Mesenchymal stem cells-derived exosomes: A new possible therapeutic strategy for Parkinson’s disease?

Parkinson’s disease (PD) is the second most prevalent neurodegenerative disorder worldwide. Clinically, it is characterized by severe motor complications caused by a progressive degeneration of dopaminergic neurons (DAn) and dopamine loss. Current treatment is focused on mitigating the symptoms through administration of levodopa, rather than on preventing DAn damage. Therefore, the use and development of neuroprotective/disease-modifying strategies is an absolute need, which can lead to promising gains on PD translational research. Mesenchymal stem cells (MSCs)–derived exosomes have been proposed as a promising therapeutic tool, since it has been demonstrated that they can act as biological nanoparticles with beneficial effects in different pathological conditions, including PD. Thus, considering their potential protective action in lesioned sites, MSCs-derived exosomes might also be active modulators of the neuroregeneration processes, opening a door for their future use as therapeutical strategies in human clinical trials. Therefore, in this review, we analyze the current understanding of MSCs-derived exosomes as a new possible therapeutic strategy for PD, by providing an overview about the potential role of miRNAs in the cellular and molecular basis of PD.This research was funded by Portuguese Foundation for Science and Technology(FCT): IF Development Grant (IF/00111/2013) to AJ Salgado) and Post-Doctoral Fellowship to F.G. Teixeira (SFRH/BPD/118408/2016). This article has been developed under the scope of the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). This work has been funded by FEDER funds, through the Competitiveness Internationalization Operational Programme (POCI), and by National funds, through FCT, under the scope of the projects POCI-01-0145-FEDER-007038 and POCI-01-0145-FEDER-029751.info:eu-repo/semantics/publishedVersio

Preclinical comparison of stem cells secretome and levodopa application in a 6-hydroxydopamine rat model of Parkinson’s Disease

Parkinson’s Disease (PD) is characterized by the massive loss of dopaminergic neurons, leading to the appearance of several motor impairments. Current pharmacological treatments, such as the use of levodopa, are yet unable to cure the disease. Therefore, there is a need for novel strategies, particularly those that can combine in an integrated manner neuroprotection and neuroregeneration properties. In vitro and in vivo models have recently revealed that the secretome of mesenchymal stem cells (MSCs) holds a promising potential for treating PD, given its effects on neural survival, proliferation, differentiation. In the present study, we aimed to access the impact of human bone marrow MSCs (hBM-MSCs) secretome in 6-hydroxydopamine (6-OHDA) PD model when compared to levodopa administration, by addressing animals’ motor performance, and substantia nigra (SN), and striatum (STR) histological parameters by tyrosine hydroxylase (TH) expression. Results revealed that hBM-MSCs secretome per se appears to be a modulator of the dopaminergic system, enhancing TH-positive cells expression (e.g., dopaminergic neurons) and terminals both in the SN and STR when compared to the untreated group 6-OHDA. Such finding was positively correlated with a significant amelioration of the motor outcomes of 6-OHDA PD animals (assessed by the staircase test). Thus, the present findings support hBM-MSCs secretome administration as a potential therapeutic tool in treating PD, and although we suggest candidate molecules (Trx1, SEMA7A, UCHL1, PEDF, BDNF, Clusterin, SDF-1, CypA, CypB, Cys C, VEGF, DJ-1, Gal-1, GDNF, CDH2, IL-6, HSP27, PRDX1, UBE3A, MMP-2, and GDN) and possible mechanisms of hBM-MSCs secretome-mediated effects, further detailed studies are needed to carefully and clearly define which players may be responsible for its therapeutic actions. By doing so, it will be reasonable to presume that potential treatments that can, per se, or in combination modulate or slow PD may lead to a rational design of new therapeutic or adjuvant strategies for its functional modeling and repair.This work was supported by the European Regional Development Fund (FEDER), through the Competitiveness Internationalization Operational Programme (POCI), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the projects POCI-01-0145-FEDER-029751 and POCI-01-0145-FEDER-007038. This article has also been developed under the scope of the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER)

Innovation of Textiles through Natural By-Products and Wastes

Nowadays, the competitiveness of the textile industry and the consumers’ interest have been increasing the demand for innovative and functional textiles. Allied to this, sustainable developments are playing an increasingly important role in the textile industry. Such concerns led to a new development strategy based on the valorization of bio-based wastes and by-products of different industries, inserting this in the circular economy paradigm. These bio-based wastes and by-products come from several industries, as the agri-food industry. These resources present an enormous potential for valorization in the textile finish due to their intrinsic properties (antimicrobial, prebiotic, antioxidant activity, among others). This chapter will review the latest innovation and textile product development through different by-products and wastes, their main properties and characteristics and the advantages that they offer to the textile industry

Mesenchymal stem cells secretome – derived exosomes: biological nanovesicles for the treatment of Parkinson’s Disease?

Dissertação de mestrado em Ciências da SaúdeA doença de Parkinson (DP) é a segunda doença neurodegenerativa mais comum no mundo. É caracterizada clinicamente por complicações motoras severas causadas pela degeneração progressiva de neurónios dopaminérgicos (DAn) e pelo declínio de dopamina. Os tratamentos atuais focam-se no uso de estratégias farmacológicas como a administração de levodopa, focando se apenas na atenuação dos sintomas motores e não na regeneração dos DAn. Portanto, o desenvolvimento de estratégias regenerativas é essencial, as quais podem levar a ganhos promissores na investigação translacional da DP. O secretoma de células estaminais mesenquimatosas humanas (hMSCs) tem sido proposto como uma ferramenta terapêutica promissora para a DP, dado a sua capacidade de modular a sobrevivência de DAn. Enquanto que com a fração proteica, o nosso laboratório já identificou proteínas promissoras com ações terapêuticas na DP, levando a melhorias histológicas e comportamentais, o potencial da sua fração vesicular/exossomal ainda permanece pouco explorada. As vesículas derivadas das hMSCs são capazes de atuar como nanopartículas biológicas com efeitos benéficos em diferentes condições patológicas, incluindo na DP. Assim sendo, o objetivo deste trabalho foi isolar e caracterizar os exossomas derivados do secretoma de hMSCs, assim como avaliar o seu impacto na sobrevivência, e na diferenciação neuronal através do uso de modelos in vitro e in vivo da DP. Os resultados demonstraram que foi possível o isolamento de exossomas por um protocolo de ultracentrifugação diferencial, ao caracterizar as amostras através da técnica Dynamic Light Scattering (DLS). Adicionalmente, ensaios in vitro, com o objetivo de avaliar a diferenciação neuronal para marcadores DCX e MAP-2, revelaram que a fração exossomal foi capaz de induzir diferenciação de células progenitoras neurais (NPCs) ao mesmo nível que o secretoma total, enquanto que a fração proteica não foi capaz de induzir tal efeito. Num modelo de DP de 6-OHDA, foi possível observar que as diferentes frações do secretoma induziram um efeito positivo na performance motora e na análise histológica, apesar de apresentarem efeitos distintos, indicando assim que o secretoma e as suas diferentes frações podem influenciar diferentes mecanismos e vias. Em suma, é possível concluir que o secretoma de hMSCs e as suas frações podem ser moduladores de diferentes mecanismos de neuroregeneração, abrindo uma oportunidade para o seu uso como estratégia terapêutica no tratamento da DP.Parkinson’s Disease (PD) is the second most prevalent neurodegenerative disorder in the world. It is clinically characterized by severe motor complications caused by the progressive degeneration of dopaminergic neurons (DAn) and dopamine loss. Current treatment focus on mitigating the symptoms through levodopa administration, rather than preventing DAn damage. Therefore, the development of regenerative strategies is essential, which can lead to promising gains on PD translational research. Human mesenchymal stem cells (hMSCs) secretome has been proposed as a promising therapeutic tool, given their ability to modulate DAn survival. While with the protein fraction, our lab has already identified promising proteins with therapeutic actions on PD, leading to histological and behavioral improvements, the potential of its vesicular/exosomal fraction still remains not fully understood. hMSCs – derived vesicles are able to act as biological nanoparticles with beneficial effects in different pathological conditions, including PD. Therefore, the aim of this work was to isolate hMSCs secretome-derived exosomes and characterize its fraction, as well as assess their impact on neuronal survival, and differentiation through the use of in vitro and in in vivo models of PD. Our results have demonstrated that we were able to isolate hMSCs-derived exosomes by a differential ultracentrifugation protocol, when characterizing exosomal samples through Dynamic Light Scattering (DLS) method. Concerning in vitro assays assessing neuronal differentiation to DCX and MAP-2 markers, results revealed that the exosomal fraction was able to induce neural progenitor cells (NPCs) differentiation at the same levels as the whole secretome, while the protein separated fraction was not able to induce such effect. In a 6-OHDA rat model of PD, we have observed that hBM-MSCs secretome and its derived fractions displayed a positive impact on animals’ motor and histological performance, although presenting distinct effects, thereby indicating that the secretome and its different fractions may impact different mechanisms and pathways. Overall, we concluded that the use of the secretome collected from hBM-MSCs and its different fractions might be active modulators of different neuroregeneration mechanisms, opening a door for their future use as therapeutical strategies in the treatment of PD.This work was supported by the Portuguese Foundation for Science and Technology (FCT): IF Development Grant (IF/00111/2013) to AJ Salgado and Post-Doctoral Fellowship to FG Teixeira (SFRH/BPD/118408/2016). This work was funded by FEDER, through the Competitiveness Internationalization Operational Programme (POCI), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the project POCI-01-0145-FEDER-029751. This article has also been developed under the scope of the project NORTE-01-0145-FEDER 000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). This work has been funded by FEDER funds, through the Competitiveness Factors Operational Programme (COMPETE), and by National funds, through FCT, under the scope of the project POCI-01-0145-FEDER-007038

Extracellular matrix-derived materials for tissue engineering and regenerative medicine: A journey from isolation to characterization and application

Biomaterial choice is an essential step during the development tissue engineering and regenerative medicine (TERM) applications. The selected biomaterial must present properties allowing the physiological-like recapitulation of several processes that lead to the reestablishment of homeostatic tissue or organ function. Biomaterials derived from the extracellular matrix (ECM) present many such properties and their use in the field has been steadily increasing. Considering this growing importance, it becomes imperative to provide a comprehensive overview of ECM biomaterials, encompassing their sourcing, processing, and integration into TERM applications. This review compiles the main strategies used to isolate and process ECM-derived biomaterials as well as different techniques used for its characterization, namely biochemical and chemical, physical, morphological, and biological. Lastly, some of their applications in the TERM field are explored and discussed

Impact of aging on the 6-OHDA-induced rat model of Parkinson’s disease

Parkinson’s disease (PD) is the second most common age-related neurodegenerative disorder. The neurodegeneration leading to incapacitating motor abnormalities mainly occurs in the nigrostriatal pathway due to the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Several animal models have been developed not only to better understand the mechanisms underlying neurodegeneration but also to test the potential of emerging disease-modifying therapies. However, despite aging being the main risk factor for developing idiopathic PD, most of the studies do not use aged animals. Therefore, this study aimed at assessing the effect of aging in the unilateral 6-hydroxydopamine (6-OHDA)-induced animal model of PD. For this, female young adult and aged rats received a unilateral injection of 6-OHDA into the medial forebrain bundle. Subsequently, the impact of aging on 6-OHDA-induced effects on animal welfare, motor performance, and nigrostriatal integrity were assessed. The results showed that aging had a negative impact on animal welfare after surgery. Furthermore, 6-OHDA-induced impairments on skilled motor function were significantly higher in aged rats when compared with their younger counterparts. Nigrostriatal histological analysis further revealed an increased 6-OHDA-induced dopaminergic cell loss in the SNpc of aged animals when compared to young animals. Overall, our results demonstrate a higher susceptibility of aged animals to 6-OHDA toxic insult.This research was funded by Prémios Santa Casa Neurociências—Prize Mantero Belard for Neurodegenerative Diseases Research (MB-28-2019); Portuguese Foundation for Science and Technology [Ph.D. fellowship to S.B.-A. (PD/BDE/135568/2018), B.M.-P. (SFRH/BD/120124/2016), A.M. (PDE/BDE/113598/2015), and D.S. (PD/BDE/135567/2018)]. This work was also funded by FEDER, through the Competitiveness Internationalization Operational Programme (POCI), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of the projects POCI-01-0145-FEDER-007038; POCI-01-0145-FEDER-029751 and POCI-01-0145-FEDER-032619. This article has also been developed under the scope of the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER)

Fractionating stem cells secretome for Parkinson's disease modeling: Is it the whole better than the sum of its parts?

Human mesenchymal stem cells (hMSCs) secretome has been have been at the forefront of a new wave of possible therapeutic strategies for central nervous system neurodegenerative disorders, as Parkinson's disease (PD). While within its protein fraction, several promising proteins were already identified with therapeutic properties on PD, the potential of hMSCs-secretome vesicular fraction remains to be elucidated. Such highlighting is important, since hMSCs secretome-derived vesicles can act as biological nanoparticles with beneficial effects in different pathological contexts. Therefore, in this work, we have isolated hMSCs secretome vesicular fraction, and assessed their impact on neuronal survival, and differentiation on human neural progenitors' cells (hNPCs), and in a 6-hydroxydopamine (6-OHDA) rat model of PD when compared to hMSCs secretome (as a whole) and its protein derived fraction. From the results, we have found hMSCs vesicular fraction as polydispersity source of vesicles, which when applied in vitro was able to induce hNPCs differentiation at the same levels as the whole secretome, while the protein separated fraction was not able to induce such effect. In the context of PD, although distinct effects were observed, hMSCs secretome and its derived fractions displayed a positive impact on animals' motor and histological performance, thereby indicating that hMSCs secretome and its different fractions may impact different mechanisms and pathways. Overall, we concluded that the use of the secretome collected from hMSCs and its different fractions might be active modulators of different neuroregeneration mechanisms, which could open new therapeutical opportunities for their future use as a treatment for PD.Prémios Santa Casa Neurociências Prize Mantero Belard for Neurodegenerative Diseases Research (MB-28-2019). This work was supported by the European Regional Development Fund (FEDER), through the Competitiveness Internationalization Operational Programme (POCI), and by National funds, through the Foundation for Science and Technology (FCT), under the scope of projects UIDB/50026/2020; UIDP/50026/2020 and POCI-01-0145-FEDER-029751. This article has also been developed under the scope of the project NORTE-01-0145-FEDER-000023, supported by the Northern Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). This work has been funded by ICVS Scientific Microscopy Platform, member of the national infrastructure PPBI - Portuguese Platform of Bioimaging (PPBI–POCI-01-0145-FEDER-022122)info:eu-repo/semantics/publishedVersio

New self-assembled supramolecular hydrogels based on dehydropeptides

Supramolecular hydrogels rely on small molecules that self-assemble in water as a result of the cooperative effect of several relatively weak intermolecular interactions. Peptide-based low molecular weight hydrogelators have attracted enormous interest owing to the simplicity of small molecules combined with the versatility and biocompatibility of peptides. In this work, naproxen, a well known non-steroidal anti-inflammatory drug, was N-conjugated with various dehydrodipeptides to give aromatic peptide amphiphiles that resist proteolysis. Molecular dynamics simulations were used to obtain insight into the underlying molecular mechanism of self-assembly and to rationalize the design of this type of hydrogelators. The results obtained were in excellent agreement with the experimental observations. Only dehydrodipeptides having at least one aromatic amino acid gave hydrogels. The characterization of the hydrogels was carried out using transmission electron microscopy (TEM), circular dichroism (CD), fluorescence spectroscopy and also rheological assays.Thanks are due to Foundation for Science and Technology (FCT) – Portugal, QREN and program FEDER/COMPETE for financial support through Centre of Chemistry (CQ-UM) of University of Minho. FCT is also acknowledged for PhD grants of G. Pereira (SFRH/BD/38766/2007), H. Vilaça (SFRH/BD/72651/2010) and T. G. Castro (SFRH/BD/79195/2011), co-funded by the European Social Fund. The NMR spectrometer Bruker Avance III 400 is part of the Portuguese NMR Network (Rede/1517/RMN/2005) which is also supported by the FCT

Chapter Innovation of Textiles through Natural By-Products and Wastes

Nowadays, the competitiveness of the textile industry and the consumers’ interest have been increasing the demand for innovative and functional textiles. Allied to this, sustainable developments are playing an increasingly important role in the textile industry. Such concerns led to a new development strategy based on the valorization of bio-based wastes and by-products of different industries, inserting this in the circular economy paradigm. These bio-based wastes and by-products come from several industries, as the agri-food industry. These resources present an enormous potential for valorization in the textile finish due to their intrinsic properties (antimicrobial, prebiotic, antioxidant activity, among others). This chapter will review the latest innovation and textile product development through different by-products and wastes, their main properties and characteristics and the advantages that they offer to the textile industry