Respiratory Viral Infections from 2015 to 2022 in the HIVE Cohort of American Households: Incidence, illness characteristics, and seasonality.

BACKGROUND: Viral respiratory illnesses are the most common acute illnesses experienced and generally follow a predicted pattern over time. The SARS-CoV-2 pandemic interrupted that pattern. METHODS: The HIVE (Household Influenza Vaccine Evaluation) study was established in 2010 to follow a cohort of Southeast Michigan households over time. Initially focused on influenza, surveillance was expanded to include other major respiratory pathogens, and, starting in 2015, the population was followed year-round. Symptoms of acute illness were reported, and respiratory specimens were collected and tested to identify viral infections. Based on the known population being followed, virus-specific incidence was calculated. RESULTS: From 2015 to 2022, 1755 participants were followed in HIVE for 7785 person-years with 7833 illnesses documented. Before the pandemic, rhinovirus (RV) and common cold human coronaviruses (HCoVs) were the viruses most frequently identified, and incidence decreased with increasing age. Type A influenza was next but with comparable incidence by age. Parainfluenza and respiratory syncytial viruses were less frequent overall, followed by human metapneumoviruses. Incidence was highest in young children, but infections were frequently documented in all age groups. Seasonality followed patterns established decades ago. The SARS-CoV-2 pandemic disrupted these patterns, except for RV and, to a lesser extent, HCoVs. In the first two years of the pandemic, RV incidence far exceeded that of SARS-CoV-2. CONCLUSION: Longitudinal cohort studies are important in comparing the incidence, seasonality, and characteristics of different respiratory viral infections. Studies documented the differential effect of the pandemic on the incidence of respiratory viruses in addition to SARS-CoV-2.

Comparison of a Scaled Cadaver-Based Musculoskeletal Model With a Clinical Upper Extremity Model.

Reliably and accurately estimating joint/segmental kinematics from optical motion capture data has remained challenging. Studies objectively characterizing human movement patterns have typically involved inverse kinematics and inverse dynamics techniques. Subsequent research has included scaled cadaver-based musculoskeletal (MSK) modeling for noninvasively estimating joint and muscle loads. As one of the ways to enhance confidence in the validity of MSK model predictions, the kinematics from the preceding step that drives such a model needs to be checked for agreement or compared with established/widely used models. This study rigorously compares the upper extremity (UE) joint kinematics calculated by the Dutch Shoulder Model implemented in the AnyBody Managed Model Repository (involving multibody kinematics optimization (MKO)) with those estimated by the Vicon Plug-in Gait model (involving single-body kinematics optimization (SKO)). Ten subjects performed three trials of (different types of) reaching tasks in a three-dimensional marker-based optical motion capture laboratory setting. Joint angles, processed marker trajectories, and reconstruction residuals corresponding to both models were compared. Scatter plots and Bland-Altman plots were used to assess the agreement between the two model outputs. Results showed the largest differences between the two models for shoulder, followed by elbow and wrist, with all root-mean-squared differences less than 10 deg (although this limit might be unacceptable for clinical use). Strong-to-excellent Spearman's rank correlation coefficients were found between the two model outputs. The Bland-Altman plots showed a good agreement between most of the outputs. In conclusion, results indicate that these two models with different kinematic algorithms broadly agree with each other, albeit with few key differences.

Effectiveness of Influenza Vaccines in the HIVE Household Cohort Over 8 Years: Is There Evidence of Indirect Protection?

BACKGROUND: The evidence that influenza vaccination programs regularly provide protection to unvaccinated individuals (ie, indirect effects) of a community is lacking. We sought to determine the direct, indirect, and total effects of influenza vaccine in the Household Influenza Vaccine Evaluation (HIVE) cohort. METHODS: Using longitudinal data from the HIVE cohort from 2010-11 through 2017-18, we estimated direct, indirect, and total influenza vaccine effectiveness (VE) and the incidence rate ratio of influenza virus infection using adjusted mixed-effect Poisson regression models. Total effectiveness was determined through comparison of vaccinated members of full or partially vaccinated households to unvaccinated individuals in completely unvaccinated households. RESULTS: The pooled, direct VE against any influenza was 30.2% (14.0-43.4). Direct VE was higher for influenza A/H1N1 43.9% (3.9 to 63.5) and B 46.7% (17.2 to 57.5) than A/H3N2 31.7% (10.5 to 47.8) and was higher for young children 42.4% (10.1 to 63.0) than adults 18.6% (-6.3 to 37.7). Influenza incidence was highest in completely unvaccinated households (10.6 per 100 person-seasons) and lower at all other levels of household vaccination coverage. We found little evidence of indirect VE after adjusting for potential confounders. Total VE was 56.4% (30.1-72.9) in low coverage, 43.2% (19.5-59.9) in moderate coverage, and 33.0% (12.1 to 49.0) in fully vaccinated households. CONCLUSIONS: Influenza vaccines may have a benefit above and beyond the direct effect but that effect in this study was small. Although there may be exceptions, the goal of global vaccine recommendations should remain focused on provision of documented, direct protection to those vaccinated.

Ion current and action potential alterations in peripheral neurons subject to uniaxial strain.

Peripheral nerves, subject to continuous elongation and compression during everyday movement, contain neuron fibers vital for movement and sensation. At supraphysiological strains resulting from trauma, chronic conditions, aberrant limb positioning, or surgery, conduction blocks occur which may result in chronic or temporary loss of function. Previous in vitro stretch models, mainly focused on traumatic brain injury modelling, have demonstrated altered electrophysiological behavior during localized deformation applied by pipette suction. Our aim was to evaluate the changes in voltage-activated ion channel function during uniaxial straining of neurons applied by whole-cell deformation, more physiologically relevant model of peripheral nerve trauma. Here, we quantified experimentally the changes in inwards and outwards ion currents and action potential (AP) firing in dorsal root ganglion-derived neurons subject to uniaxial strains, using a custom-built device allowing simultaneous cell deformation and patch clamp recording. Peak inwards sodium currents and rectifying potassium current magnitudes were found to decrease in cells under stretch, channel reversal potentials were found to be left-shifted, and half-maximum activation potentials right-shifted. The threshold for AP firing was increased in stretched cells, although neurons retained the ability to fire induced APs. Overall, these results point to ion channels being damaged directly and immediately by uniaxial strain, affecting cell electrophysiological activity, and can help develop prevention and treatment strategies for peripheral neuropathies caused by mechanical trauma.

Elastic fibres are broadly distributed in tendon and highly localized around tenocytes.

Elastic fibres have the unique ability to withstand large deformations and are found in numerous tissues, but their organization and structure have not been well defined in tendon. The objective of this study was to characterize the organization of elastic fibres in tendon to understand their function. Immunohistochemistry was used to visualize elastic fibres in bovine flexor tendon with fibrillin-1, fibrillin-2 and elastin antibodies. Elastic fibres were broadly distributed throughout tendon, and highly localized longitudinally around groups of cells and transversely between collagen fascicles. The close interaction of elastic fibres and cells suggests that elastic fibres are part of the pericellular matrix and therefore affect the mechanical environment of tenocytes. Fibres present between fascicles are likely part of the endotenon sheath, which enhances sliding between adjacent collagen bundles. These results demonstrate that elastic fibres are highly localized in tendon and may play an important role in cellular function and contribute to the tissue mechanics of the endotenon sheath.

Tendon mechanobiology: experimental models require mathematical underpinning.

Mathematical and computational modeling is in demand to help address current challenges in mechanobiology of musculoskeletal tissues. In particular for tendon, the high clinical importance of the tissue, the huge mechanical demands placed on it and its ability to adapt to these demands, require coupled, multiscale models incorporating complex geometrical and microstructural information as well as time-based descriptions of cellular activity and response.This review introduces the information sources required to develop such multiscale models. It covers tissue structure and biomechanics, cell biomechanics, the current understanding of tendon's ability in health and disease to update its properties and structure and the few already existing multiscale mechanobiological models of the tissue. Finally, a sketch is provided of what such models could achieve ideally, pointing out where experimental data and knowledge are still missing.

A novel method for the accurate evaluation of Poisson's ratio of soft polymer materials.

A new method with a simple algorithm was developed to accurately measure Poisson's ratio of soft materials such as polyvinyl alcohol hydrogel (PVA-H) with a custom experimental apparatus consisting of a tension device, a micro X-Y stage, an optical microscope, and a charge-coupled device camera. In the proposed method, the initial positions of the four vertices of an arbitrarily selected quadrilateral from the sample surface were first measured to generate a 2D 1st-order 4-node quadrilateral element for finite element numerical analysis. Next, minimum and maximum principal strains were calculated from differences between the initial and deformed shapes of the quadrilateral under tension. Finally, Poisson's ratio of PVA-H was determined by the ratio of minimum principal strain to maximum principal strain. This novel method has an advantage in the accurate evaluation of Poisson's ratio despite misalignment between specimens and experimental devices. In this study, Poisson's ratio of PVA-H was 0.44 ± 0.025 (n = 6) for 2.6-47.0% elongations with a tendency to decrease with increasing elongation. The current evaluation method of Poisson's ratio with a simple measurement system can be employed to a real-time automated vision-tracking system which is used to accurately evaluate the material properties of various soft materials.

Development and Optimisation of Hydrogel Scaffolds for Microvascular Network Formation.

Traumatic injuries are a major cause of morbidity and mortality worldwide; however, there is limited research on microvascular traumatic injuries. To address this gap, this research aims to develop and optimise an in vitro construct for traumatic injury research at the microvascular level. Tissue engineering constructs were created using a range of polymers (collagen, fibrin, and gelatine), solvents (PBS, serum-free endothelial media, and MES/NaCl buffer), and concentrations (1-5% w/v). Constructs created from these hydrogels and HUVECs were evaluated to identify the optimal composition in terms of cell proliferation, adhesion, migration rate, viability, hydrogel consistency and shape retention, and tube formation. Gelatine hydrogels were associated with a lower cell adhesion, whereas fibrin and collagen ones displayed similar or better results than the control, and collagen hydrogels exhibited poor shape retention; fibrin scaffolds, particularly at high concentrations, displayed good hydrogel consistency. Based on the multipronged evaluation, fibrin hydrogels in serum-free media at 3 and 5% w/v were selected for further experimental work and enabled the formation of interconnected capillary-like networks. The networks formed in both hydrogels displayed a similar architecture in terms of the number of segments (10.3 ± 3.21 vs. 9.6 ± 3.51) and diameter (8.6446 ± 3.0792 µm vs. 7.8599 ± 2.3794 µm).

Tensile and shear mechanical properties of rotator cuff repair patches.

BACKGROUND: Augmentation of rotator cuff tears aims to strengthen the repair and reduce rerupture, yet studies still report high failure rates. This study determines key mechanical properties of rotator cuff repair patches, including establishing values for toughness and measuring the shear properties of repair patches and human rotator cuff tendons. We hypothesized that different repair grafts would (1) have varying material parameters, and (2) not all have mechanical properties similar to human rotator cuff tendons. MATERIALS AND METHODS: Eight specimens each from the Restore, GraftJacket, Zimmer Collagen Repair, and SportsMesh repair patches were tested to failure in tension and for suture pullout. We assessed ultimate tensile strength, tensile (Young's) modulus, and failure strain. This study also established toughness values and shear data. Storage modulus was calculated using dynamic shear analysis for the patches and 18 samples of normal rotator cuff tendon. RESULTS: We report significant variability in important mechanical properties of repair patches, with the mechanical parameters of the patches diverting variously-and often significantly-from values for human rotator cuff tendon. CONCLUSIONS: The repair grafts tested all displayed significant variation in their mechanical properties and had at least some reduced parameters compared with human rotator cuff tendons. This study offers experimentally derived information of value to surgeons when selecting rotator cuff repair grafts. A better understanding of the mechanical suitability of repair grafts for supporting human rotator cuffs is needed if repair patches are to provide a solution for the clinical problem of failure of rotator cuff repairs.

Comparing thermal discomfort with skin temperature response of lower-limb prosthesis users during exercise.

BACKGROUND: Thermal discomfort is prevalent among prosthesis users. This observational study of thirty unilateral lower-limb prosthesis users compared their skin temperatures and the thermal discomfort experienced during exercise between their residual and contralateral limbs. METHODS: Participants performed a 2-minute interval cycling exercise test. Skin temperature was measured at matched locations on each leg during the 1-minute rest intervals. Average rate-of-change in skin temperature was compared between legs using a repeated measures analysis of variance. Participants rated thermal discomfort on each leg before and after exercise, and a Wilcoxon signed-rank test was used to compare legs. Ordinal regression evaluated the relationship between the rate-of-change in temperature on the residual limb and the perceived thermal discomfort. FINDINGS: After exercise, thermal discomfort ranked higher on the amputated side (P = 0.007). On average, both legs cooled during exercise (P = 0.002), but the difference between legs was not significant. The rate-of change in skin temperature on the residual limb during exercise did not relate to the thermal discomfort experienced (odds ratio of 0.357). INTERPRETATION: These findings indicate that in this patient population, skin temperature does not explain the thermal discomfort experienced, and subjective thermal discomfort is inadequate for detecting thermoregulatory issues, with potential implications for long-term tissue health.

Engineering a uniaxial substrate-stretching device for simultaneous electrophysiological measurements and imaging of strained peripheral neurons.

Peripheral nerves are continuously subjected to mechanical strain during everyday movements, but excessive stretch can lead to damage and neuronal cell functionality can also be impaired. To better understand cellular processes triggered by stretch, it is necessary to develop in vitro experimental methods that allow multiple concurrent measurements and replicate in vivo mechanical conditions. Current commercially available cell stretching devices do not allow flexible experimental design, restricting the range of possible multi-physics measurements. Here, we describe and characterise a custom-built uniaxial substrate-straining device, with which neurons cultured on aligned patterned surfaces (50 µm wide grooves) can be strained up to 70% and simultaneously imaged with widefield and confocal imaging (up to 100x magnification). Furthermore, direct and indirect electrophysiological measurements by patch clamping and calcium imaging can be made during strain application. We characterise the strain applied to cells cultured in deformable wells by using finite element method simulations and experimental data, showing local surface strains of up to 60% with applied strains of up to 25%. We also show how patterned substrates do not alter the mechanical properties of the system compared to unpatterned surfaces whilst still inducing a homogeneous cell response to strain. The characterisation of this device will be useful for research into investigating the effect of whole-cell mechanical stretch on neurons at both single cell and network scales, with applications found in peripheral neuropathy modelling and in platforms for preventive and regenerative studies.

Membrane Mechanical Properties Regulate the Effect of Strain on Spontaneous Electrophysiology in Human iPSC-Derived Neurons.

Peripheral nerves contain neuron fibers vital for movement and sensation and are subject to continuous elongation and compression during everyday movement. At supraphysiological strains conduction blocks occur, resulting in permanent or temporary loss of function. The mechanisms underpinning these alterations in electrophysiological activity remain unclear; however, there is evidence that both ion channels and network synapses may be affected through cell membrane transmitted strain. The aim of this work was to quantify the changes in spontaneous activity resulting from application of uniaxial strain in a human iPS-derived motor neuron culture model, and to investigate the role of cell membrane mechanical properties during cell straining. Increasing strain in a custom-built cell-stretching device caused a linear decrease in spontaneous activity, and no immediate recovery of activity was observed after strain release. Imaging neuronal membranes with c-Laurdan showed changes to the lipid order in neural membranes during deformation with a decrease in lipid packing. Neural cell membrane stiffness can be modulated by increasing cholesterol content, resulting in reduced stretch-induced decrease of membrane lipid packing and in a reduced decrease in spontaneous activity caused by mechanical strain. Together these results indicate that the mechanism whereby cell injury causes impaired transmission of neural impulses may be governed by the mechanical state of the cell membrane, and contribute to establishing a direct relationship between neural uniaxial straining and loss of spontaneous neural activity.

Mechanical induction of critically delayed bone healing in sheep: radiological and biomechanical results.

This study aimed to mechanically produce a standardized ovine model for a critically delayed bone union. A tibial osteotomy was stabilized with either a rigid (group I) or mechanically critical (group II) external fixator in sheep. Interfragmentary movements and ground reaction forces were monitored throughout the healing period of 9 weeks. After sacrifice at 6 weeks, 9 weeks and 6 months, radiographs were taken and the tibiae were examined mechanically. Interfragmentary movements were considerably larger in group II throughout the healing period. Unlike group I, the operated limb in group II did not return to full weight bearing during the treatment period. Radiographic and mechanical observations showed significantly inferior bone healing in group II at 6 and 9 weeks compared to group I. After 6 months, five sheep treated with the critical fixator showed radiological bridging of the osteotomy, but the biomechanical strength of the repair was still inferior to group I at 9 weeks. The remaining three animals had even developed a hypertrophic non-union. In this study, mechanical instability was employed to induce a critically delayed healing model in sheep. In some cases, this approach even led to the development of a hypertrophic non-union. The mechanical induction of critical bone healing using an external fixation device is a reasonable attempt to investigate the patho-physiological healing cascade without suffering from any biological intervention. Therefore, the presented ovine model provides the basis for a comparative evaluation of mechanisms controlling delayed and standard bone healing.

Strain partitioning between nerves and axons: Estimating axonal strain using sodium channel staining in intact peripheral nerves.

BACKGROUND: Peripheral nerves carry afferent and efferent signals between the central nervous system and the periphery of the body. When nerves are strained above physiological levels, conduction blocks occur, resulting in debilitating loss of motor and sensory function. Understanding the effects of strain on nerve function requires knowledge of the multi-scale mechanical behaviour of the tissue, and how this is transferred to the cellular environment. NEW METHOD: The aim of this work was to establish a technique to measure the partitioning of strain between tissue and axons in axially loaded peripheral nerves. This was achieved by staining extracellular domains of sodium channels clustered at nodes of Ranvier, without altering tissue mechanical properties by fixation or permeabilisation. RESULTS: Stained nerves were imaged by multi-photon microscopy during in situ tensile straining, and digital image correlation was used to measure axonal strain with increasing tissue strain. Strain was partitioned between tissue and axon scales by an average factor of 0.55. COMPARISONS WITH EXISTING METHODS: This technique allows non-invasive probing of cell-level strain within the physiological tissue environment. CONCLUSIONS: This technique can help understand the mechanisms behind the onset of conduction blocks in injured peripheral nerves, as well as to evaluate changes in multi-scale mechanical properties in diseased nerves.

Three-dimensional printed upper-limb prostheses lack randomised controlled trials: A systematic review.

BACKGROUND: Three-dimensional printing provides an exciting opportunity to customise upper-limb prostheses. OBJECTIVE: This review summarises the research that assesses the efficacy and effectiveness of three-dimensional printed upper-limb prostheses. STUDY DESIGN: Systematic review. METHODS: PubMed, Web of Science and OVID were systematically searched for studies that reported human trials of three-dimensional printed upper-limb prostheses. The studies matching the language, peer-review and relevance criteria were ranked by level of evidence and critically appraised using the Downs and Black Quality Index. RESULTS: After removing duplicates, 321 records were identified. Eight papers met the inclusion criteria. No studies used controls; five were case studies and three were small case-series studies. All studies showed promising results, but none demonstrated external validity, avoidance of bias or statistically significant improvements over conventional prostheses. The studies demonstrated proof-of-concept rather than assessing efficacy, and the devices were designed to prioritise reduction of manufacturing costs, not customisability for comfort and function. CONCLUSION: The potential of three-dimensional printing for individual customisation has yet to be fully realised, and the efficacy and effectiveness to be rigorously assessed. Until randomised controlled trials with follow-up are performed, the comfort, functionality, durability and long-term effects on quality of life remain unknown. Clinical relevance Initial studies suggest that three-dimensional printing shows promise for customising low-cost upper-limb prosthetics. However, the efficacy and effectiveness of these devices have yet to be rigorously assessed. Until randomised controlled trials with follow-up are performed, the comfort, functionality, durability and long-term effects on patient quality of life remain unknown.

Probing multi-scale mechanics of peripheral nerve collagen and myelin by X-ray diffraction.

Peripheral nerves are continuously subjected to mechanical forces, both during everyday movement and as a result of traumatic events. Current mechanical models focus on explaining the macroscopic behaviour of the tissue, but do not investigate how tissue strain translates to deformations at the microstructural level. Predicting the effect of macro-scale loading can help explain changes in nerve function and suggest new strategies for prevention and therapy. The aim of this study was to determine the relationship between macroscopic tensile loading and micro scale deformation in structures thought to be mechanically active in peripheral nerves: the myelin sheath enveloping axons, and axially aligned epineurial collagen fibrils. The microstructure was probed using X-ray diffraction during in situ tensile loading, measuring the micro-scale deformation in collagen and myelin, combined with high definition macroscopic video extensiometry. At a tissue level, tensile loading elongates nerves axially, whilst simultaneously compressing circumferentially. The non-linear behaviour observed in both directions is evidence, circumferentially, that the nerve core components have the ability to rearrange before bearing load and axially, of a recruitment process in epineurial collagen. At the molecular level, axially aligned epineurial collagen fibrils are strained, whilst the myelin sheath enveloping axons is compressed circumferentially. During induced compression, the myelin sheath shows high circumferential stiffness, indicating a possible role in mechanical protection of axons. The myelin sheath is deformed from low loads, despite the non-linearity of whole tissue compression, indicating more than one mechanism contributing to myelin compression. Epineurial collagen shows similar load-bearing characteristics to those of other collagenous connective tissues. This new microstructural knowledge is key to understand peripheral nerve mechanical behaviour, and will support new regenerative strategies for traumatic and repetitive injury.

A biomechanical model for fibril recruitment: Evaluation in tendons and arteries.

Simulations of soft tissue mechanobiological behaviour are increasingly important for clinical prediction of aneurysm, tendinopathy and other disorders. Mechanical behaviour at low stretches is governed by fibril straightening, transitioning into load-bearing at recruitment stretch, resulting in a tissue stiffening effect. Previous investigations have suggested theoretical relationships between stress-stretch measurements and recruitment probability density function (PDF) but not derived these rigorously nor evaluated these experimentally. Other work has proposed image-based methods for measurement of recruitment but made use of arbitrary fibril critical straightness parameters. The aim of this work was to provide a sound theoretical basis for estimating recruitment PDF from stress-stretch measurements and to evaluate this relationship using image-based methods, clearly motivating the choice of fibril critical straightness parameter in rat tail tendon and porcine artery. Rigorous derivation showed that the recruitment PDF may be estimated from the second stretch derivative of the first Piola-Kirchoff tissue stress. Image-based fibril recruitment identified the fibril straightness parameter that maximised Pearson correlation coefficients (PCC) with estimated PDFs. Using these critical straightness parameters the new method for estimating recruitment PDF showed a PCC with image-based measures of 0.915 and 0.933 for tendons and arteries respectively. This method may be used for accurate estimation of fibril recruitment PDF in mechanobiological simulation where fibril-level mechanical parameters are important for predicting cell behaviour.

Rapid and efficient differentiation of functional motor neurons from human iPSC for neural injury modelling.

Primary rodent neurons and immortalised cell lines have overwhelmingly been used for in vitro studies of traumatic injury to peripheral and central neurons, but have some limitations of physiological accuracy. Motor neurons (MN) derived from human induced pluripotent stem cells (iPSCs) enable the generation of cell models with features relevant to human physiology. To facilitate this, it is desirable that MN protocols both rapidly and efficiently differentiate human iPSCs into electrophysiologically active MNs. In this study, we present a simple, rapid protocol for differentiation of human iPSCs into functional spinal (lower) MNs, involving only adherent culture and use of small molecules for directed differentiation, with the ultimate aim of rapid production of electrophysiologically functional cells for short-term neural injury experiments. We show successful differentiation in two unrelated iPSC lines, by quantifying neural-specific marker expression, and by evaluating cell functionality at different maturation stages by calcium imaging and patch clamping. Differentiated neurons were shown to be electrophysiologically altered by uniaxial mechanical deformation. Spontaneous network activity decreased with applied stretch, indicating aberrant network connectivity. These results demonstrate the feasibility of this rapid, simple protocol for differentiating iPSC-derived MNs, suitable for in vitro neural injury studies focussing on electrophysiological alterations caused by mechanical deformation or trauma.

Endochondral ossification in vitro is influenced by mechanical bending.

Bone development is influenced by the local mechanical environment. Experimental evidence suggests that altered loading can change cell proliferation and differentiation in chondro- and osteogenesis during endochondral ossification. This study investigated the effects of three-point bending of murine fetal metatarsal bone anlagen in vitro on cartilage differentiation, matrix mineralization and bone collar formation. This is of special interest because endochondral ossification is also an important process in bone healing and regeneration. Metatarsal preparations of 15 mouse fetuses stage 17.5 dpc were dissected en bloc and cultured for 7 days. After 3 days in culture to allow adherence they were stimulated 4 days for 20 min twice daily by a controlled bending of approximately 1000-1500 microstrain at 1 Hz. The paraffin-embedded bone sections were analyzed using histological and histomorphometrical techniques. The stimulated group showed an elongated periosteal bone collar while the total bone length was not different from controls. The region of interest (ROI), comprising the two hypertrophic zones and the intermediate calcifying diaphyseal zone, was greater in the stimulated group. The mineralized fraction of the ROI was smaller in the stimulated group, while the absolute amount of mineralized area was not different. These results demonstrate that a new device developed to apply three-point bending to a mouse metatarsal bone culture model caused an elongation of the periosteal bone collar, but did not lead to a modification in cartilage differentiation and matrix mineralization. The results corroborate the influence of biophysical stimulation during endochondral bone development in vitro. Further experiments with an altered loading regime may lead to more pronounced effects on the process of endochondral ossification and may provide further insights into the underlying mechanisms of mechanoregulation which also play a role in bone regeneration.

Mechanobiological modelling of tendons: Review and future opportunities.

Tendons are adapted to carry large, repeated loads and are clinically important for the maintenance of musculoskeletal health in an increasing, actively ageing population, as well as in elite athletes. Tendons are known to adapt to mechanical loading. Also, their healing and disease processes are highly sensitive to mechanical load. Computational modelling approaches developed to capture this mechanobiological adaptation in tendons and other tissues have successfully addressed many important scientific and clinical issues. The aim of this review is to identify techniques and approaches that could be further developed to address tendon-related problems. Biomechanical models are identified that capture the multi-level aspects of tendon mechanics. Continuum whole tendon models, both phenomenological and microstructurally motivated, are important to estimate forces during locomotion activities. Fibril-level microstructural models are documented that can use these estimated forces to detail local mechanical parameters relevant to cell mechanotransduction. Cell-level models able to predict the response to such parameters are also described. A selection of updatable mechanobiological models is presented. These use mechanical signals, often continuum tissue level, along with rules for tissue change and have been applied successfully in many tissues to predict in vivo and in vitro outcomes. Signals may include scalars derived from the stress or strain tensors, or in poroelasticity also fluid velocity, while adaptation may be represented by changes to elastic modulus, permeability, fibril density or orientation. So far, only simple analytical approaches have been applied to tendon mechanobiology. With the development of sophisticated computational mechanobiological models in parallel with reporting more quantitative data from in vivo or clinical mechanobiological studies, for example, appropriate imaging, biochemical and histological data, this field offers huge potential for future development towards clinical applications.