Online Measurement System for Dynamic Flow Bioreactors to Study Barrier Integrity of hiPSC-Based Blood–Brain Barrier In Vitro Models

Electrochemical impedance spectroscopy (EIS) is a noninvasive, reliable, and efficient method to analyze the barrier integrity of in vitro tissue models. This well-established tool is used most widely to quantify the transendothelial/epithelial resistance (TEER) of Transwell-based models cultured under static conditions. However, dynamic culture in bioreactors can achieve advanced cell culture conditions that mimic a more tissue-specific environment and stimulation. This requires the development of culture systems that also allow for the assessment of barrier integrity under dynamic conditions. Here, we present a bioreactor system that is capable of the automated, continuous, and non-invasive online monitoring of cellular barrier integrity during dynamic culture. Polydimethylsiloxane (PDMS) casting and 3D printing were used for the fabrication of the bioreactors. Additionally, attachable electrodes based on titanium nitride (TiN)-coated steel tubes were developed to perform EIS measurements. In order to test the monitored bioreactor system, blood–brain barrier (BBB) in vitro models derived from human-induced pluripotent stem cells (hiPSC) were cultured for up to 7 days. We applied equivalent electrical circuit fitting to quantify the electrical parameters of the cell layer and observed that TEER gradually decreased over time from 2513 Ω·cm2 to 285 Ω·cm2, as also specified in the static control culture. Our versatile system offers the possibility to be used for various dynamic tissue cultures that require a non-invasive monitoring system for barrier integrity.

In Silico Study to Enhance Delivery Efficiency of Charged Nanoscale Nasal Spray Aerosols to the Olfactory Region Using External Magnetic Fields

Various factors and challenges are involved in efficiently delivering drugs using nasal sprays to the olfactory region to treat central nervous system diseases. In this study, computational fluid dynamics was used to simulate nasal drug delivery to (1) examine effects on drug deposition when various external magnetic fields are applied to charged particles, (2) comprehensively study effects of multiple parameters (i.e., particle aerodynamic diameter; injection velocity magnitude, angle, and position; magnetic force strength and direction), and (3) determine how to achieve the optimal delivery efficiency to the olfactory epithelium. The Reynolds-averaged Navier–Stokes equations governed airflow, with a realistic inhalation waveform implemented at the nostrils. Particle trajectories were modeled using the one-way coupled Euler–Lagrange model. A current-carrying wire generated a magnetic field to apply force on charged particles and direct them to the olfactory region. Once drug particles reached the olfactory region, their diffusion through mucus to the epithelium was calculated analytically. Particle aerodynamic diameter, injection position, and magnetic field strength were found to be interconnected in their effects on delivery efficiency. Specific combinations of these parameters achieved over 65-fold higher drug delivery efficiency compared with uniform injections with no magnetic fields. The insight gained suggests how to integrate these factors to achieve the optimal efficiency.

Alginate Hydrogel Microtubes for Salivary Gland Cell Organization and Cavitation

Understanding the different regulatory functions of epithelial and mesenchymal cell types in salivary gland development and cellular organization is essential for proper organoid formation and salivary gland tissue regeneration. Here, we demonstrate a biocompatible platform using pre-formed alginate hydrogel microtubes to facilitate direct epithelial–mesenchymal cell interaction for 3D salivary gland cell organization, which allows for monitoring cellular organization while providing a protective barrier from cell-cluster loss during medium changes. Using mouse salivary gland ductal epithelial SIMS cells as the epithelial model cell type and NIH 3T3 fibroblasts or primary E16 salivary mesenchyme cells as the stromal model cell types, self-organization from epithelial–mesenchymal interaction was examined. We observed that epithelial and mesenchymal cells undergo aggregation on day 1, cavitation by day 4, and generation of an EpCAM-expressing epithelial cell layer as early as day 7 of the co-culture in hydrogel microtubes, demonstrating the utility of hydrogel microtubes to facilitate heterotypic cell–cell interactions to form cavitated organoids. Thus, pre-formed alginate microtubes are a promising co-culture method for further understanding epithelial and mesenchymal interaction during tissue morphogenesis and for future practical applications in regenerative medicine.

Comparative Effects of Basic Fibroblast Growth Factor Delivery or Voluntary Exercise on Muscle Regeneration after Volumetric Muscle Loss

Volumetric muscle loss (VML) is associated with irreversibly impaired muscle function due to traumatic injury. Experimental approaches to treat VML include the delivery of basic fibroblast growth factor (bFGF) or rehabilitative exercise. The objective of this study was to compare the effects of spatially nanopatterned collagen scaffold implants with either bFGF delivery or in conjunction with voluntary exercise. Aligned nanofibrillar collagen scaffold bundles were adsorbed with bFGF, and the bioactivity of bFGF-laden scaffolds was examined by skeletal myoblast or endothelial cell proliferation. The therapeutic efficacy of scaffold implants with either bFGF release or exercise was examined in a murine VML model. Our results show an initial burst release of bFGF from the scaffolds, followed by a slower release over 21 days. The released bFGF induced myoblast and endothelial cell proliferation in vitro. After 3 weeks of implantation in a mouse VML model, twitch force generation was significantly higher in mice treated with bFGF-laden scaffolds compared to bFGF-laden scaffolds with exercise. However, myofiber density was not significantly improved with bFGF scaffolds or voluntary exercise. In contrast, the scaffold implant with exercise induced more re-innervation than all other groups. These results highlight the differential effects of bFGF and exercise on muscle regeneration.

Wearables for Biomechanical Performance Optimization and Risk Assessment in Industrial and Sports Applications

Wearable technologies are emerging as a useful tool with many different applications. While these devices are worn on the human body and can capture numerous data types, this literature review focuses specifically on wearable use for performance enhancement and risk assessment in industrial- and sports-related biomechanical applications. Wearable devices such as exoskeletons, inertial measurement units (IMUs), force sensors, and surface electromyography (EMG) were identified as key technologies that can be used to aid health and safety professionals, ergonomists, and human factors practitioners improve user performance and monitor risk. IMU-based solutions were the most used wearable types in both sectors. Industry largely used biomechanical wearables to assess tasks and risks wholistically, which sports often considered the individual components of movement and performance. Availability, cost, and adoption remain common limitation issues across both sports and industrial applications.

The Impact of the Extracellular Matrix Environment on Sost Expression by the MLO-Y4 Osteocyte Cell Line

Bone is a dynamic organ that can adapt its structure to meet the demands of its biochemical and biophysical environment. Osteocytes form a sensory network throughout the tissue and orchestrate tissue adaptation via the release of soluble factors such as a sclerostin. Osteocyte physiology has traditionally been challenging to investigate due to the uniquely mineralized extracellular matrix (ECM) of bone leading to the development of osteocyte cell lines. Importantly, the most widely researched and utilized osteocyte cell line: the MLO-Y4, is limited by its inability to express sclerostin (Sost gene) in typical in-vitro culture. We theorised that culture in an environment closer to the in vivo osteocyte environment could impact on Sost expression. Therefore, this study investigated the role of composition and dimensionality in directing Sost expression in MLO-Y4 cells using collagen-based ECM analogues. A significant outcome of this study is that MLO-Y4 cells, when cultured on a hydroxyapatite (HA)-containing two-dimensional (2D) film analogue, expressed Sost. Moreover, three-dimensional (3D) culture within HA-containing collagen scaffolds significantly enhanced Sost expression, demonstrating the impact of ECM composition and dimensionality on MLO-Y4 behaviour. Importantly, in this bone mimetic ECM environment, Sost expression was found to be comparable to physiological levels. Lastly, MLO-Y4 cells cultured in these novel conditions responded accordingly to fluid flow stimulation with a decrease in expression. This study therefore presents a novel culture system for the MLO-Y4 osteocyte cell line, ensuring the expression of an important osteocyte specific gene, Sost, overcoming a major limitation of this model.

Flexible Strain-Sensitive Silicone-CNT Sensor for Human Motion Detection

This article describes the manufacturing technology of biocompatible flexible strain-sensitive sensor based on Ecoflex silicone and multi-walled carbon nanotubes (MWCNT). The sensor demonstrates resistive behavior. Structural, electrical, and mechanical characteristics are compared. It is shown that laser radiation significantly reduces the resistance of the material. Through laser radiation, electrically conductive networks of MWCNT are formed in a silicone matrix. The developed sensor demonstrates highly sensitive characteristics: gauge factor at 100% elongation −4.9, gauge factor at 90° bending −0.9%/deg, stretchability up to 725%, tensile strength 0.7 MPa, modulus of elasticity at 100% 46 kPa, and the temperature coefficient of resistance in the range of 30–40 °С is −2 × 10−3. There is a linear sensor response (with 1 ms response time) with a low hysteresis of ≤3%. An electronic unit for reading and processing sensor signals based on the ATXMEGA8E5-AU microcontroller has been developed. The unit was set to operate the sensor in the range of electrical resistance 5–150 kOhm. The Bluetooth module made it possible to transfer the received data to a personal computer. Currently, in the field of wearable technologies and health monitoring, a vital need is the development of flexible sensors attached to the human body to track various indicators. By integrating the sensor with the joints of the human hand, effective movement sensing has been demonstrated.

Riboflavin and Its Effect on Dentin Bond Strength: Considerations for Clinical Applicability—An In Vitro Study

Bioactive collagen crosslinkers propose to render the dentin hybrid layer less perceptive to hydrolytic challenge. This study aims to evaluate whether bond strength of dental resin composite to dentin benefits from riboflavin (RB)-sensitized crosslinking when used in a clinically applicable protocol. A total of 300 human dentin specimens were prepared consistent with the requirements for a macro-shear bond test. RB was applied on dentin, either incorporated in the primer (RBp) of a two-step self-etch adhesive or as an aqueous solution (RBs) before applying the adhesive, and blue light from a commercial polymerization device was used for RB photoactivation. Bonding protocol executed according to the manufacturer’s information served as control. Groups (n = 20) were tested after 1 week, 1 month, 3 months, 6 months or 1 year immersion times (37 °C, distilled water). The different application methods of RB significantly influenced bond strength (p < 0.001) with a medium impact (η2p = 0.119). After 1 year immersion, post hoc analysis identified a significant advantage for RB groups compared to RBp (p = 0.018), which is attributed to a pH-/solvent-dependent efficiency of RB-sensitized crosslinking, stressing the importance of formulation adjustments. We developed an application protocol for RB-sensitized crosslinking with emphasis on clinical applicability to test its performance against a gold-standard adhesive, and are confident that, with a few adjustments to the application solution, RB-sensitized crosslinking can improve the longevity of adhesive restorations in clinics.

A Bioprinted Heart-on-a-Chip with Human Pluripotent Stem Cell-Derived Cardiomyocytes for Drug Evaluation

In this work, we show that valve-based bioprinting induces no measurable detrimental effects on human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The aim of the current study was three-fold: first, to assess the response of hiPSC-CMs to several hydrogel formulations by measuring electrophysiological function; second, to customise a new microvalve-based cell printing mechanism in order to deliver hiPSC-CMs suspensions, and third, to compare the traditional manual pipetting cell-culture method and cardiomyocytes dispensed with the bioprinter. To achieve the first and third objectives, iCell2 (Cellular Dynamics International) hiPSC-CMs were used. The effects of well-known drugs were tested on iCell2 cultured by manual pipetting and bioprinting. Despite the results showing that hydrogels and their cross-linkers significantly reduced the electrophysiological performance of the cells compared with those cultured on fibronectin, the bio-ink droplets containing a liquid suspension of live cardiomyocytes proved to be an alternative to standard manual handling and could reduce the number of cells required for drug testing, with no significant differences in drug-sensitivity between both approaches. These results provide a basis for the development of a novel bioprinter with nanolitre resolution to decrease the required number of cells and to automate the cell plating process.

Maxillary Transverse Deficit: A Retrospective Study of Two Biologically Oriented Devices through a Digital Workflow

The aim of this retrospective study was to compare the efficiency of two biologically oriented devices in achieving maxillary expansion: Rapid Palatal Expander (RPE) and Nitanium Palatal Expander-2 (NPE-2). Thirty-six subjects, divided in two equal groups, were included in this study. Maxillary dental arches were scanned using Trios 3 shape®, in order to perform a digital analysis of 3D models. The models were analyzed using Autodesk Fusion 360® and Meshmixer®. All data obtained from analysis of pre-treatment and post-treatment models were processed using Prism® software. The anterior arch width, the posterior arch width, the palate height, and palatal surface were measured to evaluate differences between the devices. A D’Agostino–Pearson normality test was done to check the data. A non-parametric t-test was used to compare the anterior and posterior arch width between the two groups, while a parametric t-test was used to compare the palatal height measurements between the two groups. The p-value was calculated. The limit value fixed was 0.05. Palatal width and surface showed a significant increase in both groups, but no significant changes in palatal height were found. The data processed showed that there were no significant differences between the devices (ΔREP−ΔNPE) in variation of anterior arch width, there were no significant differences in variation of posterior arch width and there were no significant differences in variation of palatal height. The comparison between the two groups showed that both methods were equally effective in correcting transverse defect.