Establishing guidelines for obtaining optimal cell sources to use in tendon tissue engineering strategies

Cell-based approaches in tissue engineering (TE) have been barely explored for the treatment of tendon and ligament (T/L) tissues, requiring the establishment of a widely available cell source with tenogenic potential. As T/L cells are scarce, stem cells may provide a good alternative. Understanding how resident cells behave in vitro, might be useful for recapitulating the tenogenic potential of stem cells for tendon TE applications. Therefore, we propose to isolate and characterize human T/L-derived cells (hTDCs and hLDCs) and compare their regenerative potential with stem cells from adipose tissue (hASCs) and amniotic fluid (hAFSCs)(1). T/L cells were isolated using different procedures and stem cells isolated as described elsewhere(1). Moreover, T/L cells were stimu- lated into the three mesenchymal lineages, using standard differentia- tion media. Cells were characterized for the typical stem cell markers as well as T/L related markers, namely tenascin-C, collagen I and III, decorin and scleraxis, using different complementary techniques such as real time RT-PCR, immunocytochemistry and flow cytometry. No differences were observed between T/L in gene expression and protein deposition. T/L cells were mostly positive for stem ness markers (CD73/CD90/CD105), and have the potential to differentiate towards osteogenesis, chondrogenesis and adipogenesis, demonstrated by the positive staining for AlizarinRed, SafraninO, ToluidineBlue and OilRed. hASCs and hAFSCs exhibit positive expression of all tenogenic mark- ers, although at lower levels than hTDCs and hLDCs. Nevertheless, stem cells availability is key factor in TE strategies, despite that it’s still required optimization to direct their tenogenic phenotype.

Advancing tendon regeneration through the development of bio-stimulating tissue engineering approaches

Tendon tissue engineering (TE) requires tailoring scaffolds designs and properties to the anatomical and functional requirements of tendons located in different regions of the body. Cell sourcing is also of utmost importance as tendon cells are scarce. Recently, we have found that it is possible to direct the tenogenic differentiation of Amniotic fluid and Adipose tissue derived stem cells (hAFSCs and hASCs), and also that there are hASCs subpopulations that might be more prone to tenogenic differentiation. Nevertheless, biochemical stimulation may not be enough to develop functional TE substitutes for a tissue that is known to be highly dependent on mechanical loading.

Levels of leaf Herbivory in Amazonian trees from different stages In forest regeneration

Leaves from 120 canopy trees and 60 understory tree saplings growing in primary and secondary forests near Manaus, Brazil, were collected for determination of standing levels of herbivory (percent leaf area lost). Overall, levels of herbivory on leaves of central Amazonian trees were low. About one quarter of the leaves examined (n = 855) had no damage at all. In most other Neotropical sites studied the mean percentage of herbivory was found to vary between 5.7 and 13.1%, whereas in Manaus it was only 3.1%. The data presented here support the contention that levels of herbivore damage are positively related to soil fertility. No significant difference was found in herbivory levels between canopy trees and understory saplings. Also, there was no difference in damage between leaves from pioneer and late successional trees. Field assays of preference, however, revealed that leaves from pioneer trees are more palatable to leaf-cutting ants (Atta laevigata). This effect was dependent upon leaf age, being observed in mature leaves, but not in young leaves. The greater rate of leaf production in secondary forests may be a factor accounting for the greater abundance of leaf-cutting ants in secondary compared to primary forests.

Strategies of the small-scale fisheries on the Central Amazon floodplain

A multivariate approach was applied to data of small-scale fisheries developed in Central Amazon, using information about catch composition, environment, fishing gear and season of the hydrological cycle. The correspondence analysis demonstrated to be a good tool for the analysis related multispecies fisheries. The analysis identified patterns of use of fisheries resources by the riverine communities, showing the correlation between the environmental factors and the fishing strategy for the capture of target fish species, indicating the high level of empiric knowledge about the environment and fisheries.

Contribution of portuguese vernacular building strategies to indoor thermal comfort and occupants’ perception

Solar passive strategies that have been developed in vernacular architecture from different regions are a response to specific climate effects. These strategies are usually simple, low-tech and have low potential environmental impact. For this reason, several studies highlight them as having potential to reduce the demands of non-renewable energy for buildings operation. In this paper, the climatic contrast between northern and southern parts of mainland Portugal is presented, namely the regions of Beira Alta and Alentejo. Additionally, it discusses the contribution of different climate-responsive strategies developed in vernacular architecture from both regions to assure thermal comfort conditions. In Beira Alta, the use of glazed balconies as a strategy to capture solar gains is usual, while in Alentejo the focus is on passive cooling strategies. To understand the effectiveness of these strategies, thermal performances and comfort conditions of two case studies were evaluated based on the adaptive comfort model. Field tests included measurement of hygrothermal parameters and surveys on occupants’ thermal sensation. From the results, it has been found that the case studies have shown a good thermal performance by passive means alone and that the occupants feel comfortable, except during winter where there is the need to use simple heating systems.

Piezoelectric polymers as biomaterials for tissue engineering applications

Tissue engineering often rely on scaffolds for supporting cell differentiation and growth. Novel paradigms for tissue engineering include the need of active or smart scaffolds in order to properly regenerate specific tissues. In particular, as electrical and electromechanical clues are among the most relevant ones in determining tissue functionality in tissues such as muscle and bone, among others, electroactive materials and, in particular, piezoelectric ones, show strong potential for novel tissue engineering strategies, in particular taking also into account the existence of these phenomena within some specific tissues, indicating their requirement also during tissue regeneration. This referee reports on piezoelectric materials used for tissue engineering applications. The most used materials for tissue engineering strategies are reported together with the main achievements, challenges and future needs for research and actual therapies. This review provides thus a compilation of the most relevant results and strategies and a start point for novel research pathways in the most relevant and challenging open questions.

Piezoelectric poly(vinylidene fluoride) microstructure and poling state in active tissue engineering

Tissue engineering often rely on scaffolds for supporting cell differentiation and growth. Novel paradigms for tissue engineering include the need of active or smart scaffolds in order to properly regenerate specific tissues. In particular, as electrical and electromechanical clues are among the most relevant ones in determining tissue functionality in tissues such as muscle and bone, among others, electroactive materials and, in particular, piezoelectric ones, show strong potential for novel tissue engineering strategies, in particular taking also into account the existence of these phenomena within some specific tissues, indicating their requirement also during tissue regeneration. This referee reports on piezoelectric materials used for tissue engineering applications. The most used materials for tissue engineering strategies are reported together with the main achievements, challenges and future needs for research and actual therapies. This review provides thus a compilation of the most relevant results and strategies and a start point for novel research pathways in the most relevant and challenging open questions.

Poly(vinylidene fluoride-trifluoroethylene) porous films: tailoring microstructure and physical properties by solvent casting strategies

A systematic study for the production of porous poly(vinylidene fluoride-trifluoroethylene), P(VDF-TrFE), films using solvent evaporation and non-solvent induced phase separation techniques is presented. Processing parameters such as copolymer volume fraction, solvent, preset exposure time to air before immersion, and non-solvent and temperature of the coagulation bath were varied and the corresponding sample morphology, hydrophobicity, thermal and mechanical properties were determined. Film morphologies including homogeneous pore distributions, micropores, microvoids, spherulites and non-porous films were obtained. The morphology variations strongly influence sample hydrophobicity and mechanical properties. All samples crystallize in the electroactive β-phase with a degree of crystallinity around 30 %.

Development of a dextrin-based hydrogel for bone regeneration

[Excerpt] Bone tissue engineering is a very challenging and promising field, which handles with the limitations of bone regenerative capacity and the failure of current orthopedic implants [1]. This work describes the preparation and characterization of an injectable dextrin-based hydrogel (oDex) able to incorporate nanoparticles, cells, biomolecules or Bonelike~ granules [2]. (...)

Cost optimal strategies for the renovation of residential neighbourhoods towards energy and emissions neutrality – Rainha Dona Leonor case study

Buildings are one of the major consumers of energy in Europe. This makes them an important target when aiming to reduce the energy consumptions and carbon emissions. The majority of the European building stock has already some decades and so it needs renovation in order to keep its functionality. Taking advantage of these interventions, the energy performance of the buildings may also be improved. In Portugal the renovation techniques, both regarding energy efficiency measures as well as measures for the use of renewable energy sources, are normally planned at the building scale. It is important to explore the possibility of having large scale interventions, has it has been done in other countries, namely at neighbourhood scale with district energy system in order to optimize the results in terms of costs and environmental impact.

Environmental determinants for natural regeneration of gallery forest at the Cerrado/Amazonia boundaries in Brazil

Natural regeneration and structure and their relationship to environmental variables were studied in three sections of a gallery forest, in Eastern Mato Grosso, Brazil (14º43′S and 52º21′W). The assumption was that natural regeneration is constrained by environmental determinants at all stages of development of the tree community. The objective was to analyse the forest structure and to verify the relationship between species distribution and abundance at different stages of regeneration and environmental variables. In each section, 47 contiguous (10x10m) permanent plots were established to sample trees (gbh≥15cm), following a systematic design. Seedlings (0.01 to 1m height), saplings (1.01 to 2m) and poles (from 2.01m height to gbh<15cm) were sampled in sub-plots of 1x1m, 2x2m and 5x5m, respectively. In each plot, soil properties, gaps projection, bamboos, rocky cover, declivity and depth of ground watertable were determined. The relationships between the environmental variables with trees and seedling communities were assessed by canonical correspondence analysis. In spite of the sections being near to each other, they presented large differences in floristics, structure and site conditions. The forest soil presented a low cation exchange capacity and a high level of Al saturation. The occurrence of bamboos and gaps and the depth of ground watertable limited the occurrence of poles and trees. The high degree of structural heterogeneity for each regeneration category was related primarily to a humidity gradient; but soil fertility (Ca+Mg) was also a determinant of seedling and sapling communities.

Strategies to regulate myopia progression with contact lenses: a review

Purpose: Higher myopic refractive errors are associated with serious ocular complications that can put visual function at risk. There is respective interest in slowing and if possible stopping myopia progression before it reaches a level associated with increased risk of secondary pathology. The purpose of this report was to review our understanding of the rationale(s) and success of contact lenses (CLs) used to reduce myopia progression. Methods: A review commenced by searching the PubMed database. The inclusion criteria stipulated publications of clinical trials evaluating the efficacy of CLs in regulating myopia progression based on the primary endpoint of changes in axial length measurements and published in peerreviewed journals. Other publications from conference proceedings or patents were exceptionally considered when no peer-review articles were available. Results: The mechanisms that presently support myopia regulation with CLs are based on the change of relative peripheral defocus and changing the foveal image quality signal to potentially interfere with the accommodative system. Ten clinical trials addressing myopia regulation with CLs were reviewed, including corneal refractive therapy (orthokeratology), peripheral gradient lenses, and bifocal (dual-focus) and multifocal lenses. Conclusions: CLs were reported to be well accepted, consistent, and safe methods to address myopia regulation in children. Corneal refractive therapy (orthokeratology) is so far the method with the largest demonstrated efficacy in myopia regulation across different ethnic groups. However, factors such as patient convenience, the degree of initial myopia, and non-CL treatments may also be considered. The combination of different strategies (i.e., central defocus, peripheral defocus, spectral filters, pharmaceutical delivery, and active lens-borne illumination) in a single device will present further testable hypotheses exploring how different mechanisms can reinforce or compete with each other to improve or reduce myopia regulation with CLs.

Assistive locomotion strategies for active lower limb devices

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Eletrónica Médica)

Strategies to combat infections of Acinetobacter baumannii biofilms

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Biomimetic strategies to engineer mineralised human tissues

In the last few years, many reports have been describing promising biocompatible and biodegradable materials that can mimic in a certain extent the multidimensional hierarchical structure of bone, while are also capable of releasing bioactive agents or drugs in a controlled manner. Despite these great advances, new developments in the design and fabrication technologies are required to address the need to engineer suitable biomimetic materials in order tune cells functions, i.e. enhance cell-biomaterial interactions, and promote cell adhesion, proliferation, and differentiation ability. Scaffolds, hydrogels, fibres and composite materials are the most commonly used as biomimetics for bone tissue engineering. Dynamic systems such as bioreactors have also been attracting great deal of attention as it allows developing a wide range of novel in vitro strategies for the homogeneous coating of scaffolds and prosthesis with ceramics, and production of biomimetic constructs, prior its implantation in the body. Herein, it is overviewed the biomimetic strategies for bone tissue engineering, recent developments and future trends. Conventional and more recent processing methodologies are also described.