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In this work, we propose a simple, reliable, and versatile strategy to create 3D electroconductive scaffolds suitable for bone tissue engineering (TE) applications with electrical stimulation (ES). The proposed scaffolds are made of 3D-extruded poly(ε-caprolactone) (PCL), subjected to alkaline treatment, and of poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS), anchored to PCL with one of two different crosslinkers: (3-glycidyloxypropyl)trimethoxysilane (GOPS) and divinyl sulfone (DVS). Both cross-linkers allowed the formation of a homogenous and continuous coating of PEDOT:PSS to PCL. We show that these PEDOT:PSS coatings are electroconductive (11.3-20.1 S cm -1 ), stable (up to 21 days in saline solution), and allow the immobilization of gelatin (Gel) to further improve bioactivity. In vitro mineralization of the corresponding 3D conductive scaffolds was greatly enhanced (GOPS(NaOH)-Gel - 3.1 fold, DVS(NaOH)-Gel - 2.0 fold) and cell colonization and proliferation were the highest for the DVS(NaOH)-Gel scaffold. In silico modelling of ES application in DVS(NaOH)-Gel scaffolds indicates that the electrical field distribution is homogeneous, which reduces the probability of formation of faradaic products. Osteogenic differentiation of human bone marrow derived mesenchymal stem/stromal cells (hBM-MSCs) was performed under ES. Importantly, our results clearly demonstrated a synergistic effect of scaffold electroconductivity and ES on the enhancement of MSC osteogenic differentiation, particularly on cell-secreted calcium deposition and the upregulation of osteogenic gene markers such as COL I , OC and CACNA1C . These scaffolds hold promise for future clinical applications, including manufacturing of personalized bone TE grafts for transplantation with enhanced maturation/functionality or bioelectronic devices.

This work reports the rational design and fabrication of magneto-active microfiber meshes with controlled hexagonal microstructures via melt electrowriting (MEW) of a magnetized polycaprolactone-based composite. In situ iron oxide nanoparticle deposition on oxidized graphene yields homogeneously dispersed magnetic particles with sizes above 0.5 µm and low aspect ratio, preventing cellular internalization and toxicity. With these fillers, homogeneous magnetic composites with high magnetic content (up to 20 weight %) are obtained and processed in a solvent-free manner for the first time. MEW of magnetic composites enabled the creation of skeletal muscle-inspired design of hexagonal scaffolds with tunable fiber diameter, reconfigurable modularity, and zonal distribution of magneto-active and nonactive material, with elastic tensile deformability. External magnetic fields below 300 mT are sufficient to trigger out-of-plane reversible deformation. In vitro culture of C2C12 myoblasts on three-dimensional (3D) Matrigel/collagen/MEW scaffolds showed that microfibers guided the formation of 3D myotube architectures, and the presence of magnetic particles does not significantly affect viability or differentiation rates after 8 days. Centimeter-sized skeletal muscle constructs allowed for reversible, continued, and dynamic magneto-mechanical stimulation. Overall, these innovative microfiber scaffolds provide magnetically deformable platforms suitable for dynamic culture of skeletal muscle, offering potential for in vitro disease modeling., (© 2023 The Authors. Small published by Wiley‐VCH GmbH.)

Capacitively Coupled (CCoupled) electric fields are used to stimulate cell cultures in Tissue Engineering. Knowing the electric field (E-Field) magnitude in the culture medium is fundamental to establish a relationship between stimulus strength and cellular effects. We analysed eight CCoupled studies and sought to corroborate the reported estimates of the E-Field in the culture medium. First, we reviewed the basic physics underlying CCoupled stimulation and delineated three approaches to estimate the E-field. Using these approaches, we found that the reported values were overestimated in five studies, four of which were based on incorrect assumptions. In all studies, insufficient information was provided to reproduce the setup exactly. Creating electrical models of the experimental setup should improve the accuracy of the E-field estimates and enhance reproducibility. For this purpose, we developed a free open-source tool, the E-field Calculator for CCoupled systems, which is available for download from an internet hosting platform., (© 2022. The Author(s).)

Electrical stimulation (ES) has been described as a promising tool for bone tissue engineering, being known to promote vital cellular processes such as cell proliferation, migration, and differentiation. Despite the high variability of applied protocol parameters, direct coupled electric fields have been successfully applied to promote osteogenic and osteoinductive processes in vitro and in vivo. Our work aims to study the viability, proliferation, and osteogenic differentiation of human bone marrow-derived mesenchymal stem/stromal cells when subjected to five different ES protocols. The protocols were specifically selected to understand the biological effects of different parts of the generated waveform for typical direct-coupled stimuli. In vitro culture studies evidenced variations in cell responses with different electric field magnitudes (numerically predicted) and exposure protocols, mainly regarding tissue mineralization (calcium contents) and osteogenic marker gene expression while maintaining high cell viability and regular morphology. Overall, our results highlight the importance of numerical guided experiments to optimize ES parameters towards improved in vitro osteogenesis protocols. [ABSTRACT FROM AUTHOR]

Osteochondral (OC) defects affect both articular cartilage and the underlying subchondral bone. Due to limitations in the cartilage tissue's self-healing capabilities, OC defects exhibit a degenerative progression to which current therapies have not yet found a suitable long-term solution. Tissue engineering (TE) strategies aim to fabricate tissue substitutes that recreate natural tissue features to offer better alternatives to the existing inefficient treatments. Scaffold design is a key element in providing appropriate structures for tissue growth and maturation. This study presents a novel method for designing scaffolds with a mathematically defined curvature, based on the geometry of a sphere, to obtain TE constructs mimicking native OC tissue shape. The lower the designed radius, the more curved the scaffold obtained. The printability of the scaffolds using fused filament fabrication (FFF) was evaluated. For the case-study scaffold size (20.1 mm × 20.1 mm projected dimensions), a limit sphere radius of 17.064 mm was determined to ensure printability feasibility, as confirmed by scanning electron microscopy (SEM) and micro-computed tomography (μ-CT) analysis. The FFF method proved suitable to reproduce the curved designs, showing good shape fidelity and replicating the expected variation in porosity. Additionally, the mechanical behavior was evaluated experimentally and by numerical modelling. Experimentally, curved scaffolds showed strength comparable to conventional orthogonal scaffolds, and finite element analysis was used to identify the scaffold regions more susceptible to higher loads. [ABSTRACT FROM AUTHOR]

These authors jointly supervised this work: Paula Pascoal-Faria and Pedro Cavaleiro Miranda. Correction to: I Scientific Reports i https://doi.org/10.1038/s41598-022-14834-2, published online 30 June 2022 In the original version of this Article Paula Pascoal-Faria and Pedro Cavaleiro Miranda were omitted as jointly supervising authors. [Extracted from the article]

Graphene-based materials (GBM) are considered one of the 21st century's most promising materials, as they are incredibly light, strong, thin and have remarkable electrical and thermal properties. As a result, over the past decade, their combination with a diverse range of synthetic polymers has been explored in tissue engineering (TE) and regenerative medicine (RM). In addition, a wide range of methods for fabricating polymer/GBM scaffolds have been reported. This review provides an overview of the most recent advances in polymer/GBM composite development and fabrication, focusing on methods such as electrospinning and additive manufacturing (AM). As a future outlook, this work stresses the need for more in vivo studies to validate polymer/GBM composite scaffolds for TE applications, and gives insight on their fabrication by state-of-the-art processing technologies. [ABSTRACT FROM AUTHOR]

The use of digital twins in tissue engineering (TE) applications is of paramount importance to reduce the number of in vitro and in vivo tests. To pursue this aim, a novel multimodal bioreactor is developed, combining 3D design with numerical stimulation. This approach will facilitate the reproducibility between studies and the platforms optimisation (physical and digital) to enhance TE. The new bioreactor was specifically designed to be additive manufactured, which could not be reproduced with conventional techniques. Specifically, the design suggested allows the application of dual stimulation (electrical and mechanical) of a scaffold cell culture. For the selection of the most appropriate material for bioreactor manufacturing several materials were assessed for their cytotoxicity. Numerical modelling methods were then applied to the new bioreactor using one of the most appropriate material (Polyethylene Terephthalate Glycol-modified (PETG)) to find the optimal stimulation input parameters for bone TE based on two reported in vitro studies. [ABSTRACT FROM AUTHOR]