Gait Compensation among Children with Non-Operative Legg-Calvé-Perthes Disease: A Systematic Review.

Perthes disease is a condition that affects walking patterns in young children due to poor blood circulation in the hip joint. Understanding the gait strategies of affected children is of great importance for an objective assessment and better management of this condition. The aim of this systematic review was to evaluate the current literature to identify gait compensation patterns in non-operative children with Perthes disease. METHODS: A systematic electronic search was performed using MEDLINE, CINAHL, Embase, BIOSIS, and the Cochrane Library to identify studies published from inception up until December 2023. An adapted Downs and Black checklist was utilised to assess methodological quality and project risk of bias. Percentage agreement and nominal kappa statistics with bootstrapped bias-corrected 95% confidence intervals (CIs) were used. RESULT: A comprehensive literature search revealed 277 citations for review, of which 210 studies entered full-text screening. In total, eight studies met the inclusion criteria for quality assessment by two independent reviewers. The results revealed variations in data quality, with scores ranging from 12 to 17 due to missing information related to subject characteristics, biomechanical model, and power calculation. CONCLUSIONS: This review reveals common compensation strategies associated with walking among non-operative children with Perthes disease such as Trendelenburg gait due to weakness of the hip abductor muscle.

Dielectrophoresis as a Tool to Reveal the Potential Role of Ion Channels and Early Electrophysiological Changes in Osteoarthritis.

Diseases such as osteoarthritis (OA) are commonly characterized at the molecular scale by gene expression and subsequent protein production; likewise, the effects of pharmaceutical interventions are typically characterized by the effects of molecular interactions. However, these phenomena are usually preceded by numerous precursor steps, many of which involve significant ion influx or efflux. As a consequence, rapid assessment of cell electrophysiology could play a significant role in unravelling the mechanisms underlying drug interactions and progression of diseases, such as OA. In this study, we used dielectrophoresis (DEP), a technique that allows rapid, label-free determination of the dielectric parameters to assess the role of potassium ions on the dielectric characteristics of chondrocytes, and to investigate the electrophysiological differences between healthy chondrocytes and those from an in vitro arthritic disease model. Our results showed that DEP was able to detect a significant decrease in membrane conductance (6191 ± 738 vs. 8571 ± 1010 S/m2), membrane capacitance (10.3 ± 1.47 vs. 14.5 ± 0.01 mF/m2), and whole cell capacitance (5.4 ± 0.7 vs. 7.5 ± 0.3 pF) following inhibition of potassium channels using 10 mM tetraethyl ammonium, compared to untreated healthy chondrocytes. Moreover, cells from the OA model had a different response to DEP force in comparison to healthy cells; this was seen in terms of both a decreased membrane conductivity (782 S/m2 vs. 1139 S/m2) and a higher whole cell capacitance (9.58 ± 3.4 vs. 3.7 ± 1.3 pF). The results show that DEP offers a high throughput method, capable of detecting changes in membrane electrophysiological properties and differences between disease states.

DEP-Dots for 3D cell culture: low-cost, high-repeatability, effective 3D cell culture in multiple gel systems.

It is known that cells grown in 3D are more tolerant to drug treatment than those grown in dispersion, but the mechanism for this is still not clear; cells grown in 3D have opportunities to develop inter-cell communication, but are also closely packed which may impede diffusion. In this study we examine methods for dielectrophoresis-based cell aggregation of both suspension and adherent cell lines, and compare the effect of various drugs on cells grown in 3D and 2D. Comparing viability of pharmacological interventions on 3D cell clusters against both suspension cells and adherent cells grown in monolayer, as well as against a unicellular organism with no propensity for intracellular communication, we suggest that 3D aggregates of adherent cells, compared to suspension cells, show a substantially different drug response to cells grown in monolayer, which increases as the IC50 is approached. Further, a mathematical model of the system for each agent demonstrates that changes to drug response are due to inherent changes in the system of adherent cells from the 2D to 3D state. Finally, differences in the electrophysiological membrane properties of the adherent cell type suggest this parameter plays an important role in the differences found in the 3D drug response.

Dual-Task Effects on Performance of Gait and Balance in People with Knee Pain: A Systematic Scoping Review.

Dual-task paradigms have been increasingly used to assess the interaction between cognitive demands and the control of balance and gait. The interaction between functional and cognitive demands can alter movement patterns and increase knee instability in individuals with knee conditions, such as knee anterior cruciate ligament (ACL) injury or osteoarthritis (OA). However, there is no consensus on the effects of dual-task on gait mechanics and balance in those individuals. This systematic scoping review aims to examine the impact of dual-task gait and standing balance on motor and cognitive performance in individuals with knee OA or ACL injury. A comprehensive search of MEDLINE, PubMed, Web of Science, and EMBASE electronic databases up until December 2019 was carried out. Inclusion criteria was limited to include dual-task studies that combined cognitive tasks performed simultaneously with gait or standing balance in individuals with knee OA or ACL injuries. In total, fifteen studies met the inclusion criteria, nine articles examined dual-task effects on balance, and six articles reported the effects of dual-task on gait. The total number of individuals included was 230 individuals with ACL injuries, and 168 individuals with knee OA. A decline in gait and balance performance during dual-task testing is present among individuals with ACL injury and/or ACL reconstruction and knee OA. Further research is required, but dual taking assessment could potentially be used to identify individuals at risk of falling or further injury and could be used to develop targeted rehabilitation protocols. A variety of outcome measures have been used across the studies included, making comparisons difficult. The authors, therefore, recommend developing a standardized set of biomechanical balance variables.

Ten-Second Electrophysiology: Evaluation of the 3DEP Platform for high-speed, high-accuracy cell analysis.

Electrical correlates of the physiological state of a cell, such as membrane conductance and capacitance, as well as cytoplasm conductivity, contain vital information about cellular function, ion transport across the membrane, and propagation of electrical signals. They are, however, difficult to measure; gold-standard techniques are typically unable to measure more than a few cells per day, making widespread adoption difficult and limiting statistical reproducibility. We have developed a dielectrophoretic platform using a disposable 3D electrode geometry that accurately (r2 > 0.99) measures mean electrical properties of populations of ~20,000 cells, by taking parallel ensemble measurements of cells at 20 frequencies up to 45 MHz, in (typically) ten seconds. This allows acquisition of ultra-high-resolution (100-point) DEP spectra in under two minutes. Data acquired from a wide range of cells - from platelets to large cardiac cells - benchmark well with patch-clamp-data. These advantages are collectively demonstrated in a longitudinal (same-animal) study of rapidly-changing phenomena such as ultradian (2-3 hour) rhythmicity in whole blood samples of the common vole (Microtus arvalis), taken from 10 µl tail-nick blood samples and avoiding sacrifice of the animal that is typically required in these studies.

Rhythmic potassium transport regulates the circadian clock in human red blood cells.

Circadian rhythms organize many aspects of cell biology and physiology to a daily temporal program that depends on clock gene expression cycles in most mammalian cell types. However, circadian rhythms are also observed in isolated mammalian red blood cells (RBCs), which lack nuclei, suggesting the existence of post-translational cellular clock mechanisms in these cells. Here we show using electrophysiological and pharmacological approaches that human RBCs display circadian regulation of membrane conductance and cytoplasmic conductivity that depends on the cycling of cytoplasmic K+ levels. Using pharmacological intervention and ion replacement, we show that inhibition of K+ transport abolishes RBC electrophysiological rhythms. Our results suggest that in the absence of conventional transcription cycles, RBCs maintain a circadian rhythm in membrane electrophysiology through dynamic regulation of K+ transport.

Process development for cell aggregate arrays encapsulated in a synthetic hydrogel using negative dielectrophoresis.

Spatial patterning of cells is of great importance in tissue engineering and biotechnology, enabling, for example the creation of bottom-up histoarchitectures of heterogeneous cells, or cell aggregates for in vitro high-throughput toxicological and therapeutic studies within 3D microenvironments. In this paper, a single-step process for creating peelable and resilient hydrogels, encapsulating arrays of biological cell aggregates formed by negative DEP has been devised. The dielectrophoretic trapping within low-energy regions of the DEP-dot array reduces cell exposure to high field stresses while creating distinguishable, evenly spaced arrays of aggregates. In addition to using an optimal combination of PEG diacrylate pre-polymer solution concentration and a novel UV exposure mechanism, total processing time was reduced. With a continuous phase medium of PEG diacrylate at 15% v/v concentration, effective dielectrophoretic cell patterned arrays and photo-polymerisation of the mixture was achieved within a 4 min period. This unique single-step process was achieved using a 30 s UV exposure time frame within a dedicated, wide exposure area DEP light box system. To demonstrate the developed process, aggregates of yeast, human leukemic (K562) and HeLa cells were immobilised in an array format within the hydrogel. Relative cell viability for both cells within the hydrogels, after maintaining them in appropriate iso-osmotic media, over a week period was greater than 90%.