Oxygen conditioning effect on an in vitro co-culture model of tendon-to-bone interface

Tendon-to-bone interface comprises a heterotypic cellular niche. The native interface is hypovascular, suggesting that the junction is physiologically hypoxic. As it bridges tendon and bone, which require different oxygen concentrations, a tight coordination of different oxygen concentrations along the junction must be considered when trying to mimic and understand biological events occurring within the tissue. Herein, an optimized in vitro co-culture model of tendon-derived cells (hTDCs) and pre-osteoblasts (pre-OBs) [1] was used to study the effect of a restricted oxygen environment on cell behavior.Hospital da Prelada (Porto, Portugal) for lipoaspirates and tendon samples; ERC Grant CoG MagTendon nr 772817, the Project NORTE-01-0145-FEDER-000021, HORIZON 2020 under the TEAMING Grant agreement No 739572 - The Discoveries CTR and FCT - PhD grant of IC (PB/DB/128088/2016

Optimization and establishment of a co-culture model to study cellular interactions in tendon-to-bone interface

Tendon detachment from its bony insertion is one of the most frequent injuries occurring in the musculoskeletal interface, constituting an unmet challenge in orthopaedics. Tendon-to-bone integration occurs at the enthesis, which is characterized by a complex structure organized in a gradient of cells and microenvironments. Hence, the maintenance of a heterotypic cellular niche is critical for tissue functionality and homeostasis. Replicating this unique complexity constitutes a challenge when addressing tendon-to-bone regeneration and interfacial tissue engineering strategies. Currently, mechanisms presiding to tendon-to-bone interface healing are not yet fully understood, particularly the interactions between tendon and bone cells in the orchestration of interfacial repair versus regeneration. Therefore, this study focused on the hypothesis that interactions between human tendon-derived cells (hTDCs) and pre-osteoblasts (pre-OB) can initiate a cascade of events, potentially leading to interfacial regeneration. Thus, hTDCs and pre-OB (pre-differentiated human adipose-derived stem cells) were used. Herein, five different ratios between basal and osteogenic media (100:0,75:25,50:50,25:75,0:100) were assessed to estimate their influence on cell behaviour and identify the ideal parameters for simultaneously supporting tenogenic and osteogenic differentiation before establishing a co-culture. Tenogenic and osteogenic differentiation were assessed through the expression of tendon and bone markers, mineralization (alizarin red, AZ) and alkaline phosphatase (ALP) quantification. Results showed that hTDCs exhibited osteogenic differentiation potential when cultured in the presence of osteogenic media, as demonstrated by an increase in ALP activity and mineralization. Pre-OB expressed osteogenic markers (OCN, OPN) in all media conditions confirming osteogenic commitment, which was simultaneously confirmed by ALP levels and AZ staining. Thus, three different conditions (100:0, 50:50, 0:100) were chosen for further studies in a direct contact co-culture system. Similarly to single cultures, a significant proliferation was observed in all conditions and mineralization was increased as soon as 7 days of culture. Additionally, osteogenic, tenogenic and interface-relevant markers will be assessed to study the effect of co-culture on phenotype maintenance. In summary, the present work addresses major limitations to clinical translation of cell-based therapies aiming at promoting interfacial regeneration. Particularly, we explored the influence of culture media on the maintenance of tenogenic and osteogenic niches, taking a basic and critical step towards the establishment of more complex cell-based systems. Acknowledgements Authors thank Fundação para a Ciência e Tecnologia in the framework of FCT-POPH-FSE, SFRH/BD/96593/2013 (RCA) and IF/00593/2015 (MEG); and to FCT/MCTES and the FSE/POCH, PD/59/2013 for PD/BD/128088/2016 (IC)Fundação para a Ciência e Tecnologia in the framework of FCT-POPH-FSE, SFRH/BD/96593/2013 (RCA) and IF/00593/2015(MEG); and to FCT/MCTES and the FSE/POCH, PD/59/2013 for PD/BD/128088/2016 (IC)info:eu-repo/semantics/publishedVersio

LSP in theory and practice

UIDB/03213/2020 UIDP/03213/2020Book of abstracts and Program of the 23rd International Conference on Languages for Specific Purposes (LSP 2022: Shaping knowledge through language: LSP in theory and practice), which took place at NOVA University Lisbon, Portugal (CAN – Colégio Almada Negreiros, 1099 – 032 Lisboa).publishersversionpublishe

Platelet lysate cell-laden hydrogel-coated suture threads for tendon repair

Tendon injuries constitute a major healthcare burden owing to the limited healing ability of these tissues and the poor clinical outcomes of surgical repair treatments. Recent advances in tendon tissue engineering (TTE) strategies, particularly through the use of biotextile technologies, hold great promise toward the generation of artificial living tendon constructs. We have previously developed a braided construct based on suture threads coated with gelMA:alginate hydrogel encapsulating human tendon cells. These cell-laden composite fibers enabled the replication of cell and tissue-level properties simultaneously. Based on this concept, in this study we explored the use of platelet lysate (PL), a pool of supra-physiological concentrations of growth factors (GFs), to generate a hydrogel layer, which is envisioned to act as a depot of therapeutic factors to induce tenogenic differentiation of encapsulated human adipose stem cells (hASCs). For this purpose, commercially available suture threads were first embedded in a thrombin solution and then incubated in PL containing hASCs. Herein, thrombin induces the gelation of PL and consequent hydrogel formation. After coating suture threads with the mixture of PL-ASCs, cells were found to be viable and homogeneously distributed along the fibers. Strikingly, hASCs encapsulated within the PL hydrogel layer around the suture thread were able to sense chemotactic factors present in PL and to establish connections between adjacent independent fibers, suggesting a tremendous potential of PL cell-laden hydrogel fibers as building blocks in the development of living constructs aimed at tendon repair applications.info:eu-repo/semantics/publishedVersio

Simulated hypergravity induces changes in human tendon-derived cells: from cell morphology to gene expression

Gravity influences physical and biological processes, having an impact on development, as well as homeostasis of living systems. The musculoskeletal system is comprised of several mechano- responsive tissues and altered gravitational forces are known to influence distinct properties, including bone mineral density and skeletal muscle mass. This is particularly relevant in a near- weightlessness (microgravity) environment, which is found during spaceflight and, not less importantly, during bed resting. Over the years, several studies have been conducted under simulated conditions of altered gravity owing to the advances on ground-based facilities, such as bioreactors for microgravity / hypo-gravity (1g) studies. Interestingly, microgravity-induced alterations are comparable to tissue degeneration caused by disuse and ageing. In turn, exposing musculoskeletal tissues to hypergravity may constitute a way of simulating (over)loading or, eventually, to be used as a measure to rescue cell phenotype after exposure to near-weightlessness conditions. Different studies have focused on bone, cartilage and skeletal muscle, but effects on tendons and ligaments have been underappreciated. Therefore, we evaluated the influence of increasing g-levels (5g, 10g, 15g and 20g) and different hypergravity exposure periods (4 and 16 h) on the behaviour of human tendon- derived cells (hTDCs). For this purpose, hTDCs were exposed to simulated hypergravity conditions using the Large Diameter Centrifuge (LDC) from the European Space Research and Technology Centre (ESTEC, ESA, The Netherlands). Human TDCs cultured under standard conditions (1g, normogravity, Earth gravity force) were used as controls. The effects of hypergravity on the viability of hTDCs, as well as on the expression of tendon related markers at the gene level were evaluated. Simulated hypergravity resulted in a reduced cell content after 16 h independently of g-level, as determined by DNA quantification. Additionally, the different g-levels studied led to changes in cell and cytoskeleton morphology. Strikingly, a 16-hour period of exposure resulted in alterations of gene expression profiles. Overall, gene expression of tendon-related markers, including collagen types I (col1a1) and III (col3a1), scleraxis (scx), tenomodulin (tnmd), decorin (dcn) and tenascin (tnc), seemed to be increased upon hypergravity stimulation and in comparison to cells cultured under control conditions. Altogether, these results highlight that altered gravity, particularly simulated hypergravity, has an influence on the phenotype of tendon cells, opening new avenues for research focused on using altered gravity as a model for overloading-induced tendon tissue injury or as measure to rescue the phenotype of degenerated tendon cells. Acknowledgements The authors would like to thank ESA Education Office for Spin Your Thesis! 2016 programme. R.C-A acknowledges the PhD grant SFRH/BD/96593/2013 from FCT â Fundação para a Ciência e a Tecnologia. SFRH/BD/96593/2013 from FCT –Fundação para a Ciência e a Tecnologiainfo:eu-repo/semantics/publishedVersio

Human-based nanocomposite cryogels for hemostatic and wound healing applications

In trauma surgery, a fast and effective hemostatic agent is crucial to prevent death. The current used hemostatic sponges are highly effective in stopping the hemorrhages, however they have a limited stability, shape memory, and biological functionality to induce an efficient regenerative healing after injury. Blood derivatives have attracted great attention as an inexpensive milieu of bioactive molecules (e.g., growth factors, cytokines), self-assembling scaffolding proteins (e.g., fibrinogen, fibronectin, vitronectin), and antimicrobial peptides (e.g., platelet factor-4) that have the ability to enhance angiogenesis, stem cell recruitment, and tissue regeneration. Among those, platelet lysate (PL) has attracted great attention as a milieu of supra-physiological doses of biomolecules that can be easily standardized. However, the current PL scaffolds showed limited stability and weak mechanical strength, which severely limits its performance as a bioinstructive and hemostatic biomaterial. Herein, we propose the use of aldehyde-functionalized CNC (a-CNC) that will be crosslinked through reversible Schiff base bonds established with the amine groups of PL proteins to produce a stable hemostatic cryogel for wound healing applications.EU’s H2020 programme for grant agreement 706996 and 739572 - The Discoveries CTR; FCT for SFRH/BPD/112459/2015, PD/BD/113807/2015, FOOD4CELLS (PTDC/CTM-BIO/4706/2014-POCI-01- 0145- FEDER 016716) and project NORTE-01-0145- FEDER-000021

Patient-physician discordance in assessment of adherence to inhaled controller medication: a cross-sectional analysis of two cohorts

We aimed to compare patient's and physician's ratings of inhaled medication adherence and to identify predictors of patient-physician discordance.(SFRH/BPD/115169/2016) funded by Fundação para a Ciência e Tecnologia (FCT); ERDF (European Regional Development Fund) through the operations: POCI-01-0145-FEDER-029130 ('mINSPIRERS—mHealth to measure and improve adherence to medication in chronic obstructive respiratory diseases—generalisation and evaluation of gamification, peer support and advanced image processing technologies') cofunded by the COMPETE2020 (Programa Operacional Competitividade e Internacionalização), Portugal 2020 and by Portuguese Funds through FCT (Fundação para a Ciência e a Tecnologia).info:eu-repo/semantics/publishedVersio

Morphological and Postural changes in the foot during pregnancy and puerperium : a longitudinal study

The aim of this study is to observe the morphological and postural changes to the foot that take place during pregnancy and the puerperium. Method: In this descriptive, observational, longitudinal study, we analysed 23 pregnant women, with particular attention to morphological and postural aspects of the foot, at three time points during and after pregnancy: in weeks 9-13 of gestation, weeks 32-35 of gestation and weeks 4-6 after delivery. The parameters considered were changes in foot length, the Foot Posture Index (FPI) and the Hernández Corvo Index, which were analysed using a pedigraph and taking into account the Body Mass Index (BMI). The same procedure was conducted in each review. Results: The statistical analyses obtained for each foot did not differ significantly between the three measurement times. A pronator-type footprint was most frequently observed during the third trimester of pregnancy; it was predominantly neutral during the postpartum period. Statistically significant differences between the measurement times were obtained in the right foot for cavus vs. neutral foot type (between the first and third trimesters and also between the first trimester and the puerperium) (in both cases, p < 0.0001). Conclusions: Foot length increases in the third trimester and returns to normal in the puerperium. According to FPI findings, the third trimester of pregnancy is characterised by pronation, while the posture returns to neutrality during the postpartum period. During pregnancy, the plantar arch flattens, and this persists during the puerperium. The incidence of cavus foot increases significantly in the third trimester and in the puerperium

A ATEROSCLEROSE E O INFARTO AGUDO DO MIOCÁRDIO

Atherosclerosis is a slow and progressive disease caused by the inflammatory response from the presence and accumulation of fatty plaques, especially low-density lipoprotein (LDL) in the vascular endothelium, which can result in obstruction and reduction of vessel lumen. Among these vessels, the main ones are the coronary arteries, responsible for relating the presence of atherosclerosis with Acute Myocardial Infarction (AMI). Objective: This literature review aims to relate the presence of atherosterotic plaque with the development of AMI. Methodology: This is a literature review whose articles were selected from the LILACs, SciELO and Google Scholar databases and Brazilian guideline on dyslipidemias and prevention of atherosclerosis in Portuguese, Spanish and English with the following descriptors: dyslipidemia, coronary vessels, Myocardial infarction and atherosclerosis. Final considerations: Atherosclerosis, a multifactorial and progressive disease, is related to hyperlipidemia, obesity and hypertension, and its morbidity and mortality are related to the high level of cardiovascular diseases present worldwide. There are risk factors that can be changed, such as dietary reeducation to reduce cholesterol levels and medical support for lipid and cardiovascular monitoring and lifestyle changes and non-modifiable factors such as age.La aterosclerosis es una enfermedad lenta y progresiva causada por la respuesta inflamatoria de la presencia y acumulación de placas grasas, especialmente lipoproteínas de baja densidad (LDL) en el endotelio vascular, lo que puede resultar en obstrucción y reducción de la luz del vaso. Entre estos vasos, los principales son las arterias coronarias, encargadas de relacionar la presencia de aterosclerosis con el Infarto Agudo de Miocardio (IAM). Objetivo: Esta revisión bibliográfica pretende relacionar la presencia de placa aterosterótica con el desarrollo de IAM. Metodología: Se trata de una revisión bibliográfica cuyos artículos fueron seleccionados de las bases de datos LILACs, SciELO y Google Scholar y de la guía brasileña sobre dislipidemias y prevención de la aterosclerosis en portugués, español e inglés con los siguientes descriptores: dislipidemia, vasos coronarios, infarto de miocardio y aterosclerosis. Consideraciones finales: La aterosclerosis, una enfermedad multifactorial y progresiva, está relacionada con la hiperlipidemia, la obesidad y la hipertensión, y su morbilidad y mortalidad están relacionadas con el alto nivel de enfermedades cardiovasculares presentes en todo el mundo. Hay factores de riesgo que se pueden cambiar, como la reeducación dietética para reducir los niveles de colesterol y el apoyo médico para el control lipídico y cardiovascular y cambios en el estilo de vida y factores no modificables como la edad.A aterosclerose é uma doença lenta e progressiva causada pela resposta inflamatória a partir da presença e acúmulo de placas gordurosas, em especial da lipoproteína de baixa densidade (LDL) no endotélio vascular que podem resultar na obstrução e redução da luz do vaso. Dentre esses vasos, os principais são as artérias coronárias, responsável por relacionar a presença da aterosclerose com o Infarto Agudo do Miocárdio (IAM). Objetivo: Essa revisão bibliográfica objetiva relacionar a presença da placa aterosterótica com o desenvolvimento do IAM. Metodologia: Trata-se de uma revisão bibliográfica cujos artigos foram selecionados das bases de dados do LILACs, SciELO e Google Acadêmico e Diretriz brasileira de dislipidemias e prevenção da aterosclerose nas línguas portuguesa, espanhola e inglesa com os seguintes descritores: dislipidemia, vasos coronários, Infarto do Miocárdio e Aterosclerose. Considerações finais: A aterosclerose, doença multifatorial e progressiva, é relacionada à hiperlipidemia, obesidade e hipertensão, e tem sua morbidade e mortalidade relacionada ao alto nível de doenças cardiovasculares presentes a nível mundial. Há fatores de risco passíveis de mudanças, como reeducação alimentar a fim de reduzir níveis de colesterol e apoio médico para monitoramento lipídico e cardiovascular e mudanças no estilo de vida e fatores não modificáveis, como a idade.A aterosclerose é uma doença lenta e progressiva causada pela resposta inflamatória a partir da presença e acúmulo de placas gordurosas, em especial da lipoproteína de baixa densidade (LDL) no endotélio vascular que podem resultar na obstrução e redução da luz do vaso. Dentre esses vasos, os principais são as artérias coronárias, responsável por relacionar a presença da aterosclerose com o Infarto Agudo do Miocárdio (IAM). Objetivo: Essa revisão bibliográfica objetiva relacionar a presença da placa aterosterótica com o desenvolvimento do IAM. Metodologia: Trata-se de uma revisão bibliográfica cujos artigos foram selecionados das bases de dados do LILACs, SciELO e Google Acadêmico e Diretriz brasileira de dislipidemias e prevenção da aterosclerose nas línguas portuguesa, espanhola e inglesa com os seguintes descritores: dislipidemia, vasos coronários, Infarto do Miocárdio e Aterosclerose. Considerações finais: A aterosclerose, doença multifatorial e progressiva, é relacionada à hiperlipidemia, obesidade e hipertensão, e tem sua morbidade e mortalidade relacionada ao alto nível de doenças cardiovasculares presentes a nível mundial. Há fatores de risco passíveis de mudanças, como reeducação alimentar a fim de reduzir níveis de colesterol e apoio médico para monitoramento lipídico e cardiovascular e mudanças no estilo de vida e fatores não modificáveis, como a idade