Browsing by Author "Fernandes, Ana"
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- Design and operation of a fully controlled platform for the production and purification of well-defined mesenchymal stromal cell (MSC)-derived extracellular vesiclesPublication . Fernandes, Ana; Rosa, Sara; Silva, Ricardo; Cunha, Raquel; Fuzeta, Miguel Almeida; Calado, Cecília; Carvalho, Carla; Cabral, Joaquim M.S.; Azevedo, AnaCurrently, the production of extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSC) is performed in laborious and time-consuming static culture systems using serum-containing media, and their purification is through ultracentrifugation based methods. The whole process lacks on resolution, selectivity and capacity. Additionally, several differences were observed, in terms of the cargo of EVs, between EVs isolated from culture supernatants of MSC expanded under different culture conditions, stressing the importance of controlling all culture process parameters. A robust and indepth process to manufacture MSC-EVs may provide a new therapeutic paradigm for cell-free MSC-based therapies.
- Development of scalable processes to produce therapeutic mesenchymal stromal cell-derived extracellular vesicles and their characterizationPublication . Cunha, Raquel M. S.; Vargas, Elga; Pires, Filipa; Calado, Cecília; Cabral, Joaquim M.S.; Silva, Cláudia; Fernandes, AnaIn summary, this study contributes to the establishment of a scalable process for MSC-EV production.
- Evaluating the impact of culture conditions on human mesenchymal stromal cell-derived extracellular vesicles molecular fingerprint through FTIR spectroscopyPublication . Fernandes, Ana; Pereira, Maria; Ramalhete, Luís; Aleixo, Sandra; Silva, Cláudia; Cabral, Joaquim M.S.; Calado, CecíliaThis work is a step forward into understanding how different culture conditions affect MSC-derived EVs characteristics.
- Impact of the human mesenchymal stem cells donor on conditional medium compositionPublication . Pereira, Maria João Canha; Ramalhete, Luís; Aleixo, Sandra; da Silva, Cláudia L.; Cabral, Joaquim M.S.; Calado, Cecília; Fernandes, AnaExosomes produced by Mesenchymal Stem Cells (MSCs) can represent a very appealing strategy for cell-free therapies. However, to achieve this reality it is necessary to further understand the process associated to the MSC culture when conditioned to produce exosomes. In the present work, it was evaluated how different MSC obtained from different donors may affect the conditioned media composition and how this can be influenced by the conditioned media type (DMEM versus Xeno-Free medium, XF). The molecular fingerprint of the conditioned media composition was obtained by mid-infrared (MIR) spectroscopy, as this technique reflects fundamental vibrations of a high diversity of functional chemical groups present in biological samples. It was observed by principal component analysis of the second derivative spectra of conditioned media that the media chemical composition depends more from the MSCs donor than the conditioning days. Diverse spectral regions, characteristic of defined chemical groups, enabled to discriminate the chemical composition of the media according to the MSC donor. All of this was observed in both types of media (DMEM and XF). This work is a step forward to understand how different MSC donors and conditioned media may affect the exosomes characteristics.
- Scaling-up the manufacturing of well-characterized mesenchymal stromal cell-derived extracellular vesicles for biomedical applicationsPublication . Fernandes, Ana; Rosa, Sara; Silva, Ricardo; Cunha, Raquel M. S.; Fuzeta, Miguel de Almeida; Calado, Cecília; Carvalho, Carla; Cabral, Joaquim M.S.; Azevedo, Ana; Silva, CláudiaThe platform established herein could be applied to the production of well-characterized SPC-EVs targeting their biomedical use in different settings (e.g. as drug delivery systems), as well as EVs from other parental cells lines (i.e. dendritic cells) in therapeutic settings as cancer.
- Treino propriocetivo como estratégia de intervenção para melhoria do equilíbrio em indivíduos pós-acidente vascular cerebral: uma revisão sistemáticaPublication . Fernandes, Ana; Fernandes, Sofia Rita; Fernandes, BeatrizIntrodução – As alterações no equilíbrio correspondem a mais de metade das consequências provocadas pelo AVC, por isso, continua a ser de extrema importância investigar qual a melhor estratégia de intervenção nestas alterações de acordo com a atual evidência. Apesar de o treino propriocetivo ser considerado um treino válido em indivíduos pós-AVC, o seu contributo para a melhoria do equilíbrio nestes indivíduos permanece por esclarecer. Objetivo – Verificar a eficácia do treino propriocetivo na melhoria do equilíbrio em indivíduos pós-AVC. Métodos – Foi efetuada uma revisão sistemática seguindo as recomendações PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis). A pesquisa foi realizada nas bases de dados PubMed, PEDro e ScienceDirect, utilizando termos de pesquisa específicos. A Escala PEDro foi utilizada para avaliar a qualidade metodológica dos estudos. Critérios de inclusão: estudos randomizados controlados (RCT) que incluíssem indivíduos com idade igual ou superior a 18 anos, com diagnóstico de AVC e alterações no equilíbrio, cuja intervenção contemplasse treino propriocetivo e a avaliação tivesse medidas objetivas do equilíbrio. Critérios de exclusão: estudos em indivíduos com outras morbilidades neurológicas, com patologia músculo-esquelética nos membros inferiores ou outro tipo de patologias suscetíveis de influenciar o sistema propriocetivo. Resultados – A pesquisa identificou um total de 855 estudos, dos quais 14 foram incluídos na revisão após a aplicação dos critérios de elegibilidade. Estes envolveram 677 indivíduos com diagnóstico de AVC crónico (sete estudos) e agudo/subagudo (sete estudos). Os tipos de treino propriocetivo utilizados nos grupos de intervenção (GI) consistiram em: equilíbrio; estimulação somatossensorial com vibração ou estimulação elétrica funcional; reposicionamento articular, combinado com fisioterapia convencional, aplicada também nos grupos de controlo (GC). As medidas de avaliação objetiva do equilíbrio utilizadas consistiram nas escalas de equilíbrio de Berg (BBS) e/ou Tinneti (TBS) e Mini-Balance Evaluation Systems Test (Mini-Best). Houve diferenças estatisticamente significativas (p<0,05) entre GI e GC em 10 dos 14 estudos. Conclusão – De acordo com a presente revisão, apesar da heterogeneidade das intervenções (tipo de exercício e duração) e os instrumentos de avaliação usados na análise dos resultados, regista-se uma tendência para a maioria dos estudos incluídos apresentarem resultados positivos nas medidas que avaliam o equilíbrio e o treino propriocetivo.
- Wasteful azo dyes as a source of biologically active building blocksPublication . Fernandes, Ana; Pinto, Bruna; Bonardo, Lorenzo; Royo, Beatriz; Robalo, M. Paula; Martins, Lígia O.In this work, an environment-friendly enzymatic strategy was developed for the valorisation of dye-containing wastewaters. We set up biocatalytic processes for the conversion of azo dyes representative of the main classes used in the textile industry into valuable aromatic compounds: aromatic amines, phenoxazinones, phenazines, and naphthoquinones. First, purified preparations of PpAzoR azoreductase efficiently reduced mordant, acid, reactive, and direct azo dyes into aromatic amines, and CotA-laccase oxidised these compounds into phenazines, phenoxazinones, and naphthoquinones. Second, whole cells containing the overproduced enzymes were utilised in the two-step enzymatic conversion of the model mordant black 9 dye into sodium 2-amino-3-oxo-3H-phenoxazine-8-sulphonate, allowing to overcome the drawbacks associated with the use of expensive purified enzymes, co-factors, or exquisite reaction conditions. Third, cells immobilised in sodium alginate allowed recycling the biocatalysts and achieving very good to excellent final phenoxazine product yields (up to 80%) in water and with less impurities in the final reaction mixtures. Finally, one-pot systems using recycled immobilised cells co-producing both enzymes resulted in the highest phenoxazinone yields (90%) through the sequential use of static and stirring conditions, controlling the oxygenation of reaction mixtures and the successive activity of azoreductase (anaerobic) and laccase (aerobic).