IPSCs, a promising tool to restore muscle atrophy

Induced pluripotent stem cells (iPSCs) are a promising tool for regenerative medicine in chronic conditions associated with muscle atrophy since iPSCs are easier to obtain, pose less ethical limitations and can better capture human genetic diversity compared with human embryonic stem cells. We highlight the potentiality of iPSCs for treating muscle-affecting conditions for which no effective cure is yet available, notably aging sarcopenia and inherited neurometabolic conditions

New insight on obesity and adipose-derived stem cells by comprehensive metabolomics

Obesity affects the functional capability of adipose-derived stem cells (ASCs) and their effective use in regenerative medicine through mechanisms still poorly understood. Here we employed a multiplatform (LC/MS, CE/MS, GC/MS) metabolomics untargeted approach to investigate the metabolic alteration underlying the inequalities observed in obese-derived ASCs. The metabolic fingerprint (metabolites within the cells) and footprint (metabolites secreted in the culture medium) from humans or mice, obese and non-obese derived ASCs, were characterized by providing valuable information. Metabolites associated to glycolysis, TCA, pentose phosphate pathway and polyol pathway were increased in the footprint of obese-derived human ASCs indicating alterations in the carbohydrate metabolism; whereas from the murine model, deep differences in lipid and amino acid catabolism were highlighted. Therefore, new insights on the ASCs metabolome were provided that enhance our understanding of the processes underlying the ASCs stemness capacity and its relationship with obesity, in different cell models.

Método LNEC para avaliação e classificação da qualidade acústica de edifícios habitacionais: ferramenta de estimação de custos médios de transição entre classes acústicas

O conforto acústico na habitação é essencial para permitir um repouso tranquilo e regenerativo. Este conforto é obtido principalmente pela redução do ruído e aumento do isolamento sonoro dos elementos de compartimentação. Para a qualificação deste conforto é necessário efetuar uma análise global do edifício, onde são considerados os fatores internos e externos à habitação, ou seja, considerando a acústica da envolvente (Vizinhança), do edifício (Edifício) e da fração (Habitação). Para este efeito foi produzido o “Método LNEC para avaliação e classificação da qualidade acústica de edifícios habitacionais”, o qual permite fazer uma avaliação global do conforto acústico na habitação. Este método inovador em Portugal origina uma Classe Acústica LNEC que permite representar com razoável fiabilidade o conforto acústico realmente sentido e a qualidade acústica da habitação. Este método pode ser aplicado a edifícios novos e a edifícios a reabilitar. De modo a permitir uma estimação dos custos médios necessários para transitar entre determinadas classes (e alcançar respetivo conforto acústico) é necessário possuir uma ferramenta de cálculo apropriada. Deste modo apresenta-se nesta comunicação uma ferramenta para estimar os custos de transição entre classes acústicas. Esta metodologia permite fazer escolhas mais fundamentadas nos processos de obtenção de determinado conforto acústico. Abstract The acoustic comfort in dwellings is essential to a peaceful and regenerative sleep. The achievement of this comfort is primarily obtained by reducing noise and increasing sound insulation of separating elements. To classify this comfort is necessary to conduct a comprehensive analysis of the habitation. This global analysis assesses the internal and external factors of the habitation, evaluating the acoustics of the environment (Vicinity), of the building (Building) and of the dwelling place (Lodging). Recently in Portugal was developed the “LNEC method for evaluation and acoustic quality classification of residential buildings” that allows an overall evaluation of the acoustic comfort in dwellings, resulting also in a “LNEC Acoustic Class” that portrays the real acoustic comfort sensed. Since this method can be applied to evaluate the acoustic comfort of new and of restored buildings, it is necessary a tool that gives an estimation of the needed investment to upgrade to a specific “LNEC Acoustic Class” (and achieve the respective acoustic comfort). In this communication is presented a tool that allows the estimation of that upgrade costs.

Role of nuclear redox control, intra-population heterogeneity and oxygen tension in human amniotic fluid stem cells aging

Amniotic fluid stem cells (hAFSC) are emerging as a potential therapeutic approach for various disorders. The low number of available hAFSC requires their ex vivo expansion prior to clinical use, however, during their in vitro culture, hAFSC quickly reach replicative senescence. The principal aim of this study was to investigate the aging process occurring during in vitro expansion of hAFSC, focusing on the redox control that has been reported to be affected in premature and physiological aging. My results show that a strong heterogeneity is present among samples that reflects their different behaviour in culture. I identified three proteins, namely Nox4, prelamin A and PML, which expression increases during hAFSC aging process and could be used as new biomarkers to screen the samples. Furthermore, I found that Nox4 degradation is regulated by sumoylation via proteasome and involves interactions with PML bodies and prelamin A. Since various studies revealed that donor-dependent differences could be explained by cell-to-cell variation within each patient, I studied in deep this phenomenon. I showed that the heterogeneity among samples is also accompanied by a strong intra-population heterogeneity. Separation of hAFSC subpopulations from the same donor, using Celector® technology, showed that an enrichment in the last eluted fraction could improve hAFSC application in regenerative medicine. One of the other problems is that nowadays hAFSC are expanded under atmospheric O2 concentration, which is higher than the O2 tension in their natural niches. This higher O2 concentration might cause environmental stress to the in vitro cultured hAFSCs and accelerate their aging process. Here, I showed that prolonged low oxygen tension exposure preserves different hAFSC stemness properties. In conclusion, my study pointed different approaches to improve in vitro hAFSC expansion and manipulation with the purpose to land at stem cell therapy.

A preclinical study of spinal cord injury focused on cellular and molecular modifications as potential targets for innovative therapies

Spinal Cord Injury (SCI) is a devastating condition for human and animal health. In SCI particularly, neurons, oligodendrocytes precursor cells, and mature oligodendrocytes are highly vulnerable to the toxic microenvironment after the lesion and susceptible to the elevated levels of noxious stimuli. Thus the regenerative response of the organism in case of SCI is significantly reduced, and only little spontaneous amelioration is observed in lesioned patients during the early phases. This work mainly focuses on studying and characterizing the modification induced by the SCI in a preclinical animal model. We investigated the ECM composition in the spinal cord segments surrounding the primary lesion site at a gene expression level. We found Timp1 and CD44 as a crucial hub in the secondary cascade of SCI in both spinal cord segments surrounding the lesion site. Interestingly, a temporal and anatomical difference in gene expression, indicating a complex regulation of ECM genes after SCI that could be used as a tool for regenerative medicine. We also investigated the modification in synaptic plasticity-related gene expression in spinal and supraspinal areas involved in motor control. We confirmed the anatomical and temporal difference in gene expression in spinal cord tissue. This analysis suggests that a molecular mapping of the lesion-induced modification could be a useful tool for regenerative medicine. In the last part, we evaluated the efficacy of an implantable biopolymer loaded with an anti-inflammatory drug and a pro-myelinating agent on the acute phase of SCI in our preclinical model. We found a consistent reduction of the inflammatory state in the spinal lesion site and the cord's surrounding segments. Moreover, we found increased preservation of the spinal cord tissue with a related upregulation of neuronal and oligodendroglial markers after lesion. Our treatment showed effective ameliorating functional outcome and reducing the lesion extension in the chronic phase.

Applied biomaterials from sustainable sources

If we look back in time at the history of humanity, we can state that our generation is living an era of outstanding efficiency and progress because of globalization and global competition, even if this is resulting in the rapid depletion of energy sources and raw materials. The environmental impact of non-biodegradable plastic wastes is of increasing global concern: nowadays, imagining a world without synthetic plastics seems impossible, though their large-scale production and their extensive use have only spread since the end of the World War II. In recent years, the demand for sustainable materials has increased significantly and, with a view to circular economy, research has also focused on the enhancement and subsequent reuse of waste materials produced by industrial processing, intensive farming and the agricultural sector. Plastic polymers have been the most practical and economical solution for decades due to their low cost, prompt availability and excellent optical, mechanical and barrier properties. Biodegradable polymers could replace them in many applications, thus reducing the problems of traditional plastics disposability and the dependence on petroleum. Natural biopolymers are in fact characterized by a high biocompatibility and biodegradability and have already prompted research in the field of regenerative medicine. During my PhD, my goal was to use natural polymers from sustainable sources as raw materials to produce biomaterials, which are materials designed to interface with biological systems to evaluate, support or replace any tissue, organ, or function of the body. I focused on the use of the most abundant biopolymers in nature to produce biomaterials in the form of films, scaffolds and cements. After a complete characterization, the materials were proposed for suitable applications in different fields, from tissue engineering to cosmetics and food packaging. Some of the obtained results were published on international scientific and peer-reviewed journals.

3D Printing and bioprinting of osteomimetic materials for 3D in vitro models and tissue repair applications

Bone disorders have severe impact on body functions and quality life, and no satisfying therapies exist yet. The current models for bone disease study are scarcely predictive and the options existing for therapy fail for complex systems. To mimic and/or restore bone, 3D printing/bioprinting allows the creation of 3D structures with different materials compositions, properties, and designs. In this study, 3D printing/bioprinting has been explored for (i) 3D in vitro tumor models and (ii) regenerative medicine. Tumor models have been developed by investigating different bioinks (i.e., alginate, modified gelatin) enriched by hydroxyapatite nanoparticles to increase printing fidelity and increase biomimicry level, thus mimicking the organic and inorganic phase of bone. High Saos-2 cell viability was obtained, and the promotion of spheroids clusters as occurring in vivo was observed. To develop new syntethic bone grafts, two approaches have been explored. In the first, novel magnesium-phosphate scaffolds have been investigated by extrusion-based 3D printing for spinal fusion. 3D printing process and parameters have been optimized to obtain custom-shaped structures, with competent mechanical properties. The 3D printed structures have been combined to alginate porous structures created by a novel ice-templating technique, to be loaded by antibiotic drug to address infection prevention. Promising results in terms of planktonic growth inhibition was obtained. In the second strategy, marine waste precursors have been considered for the conversion in biogenic HA by using a mild-wet conversion method with different parameters. The HA/carbonate ratio conversion efficacy was analysed for each precursor (by FTIR and SEM), and the best conditions were combined to alginate to develop a composite structure. The composite paste was successfully employed in custom-modified 3D printer for the obtainment of 3D printed stable scaffolds. In conclusion, the osteomimetic materials developed in this study for bone models and synthetic grafts are promising in bone field.

Implementazione di un sistema di controllo atto a ottimizzare l’efficienza di convertitori DC-DC impiegati in ambito Automotive e realizzati con tecnologia GaN

Lo scopo del presente progetto di ricerca è quello di dimostrare quelli che potrebbero essere i miglioramenti nell’adottare tecnologie GaN nei convertitori di potenza, soffermandosi sull’ambito Automotive. Si è dunque progettato, simulato ed infine analizzato un sistema di conversione bidirezionale DC-DC. Le prestazioni ottime sono state poi raggiunte attraverso l’introduzione di un sistema di controllo in grado di garantire, in ogni condizione, il miglior funzionamento possibile che garantisca il soddisfacimento delle richieste da parte del veicolo. Questo consentirà quindi di gestire il funzionamento del convertitore sia nelle fasi di scarica che in quelle di rigenerazione (regenerative braking). Al convertitore, è stato introdotto anche un controllo adattivo per la ricerca del dead-time ottimo dei segnali di pilotaggio dei transistor.