Living Bacterial Sacrificial Porogens to Engineer Decellularized Porous Scaffolds

Decellularization and cellularization of organs have emerged as disruptive methods in tissue engineering and regenerative medicine. Porous hydrogel scaffolds have widespread applications in tissue engineering, regenerative medicine and drug discovery as viable tissue mimics. However, the existing hydrogel fabrication techniques suffer from limited control over pore interconnectivity, density and size, which leads to inefficient nutrient and oxygen transport to cells embedded in the scaffolds. Here, we demonstrated an innovative approach to develop a new platform for tissue engineered constructs using live bacteria as sacrificial porogens. E.coli were patterned and cultured in an interconnected three-dimensional (3D) hydrogel network. The growing bacteria created interconnected micropores and microchannels. Then, the scafold was decellularized, and bacteria were eliminated from the scaffold through lysing and washing steps. This 3D porous network method combined with bioprinting has the potential to be broadly applicable and compatible with tissue specific applications allowing seeding of stem cells and other cell types

Dementia in Latin America : paving the way towards a regional action plan

Regional challenges faced by Latin American and Caribbean countries (LACs) to fight dementia, such as heterogeneity, diversity, political instabilities, and socioeconomic disparities, can be addressed more effectively grounded in a collaborative setting based on the open exchange of knowledge. In this work, the Latin American and Caribbean Consortium on Dementia (LAC-CD) proposes an agenda for integration to deliver a Knowledge to Action Framework (KtAF). First, we summarize evidence-based strategies (epidemiology, genetics, biomarkers, clinical trials, nonpharmacological interventions, networking and translational research) and align them to current global strategies to translate regional knowledge into actions with transformative power. Then, by characterizing genetic isolates, admixture in populations, environmental factors, and barriers to effective interventions and mapping these to the above challenges, we provide the basic mosaics of knowledge that will pave the way towards a KtAF. We describe strategies supporting the knowledge creation stage that underpins the translational impact of KtAF

Process Optimization for the 3D Printing of PLA and HNT Composites with Arburg Plastic Freeforming

The industrial use of additive manufacturing continues to rapidly increase as new technology developments become available. The Arburg plastic freeforming (APF) process is designed to utilize standard polymeric granules in order to print parts with properties similar to those of molded parts. Despite the emerging industrial importance of APF, the current body of knowledge regarding this technology is still very limited, especially in the field of biodegradable polymer composites. To this end, poly(lactic acid) (PLA) was reinforced with halloysite nanotubes (HNTs) by hot melt extrusion. The PLA/HNT (0–10 wt%.) composites were analyzed in terms of their rheology, morphology, and thermal and mechanical properties. A study of the processing properties of these composites in the context of APF was performed to ensure the consistency of 3D-printed, high-quality components. The optimized machine settings were used to evaluate the tensile properties of specimens printed with different axis orientations (XY and XZ) and deposition angles (0 and 45°). Specimens printed with an XY orientation and deposition angle starting at 0° resulted in the highest mechanical properties. In this study, the use of PLA/HNT composites in an APF process was reported for the first time, and the current methodology achieved satisfactory results in terms of the 3D printing and evaluation of successful PLA/HNT composites to be used as feedstock in an APF process

Process Optimization for the 3D Printing of PLA and HNT Composites with Arburg Plastic Freeforming

The industrial use of additive manufacturing continues to rapidly increase as new technology developments become available. The Arburg plastic freeforming (APF) process is designed to utilize standard polymeric granules in order to print parts with properties similar to those of molded parts. Despite the emerging industrial importance of APF, the current body of knowledge regarding this technology is still very limited, especially in the field of biodegradable polymer composites. To this end, poly(lactic acid) (PLA) was reinforced with halloysite nanotubes (HNTs) by hot melt extrusion. The PLA/HNT (0–10 wt%.) composites were analyzed in terms of their rheology, morphology, and thermal and mechanical properties. A study of the processing properties of these composites in the context of APF was performed to ensure the consistency of 3D-printed, high-quality components. The optimized machine settings were used to evaluate the tensile properties of specimens printed with different axis orientations (XY and XZ) and deposition angles (0 and 45°). Specimens printed with an XY orientation and deposition angle starting at 0° resulted in the highest mechanical properties. In this study, the use of PLA/HNT composites in an APF process was reported for the first time, and the current methodology achieved satisfactory results in terms of the 3D printing and evaluation of successful PLA/HNT composites to be used as feedstock in an APF process

Role of Yap/Taz in mechanotransduction

Cells perceive their microenvironment not only through soluble signals but also through physical and mechanical cues, such as extracellular matrix (ECM) stiffness or confined adhesiveness. By mechanotransduction systems, cells translate these stimuli into biochemical signals controlling multiple aspects of cell behaviour, including growth, differentiation and cancer malignant progression, but how rigidity mechanosensing is ultimately linked to activity of nuclear transcription factors remains poorly understood. Here we report the identification of the Yorkie-homologues YAP (Yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif, also known as WWTR1) as nuclear relays of mechanical signals exerted by ECM rigidity and cell shape. This regulation requires Rho GTPase activity and tension of the actomyosin cytoskeleton, but is independent of the Hippo/LATS cascade. Crucially, YAP/TAZ are functionally required for differentiation of mesenchymal stem cells induced by ECM stiffness and for survival of endothelial cells regulated by cell geometry; conversely, expression of activated YAP overrules physical constraints in dictating cell behaviour. These findings identify YAP/TAZ as sensors and mediators of mechanical cues instructed by the cellular microenvironment

Microwrinkled conducting polymer interface for anisotropic multicellular alignment

Surfaces with controlled micro and nanoscale topographical cues are useful as smart scaffolds and biointerfaces for cell culture. Recently, use of thin-film and surface wrinkling is emerging as a rapid unconventional method for preparing topographically patterned surfaces, especially suited for the production of smart patterns over large area surfaces. On the other hand, there is an increasing interest in employing conducting polymers as soft, biocompatible, conductive biointerfaces or as parts of bioelectronic devices. A novel convenient and versatile method is presented for producing anisotropic topographical cues at the micro- and nanoscale on conducting polymer surfaces. Micro and nanowrinkles were formed during the heat-shrinking process of a thermo-retractable polystyrene substrate. Surface wrinkling was due to the mismatch between the mechanical properties of a conducting polymer ultrathin film and the substrate. Various geometries of wrinkled structures were prepared, demonstrating the tunability of topography depending on the thickness of the conductive film. A method for patterning the conductive properties of the wrinkled substrates was also presented. Such surfaces acted as smart scaffolds for the functional alignment of cells, envisioning their electrical stimulation. Cell adhesion and proliferation were evaluated, comparing different topographies, and a preferential anisotropic alignment of C2C12 murine skeletal muscle cells along wrinkles was demonstrated. The observed trends were also confirmed concerning the formation of aligned myotubes in C2C12 differentiation stage. Furthermore, improved results in terms of aligned and mature myotube formation were obtained by co-culturing C2C12 cells with a fibroblasts feeder layer. The combination of living cells and tunable conductive nanowrinkles will represent a unique tool for the development of innovative biomedical devices