monitoring and quantification of these multi-layer sandwich materials.

Sumac (Rhus coriaria L.) is a commonly used spice in the Mediterranean region and considered as healthy food ingredients. The beneficial value of sumac is well documented in folk medicine. Accumulating data explored the phytochemical, nutritional and therapeutic proprieties suggesting sumac as a potential functional food. Here, we discuss the general and scientific aspects of sumac. Sumac is rich in different polyphenolic compounds such as flavonoids, tannins, and phenolic acids. The potential therapeutic effects of sumac have been studied in various cellular and animal models, as well as in human. These reports suggest that Sumac has potential effect against oxidative stress, inflammation, obesity, hyperglycemia, hypercholesterolemia, and hyperlipidemia, which represent key pathogenic mechanisms contributing to cardio-metabolic, liver, and cancer diseases. Clinical studies using sumac or its major compounds, suggest that this herbal product may represent a useful therapeutic tool in the management of metabolic-related conditions such as liver-atherosclerosis complications.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disorder in industrialized countries and is associated with increased risk of cardiovascular, hepatic and metabolic diseases. Molecular mechanisms on the root of the disrupted lipid homeostasis in NAFLD and potential therapeutic strategies can benefit of in vivo and in vitro experimental models of fatty liver. Here, we describe the high fat diet (HFD)-fed rat in vivo model, and two in vitro models, the primary cultured rat fatty hepatocytes or the FaO rat hepatoma fatty cells, mimicking human NAFLD. Liver steatosis was invariably associated with increased number/size of lipid droplets (LDs) and modulation of expression of genes coding for key genes of lipid metabolism such as peroxisome proliferator-activated receptors (Ppars) and perilipins (Plins). In these models, we tested the anti-steatotic effects of 3,5-L-diiodothyronine (T2), a metabolite of thyroid hormones. T2 markedly reduced triglyceride content and LD size acting on mRNA expression of both Ppars and Plins. T2 also stimulated mitochondrial oxidative metabolism of fatty acids. We conclude that in vivo and especially in vitro models of NAFLD are valuable tools to screen a large number of compounds counteracting the deleterious effect of liver steatosis. Because of the high and negative impact of liver steatosis on human health, ongoing experimental studies from our group are unravelling the ultimate translational value of such cellular models of NAFLD.

Phenolic compounds (PC) of virgin olive oil exert several biochemical and pharmacological beneficial effects. Some dietary PC seem to prevent/improve obesity and metabolic-related disorders such as non-alcoholic fatty liver disease (NAFLD). We investigated the possible effects of PC extracted from olive pomace (PEOP) and of the main single molecules present in the extract (tyrosol, apigenin, oleuropein, p-coumaric and caffeic acid) in protecting hepatocytes and endothelial cells against triglyceride accumulation and oxidative stress.Rat hepatoma and human endothelial cells were exposed to a mixture of oleate/palmitate to mimic the condition of NAFLD and atherosclerosis, respectively. Then, cells were incubated for 24 h in the absence or in the presence of PC or PEOP. Different parameters were evaluated, such as lipid accumulation and oxidative stress-related markers.In hepatic cells, expression of peroxisome proliferator-activated receptors (PPARs) and of stearoyl-CoA desaturase 1 (SCD-1) were assessed as index of lipid metabolism. In endothelial cells, expression of intercellular adhesion molecule-1 (ICAM-1), activation of nuclear factor kappa-B (NF-kB), release of nitric oxide (NO), and wound-healing rate were assessed as index of inflammation.PEOP extract ameliorated hepatic lipid accumulation and lipid-dependent oxidative imbalance thus showing potential applications as therapeutic agent tuning down hepatosteatosis and atherosclerosis.

A finite element cohesive zone model able to reproduce the effect of hydrogen exposition on the fracture toughness properties of a low alloy Cr–Mo steel, AISI 4130, is here presented. The model simulates the embrittlement effect of hydrogen by decreasing the constitutive law of cohesive elements that reproduces the crack propagation behaviour of the material according to the calculated total hydrogen concentration. The model is validated by comparison with experimental data. Finally, a sensitivity analysis of the model that mimics both different material parameters and environment conditions proves its efficiency as a predictive tool for the mechanical response of the material.

Background Bone strain index (BSI) is a tool measuring bone strain, derived from dual x-ray photon absorptiometry. It is able to characterise an aspect of bone quality that, joined to the quantity and quality parameters of bone mineral density (BMD) and trabecular bone score (TBS), permits an accurate definition of fracture risk. As no data are available about BSI precision, our aim was to assess its in vitro reproducibility. Methods A Hologic spine phantom was used to perform BSI scans with three different scan modes: fast array (FA), array (A), and high definition (HD). Different soft tissue thicknesses (1, 3, 6 cm) of fresh pork rind layers as a surrogate of abdominal fat were interposed. For each scan mode, the phantom was consecutively scanned 25 times without repositioning. Results In all scan modes (FA, A, HD) and at every fat thickness, BSI reproducibility was lower than that of BMD. The highest reproducibility was found using HD-mode with 1 cm of pork rind and the lowest one using HD-mode with 6 cm of pork rind. Increasing fat thickness, BSI reproducibility tended to decrease. BSI least significant change appeared to be about three times that of BMD in all modalities and fat thicknesses. Without pork rind superimposition and with 1-cm fat layer, BSI reproducibility was highest with HD-mode; with 3 or 6 cm fat thickness, it was higher with A-mode. Conclusions BSI reproducibility was worse than that of BMD, but it is less sensitive to fat thickness increase, similarly to TBS.

Bone strain index (BSI) is a dual-energy X-ray absorptiometry (DXA)-derived index of bone strength obtained from lumbar densitometric scan. We estimated the reproducibility of BSI in healthy women with different body mass index. We enrolled postmenopausal women (mean age ± SD: 66 ± 10 years) divided into three groups (A, B and C) according to body mass index (BMI: 88 cm), each of 30 subjects. They underwent two DXA examinations with in-between repositioning, according to the International Society for Clinical Densitometry guidelines for precision estimation. Bone mineral density (BMD) and BSI were expressed as g/cm2 and absolute value, respectively. The coefficient of variation (CoV) was calculated as the ratio between root-mean-square standard deviation and mean; least significant change percentage (LSC%) as 2.77 × CoV; reproducibility as the complement to 100% LSC. BSI increased proportionally to BMI and WC and significantly in group C compared to B and A (p = 0.032 and 0.006, respectively). BSI was significantly higher in E compared to D (p = 0.017), whereas no differences were observed in BMD. Although BSI reproducibility was slightly lower in group C (89%), the differences were not significant between all groups. BMD reproducibility did not significantly differ between all groups. BSI reproducibility was significantly lower than that of BMD and decreased proportionally to BMI and WC increase. This reduction of BSI reproducibility was more pronounced in patients with BMI ≥ 30 and WC > 88, as expected, being BSI a parameter sensible to weight.

Overconsumption of fats and sugars is a major cause of development of non‑alcoholic fatty liver disease (NAFLD). The main objectives of the present study were to explore the pathways sustaining the interfering metabolic effects of excess fructose and fatty acids in hepatocytes, and to clarify the mechanisms through which the nutraceutical silybin rescues the functional and metabolic alterations that are associated with the NALFD progression. Cultured hepatocytes were exposed to fructose and fatty acids, alone or in combination, to induce different grades of steatosis in vitro. Cell viability, apoptosis, free radical production, lipid content, lipid peroxidation and activity of lipogenic enzymes were assessed by spectrophotometric assays. Oxygen consumption and mitochondrial respiration parameters were measured using a Seahorse analyzer. Expression of markers for liver steatosis and dysfunction were also evaluated by reverse transcription‑quantitative polymerase chain reaction. The data revealed that fructose and fatty acid combination in vitro had a positive interference on lipogenic pathways, leading to more severe steatosis and liver dysfunction, reduced cell viability, increased apoptosis, oxidative stress and mitochondrial respiration. Hepatic cell abnormalities were almost completely alleviated by silybin treatment. These findings suggest that silybin may serve as a novel and cost‑effective dietary supplement for treatment and/or prevention of hepatosteatosis associated with NAFLD progression.

Patient empowerment is a multi-factorial concept and its relevance has led to a growing body of literature; despite this attention, there is still no agreement regarding the elements that define its expression. While several studies have already investigated the positive effect of empowerment interventions on the care process outcome, the aim of this study is to investigate which factors can foster an empowered management of the cancer condition from the patient's perspective. To examine patients' perception of empowerment enablers, we asked for participants' input on the role of three factors frequently cited as positively affected by empowerment: care quality, perception of direct control and relationships within the care context, during the care process. Three focus groups were conducted with 34 cancer patients. The results highlight the perception of direct control on their treatment as the least valued element (2.87, SD 0.566) when compared with care quality (3.75, SD 0.649) and relational support in the care context (3.91, SD 0.274). Unlike traditional approaches to empowerment, patient's expression of empowerment does not mainly reside in the direct control of their condition as much as in an active role within the relationship with caretakers, such as the ability to choose the doctor, the care team or the health organisation in charge of their healthcare. Emerging aspects from this analysis of patient's perspective are central in order to adequately consider empowerment in the care process and to provide more effective care strategies.

Amplification in toughness and balance with stiffness and strength are fundamental characteristics of biological structural composites, and a long sought-after objective for engineering design. Nature achieves these properties through a combination of multiscale key features. Yet, emulating all these features into synthetic de novo materials is rather challenging. Here, we fine-tune manual lamination, to implement a newly designed bone-inspired structure into fiber-reinforced composites. An integrated approach, combining numerical simulations, ad hoc manufacturing techniques, and testing, yields a novel composite with enhanced fracture toughness and balance with stiffness and strength, offering an optimal lightweight material solution with better performance than conventional materials such as metals and alloys. The results also show how the new design significantly boosts the fracture toughness compared to a classic laminated composite, made of the same building blocks, also offering an optimal tradeoff with stiffness and strength. The predominant mechanism, responsible for the enhancement of fracture toughness in the new material, is the continuous deviation of the crack from a straight path, promoting large energy dissipation and preventing a catastrophic failure. The new insights resulting from this study can guide the design of de novo fiber-reinforced composites toward better mechanical performance to reach the level of synergy of their natural counterparts.

Ethnopharmacological relevance Thymbra spicata, a member of the Lamiaceae family, is native to eastern Mediterranean area. Leaves of this plant are rich in phenolic compounds and are a popular remedy of traditional medicine in Lebanon to prevent and/or counteract hyperlipidemia and hyperglycemia. Aim of the study To evaluate the antisteatotic and antioxidant activities of extracts from leaves of Thymbra spicata L. using in vitro models of non-alcoholic fatty liver disease (NAFLD), a leading cause of liver-related morbidity and mortality worldwide, for whom no effective treatments are still available. Materials and methods Two different extracts from Thymbra spicata L. aerial parts were prepared using water (TW) or ethanol (TE) as solvent. Their chemical composition was characterized by gas and liquid chromatography coupled with mass spectrometry. Both extracts were tested on cultured hepatic and endothelial cells treated to mimic in vitro a multisistemic pathology such as NAFLD. We assayed the effects on lipid accumulation, free radical production, lipid peroxidation, cell migration. Results Both the total phenolic and the total flavonoid contents were higher in the ethanolic extract. Rosmarinic acid was the most abundant polyphenol in TW, while TE was richer in carvacrol. Our findings demonstrated that both extracts ameliorated lipid accumulation, oxidative stress and inflammation in the NAFLD cellular models. However, the aqueous extract was more effective to reduce hepatic steatosis, and the ethanolic extract had higheranti-oxidant potential and wound healing activity. Conclusions T. spicata extracts could be promising bioactive products to develop natural therapeutic agents or dietary supplements to treat NAFLD and obesity-related metabolic disease. Our findings suggest that while the ethanolic extract might be used in preventing endothelium dysfunction, the aqueous extract would act better as lipid-lowering agent.

Dual Energy X-Ray Absorptiometry (DXA) is currently the most widely adopted non-invasive clinical technique to assess bone mineral density and bone mineral content in human research and represents the primary tool for the diagnosis of osteoporosis. DXA measures areal bone mineral density, BMD, which does not account for the three-dimensional structure of the vertebrae and for the distribution of bone mass. The result is that longitudinal DXA can only predict about 70% of vertebral fractures. This study proposes a complementary tool, based on Finite Element (FE) models, to improve the DXA accuracy. Bone is simulated as elastic and inhomogeneous material, with stiffness distribution derived from DXA greyscale images of density. The numerical procedure simulates a compressive load on each vertebra to evaluate the local minimum principal strain values. From these values, both the local average and the maximum strains are computed over the cross sections and along the height of the analysed bone region, to provide a parameter, named Strain Index of Bone (SIB), which could be considered as a bone fragility index. The procedure is initially validated on 33 cylindrical trabecular bone samples obtained from porcine lumbar vertebrae, experimentally tested under static compressive loading. Comparing the experimental mechanical parameters with the SIB, we could find a higher correlation of the ultimate stress, σULT, with the SIB values (R2adj = 0.63) than that observed with the conventional DXA-based clinical parameters, i.e. Bone Mineral Density, BMD (R2adj = 0.34) and Trabecular Bone Score, TBS (R2adj = -0.03). The paper finally presents a few case studies of numerical simulations carried out on human lumbar vertebrae. If our results are confirmed in prospective studies, SIB could be used-together with BMD and TBS-to improve the fracture risk assessment and support the clinical decision to assume specific drugs for metabolic bone diseases.

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease often associated with overnutrition. Number and morphometry of lipid droplets (LDs) define micro vs macrovesicular steatosis, influence the morphology and function of hepatocytes and possibly their stiffness. The link between grade and features of steatosis and biomechanical properties of single hepatocytes requires deeper investigations. In vitro NAFLD models with distinct steatosis conditions were set by exposing FaO hepatoma cells to single or combined fructose (Fru), fatty acids (FA), and tumor necrosis factor (TNF)α. Single Cell Force Spectroscopy and Quantitative Phase Microscopy quantified the single cell stiffness and a series of morphometric parameters; the mRNA expression of genes involved in lipid metabolism was quantified by real-time PCR. In our models, LD size and number increased with Fru and FA as single agents, and more with combined Fru/FA (macrovesicular steatosis), while FA/TNFα combination increased LD number with a reduction in their size (microvesicular steatosis). We found that the changes in LD size and number influenced cell stiffness and morphometry as follows: (i) single cell elasticity increased in macrovesicular steatosis (maximally with combined Fru/FA); (ii) FA-induced steatosis resulted in cells thinner and larger, whereas combined FA/TNFα shrunk the hepatocytes. Taken together the data on hepatocyte biomechanics show that, in addition to extent of lipid accumulation, cell stiffness is mainly influenced by LD size, while cell morphometry directly relates to LD number. Our findings suggest that a novel mechanobiology perspective might provide future contributions in NAFLD research.

Background:Fatty liver, or steatosis, is a condition of excess accumulation of lipids, mainly under form of triglycerides (TG), in the liver, and it is the hallmark of non-alcoholic fatty liver disease (NAFLD). NAFLD is the most common liver disorder world-wide and it has frequently been associated with obesity, hyperlipidemia and insulin resistance. Free fatty acids (FA) are the major mediators of hepatic steatosis; patients with NAFLD have elevated levels of circulating FA that correlate with disease severity.Methods:Steatosis is a reversible condition that can be resolved with changed behaviors, or that can progress towards more severe liver damages such as steatohepatitis (NASH), fibrosis and cirrhosis. In NAFLD, FA of exogenous or endogenous origin accumulate in the hepatocytes and trigger liver damages. Excess TG are stored in cytosolic lipid droplets (LDs) that are dynamic organelles acting as hubs for lipid metabolism.Results:In the first part of this review, we briefly reassumed the main classes of FA and their chemical classification as a function of the presence and number of double bonds, their metabolic pathways and effects on human health. Then, we summarized the main genetic and diet-induced animal models of NAFLD, as well as the cellular models of NAFLD.Conclusions:In recent years, both the diet-induced animal models of NAFLD as well as the cellular models of NAFLD have found ever more application to investigate the mechanisms involved in NAFLD, and we referred to their advantages and disadvantages.

Objectives: To investigate stakeholder perspectives on how patient preference studies (PPS) should be designed and conducted to allow for inclusion of patient preferences in decision-making along the medical product life cycle (MPLC), and how patient preferences can be used in such decision-making. Methods: Two literature reviews and semi-structured interviews (n = 143) with healthcare stakeholders in Europe and the US were conducted; results of these informed the design of focus group guides. Eight focus groups were conducted with European patients, industry representatives and regulators, and with US regulators and European/Canadian health technology assessment (HTA) representatives. Focus groups were analyzed thematically using NVivo. Results: Stakeholder perspectives on how PPS should be designed and conducted were as follows: 1) study design should be informed by the research questions and patient population; 2) preferred treatment attributes and levels, as well as trade-offs among attributes and levels should be investigated; 3) the patient sample and method should match the MPLC phase; 4) different stakeholders should collaborate; and 5) results from PPS should be shared with relevant stakeholders. The value of patient preferences in decision-making was found to increase with the level of patient preference sensitivity of decisions on medical products. Stakeholders mentioned that patient preferences are hardly used in current decision-making. Potential applications for patient preferences across industry, regulatory and HTA processes were identified. Four applications seemed most promising for systematic integration of patient preferences: 1) benefit-risk assessment by industry and regulators at the marketing-authorization phase; 2) assessment of major contribution to patient care by European regulators; 3) cost-effectiveness analysis; and 4) multi criteria decision analysis in HTA. Conclusions: The value of patient preferences for decision-making depends on the level of collaboration across stakeholders; the match between the research question, MPLC phase, sample, and preference method used in PPS; and the sensitivity of the decision regarding a medical product to patient preferences. Promising applications for patient preferences should be further explored with stakeholders to optimize their inclusion in decision-making. ispartof: FRONTIERS IN PHARMACOLOGY vol:10 ispartof: location:Switzerland status: published

Delamination is the major failure mechanism in composite laminates and eventually leads to material failure. An early-detection and a better understanding of this phenomenon, through non-destructive assessment, can provide a proper in situ repair and allow a better evaluation of its effects on residual strength of lightweight structural components. Here we adopt a joint numerical-experimental approach to study the effect of delamination on the fatigue life of glass/epoxy composites. To identify and monitor the evolution of the delamination during loading, we carried out stepwise cyclic tests coupled with IR-thermography on both undamaged and artificially-damaged samples. The outcome of the tests shows that IR-thermography is able to identify a threshold stress, named damage stress σ D , which is correlated to the damage initiation and the fatigue performance of the composite. Additionally, we performed FE-simulations, implementing the delamination by cohesive elements . Such models, calibrated on the basis of the experimental fatigue results, can provide a tool to assess the effect of parameters, such as the delamination size and location and composite stacking sequence, on the residual strength and fatigue life of the composite material.

The AIAS group studying Energetic Methods for Experimental Analysis, MEAS, is performing round robin experimental tests for the rapid determination of fatigue limit on steels by different thermographic techniques. This work is part of the project and describes the experimental activity performed at Politecnico di Milano, based on stepwise cyclic tests. Thermograms are processed in terms of: 1) amplitude of the first order harmonic, in-phase with respect to the loading signal; 2) amplitude of the second harmonic, out-of-phase; 3) slope of the thermal signal with respect to the number of cycles. These values typically show bilinear trends, allowing to define a breakup point and a corresponding stress which is the thermographic estimation of the fatigue limit. The paper presents and discusses the results of tests with different stress ratios, i.e. fully reversed cycling with R=-1 and tensile-tensile cycling with R=0.1.

Machines for cold wire drawing are widely spread in the industrial field to reduce wires’ section. The system is complex, because the axial pulling force has to correctly deform wires into the drawing dies. The present work focuses on a continuous press and pulling system, with a track-chain. Normal force at the steel clamps can be generated independently on the axial force, but avoiding slipping and local yielding at the external surface of the wire. The work presents a numerical model of wire-clamp contact, focusing on the yielded area at the wire surface, and aimed at the identification of the limit clamping force that avoids slipping and yielding. Reliability of results is critically discussed, based on the choice of element formulation and type, and mesh refinement. Based on this analysis, control parameters of the drawing machine are modified to experimentally verify this limit normal force estimated by the numerical model.

The connecting rod (or conrod) is an important component of an internal combustion engine. An eventual failure of the bolts clamping the big end of conrods can cause failure of the engine; for this reason, they are addressed as “critical fasteners”. Bolts manufacturers must guarantee steel purity and avoid any surface defects, thread forming laps, surface decarburization, surface phosphate diffusion or hydrogen embrittlement risk in order to increase their reliability. This paper is focused on coating treatments of bolts and on their effect on fatigue life. An experimental testing campaign has been performed to investigate the effect of Mn-phosphate coating and micro Zn- phosphate coating on fatigue performances of the bolts, compared with the not coated ones. Particular attention is paid to the surface preparation of the bolt (i.e. by acid pickling or sandblasting) before the final phosphate coating operation. Diffusible and total hydrogen has been measured on all the bolts to verify the possible influence of hydrogen embrittlement.

Quartz is a well-known occupational fibrogenic agent able to cause fibrosis and other severe pulmonary diseases such as silicosis and lung cancer. The silicotic pathology owes its severity to the structural and chemo-physical properties of the particles such as shape, size and abundance of surface radicals. In earlier studies, we reported that significant amounts of surface radicals can be generated on crystalline silica by chemical aggression with ascorbic acid (AA), a vitamin naturally abundant in the lung surfactant, and this reaction led to enhanced cytotoxicity and production of inflammatory mediators in a macrophage cell line. However in the lung, other cells acting in the development of silicosis, like fibroblasts and endothelial cells, can come to direct contact with inhaled quartz. We investigated the cytotoxic/pro-inflammatory effects of AA-treated quartz microcrystals (QA) in human primary fibroblasts and endothelial cells as compared to unmodified microcrystals (Q). Our results show that, in fibroblasts, the abundance of surface radicals on quartz microcrystals (Q vs QA) significantly enhanced cell proliferation (with or without co-culture with macrophages), reactive oxygen species (ROS) production, NF-κB nuclear translocation, smooth muscle actin, fibronectin, Bcl-2 and tissue inhibitor of metalloproteinase-1 expression and collagen production. Contrariwise, endothelial cells reacted to the presence of quartz microcrystals independently from the abundance of surface radicals showing similar levels of cytotoxicity, ROS production, cell migration, MCP-1, ICAM-I and fibronectin gene expression when challenged with Q or QA. In conclusion, our in vitro experimental model demonstrates an important and quite unexplored direct contribute of silica surface radicals to fibroblast proliferation and fibrogenic responses.

Inspired by the fact that nature provides multifunctional composites by using universal building blocks, the authors design and test synthetic composites with a pattern inspired by the microstructure of cortical bone. Using a high-resolution multimaterial 3D printer, the authors are able to manufacture samples and investigate their fracture behavior in mechanical tests. The authors’ results demonstrate that the bone-inspired design is critical for toughness amplification and balance with material strength. The failure modes of the authors’ synthetic composites show similarities with the cortical bone, like crack deflection and branching, constrained microcracking, and fibril bridging. The authors’ results confirm that our design is eligible to reproduce the fracture and toughening mechanism of bone.

Aim of the present work is to study the mechanical behavior in presence of hydrogen of a quenched&tempered low alloy Cr–Mo steel, AISI 4130, used for hydrogen storage and transportation. The sensitivity of steels to hydrogen embrittlement is generally determined by means of two kinds of experimental procedures to hydrogen charge the specimens: gaseous hydrogen in environmental chambers or autoclaves or H2S solution (recommended by NACE standard). In this work experimental tests are carried out on specimens pre-charged by means of an electrochemical method. A campaign of toughness and fatigue crack growth tests is performed on specimens without and with pre-charged hydrogen. The results are compared and discussed with other, from literature, obtained by following more traditional testing procedures. By this comparison the adopted experimental procedure, that is safer and easier, seems to provide useful information.

Composite materials are widely used to build structures for their great mechanical performance combined with a low weight. However, the relatively low toughness of some composite materials is often a limitation as it can cause sudden failure. At present, there is a need for new lightweight materials with a good combination of strength and toughness, to be used for a variety of structural applications. Examples of effective design solutions can be found in natural materials, showing an optimal strength-toughness balance. Such materials can be a good source of inspiration for the design of new smart materials, by following a biomimetic approach. Among natural materials, bone tissue is an intriguing one. Bone combines few meagre constituents, hydroxyapatite and collagen, as building blocks to build up a complex hierarchical structure, reaching remarkable mechanical properties. In particualr, bone has an outstanding toughness, which makes it very attractive for research studies. Indeed, bone toughness is about three to five orders of magnitude more than that of the mineral it is made of. Until now, no conventional synthetic composite material has shown such amplification in toughness, compared to the basic constituents as in bone. For this reason, bone can be considered as a biomimetic model material that many researchers have recently tried to mimic by means of various techniques. In this study, we take inspiration from bone to design and manufacture new composite materials inspired by the microstructure of cortical bone, with the aim of mimicking some toughening mechanisms and improving the toughness of conventional composites. We focus on the microstructural level, since the fundamental toughening mechanisms occur at the microscale, and we mimic the main features involved in the fracture process in our new design. The choice of the key features to be mimicked in the biomimetic material design process is guided by a previous experimental campaign performed on bovine cortical bone. Here we describe the manufacturing process, tailored to get the designed material, and an experimental characterization to assess the mechanical performance and the failure modes. Numerical models have also been used in the design phase to choose the proper biomimetic pattern. The results of the tests allow us to confirm the promising mechanical characteristics of such materials, compared to our previous design solutions and to similar classic structural composites (e.g. laminates). Moreover, the outcome of microscopic observations confirm many similarities with some of the toughening mechanisms occurring in cortical bone, confirming the key role, played by the bone-inspired microstructural features, in determining and enhancing the fracture toughness of the composites. This study represents a successful laboratory example, which could be scaled-up in practical applications, where toughness improvement is necessary.

Ductile 3-layered sandwich panels made of metal cover sheets and a polyolefin core were produced with different layer thicknesses of core and cover sheets in laboratory scale. Because of their lightweight potential, they are interesting for the use in automotive, nautical and even aviation applications. Their impact behaviour is changing, depending on their material and geometrical conditions (quality of the metal and polymer, thickness of the different layers). Therefore, impact tests were performed to investigate this effect, using a steel-polyolefin combination with varying thicknesses of the layers and measuring the post-impact deflections. The results show that all the considered factors influence the panels’ crashworthiness. With the aim of studying the mechanical behaviour of damaged specimens, post-impact tensile tests were performed. Tests were monitored by thermography, a valid technique able to identify the presence of a damaged region in the panels before reaching the yielding stress. A discussion on the obtained experimental results proposes some parameters to identify and quantify the impact damage: the specific deflection and related area, the strain to failure and the damage stress from thermography.

Transgenic mice overexpressing spermine oxidase (SMO) in the cerebral cortex (Dach-SMO mice) showed increased vulnerability to excitotoxic brain injury and kainate-induced epileptic seizures. To investigate the mechanisms by which SMO overexpression leads to increased susceptibility to kainate excitotoxicity and seizure, in the cerebral cortex of Dach-SMO and control mice we assessed markers for astrocyte proliferation and neuron loss, and the ability of kainate to evoke glutamate release from nerve terminals and astrocyte processes. Moreover, we assessed a possible role of astrocytes in an in vitro model of epileptic-like activity in combined cortico-hippocampal slices recorded with a multi-electrode array device. In parallel, as the brain is a major metabolizer of oxygen and yet has relatively feeble protective antioxidant mechanisms, we analyzed the oxidative status of the cerebral cortex of both SMO-overexpressing and control mice by evaluating enzymatic and non-enzymatic scavengers such as metallothioneins. The main findings in the cerebral cortex of Dach-SMO mice as compared to controls are the following: astrocyte activation and neuron loss; increased oxidative stress and activation of defense mechanisms involving both neurons and astrocytes; increased susceptibility to kainate-evoked cortical epileptogenic activity, dependent on astrocyte function; appearance of a glutamate-releasing response to kainate from astrocyte processes due to activation of Ca(2+)-permeable AMPA receptors in Dach-SMO mice. We conclude that reactive astrocytosis and activation of glutamate release from astrocyte processes might contribute, together with increased reactive oxygen species production, to the vulnerability to kainate excitotoxicity in Dach-SMO mice. This mouse model with a deregulated polyamine metabolism would shed light on roles for astrocytes in increasing vulnerability to excitotoxic neuron injury.

Ethnopharmacological relevance Ephedra foeminea is a member of the Ephedraceae family which is widespread in the eastern Mediterranean area. In Lebanon, Ephedra is a popular remedy in traditional medicine to prevent and/or counteract many stress oxidative-related diseases like inflammation and bacterial infections. Aim of the study Oxidative stress leads to endothelial cell dysfunction, and is a major factor contributing to etiology of atherosclerosis and related diseases. This study aims to investigate the antioxidant and cytoprotective potential of extracts from E. foeminea fruits on human endothelial cells exposed to hydrogen peroxide (H2O2) to mimic in vitro vascular endothelium dysfunction. Materials and methods Different extracts of E. foeminea fruits were prepared using pure ethanol (EE), methanol/water (EMW), pure hexane (Ehex) or ethyl acetate/water (Epoly) as extraction solvents. The phenolome profile of each extract was characterized using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS). Total phenolic and flavonoid content, and radical scavenging properties of the extracts were assessed spectrophotometrically. Then, the effects on human endothelial cells HECV were evaluated. Results Epoly extract showed the highest phenol and flavonoid content, and the highest radical scavenging capacity. On H2O2-insulted HECV cells Epoly was able: (i) to counteract the ROS/RNS production and lipid peroxidation; (ii) to rescue the ROS-dependent decrease in the mitochondrial membrane potential; (iii) to counteract the apoptosis induction; (iv) to restore endothelial cell viability and migration. Conclusions The findings indicated that the polyphenol-enriched extract Epoly from E. foeminea fruits is endowed with in vitro anti-oxidant and anti-apoptotic effects and might be used as nutraceutical for treating ROS-related endothelium dysfunction and inflammation.

Natural materials represent ideal biomimetic models for materials design. However, the sophisticated natural hierarchical architectures are rather difficult to be implemented in synthetic FRCs and components through classic manufacturing methods. We propose a new method, called squeeze-winding (SW), specifically designed for the fabrication of FRC-tubes with a bone-inspired internal structure. SW represents a modification to the classic filament winding (FW) and is aimed at implementing a bone-like structure characterized by axially-oriented cylindrical features and achieving a high reinforcement volume fraction (~60%). The outcome of compression testing shows: a high repeatability of the mechanical characteristics achieved by SW, a higher fiber volume fraction of SW tubes compared to the conventional FW tubes, and improved mechanical properties. Moreover, we demonstrate how SW can be used to manufacture a sophisticated bio-inspired design, which mimics the cortical bone microstructure, ensuring enhanced mechanical properties compared to laminated tubes obtained by FW.

Bone quantity and quality are considered the main predictors of bone mechanical properties (i.e., strength and fracture resistance). These factors deal with the morphology and chemical composition of bone and can be assessed by non-invasive techniques such as dual-energy x-ray absorptiometry (DXA), providing the bone mineral density (BMD) and the trabecular bone score (TBS). These parameters, and in particular BMD, are currently used as clinical predictors of fracture risk but do not provide information regarding the fatigue life. Bone is continuously subjected to fatigue loading and fatigue-induced damage can be crucial in fragility fractures. To probe the effect of fatigue-induced damage on bone microarchitecture and elucidate the effect of such damage on the bone clinical parameters, we combined fatigue testing on ex-vivo porcine trabecular bone samples with DXA measurements and μCT imaging. In addition, we performed interrupted cyclic tests at different load levels and measured fatigue-induced damage accumulation in the form of stiffness degradation. We also highlighted the change of clinical and microstructural parameters during the accumulation of fatigue-induced damage in interrupted fatigue tests. Our results suggest that the parameters obtained from the current non-invasive diagnostic protocols (i.e. μCT and DXA) are not able to assess the amount of fatigue-induced damage. This can be due to the fact that such techniques provide global parameters, whereas fatigue-induced damage is a local phenomenon, closely connected to the microarchitecture.