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Cells respond to mechanical cues from their environment through a process of mechanosensing and mechanotransduction. Cell stretching devices are important tools to study the molecular pathways responsible for cellular responses to mechanobiological processes. We describe the development and testing of a uniaxial cell stretcher that has applications for microscopic as well as biochemical analyses. By combining simple fabrication techniques with adjustable control parameters, the stretcher is designed to fit a variety of experimental needs. The stretcher can be used for static and cyclic stretching. As a proof of principle, we visualize stretch induced deformation of cell nuclei via incremental static stretch, and changes in IEX1 expression via cyclic stretching. This stretcher is easily modified to meet experimental needs, inexpensive to build, and should be readily accessible for most laboratories with access to 3D printing., (© 2021. Published by The Company of Biologists Ltd.)

Quantitative assessment of soft tissue elasticity is crucial to a broad range of applications, such as biomechanical modeling, physiological monitoring, and tissue diseases diagnosing. However, the modulus measurement of soft tissues, particularly in vivo, has proved challenging since the instrument has to reach the site of soft tissue and be able to measure in a very short time. Here, we present a simple method to measure the elastic modulus of soft tissues on site by exploiting buckling of a long slender bar to quantify the applied force and a spherical indentation to extract the elastic modulus. The method is realized by developing a portable pen-sized instrument (EPen: Elastic modulus pen). The measurement accuracies are verified by independent modulus measures using commercial nanoindenter. Quantitative measurements of the elastic modulus of mouse pancreas, healthy and cancerous, surgically exposed but attached to the body further confirm the potential clinical utility of the EPen.

While many fields have contributed to biological physics, nanotechnology offers a new scale of observation. High-speed atomic force microscopy (HS-AFM) provides nanometre structural information and dynamics with subsecond resolution of biological systems. Moreover, HS-AFM allows us to measure piconewton forces within microseconds giving access to unexplored, fast biophysical processes. Thus, HS-AFM provides a tool to nourish biological physics through the observation of emergent physical phenomena in biological systems. In this review, we present an overview of the contribution of HS-AFM, both in imaging and force spectroscopy modes, to the field of biological physics. We focus on examples in which HS-AFM observations on membrane remodelling, molecular motors or the unfolding of proteins have stimulated the development of novel theories or the emergence of new concepts. We finally provide expected applications and developments of HS-AFM that we believe will continue contributing to our understanding of nature, by serving to the dialogue between biology and physics. This article is part of a discussion meeting issue 'Dynamic in situ microscopy relating structure and function'.

Background/aims: This study aimed to establish a precise and well-defined working model, assessing pharmaceutical effects on vascular smooth muscle cell monolayer in-vitro. It describes various analysis techniques to determine the most suitable to measure the biomechanical impact of vasoactive agents by using CellDrum technology., Methods: The so-called CellDrum technology was applied to analyse the biomechanical properties of confluent human aorta muscle cells (haSMC) in monolayer. The cell generated tensions deviations in the range of a few N/m² are evaluated by the CellDrum technology. This study focuses on the dilative and contractive effects of L-type Ca 2+ channel agonists and antagonists, respectively. We analyzed the effects of Bay K8644, nifedipine and verapamil. Three different measurement modes were developed and applied to determine the most appropriate analysis technique for the study purpose. These three operation modes are called, particular time mode" (PTM), "long term mode" (LTM) and "real-time mode" (RTM)., Results: It was possible to quantify the biomechanical response of haSMCs to the addition of vasoactive agents using CellDrum technology. Due to the supplementation of 100nM Bay K8644, the tension increased approximately 10.6% from initial tension maximum, whereas, the treatment with nifedipine and verapamil caused a significant decrease in cellular tension: 10nM nifedipine decreased the biomechanical stress around 6,5% and 50nM verapamil by 2,8%, compared to the initial tension maximum. Additionally, all tested measurement modes provide similar results while focusing on different analysis parameters., Conclusion: The CellDrum technology allows highly sensitive biomechanical stress measurements of cultured haSMC monolayers. The mechanical stress responses evoked by the application of vasoactive calcium channel modulators were quantified functionally (N/m²). All tested operation modes resulted in equal findings, whereas each mode features operation-related data analysis., Competing Interests: The authors have no conflicts of interest to declare., (© Copyright by the Author(s). Published by Cell Physiol Biochem Press.)

In this paper, we present a transparent mechanical stimulation device capable of uniaxial stimulation, which is compatible with standard bioanalytical methods used in cellular mechanobiology. We validate the functionality of the uniaxial stimulation system using human-induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs). The pneumatically controlled device is fabricated from polydimethylsiloxane (PDMS) and provides uniaxial strain and superior optical performance compatible with standard inverted microscopy techniques used for bioanalytics (e.g., fluorescence microscopy and calcium imaging). Therefore, it allows for a continuous investigation of the cell state during stretching experiments. The paper introduces design and fabrication of the device, characterizes the mechanical performance of the device and demonstrates the compatibility with standard bioanalytical analysis tools. Imaging modalities, such as high-resolution live cell phase contrast imaging and video recordings, fluorescent imaging and calcium imaging are possible to perform in the device. Utilizing the different imaging modalities and proposed stretching device, we demonstrate the capability of the device for extensive further studies of hiPSC-CMs. We also demonstrate that sarcomere structures of hiPSC-CMs organize and orient perpendicular to uniaxial strain axis and thus express more maturated nature of cardiomyocytes.

The higher-order structure (HOS) of protein therapeutics is a critical quality attribute directly related to their function. Traditionally, the HOS of protein therapeutics has been characterized by methods with low to medium structural resolution such as Fourier-transform infrared (FTIR), circular dichroism (CD), and intrinsic fluorescence spectroscopy, and differential scanning calorimetry (DSC). Recently, high-resolution nuclear magnetic resonance (NMR) methods have emerged as powerful tools for HOS characterization. NMR is a multi-attribute method with unique capabilities to provide information about all the structural levels of proteins in solution. We have in this study compared 1 D 1 H Profile NMR with the established biophysical methods for HOS assessments using a set of blended samples of the monoclonal antibodies belonging to the subclasses IgG1 and IgG2. The study shows that Profile NMR can distinguish between most sample combinations (93%), DSC can differentiate 61% of the sample combinations, and near-ultraviolet CD spectroscopy can differentiate 52% of the sample combinations, whereas no significant distinction could be made between any samples using FTIR or intrinsic fluorescence. Our data therefore show that NMR has superior ability to address differences in HOS, a feature that could be directly applicable in comparability and similarity assessments., (Copyright © 2020 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

The role and importance of mechanical properties of cells and tissues in cellular function, development and disease has widely been acknowledged, however standard techniques currently used to assess them exhibit intrinsic limitations. Recently, Brillouin microscopy, a type of optical elastography, has emerged as a non-destructive, label- and contact-free method that can probe the viscoelastic properties of biological samples with diffraction-limited resolution in 3D. This led to increased attention amongst the biological and medical research communities, but it also sparked debates about the interpretation and relevance of the measured physical quantities. Here, we review this emerging technology by describing the underlying biophysical principles and discussing the interpretation of Brillouin spectra arising from heterogeneous biological matter. We further elaborate on the technique's limitations, as well as its potential for gaining insights in biology, in order to guide interested researchers from various fields.

Biomechanical stimulation is a common strategy to improve the growth, maturation, and function of a variety of engineered tissues. However, identifying optimized biomechanical conditioning protocols is challenging, as cell responses to mechanical stimuli are modulated by other multifactorial microenvironmental cues, including soluble factors and biomaterial properties. Traditional bioreactors lack the throughput necessary for combinatorial testing of cell activity in mechanically stimulated engineered tissues. Microfabricated systems can improve experimental throughput, but often do not provide uniform mechanical loading, are challenging to use, lack robustness, and offer limited amounts of cells and tissue for analysis. To address the need for higher-throughput, combinatorial testing of cell activity in a tissue engineering context, we developed a hybrid approach, in which flexible polydimethylsiloxane microfabricated inserts were designed to simultaneously generate multiple tensile strains when stretched cyclically in a standard dynamic bioreactor. In the embodiment presented in this study, each insert contained an array of 35 dog bone-shaped wells in which cell-seeded microscale hydrogels can be polymerized, with up to eight inserts stretched simultaneously in the bioreactor. Uniformity of the applied strains, both along the length of a microtissue and across multiple microtissues at the same strain level, was confirmed experimentally. In proof-of-principle experiments, the combinatorial effects of dynamic strain, biomaterial stiffness, and transforming growth factor (TGF)-β1 stimulation on myofibroblast differentiation were tested, revealing both known and novel interaction effects and suggesting tissue engineering strategies to regulate myofibroblast activation. This platform is expected to have wide applicability in systematically probing combinations of mechanobiological tissue engineering parameters for desired effects on cell fate and tissue function. Impact Statement In this study, we introduce a dynamic bioreactor system incorporating microfabricated inserts to enable systematic probing of the effects of combinations of mechanobiological parameters on engineered tissues. This novel platform offers the ease of use, robustness, and well-defined mechanical strain stimuli inherent in traditional dynamic bioreactors, but significantly improves throughput (up to 280 microtissues can be tested simultaneously in the embodiment presented in this study). This platform has wide applicability to systematically probe combinations of dynamic mechanical strain, biomaterial properties, biochemical stimulation, and other parameters for desired effects on cell fate and engineered tissue development.

Three-dimensional (3D) printing represents a key technology for rapid prototyping, allowing easy, rapid, and low-cost fabrication. In this work, 3D printing was applied for the in-house production of customized components of a mechanical stretching bioreactor with potential application for cardiac tissue engineering and mechanobiology studies. The culture chamber housing and the motor housing were developed as functional permanent parts, aimed at fixing the culture chamber position and at guaranteeing motor watertightness, respectively. Innovative sample holder prototypes were specifically designed and 3D-printed for holding thin and soft biological samples during cyclic stretch culture. The manufactured components were tested in-house and in a cell biology laboratory. Moreover, tensile tests and finite element analysis were performed to investigate the gripping performance of the sample holder prototypes. All the components showed suitable performances in terms of design, ease of use, and functionality. Based on 3D printing, the bioreactor optimization was completely performed in-house, from design to fabrication, enabling customization freedom, strict design-to-prototype timing, and cost and time effective testing, finally boosting the bioreactor development process.

The study compared the centre of pressure measurements (COP) and vertical ground reaction forces (vGRF) from a pressure insole system to that from force plates (FP) during two flywheel quadriceps resistance exercises: leg press and squat. The comparison was performed using a motion capture system and simultaneous measurements of COP and vGRF from FP and insoles. At lower insole-vGRF (<250 N/insole) COP accuracy deteriorated and those data were excluded from further analysis. The insoles systematically displaced the COP slightly posteriorly and medially compared to the FP measurements. Pearson's coefficient of correlation (r) between insole- and FP-COP showed good agreement in both the anteroposterior (squat: r = 0.96, leg press: r = 0.97) and mediolateral direction (squat: r = 0.84, leg press: r = 0.90), whereas the root-mean-square errors (RMSE) were lower in the mediolateral (squat: 3.9 mm, leg press: 4.5 mm) than the anteroposterior (squat and leg press: 11.8 mm) direction. Vertical GRF was slightly overestimated by the insoles in leg press and RMSE were greater in leg press (8% of peak force) than in squat (6%). Overall, results were within the range of previous studies performed on gait. The strong agreement between insole and FP measurements indicates that insoles may replace FPs in field applications and biomechanical computations during resistance exercise, provided that the applied force is sufficient., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Commercially-available Virtual Reality (VR) systems have the potential to be effective tools for simultaneous visual manipulation and kinematic data collection. Previously, these systems have been integrated with research-grade motion capture systems to provide both functionalities; however, they are yet to be used as stand-alone systems for kinematic data collection. The present study aimed to validate the HTC VIVE VR system for kinematic data collection by evaluating the accuracy of its position and orientation signals. The VIVE controller and tracker were each compared to a Polhemus Liberty magnetic tracking system sensor for angular and translational measurement error and signal drift. A sensor from each system was mounted to opposite ends of a rigid segment which was driven through fifty rotations and fifty translations. Mean angular errors for both the VIVE tracker and controller were below 0.4°. Mean translational error for both sensors was below 3 mm. Drift in the Liberty signal components was consistently lower than drift in VIVE components. However, all mean rotational drift measures were below 0.1° and all mean translational measures were below 0.35 mm. These data indicate that the HTC VIVE system has the potential to be a valid and reliable means of kinematic data collection. However, further investigation is necessary to determine the VIVE's suitability for capturing extremely minute or high-volume movements., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Two-photon lithography is a laser writing technique that can produce 3D microstructures with resolutions below the diffraction limit. This review focuses on its applications to study mechanical properties of cells, an emerging field known as mechanobiology. We review 3D structural designs and materials in the context of new experimental designs, including estimating forces exerted by single cells, studying selective adhesion on substrates, and creating 3D networks of cells. We then focus on emerging applications, including structures for assessing cancer cell invasiveness, whose migration properties depend on the cell mechanical response to the environment, and 3D architectures and materials to study stem cell differentiation, as 3D structure shape and patterning play a key role in defining cell fates., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The Interactive Balance System (IBS), a posturography device for assessing posture control, is widely used in clinical and rehabilitation settings. However, data on the validity of the device are unavailable. Fluctuations of the center of pressure (COP) were measured in 24 healthy participants (age: 29 ± 5 (mean ± SD) years, 12 females) synchronously using the IBS, which was rigidly mounted on a Kistler platform. Four different bipedal conditions were examined: eyes open or closed on stable or soft surfaces. Time series were compared using congruity (CON, proportion of the measurement time during which values of both devices changed similarly in direction), whereas IBS-specific postural outcomes were correlated with traditional postural control outcomes of the Kistler force platform. The time-displacement curves showed similar shapes for CON (>0.9) for each of the four standing conditions without differences between male and female participants (P > 0.39). The path length results of both devices showed very high linear associations, explaining on average 92% (medio-lateral) or 96% (anterior-posterior) of the common variance. The Kistler path length of the anterior-posterior direction revealed nearly perfect linear associations with the stability index of the IBS (r 2  > 0.99). The results of this study indicate that the IBS provides valid posturographic results. Since the medial-lateral and anterior-posterior trajectories of the IBS can be used to calculate COP fluctuations, comparisons between different measurement systems are possible., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Tracking head motion in a simple, portable and accurate manner during performance of postural tasks in a virtual reality environment could have important implications for investigating normal and pathological head kinematics. We investigated concurrent validity of head tracking of two Head Mounted Displays (HMDs), Oculus Rift and HTC Vive, vs. a gold-standard motion capture system (Qualisys). Head kinematics of N = 20 healthy young adults was quantified during static and dynamic postural tasks. While wearing the Oculus Rift or HTC Vive, participants observed moving stars (static tasks) or a flying ball (dynamic task). Head kinematics were recorded simultaneously by the Rift or Vive and Qualisys camera system. We calculated head directional path, acceleration in 6 directions and volume of translation movement. Intra-Class Correlations (ICC) and 95% Limits of agreement were calculated. Most ICC values were around 0.9 with several at 0.99 indicating excellent agreement between the HMDs and Qualisys. Weaker agreement was observed for vertical displacement during a static task and moderate agreement was observed pitch and yaw displacement during a dynamic task. A negative bias of a small magnitude (indicating more movement in VR) was observed for most variables in static tasks, while a positive bias was observed for most variables in the dynamic task (indicating less movement in VR). Our results generally support the concurrent validity of Oculus Rift and HTC Vive head tracking during static and dynamic standing tasks in healthy young adults. Specific task- and direction-dependent differences should be considered when planning measurement studies using these novel tools., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Recent advancements in low-cost depth cameras may provide a clinically accessible alternative to conventional three-dimensional (3D) multi-camera motion capture systems for gait analysis. However, there remains a lack of information on the validity of clinically relevant running gait parameters such as vertical oscillation (VO). The purpose of this study was to assess the validity of measures of VO during running gait using raw depth data, in comparison to a 3D multi-camera motion capture system. Sixteen healthy adults ran on a treadmill at a standard speed of 2.7 m/s. The VO of their running gait was simultaneously collected from raw depth data (Microsoft Kinect v2) and 3D marker data (Vicon multi-camera motion capture system). The agreement between the VO measures obtained from the two systems was assessed using a Bland-Altman plot with 95% limits of agreement (LOA), a Pearson's correlation coefficient (r), and a Lin's concordance correlation coefficient (r c ). The depth data from the Kinect v2 demonstrated excellent results across all measures of validity (r = 0.97; r c  = 0.97; 95% LOA = -8.0 mm - 8.7 mm), with an average absolute error and percent error of 3.7 (2.1) mm and 4.0 (2.0)%, respectively. The findings of this study have demonstrated the ability of a low cost depth camera and a novel tracking method to accurately measure VO in running gait., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Wearable technology can be used to quantify running biomechanical patterns in a runner's natural environment, however, changes in external factors during outdoor running may influence a runner's typical gait pattern. Therefore, the purpose of this study was to determine how many runs are needed to define a stable or typical running pattern. Six biomechanical variables were recorded using a single wearable sensor placed on the lower back during ten outdoor runs for twelve runners. Univariate and multivariate distributions were created and based on the probability density function, the percent of similar data points (within 95%) from each unique run for the same runner were determined. Stability was defined when the addition of data from a new run resulted in less than a 5% change in the probability density function. To cross-validate, the percent of similar data points at stability was compared between the same and different runners using a repeated-measures MANOVA (Bonferroni-corrected α = 0.007). The maximum number of runs needed to reach stability for univariate and multivariate analyses was four and five, respectively. There was a significant overall effect on similar data points between the same and different runners (p = 0.001), with a greater percent of similar data points for the same runner compared to other runners (p < 0.007). Based on biomechanical data collected using a single wearable sensor placed on the lower back, this is the first study to show that four (univariate) to five (multivariate) runs are needed to establish a stable running pattern in real-world settings., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The goal of mechanobiology is to understand the links between changes in the physical properties of living cells and normal physiology and disease. This requires mechanical measurements that have appropriate spatial and temporal resolution within a single cell. Conventional atomic force microscopy (AFM) methods that acquire force curves pointwise are used to map the heterogeneous mechanical properties of cells. However, the resulting map acquisition time is much longer than that required to study many dynamic cellular processes. Dynamic AFM (dAFM) methods using resonant microcantilevers are compatible with higher-speed, high-resolution scanning; however, they do not directly acquire force curves and they require the conversion of a limited number of instrument observables to local mechanical property maps. We have recently developed a technique that allows commercial AFM systems equipped with direct cantilever excitation to quantitatively map the viscoelastic properties of live cells. The properties can be obtained at several widely spaced frequencies with nanometer-range spatial resolution and with fast image acquisition times (tens of seconds). Here, we describe detailed procedures for quantitative mapping, including sample preparation, AFM calibration, and data analysis. The protocol can be applied to different biological samples, including cells and viruses. The transition from dAFM imaging to quantitative mapping should be easily achievable for experienced AFM users, who will be able to set up the protocol in <30 min.

We present an ultrafast single submicron particle detection method based on a half-bowtie coplanar waveguide. The method is capable of resolving the translocation of these particles at a bandwidth greater than 30 MHz. We compare experimentally the simultaneous use of our radio-frequency technique with conventional dc based resistive pulse recordings and find that our method has a throughput that is enhanced by 2 orders of magnitude. The technique incorporates a microfluidic circuit and has the potential to be employed for screening microparticles and biological cells at frequencies in excess of 1 GHz.

One of the fundamental questions guiding research in the biological sciences is how cellular systems process complex physical and environmental cues and communicate with each other across multiple length scales. Importantly, aberrant signal processing in these systems can lead to diseases that can have devastating impacts on human lives. Biophysical studies in the past several decades have demonstrated that cells can respond to not only biochemical cues but also mechanical and electrical ones. Thus, the development of new materials that can both sense and modulate all of these pathways is necessary. Semiconducting nanostructures are an emerging class of discovery platforms and tools that can push the limits of our ability to modulate and sense biological behaviors for both fundamental research and clinical applications. These materials are of particular interest for interfacing with cellular systems due to their matched dimension with subcellular components (e.g., cytoskeletal filaments), and easily tunable properties in the electrical, optical and mechanical regimes. Rational design via traditional or new approaches, such as nanocasting and mesoscale chemical lithography, can allow us to control micro- and nanoscale features in nanowires to achieve new biointerfaces. Both processes endogenous to the target cell and properties of the material surface dictate the character of these interfaces. In this Account, we focus on (1) approaches for the rational design of semiconducting nanowires that exhibit unique structures for biointerfaces, (2) recent fundamental discoveries that yield robust biointerfaces at the subcellular level, (3) intracellular electrical and mechanical sensing, and (4) modulation of cellular behaviors through material topography and remote physical stimuli. In the first section, we discuss new approaches for the synthetic control of micro- and nanoscale features of these materials. In the second section, we focus on achieving biointerfaces with these rationally designed materials either intra- or extracellularly. We last delve into the use of these materials in sensing mechanical forces and electrical signals in various cellular systems as well as in instructing cellular behaviors. Future research in this area may shift the paradigm in fundamental biophysical research and biomedical applications through (1) the design and synthesis of new semiconductor-based materials and devices that interact specifically with targeted cells, (2) the clarification of many developmental, physiological, and anatomical aspects of cellular communications, (3) an understanding of how signaling between cells regulates synaptic development (e.g., information like this would offer new insight into how the nervous system works and provide new targets for the treatment of neurological diseases), (4) and the creation of new cellular materials that have the potential to open up completely new areas of application, such as in hybrid information processing systems.

Microfluidics has the potential to transform experimental approaches across the life sciences. In this review, we discuss recent advances enabled by the development and application of microfluidic approaches to protein biophysics. We focus on areas where key fundamental features of microfluidics open up new possibilities and present advantages beyond low volumes and short time-scale analysis, conventionally provided by microfluidics. We discuss the two most commonly used forms of microfluidic technology, single-phase laminar flow and multiphase microfluidics. We explore how the understanding and control of the characteristic physical features of the microfluidic regime, the integration of microfluidics with orthogonal systems and the generation of well-defined microenvironments can be used to develop novel devices and methods in protein biophysics for sample manipulation, functional and structural studies, detection and material processing., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The accuracy of calculations of both the degree and angle of polarization depend strongly on the noise in the measurements used. The noise in the measurements recorded by both camera based systems and spectrometers can lead to significant artefacts and incorrect conclusions about high degrees of polarization when in fact none exist. Three approaches are taken in this work: firstly, the absolute error introduced as a function of the signal to noise ratio for polarization measurements is quantified in detail. An important finding here is the reason for why several studies incorrectly suggest that black (low reflectivity) objects are highly polarized. The high degree of polarization is only an artefact of the noise in the calculation. Secondly, several simple steps to avoid such errors are suggested. Thirdly, if these points can not be followed, two methods are presented for mitigating the effects of noise: a maximum likelihood estimation method and a new denoising algorithm to best calculate the degree of polarization of natural polarization information.

High-pressure microscopy is one of the powerful techniques to visualize the effects of hydrostatic pressures on research targets. It could be used for monitoring the pressure-induced changes in the structure and function of molecular machines in vitro and in vivo. This review focuses on the dynamic properties of the assemblies and machines, analyzed by means of high-pressure microscopy measurement. We developed a high-pressure microscope that is optimized both for the best image formation and for the stability to hydrostatic pressure up to 150 MPa. Application of pressure could change polymerization and depolymerization processes of the microtubule cytoskeleton, suggesting a modulation of the intermolecular interaction between tubulin molecules. A novel motility assay demonstrated that high hydrostatic pressure induces counterclockwise (CCW) to clockwise (CW) reversals of the Escherichia coli flagellar motor. The present techniques could be extended to study how molecular machines in complicated systems respond to mechanical stimuli., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background : Wearable magneto-inertial sensors are being increasingly used to obtain human motion measurements out of the lab, although their performance in applications requiring high accuracy, such as gait analysis, are still a subject of debate. The aim of this work was to validate a gait analysis system (H-Gait) based on magneto-inertial sensors, both in normal weight (NW) and overweight/obese (OW) subjects. The validation is performed against a reference multichannel recording system (STEP32), providing direct measurements of gait timings (through foot-switches) and joint angles in the sagittal plane (through electrogoniometers). Methods : Twenty-two young male subjects were recruited for the study (12 NW, 10 OW). After positioning body-fixed sensors of both systems, each subject was asked to walk, at a self-selected speed, over a 14-m straight path for 12 trials. Gait signals were recorded, at the same time, with the two systems. Spatio-temporal parameters, ankle, knee, and hip joint kinematics were extracted analyzing an average of 89 ± 13 gait cycles from each lower limb. Intraclass correlation coefficient and Bland-Altmann plots were used to compare H-Gait and STEP32 measurements. Changes in gait parameters and joint kinematics of OW with respect NW were also evaluated. Results : The two systems were highly consistent for cadence, while a lower agreement was found for the other spatio-temporal parameters. Ankle and knee joint kinematics is overall comparable. Joint ROMs values were slightly lower for H-Gait with respect to STEP32 for the ankle (by 1.9° for NW, and 1.6° for OW) and for the knee (by 4.1° for NW, and 1.8° for OW). More evident differences were found for hip joint, with ROMs values higher for H-Gait (by 6.8° for NW, and 9.5° for OW). NW and OW showed significant differences considering STEP32 ( p = 0.0004), but not H-Gait ( p = 0.06). In particular, overweight/obese subjects showed a higher cadence (55.0 vs. 52.3 strides/min) and a lower hip ROM (23.0° vs. 27.3°) than normal weight subjects. Conclusions : The two systems can be considered interchangeable for what concerns joint kinematics, except for the hip, where discrepancies were evidenced. Differences between normal and overweight/obese subjects were statistically significant using STEP32. The same tendency was observed using H-Gait., Competing Interests: The authors declare that they have no competing interests. Ethics approval and consent to participate: This study was approved by the local Institutional Review Board and all procedures conformed to the Helsinki declaration. Written informed consent was obtained from all participants. Consent for publication: Consent for publication was obtained for the pictures in . Availability of data and material: The datasets analyzed during the current study are available from the corresponding author on reasonable request.

Background: Body movements, such as trunk flexion and rotation, are risk factors for low back pain in occupational settings, especially in healthcare workers. Wearable motion capture systems are potentially useful to monitor lower back movement in healthcare workers to help avoid the risk factors. In this study, we propose a novel system using sheet stretch sensors and investigate the system validity for estimating lower back movement., Methods: Six volunteers (female:male = 1:1, mean age: 24.8 ± 4.0 years, height 166.7 ± 5.6 cm, weight 56.3 ± 7.6 kg) participated in test protocols that involved executing seven types of movements. The movements were three uniaxial trunk movements (i.e., trunk flexion-extension, trunk side-bending, and trunk rotation) and four multiaxial trunk movements (i.e., flexion + rotation, flexion + side-bending, side-bending + rotation, and moving around the cranial-caudal axis). Each trial lasted for approximately 30 s. Four stretch sensors were attached to each participant's lower back. The lumbar motion angles were estimated using simple linear regression analysis based on the stretch sensor outputs and compared with those obtained by the optical motion capture system., Results: The estimated lumbar motion angles showed a good correlation with the actual angles, with correlation values of r = 0.68 (SD = 0.35), r = 0.60 (SD = 0.19), and r = 0.72 (SD = 0.18) for the flexion-extension, side bending, and rotation movements, respectively (all P < 0.05). The estimation errors in all three directions were less than 3°., Conclusion: The stretch sensors mounted on the back provided reasonable estimates of the lumbar motion angles. The novel motion capture system provided three directional angles without capture space limits. The wearable system possessed great potential to monitor the lower back movement in healthcare workers and helping prevent low back pain.

Background: Application of in-shoe multi-segment foot kinematic analyses currently faces a number of challenges, including: (i) the difficulty to apply regular markers onto the skin, (ii) the necessity for an adequate shoe which fits various foot morphologies and (iii) the need for adequate repeatability throughout a repeated measure condition. The aim of this study therefore was to design novel magnet based 3D printed markers for repeated in-shoe measurements while using accordingly adapted modified shoes for a specific multi-segment foot model., Methods: Multi-segment foot kinematics of ten participants were recorded and kinematics of hindfoot, midfoot and forefoot were calculated. Dynamic trials were conducted to check for intra and inter-session repeatability when combining novel markers and modified shoes in a repeated measures design. Intraclass correlation coefficients were calculated to determine reliability., Results: Both repeatability and reliability were proven to be good to excellent with maximum joint angle deviations of 1.11° for intra-session variability and 1.29° for same-day inter-session variability respectively and ICC values of >0.91., Conclusion: The novel markers can be reliably used in future research settings using in-shoe multi-segment foot kinematic analyses with multiple shod conditions.

The Radiological Research Accelerator Facility (RARAF) is in its 50th year of operation. It was commissioned on April 1, 1967 as a collaboration between the Radiological Research Laboratory (RRL) of Columbia University, and members of the Medical Research Center of Brookhaven National Laboratory (BNL). It was initially funded as a user facility for radiobiology and radiological physics, concentrating on monoenergetic neutrons. Facilities for irradiation with MeV light charged particles were developed in the mid-1970s. In 1980 the facility was relocated to the Nevis Laboratories of Columbia University. RARAF now has seven beam lines, each having a dedicated irradiation facility: monoenergetic neutrons, charged particle track segments, two charged particle microbeams (one electrostatically focused to <1 μm, one magnetically focused), a 4.5 keV soft X-ray microbeam, a neutron microbeam, and a facility that produces a neutron spectrum similar to that of the atomic bomb dropped at Hiroshima. Biology facilities are available on site within close proximity to the irradiation facilities, making the RARAF very user friendly.

The mechanical properties of the cellular microenvironment and their spatiotemporal variations are thought to play a central role in sculpting embryonic tissues, maintaining organ architecture and controlling cell behavior, including cell differentiation. However, no direct in vivo and in situ measurement of mechanical properties within developing 3D tissues and organs has yet been performed. Here we introduce a technique that employs biocompatible, magnetically responsive ferrofluid microdroplets as local mechanical actuators and allows quantitative spatiotemporal measurements of mechanical properties in vivo. Using this technique, we show that vertebrate body elongation entails spatially varying tissue mechanics along the anteroposterior axis. Specifically, we find that the zebrafish tailbud is viscoelastic (elastic below a few seconds and fluid after just 1 min) and displays decreasing stiffness and increasing fluidity toward its posterior elongating region. This method opens new avenues to study mechanobiology in vivo, both in embryogenesis and in disease processes, including cancer.

Characterization of molecular interactions in terms of basic binding parameters such as binding affinity, stoichiometry, and thermodynamics is an essential step in basic and applied science. MicroScale Thermophoresis (MST) is a sensitive biophysical method to obtain this important information. Relying on a physical effect called thermophoresis, which describes the movement of molecules through temperature gradients, this technology allows for the fast and precise determination of binding parameters in solution and allows the free choice of buffer conditions (from buffer to lysates/sera). MST uses the fact that an unbound molecule displays a different thermophoretic movement than a molecule that is in complex with a binding partner. The thermophoretic movement is altered in the moment of molecular interaction due to changes in size, charge, and hydration shell. By comparing the movement profiles of different molecular ratios of the two binding partners, quantitative information such as binding affinity (pM to mM) can be determined. Even challenging interactions between molecules of small sizes, such as aptamers and small compounds, can be studied by MST. Using the well-studied model interaction between the DH25.42 DNA aptamer and ATP, this manuscript provides a protocol to characterize aptamer-small molecule interactions. This study demonstrates that MST is highly sensitive and permits the mapping of the binding site of the 7.9 kDa DNA aptamer to the adenine of ATP.

In vitro tests are of fundamental importance for investigating cell mechanisms in response to mechanical stimuli or the impact of the genotype on cell mechanical properties. In particular, the application of controlled forces to activate specific bio-pathways and investigate their effects, mimicking the role of the cellular environment, is becoming a prominent approach in the emerging field of mechanobiology. Here, we present an on-chip device based on magnetic domain wall manipulators, which allows the application of finely controlled and localized forces on target living cells. In particular, we demonstrate the application of a magnetic force in the order of hundreds of pN on the membrane of HeLa cells cultured on-chip, via manipulation of 1 μm superparamagnetic beads. Such a mechanical stimulus produces a sizable local indentation of the cellular membrane of about 2 μm. Upon evaluation of the beads' position within the magnetic field originated by the domain wall, the force applied during the experiments is accurately quantified via micromagnetic simulations. The obtained value is in good agreement with that calculated by the application of an elastic model to the cellular membrane.

In vitro engineering systems can be powerful tools for studying tissue development in response to biophysical stimuli as well as for evaluating the functionality of engineered tissue grafts. It has been challenging, however, to develop systems that adequately integrate the application of biomimetic mechanical strain to engineered tissue with the ability to assess functional outcomes in real time. The aim of this study was to design a bioreactor system capable of real-time conditioning (dynamic, uniaxial strain, and electrical stimulation) of centimeter-long 3D tissue engineered constructs simultaneously with the capacity to monitor local strains. The system addresses key limitations of uniform sample loading and real-time imaging capabilities. Our system features an electrospun fibrin scaffold, which exhibits physiologically relevant stiffness and uniaxial alignment that facilitates cell adhesion, alignment, and proliferation. We have demonstrated the capacity for directly incorporating human adipose-derived stromal/stem cells into the fibers during the electrospinning process and subsequent culture of the cell-seeded constructs in the bioreactor. The bioreactor facilitates accurate pre-straining of the 3D constructs as well as the application of dynamic and static uniaxial strains while monitoring bulk construct tensions. The incorporation of fluorescent nanoparticles throughout the scaffolds enables in situ monitoring of local strain fields using fluorescent digital image correlation techniques, since the bioreactor is imaging compatible, and allows the assessment of local sample stiffness and stresses when coupled with force sensor measurements. In addition, the system is capable of measuring the electromechanical coupling of skeletal muscle explants by applying an electrical stimulus and simultaneously measuring the force of contraction. The packaging of these technologies, biomaterials, and analytical methods into a single bioreactor system has produced a powerful tool that will enable improved engineering of functional 3D ligaments, tendons, and skeletal muscles. Biotechnol. Bioeng. 2016;113: 1825-1837. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Wood contains a large amount of air, even in functional xylem. Air embolisms in the xylem affect water transport and can determine plant growth and survival. Embolisms are usually estimated with laborious hydraulic methods, which can be prone to several artefacts. Here, we describe a new method for estimating embolisms that is based on air flow measurements of entire branches. To calculate the amount of air flowing out of the branch, a vacuum was applied to the cut bases of branches under different water potentials. We first investigated the source of air by determining whether it came from inside or outside the branch. Second, we compared embolism curves according to air flow or hydraulic measurements in 15 vessel- and tracheid-bearing species to test the hypothesis that the air flow is related to embolism. Air flow came almost exclusively from air inside the branch during the 2.5-min measurements and was strongly related to embolism. We propose a new embolism measurement method that is simple, effective, rapid and inexpensive, and that allows several measurements on the same branch, thus opening up new possibilities for studying plant hydraulics., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)

Background: Skin, as a protective barrier to exogenous substances, can be modulated by various internal and external factors that can affect its functional state. In order to prevent the early symptoms and signs of diseases of the skin, frequent skin health assessment should be performed. The aims of the study were to evaluate four skin properties of transepidermal water loss (TEWL), hydration, elasticity, and pigmentation using a non-invasive skin assessment tool, DermaLab Combo(®) , and also to determine possible factors that may influence skin condition., Methods: DermaLab(®) Combo was used to measure TEWL, hydration, pigmentation, and elasticity on the forearm of volunteers by using different probes. In this study, four parameters were observed to reflect the health of the skin in 100 volunteers., Results: There were significant differences (P < 0.05) between TEWL, hydration, pigmentation, and elasticity in different genders on the same anatomical site of the forearm. Female subjects have a higher average value of TEWL, hydration, and elasticity compared to male subjects. The differences may be due to an individual's daily activity and use of skin care products as well as environmental factors. The use of moisturiser and drinking lots of water may keep the skin hydrated and delay the process of skin ageing as shown by the better hydration and elasticity observed (P < 0.05)., Conclusion: In this study, it can be concluded that DermaLab(®) Combo is a reliable skin analysis instrument that offers high precision, accuracy, and reproducibility for all the measuring parameters. It has also been found that daily activities and habits influence skin condition as reflected by the measurement of these biophysical skin parameters., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Our tissues consist of individual cells that respond to the elasticity of their environment, which varies between and within tissues. To better understand mechanically driven cell migration, it is necessary to manipulate the stiffness gradient across a substrate. Here, we have demonstrated a new variant of the microfabricated polymeric pillar array platform that can decouple the stiffness gradient from the ECM protein area. This goal is achieved via a "stepped" micro pillar array device (SMPAD) in which the contact area with the cell was kept constant while the diameter of the pillar bodies was altered to attain the proper mechanical stiffness. Using double-step SU-8 mold fabrication, the diameter of the top of every pillar was kept uniform, whereas that of the bottom was changed, to achieve the desired substrate rigidity. Fibronectin was immobilized on the pillar tops, providing a focal adhesion site for cells. C2C12, HeLa and NIH3T3 cells were cultured on the SMPAD, and the motion of the cells was observed by time-lapse microscopy. Using this simple platform, which produces a purely physical stimulus, we observed that various types of cell behavior are affected by the mechanical stimulus of the environment. We also demonstrated directed cell migration guided by a discrete rigidity gradient by varying stiffness. Interestingly, cell velocity was highest at the highest stiffness. Our approach enables the regulation of the mechanical properties of the polymeric pillar array device and eliminates the effects of the size of the contact area. This technique is a unique tool for studying cellular motion and behavior relative to various stiffness gradients in the environment.

In mechanobiology the study of cell response to mechanical stimuli is fundamental, and the involved processes (i.e., mechanotransduction) need to be investigated by interfacing (mechanically and electrically) with the cells in dynamic and non-invasive natural-like conditions. In this work, we present a novel soft, stretchable and conductive biointerface that allows both cell mechanical stimulation and dynamic impedance recording. The biointerface stretchability and conductivity, jointly to the biocompatibility and transparency needed to perform cell culture studies, were obtained by exploiting the formation of wrinkles on the surface of a 90 nm thick conductive layer of poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) on a pre-stretched 130 μm thick poly(dimethylsiloxane) (PDMS) substrate. Cell adhesion and proliferation of SH-SY5Y human neuroblastoma cells were evaluated, and cell differentiation on the corrugated surface was assessed. We demonstrate how the biointerface remains conductive when applying uniaxial strain up to 10%, and when cell culturing is performed. Finally, a reduction of about 30% of the relative impedance variation signal was measured, with respect to the control, as a result of the mechanical stimulation of cells.

Around 1900, several experimenters investigated turbulences in wind tunnels or water basins by creating visualizations. One of them, the German zoologist Friedrich Ahlborn (1858-1937), was familiar with the works by his contemporaries but he struck a new path. He combined three different kinds of photographs taken at the same time and showed the same situation in his water trough-but each in a different way. With this first basic operation, Ahlborn heuristically opened up a previously non-existent space for experimentation, analysis, and recombination. He generated an astonishing diversity of information by adopting the tactics of 'inversions' in which he interpreted one part of the experimental setup, or its results, in different ways. Between the variants of the 'autographs' which he developed, he defined areas of intersection to be able to translate results from individual records into each other. To this end, Ahlborn created other sets of visual artifacts such as drawn diagrams, three-dimensional wire frame constructions, and clay reliefs. His working method can be described as a cascading array of successive modeling steps, as elaborated by Eric Winsberg (1999), or of inscriptions in Bruno Latour's words (Latour, 1986). By examining Ahlborn's procedures closely we propose conceptualizations for the experimenter's various operations., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

To study the behavior of biological macromolecules and enzymatic reactions under force, advances in single-molecule force spectroscopy have proven instrumental. Magnetic tweezers form one of the most powerful of these techniques, due to their overall simplicity, non-invasive character, potential for high throughput measurements, and large force range. Drawbacks of magnetic tweezers, however, are that accurate determination of the applied forces can be challenging for short biomolecules at high forces and very time-consuming for long tethers at low forces below ∼1 piconewton. Here, we address these drawbacks by presenting a calibration standard for magnetic tweezers consisting of measured forces for four magnet configurations. Each such configuration is calibrated for two commonly employed commercially available magnetic microspheres. We calculate forces in both time and spectral domains by analyzing bead fluctuations. The resulting calibration curves, validated through the use of different algorithms that yield close agreement in their determination of the applied forces, span a range from 100 piconewtons down to tens of femtonewtons. These generalized force calibrations will serve as a convenient resource for magnetic tweezers users and diminish variations between different experimental configurations or laboratories.

The nucleus is physically distinct from the cytoplasm in ways that suggest new ideas and approaches for interrogating the operation of this organelle. Chemical bond formation and breakage underlie the lives of cells, but as this special issue of Molecular Biology of the Cell attests, the nonchemical aspects of cell nuclei present a new frontier to biologists and biophysicists., (© 2014 Pederson and Marko. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

Understanding the electrical biophysical properties of the cell membrane can be difficult for neuroscience students as it relies solely on lectures of theoretical models without practical hands on experiments. To address this issue, we developed an open-source lipid bilayer amplifier, the OpenPicoAmp, which is appropriate for use in introductory courses in biophysics or neurosciences at the undergraduate level, dealing with the electrical properties of the cell membrane. The amplifier is designed using the common lithographic printed circuit board fabrication process and off-the-shelf electronic components. In addition, we propose a specific design for experimental chambers allowing the insertion of a commercially available polytetrafluoroethylene film. We provide a complete documentation allowing to build the amplifier and the experimental chamber. The students hand-out giving step-by step instructions to perform a recording is also included. Our experimental setup can be used in basic experiments in which students monitor the bilayer formation by capacitance measurement and record unitary currents produced by ionic channels like gramicidin A dimers. Used in combination with a low-cost data acquisition board this system provides a complete solution for hands-on lessons, therefore improving the effectiveness in teaching basic neurosciences or biophysics.

The properties of substrates and extracellular matrices (ECM) are important factors governing the functions and fates of mammalian adherent cells. For example, substrate stiffness often affects cell differentiation. At focal adhesions, clustered-integrin bindings link cells mechanically to the ECM. In order to quantitate the affinity between cell and substrate, the cell adhesion force must be measured for single cells. In this study, forcible detachment of a single cell in the vertical direction using AFM was carried out, allowing breakage of the integrin-substrate bindings. An AFM tip was fabricated into an arrowhead shape to detach the cell from the substrate. Peak force observed in the recorded force curve during probe retraction was defined as the adhesion force, and was analyzed for various types of cells. Some of the cell types adhered so strongly that they could not be picked up because of plasma membrane breakage by the arrowhead probe. To address this problem, a technique to reinforce the cellular membrane with layer-by-layer nanofilms composed of fibronectin and gelatin helped to improve insertion efficiency and to prevent cell membrane rupture during the detachment process, allowing successful detachment of the cells. This method for detaching cells, involving cellular membrane reinforcement, may be beneficial for evaluating true cell adhesion forces in various cell types., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Hydrodynamic lift forces acting on cells and particles in fluid flow receive ongoing attention from medicine, mathematics, physics and engineering. The early findings of Fåhræus & Lindqvist on the viscosity change of blood with the diameter of capillaries motivated extensive studies both experimentally and theoretically to illuminate the underlying physics. We review this historical development that led to the discovery of the inertial and non-inertial lift forces and elucidate the origins of these forces that are still not entirely clear. Exploiting microfluidic techniques induced a tremendous amount of new insights especially into the more complex interactions between the flow field and deformable objects like vesicles or red blood cells. We trace the way from the investigation of single cell dynamics to the recent developments of microfluidic techniques for particle and cell sorting using hydrodynamic forces. Such continuous and label-free on-chip cell sorting devices promise to revolutionize medical analyses for personalized point-of-care diagnosis. We present the state-of-the-art of different hydrodynamic lift-based techniques and discuss their advantages and limitations., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Monitoring cell growth and measuring physical features of food-borne pathogenic bacteria are important for better understanding the conditions under which these organisms survive and proliferate. To address this challenge, buoyant masses of live and dead Escherichia coli O157:H7 and Listeria innocua were measured using Archimedes, a commercially available suspended microchannel resonator (SMR). Cell growth was monitored with Archimedes by observing increased cell concentration and buoyant mass values of live growing bacteria. These growth data were compared to optical density measurements obtained with a Bioscreen system. We observed buoyant mass measurements with Archimedes at cell concentrations between 10(5) and 10(8) cells/ml, while growth was not observed with optical density measurements until the concentration was 10(7) cells/ml. Buoyant mass measurements of live and dead cells with and without exposure to hydrogen peroxide stress were also compared; live cells generally had a larger buoyant mass than dead cells. Additionally, buoyant mass measurements were used to determine cell density and total mass for both live and dead cells. Dead E. coli cells were found to have a larger density and smaller total mass than live E. coli cells. In contrast, density was the same for both live and dead L. innocua cells, while the total mass was greater for live than for dead cells. These results contribute to the ongoing challenge to further develop existing technologies used to observe cell populations at low concentrations and to measure unique physical features of cells that may be useful for developing future diagnostics.

Time-delay of transmitted pulses with respect to the incident pulse in bacteriorhodopsin films has been studied without the use of a pump beam. Based on a modified saturable absorber model, analytical expressions of the transmitted pulse have been obtained. As a result, time delay, distortion and fractional delay have been analyzed for sinusoidal pulses with a low background. A good agreement between theory and experiences has been observed.

A multi-layered polydimethylsiloxane microfluidic device with an integrated suspended membrane has been fabricated that allows dynamic and multi-axial mechanical deformation and simultaneous live-cell microscopy imaging. The transparent membrane's strain field can be controlled independently along two orthogonal directions. Human foreskin fibroblasts were immobilized on the membrane's surface and stretched along two orthogonal directions sequentially while performing live-cell imaging. Cyclic deformation of the cells induced a reversible reorientation perpendicular to the direction of the applied strain. Cells remained viable in the microdevice for several days. As opposed to existing microfluidic or macroscale stretching devices, this device can impose changing, anisotropic and time-varying strain fields in order to more closely mimic the complexities of strains occurring in vivo.

Zero-mode waveguides provide a powerful technology for studying single-molecule real-time dynamics of biological systems at physiological ligand concentrations. We customized a commercial zero-mode waveguide-based DNA sequencer for use as a versatile instrument for single-molecule fluorescence detection and showed that the system provides long fluorophore lifetimes with good signal to noise and low spectral cross-talk. We then used a ribosomal translation assay to show real-time fluidic delivery during data acquisition, showing it is possible to follow the conformation and composition of thousands of single biomolecules simultaneously through four spectral channels. This instrument allows high-throughput multiplexed dynamics of single-molecule biological processes over long timescales. The instrumentation presented here has broad applications to single-molecule studies of biological systems and is easily accessible to the biophysical community.

Magnetic tweezers (MTW) enable highly accurate forces to be transduced to molecules to study mechanotransduction at the molecular or cellular level. We review recent MTW studies in single molecule and cell biophysics that demonstrate the flexibility of this technique. We also discuss technical advances in the method on several fronts, i.e., from novel approaches for the measurement of torque to multiplexed biophysical assays. Finally, we describe multi-component nanorods with enhanced optical and magnetic properties and discuss their potential as future MTW probes.

Background: Challenges in accurate, in vivo quantification of multi-planar knee kinematics and relevant timing sequence during high-risk injurious tasks pose challenges in understanding the relative contributions of joint loads in non-contact injury mechanisms. Biomechanical testing on human cadaveric tissue, if properly designed, offers a practical means to evaluate joint biomechanics and injury mechanisms. This study seeks to investigate the detailed interactions between tibiofemoral joint multi-planar kinematics and anterior cruciate ligament strain in a cadaveric model of landing using a validated physiologic drop-stand apparatus., Methods: Sixteen instrumented cadaveric legs, mean 45(SD 7) years (8 female and 8 male) were tested. Event timing sequence, change in tibiofemoral kinematics (position, angular velocity and linear acceleration) and change in anterior cruciate ligament strain were quantified., Findings: The proposed cadaveric model demonstrated similar tibiofemoral kinematics/kinetics as reported measurements obtained from in vivo studies. While knee flexion, anterior tibial translation, knee abduction and increased anterior cruciate ligament strain initiated and reached maximum values almost simultaneously, internal tibial rotation initiated and peaked significantly later (P<0.015 for all comparisons). Further, internal tibial rotation reached mean 1.8(SD 2.5)°, almost 63% of its maximum value, at the time that peak anterior cruciate ligament strain occurred, while both anterior tibial translation and knee abduction had already reached their peaks., Interpretation: Together, these findings indicate that although internal tibial rotation contributes to increased anterior cruciate ligament strain, it is secondary to knee abduction and anterior tibial translation in its effect on anterior cruciate ligament strain and potential risk of injury., (© 2013. Published by Elsevier Ltd on behalf of American Society of Biomechanics. All rights reserved.)