The native cell microenvironment activates signaling pathways through mechanotransduction mechanisms, influencing cells' physiological and functional outcomes. Magnetic fields are explored to manipulate these environments, and magnetic nanoparticles (MNPs) are highlighted as nano-instructive agents capable of activating key signaling pathways, presenting exciting possibilities in tissue engineering. Still, the ability to precisely control the assembly and differentiation of stem cells within a dynamically responsive microenvironment, crucial for effective tissue regeneration, remains unexplored. This study showcases a novel method wherein MNPs facilitate the precise assembly of magnetically responsive cells into complex 3D tissue structures upon internalization and exposure to temporally defined cyclic magnetic fields. By remotely stimulating these constructs, it is demonstrated for the first time the possibility of remote-controlled modulation of stem cell fate in vivo without biochemical supplementation. Notably, this approach led to ectopic bone formation, highlighting the ability of magnetic actuation to drive osteogenesis in non-bone environments. MNP-driven mechanical stimulation of implanted tissues functions as a bioresponsive system guiding osteogenic differentiation of human adipose-derived stem cells. The in vivo model further illustrates accelerated construct integration, enhanced osteogenic differentiation, and minimal local inflammation, underscoring the potential of this less invasive, remotely controllable platform to advance regenerative strategies for bone engineering.

The development of effective cell delivery therapies faces challenges regarding cell viability and retention after injection. Hydrogel-based materials, designed to mimic extracellular matrix components for cell protection during injection and to enhance local availability, often rely on animal-derived components that raise immunogenicity concerns. Alternatively, those employing polysaccharides and synthetic polymers may exhibit suboptimal cell adhesive properties. This study showcases the development of injectable human protein-derived cell carrier microgels made from methacryloyl platelet lysates. These microgels sustain cell viability by providing an enriched and cost-effective environment of growth factors and proteins while promoting the outward migration of mesenchymal stem cells through controlled enzyme-mediated degradation. Employing a solvent-free and reproducible method using superhydrophobic surfaces, human-derived microgels are successfully fabricated via light irradiation, with sizes adjustable by varying droplet volume. Additionally, the incorporation of collagenase facilitates enzyme-mediated cell migration without compromising viability. Injectability tests confirm that microgel administration preserves both size and morphology, and their effectiveness in filling irregular defects in a porcine tissue highlights their suitability for therapeutic applications. Ultimately, these microgels can be modified to include magnetic nanoparticles, enabling spatial control and fixation using an external magnetic field, and potential imaging capabilities, positioning them as promising candidates for personalized cell therapies.

One of the foremost targets in the advancement of biomaterials to engineer vascularized tissues is not only to replicate the composition of the intended tissue but also to create thicker structures incorporating a vascular network for adequate nutrients and oxygen supply. For the first time, to the best of current knowledge, a clinically relevant biomaterial is developed, demonstrating that hydrogels made from the human decellularized extracellular matrix can exhibit robust mechanical properties (in the kPa range) and angiogenic capabilities simultaneously. These properties enable the culture and organization of human umbilical vein endothelial cells into tubular structures, maintaining their integrity for 14 days in vitro without the need for additional polymers or angiogenesis-related factors. This is achieved by repurposing the placenta chorionic membrane (CM), a medical waste with an exceptional biochemical composition, into a valuable resource for bioengineering purposes. After decellularization, the CM underwent chemical modification with methacryloyl groups, giving rise to methacrylated CM (CMMA). CMMA preserved key proteins, as well as glycosaminoglycans. The resulting hydrogels rapidly photopolymerize and have enhanced strength and customizable mechanical properties. Furthermore, they demonstrate angio-vasculogenic competence in vitro and in vivo, holding significant promise as a humanized platform for the engineering of vascularized tissues.

The inclusion of hollow channels in tissue-engineered hydrogels is crucial for mimicking the natural physiological conditions and facilitating the delivery of nutrients and oxygen to cells. Although bio-fabrication techniques provide diverse strategies to create these channels, many require sophisticated equipment and time-consuming protocols. Herein, collagenase, a degrading agent for methacrylated gelatin hydrogels, and magnetic nanoparticles (MNPs) are combined and processed into enzymatically active spherical structures using a straightforward oil bath emulsion methodology. The generated microgels are then used to microfabricate channels within biomimetic hydrogels via a novel sculpturing approach that relied on the precise coupling of protein-enzyme pairs (for controlled local degradation) and magnetic actuation (for directional control). Results show that the sculpting velocity can be tailored by adjusting the magnetic field intensity or concentration of MNPs within the microgels. Additionally, varying the magnetic field position or microgel size generated diverse trajectories and channels of different widths. This innovative technology improves the viability of encapsulated cells through enhanced medium transport, outperforming non-sculpted hydrogels and offering new perspectives for hydrogel vascularization and drug/biomolecule administration. Ultimately, this novel concept can help design fully controlled channels in hydrogels or soft materials, even those with complex tortuosity, in a single wireless top-down biocompatible step.

The optimized physical adhesion between bees' leg hairs and pollen grains-whereby the latter's diameter aligns with the spacing between the hairs-has previously inspired the development of a biomimetic drug dressing. Combining this optimized process with the improved natural mussels' adhesion in wet environments in a dual biomimetic process, it is herein proposed the fabrication of a natural-derived micropatterned hydrogel patch of methacrylated laminarin (LAM-MET), with enriched drug content and improved adhesiveness, suitable for applications like wound healing. Enhanced adhesion is accomplished by modifying LAM-MET with hydroxypyridinone groups, following the patch microfabrication by soft lithography and UV/vis-irradiation, resulting in a membrane with micropillars with a high aspect ratio. Following the biomimetics rational, a drug patch is engineered by combining the microfabricated dressing with drug particles milled to fit the spaces between pillars. Controlled drug release is achieved, together with inherent antibacterial activity against Escherichia coli and Pseudomonas aeruginosa, and enhanced biocompatibility using the bare micropatterned patches. This new class of biomimetic dressings overcomes the challenges of current patches, like poor mechanical properties and biocompatibility, limited adhesiveness and drug dosage, and lack of prolonged antimicrobial activity, opening new insights for the development of high drug-loaded dressings with improved patient compliance.

The fabrication of biological substitutes to repair, replace, or enhance tissue- and organ-level functions is a long-sought goal of tissue engineering (TE). However, the clinical translation of TE is hindered by several challenges, including the lack of suitable mechanical, chemical, and biological properties in one biomaterial, and the inability to generate large, vascularized tissues with a complex structure of native tissues. Over the past decade, a new generation of "smart" materials has revolutionized the conventional medical field, transforming TE into a more accurate and sophisticated concept. At the vanguard of scientific development, magnetic nanoparticles (MNPs) have garnered extensive attention owing to their significant potential in various biomedical applications owing to their inherent properties such as biocompatibility and rapid remote response to magnetic fields. Therefore, to develop functional tissue replacements, magnetic force-based TE (Mag-TE) has emerged as an alternative to conventional TE strategies, allowing for the fabrication and real-time monitoring of tissues engineered in vitro. This review addresses the recent studies on the use of MNPs for TE, emphasizing the in vitro, in vivo, and clinical applications. Future perspectives of Mag-TE in the fields of TE and regenerative medicine are also discussed.

Fabrication of vascularized large-scale constructs for regenerative medicine remains elusive since most strategies rely solely on cell self-organization or overly control cell positioning, failing to address nutrient diffusion limitations. We propose a modular and hierarchical tissue-engineering strategy to produce bonelike tissues carrying signals to promote prevascularization. In these 3D systems, disc-shaped microcarriers featuring nanogrooved topographical cues guide cell behavior by harnessing mechanotransduction mechanisms. A sequential seeding strategy of adipose-derived stromal cells and endothelial cells is implemented within compartmentalized, liquefied-core macrocapsules in a self-organizing and dynamic system. Importantly, our system autonomously promotes osteogenesis and construct's mineralization while promoting a favorable environment for prevascular-like endothelial organization. Given its modular and self-organizing nature, our strategy may be applied for the fabrication of larger constructs with a highly controlled starting point to be used for local regeneration upon implantation or as drug-screening platforms.

The goal of this work was to examine whether elevated iodine intake was associated with adverse effects on IQ among school-age children in Portugal. In a representative sample of children from the north of the country, IQ percentiles by age (assessed with Raven’s Colored Progressive Matrices) were dichotomized to <50 (“below-average” IQs) and ≥50. Morning urine iodine concentrations, corrected for creatinine, were dichotomized to <250 µg/g and ≥250 µg/g, according to the European Commission/Scientific Committee on Food’s tolerable upper level of daily iodine intake for young children. Data were examined with Chi-square tests, logistic regression, and GLM univariate analysis. The sample (N = 1965) was classified as generally iodine-adequate (median urinary iodine concentration = 129 µg/L; median iodine-to-creatinine ratio = 126 µg/g) according to the WHO’s criteria. A greater proportion of children in the ≥250 µg/g group had below-average IQs, compared to children with less than 250 µg/g (p = 0.037), despite a sizable (though non-significant) proportion of children in the less-than-250 µg/g group also presenting below-average IQs, at the bottom of the iodine distribution (<50 µg/g). The proportion of below-average IQs increased with increasingly elevated iodine concentrations (p = 0.047). The association remained significant after the adjustment for confounders, with the elevated iodine group showing increased odds of having below-average IQs when compared with the non-elevated iodine group (OR 1.55; 95% CI 1.11–2.17; p = 0.011). Consistently, the former group presented a lower mean IQ than the latter (p = 0.006). High iodine intake was associated with lower IQs even in a population classified as iodine-adequate. These results bear on child cognition and on initiatives involving iodine supplementation.

A long-sought goal in tissue engineering (TE) is the development of tissues able to recapitulate the complex architecture of the native counterpart. Microtissues, by resembling the functional units of living structures, can be used to recreate tissues' architecture. Howbeit, microfabrication methodologies fail to reproduce cell-based tissues with uniform shape. At the macroscale, complex tissues are already produced by magnetic-TE using solely magnetized cells as building materials. The enhanced extracellular matrix (ECM) deposition guaranties the conservation of tissues' architecture, leading to a successful cellular engraftment. Following the same rational, now the combination of a versatile microfabrication-platform is proposed with magnetic-TE to generate robust micro-tissues with complex architecture for TE purposes. Small tissue units with circle, square, and fiber-like shapes are designed with high fidelity acting as building blocks for engineering complex tissues. Notably, freestanding microtissues maintain their geometry after 7 days post-culturing, overcoming the challenges of microtissues fabrication. Lastly, the ability of microtissues in invading distinct tissue models while releasing trophic factors is substantiated in methacryloyl laminarin (LAM) and platelet lysates (PLMA) hydrogels. By simply using cells as building units and such microfabrication-platform, the fabrication of complex multiscale and multifunctional tissues with clinical relevance is envisaged, including for therapies or disease models.

In the last decades, the transdermal drug delivery systems (DDS) have been recognized as a promising technology that offers significant benefits over other dosages forms. De facto, their painless application, less frequent replacement, and greater flexibility of dosing are triggering the research and development of such devices. Biopolymers are being widely explored in pharmaceutics for the manufacturing of transdermal DDS mostly due to their safety, biocompatibility, low toxicity, and controlled degradation by human enzymes. This chapter highlights the most recent advances in the engineering of patches for transdermal drug delivery, based on natural biopolymers, emphasizing their combination with drug carriers that enable enhanced drug solubility and a more controlled release of the drug over time. The benefits and limitations of different patch formulations are considered with reference to their appliance to transdermal drug delivery. Critical insights on the current state of patches for transdermal drug delivery are also provided.

Hydrogels have been used in combination with cells for several biomedical and biotechnological applications. Nevertheless, the use of bulk hydrogels has exhibited severe limitations in diffusion of oxygen, nutrients, and metabolites. Here, a support for cell culture is reported where glucose is generated in situ by the own hydrogel degradation, allowing cell survival and function while promoting tissue growth. For this purpose, laminaran (or laminarin)-based hydrogels were fabricated, immobilizing the adequate enzymes to obtain structural platforms for 3D cell culture and providing glucose feeding for metabolic activity of cells through polysaccharide degradation. We demonstrate that tumor A549 cells and human mesenchymal stem cells (hMSCs) can use the glucose resultant from the hydrogel degradation to survive and grow in non-added glucose cell culture medium. Additionally, in vivo biocompatibility and biodegradability of laminaran-based hydrogels were explored for the first time. The self-feeding hydrogels exhibited high potential in cell survival compared to native cell-laden laminaran hydrogels over two weeks of sub-cutaneous implantation. Such bioscaffolds with enzyme-empowered degradation capacity can be applied in diverse biotechnological contexts such as tissue regeneration devices, biofactories, disease models, and cell delivery systems.

Cell-based tissue engineering (TE) has been rapidly evolving to meet the demand of scaffold-free regenerative tissues. Commonly used scaffold-based strategies are often associated with delayed tissue formation, rejection and tissue failure. Scaffold-free TE is a bottom-up methodology that makes use of cell-based soft biomaterials such as cell sheets(CS), spheroids or multishaped microtissues and fiber-shaped multicellular aggregates as building blocks for the construction of functional biomimetic 3D heterogeneous tissues(i.e. consisting of different cell types). Cell-based soft biomaterials are a type of scaffold-free tissues where biomaterials may be used to stimulate and help to orchestrate predetermined functions of natural tissues. The following sections will explore the different cell-based or scaffold-free systems, their advantages, challenges, methods and applications. Major advances in the scaffold-free field resulting from the well-architectured combination of the different methodologies are also reported.

This short review covers the most recent literature on the bottom-up assembling of composite dry powder formulations using supercritical carbon dioxide (scCO2)-based technologies for pulmonary delivery. Special attention is given to the integration of scCO2–assisted spray drying (SASD) with different technologies, namely living polymerization, layer-by-layer deposition and quality-by-design (QbD). Examples of inhalation drug delivery systems are provided to highlight specific advantages of the use of SASD versus traditional particle engineering technologies. The opportunities to generate release particles from small molecule therapeutics to biologic drugs and nucleic acids, the scale-up of processes and perspectives are examined.

Algae are an underexploited source of natural bioactive compounds in Western countries, so an increasing interest in the valorization of these marine organisms has emerged in recent years. In this work, the effect of extracting solvent on the extraction yield, phenolic content, antioxidant capacity, and antimicrobial activity of nine brown macroalgae species (Ascophyllum nodosum, Himanthalia elongata, Undaria pinnatifida, Pelvetia canaliculata, Saccharina latissima, Bifurcaria bifurcata, Laminaria ochroleuca, Sargassum muticum, and Fucus spiralis) was assessed. Total phenolic content (TPC) and the antioxidant properties of extracts by different assays: radical scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP) were performed. The antimicrobial activity of extracts was studied against six different foodborne microorganisms: Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Escherichia coli, Salmonella enteritidis, and Pseudomonas aeruginosa. The highest extraction yield was achieved in ethanolic extracts. However, the highest TPC and FRAP values were obtained on the ethyl acetate extracts, especially from A. nodosum. Concerning algal species, the highest TPC and FRAP values were found in A. nodosum, while the highest DPPH-RSA values were achieved in the hexane extracts of B. bifurcata. The antimicrobial activity of algal extracts varied according to the solvent and alga selected, suggesting the species- and solvent-dependent behavior of this property, with B. bifurcata extracts showing the highest results for a wide range of bacteria. Our results provide insight on the characterization of widespread brown algae in the coasts of the North-Western region of the Iberian Peninsula, reflecting multiple health-enhancing properties which may lead to their exploitation in food, pharmacological, and cosmetic industries.

Small interfering RNA (siRNA) therapy has significant potential for the treatment of myriad diseases, including cancer. While intravenous routes of delivery have been found to be effective for efficient targeting to the liver, achieving high accumulations selectively in other organs, including lung tissues, can be a challenge. We demonstrate the rational design and engineering of a layer-by-layer (LbL) nanoparticle-containing aerosol that is able to achieve efficient, multistage delivery of siRNA in vitro. For the purpose, LbL nanoparticles were, for the first time, encapsulated in composite porous micro scale particles using a supercritical CO2-assisted spray drying (SASD) apparatus using chitosan as an excipient. Such particles exhibited aerodynamic properties highly favorable for pulmonary administration, and effective silencing of mutant KRAS in lung cancer cells derived from tumors of a non-small cell lung cancer (NSCLC) autochthonous model. Furthermore, efficient alveolar accumulation following inhalation in healthy mice was also observed, corroborating in vitro aerodynamic results, and opening new perspectives for further studies of effective lung therapies These results show that multistage aerosols assembled by supercritical CO2-assisted spray drying can enable efficient RNA interference therapy of pulmonary diseases including lung cancer.

The goal of this study was to examine how IQ changes (the Flynn effect) among school-aged children in Portugal behaved over a period of 25 years (1991, 2008 and 2016) that registered major economic transformations in the country. The country's initial move to prosperity was followed by a recent move into recession, which might affect IQ gains. Changes in IQ by level of intellectual ability were also inspected. Children's scores on Raven's Coloured Progressive Matrices in Portugal were compared between the years 1991, 2008 and 2016. Differences in IQs were calculated according to Flynn's (1987) procedures. In addition, percentile distributions of children's scores in the period of economic growth (between 1991 and 2008) were compared for inspection of IQ changes across levels of intellectual ability. Results showed an IQ gain of 2.6 points per decade from 1991 to 2008, followed by a smaller increase (of 1.7 points per decade) from 2008 to 2016, with negative gains observed for seven-, eight- and 10-year-old children. The gains in IQ from 1991 to 2008 have occurred mainly for lower levels of intellectual ability. IQ relative gains accompanied the country's economic fluctuations, and children in lower levels of intellectual ability seemed to have benefitted the most from the economic growth. The trends in the country are suggestive of the influence of economic fluctuations on family dynamics, which, although moderated by education, have consequences for children's cognitive development.

The lack of effective strategies to produce vascularized 3D bone transplants in vitro, hampers the development of thick-constructed bone, limiting the translational of lab-based engineered system to clinical practices. Cell sheet (CS) engineering techniques provide an excellent microenvironment for vascularization since the technique can maintain the intact cell matrix, crucial for angiogenesis. In an attempt to develop hierarchical vascularized 3D cellular constructs, we herein propose the construction of stratified magnetic responsive heterotypic CSs by making use of iron oxide nanoparticles previously internalized within cells. Magnetic force-based CS engineering allows for the construction of thick cellular multilayers. Results show that osteogenesis is achieved due to a synergic effect of human umbilical vein endothelial cells (HUVECs) and adipose-derived stromal cells (ASCs), even in the absence of osteogenic differentiating factors. Increased ALP activity, matrix mineralization, osteopontin and osteocalcin detection were achieved over a period of 21 days for the heterotypic CS conformation (ASCs/HUVECs/ASCs), over the homotypic one (ASCs/ASCs), corroborating our findings. Moreover, the validated crosstalk between BMP-2 and VEGF releases triggers not only the recruitment of blood vessels, as demonstrated in an in vivo CAM assay, as well as the osteogenesis of the 3D cell construct. The in vivo angiogenic profile also demonstrated preserved human vascular structures and human cells showed the ability to migrate and integrate within the chick vasculature.