Annals of Biomedical Engineering

BackgroundConventional left ventricular assist device ramp metrics are load dependent, obscuring intrinsic myocardial recovery. A mechanistic, patient-specific representation of ventricular mechanics, identifiable from routine clinical data, could provide quantitative indices of intrinsic left ventricular (LV) function for longitudinal recovery surveillance. ObjectiveTo develop and verify a ramp-integrated, patient-specific model of HeartMate 3-assisted LV function that can yield indices of intrinsic myocardial contractility and remodeling. MethodsWe represented LV pressure-volume (PV) behavior with a PV envelope composed of a monotonic passive PV relation (pPVR) and a unimodal active PV relation (aPVR). We developed a parameterization procedure to infer the patient-specific shape of this envelope directly from routine ramp-test data. We then embedded the parameterized envelope within the PSCOPE framework, a hybrid platform that couples a lumped-parameter network to a physical HeartMate 3 flow loop, to reproduce clinical ramp hemodynamics. Percent residuals between simulated outputs and the corresponding clinical measurements verified the implementation of the PV envelope within PSCOPE. ResultsIn three HeartMate 3 recipients, the PSCOPE models reproduced ramp hemodynamics with residuals generally [≤] 20% across pump speeds and measured variables. Cardiac index residuals ranged from 0-18.5%, systemic and pulmonary arterial pressure residuals remained [≤] 18.4%, and pulmonary arterial wedge pressure residuals remained [≤] 20%. The PSCOPE models matched central venous pressure within [≤] 3 mmHg in all cases, although one setting yielded a 33.3% residual due to a low reference pressure. For one patient, the model reproduced ramp hemodynamics at a speed deliberately withheld from PV-envelope parameterization with residuals [≤] 10%, supporting cross-speed generalizability. Patient-specific PV envelopes also revealed clinically meaningful heterogeneity in LV diastolic stiffness, volume threshold for declining systolic function, operating PV points for peak systolic function, and contractile reserve. ConclusionsRamp-integrated parameterization of the monotonic pPVR and unimodal aPVR yields a compact, mechanistic PV envelope that is identifiable from routine clinical data and verifiable within PSCOPE. The resulting indices characterize intrinsic LV function and may enhance longitudinal recovery surveillance and inform LVAD management. Prospective multicenter validation is warranted to confirm the generalizability and clinical utility of this approach.

IntroductionStereological estimates of villous membrane thickness and surface area are widely used to infer the diffusive exchange capacity of the human placenta. A key geometric determinant of exchange capacity can be expressed as an effective diffusive length scale. Here we combine virtual histological sections with computational modelling in realistic villous geometries to assess the accuracy of classical stereological estimates of this diffusive length scale. MethodsTwo terminal villi, reconstructed from three-dimensional imaging, were digitally sectioned to generate random two-dimensional geometries containing fetal capillaries and surrounding villous tissue. For each section, we simulated steady diffusive transport between the fetal capillary and intervillous space boundaries to obtain a physics-based diffusive length scale as a reference case. Using the same geometries, we applied standard line-intercept stereology to measure harmonic-mean barrier thickness and boundary-length densities, from which a stereological estimate of diffusive length scale was derived. ResultsAcross both villi, stereology systematically overestimated the diffusive length scale by approximately 15-25%, depending on villus and section. We identified sources of this discrepancy, including interface curvature and assumptions underpinning the stereological correction factors, using idealised models of villus structure. ConclusionThese findings highlight the need for stereological approaches that account for curvature when interpreting placental structure-function relationships.

PurposeTo characterize the frequency-dependent bioimpedance properties of major ocular tissues in intact ex vivo porcine eyes under simulated surgical conditions and evaluate tissue separability at discrete frequencies. MethodsBioimpedance spectra were acquired from sclera, corneal epithelium, iris, lens, vitreous, and retina in intact ex vivo porcine eyes using a two-electrode probe and a precision LCR meter over 5 kHz to 1 MHz. Measurements were obtained under balanced salt solution and ophthalmic viscosurgical device conditions. Probe-tissue contact was verified by microscope visualization and optical coherence tomography. Tissue separability at 5, 50, 100, and 900 kHz was evaluated using global and pairwise statistical comparisons, effect sizes, and ROC-based separability metrics. Robotic-stabilized and handheld measurements were also compared. ResultsOcular tissues demonstrated distinct, frequency-dependent impedance magnitude distributions. Across sampled frequencies, 60% to 80% of tissue pairs showed significant differences after multiplicity correction. Median pairwise effect sizes ranged from Cohens d = 0.48 at 5 kHz to 1.04 to 1.06 at 50 to 100 kHz. Median ROC-based separability was 0.91 at 5 kHz and 0.76 to 0.77 at 50 to 900 kHz. Robotic-stabilized measurements showed lower variance than handheld measurements, although tissue-specific impedance ranges and frequency-dependent trends were preserved across acquisition modes. ConclusionsMajor ocular tissues exhibit reproducible, frequency-dependent bioimpedance signatures in intact ex vivo eyes under simulated surgical preparation. These findings establish a physiologically relevant ocular impedance reference dataset and support bioimpedance as a complementary modality for tissue differentiation in ophthalmic microsurgery.

Background and ObjectiveThe foot-ankle complex is a highly articulated and mechanically constrained system, often simplified as a chain of few rigid segments, neglecting many bone-to-bone motions and raising questions about the accurate representation of interaction with ground. This study proposes a new reduced-order multibody formulation that captures intrinsic kinematic constraints of the foot through motion synergies. MethodsBones kinematic coupling, or motion synergies, were experimentally derived from weight-bearing CT scans using principal component analysis. These couplings were embedded in a synergy-based multibody kinematic optimization framework describing the foot-ankle with five degrees of freedom: ankle flexion; foot adduction, pronation, and arching; and toe flexion. Model accuracy was evaluated against bone-level experimental kinematics. The model was applied to gait data from healthy, flat, and diabetic feet and compared with a standard multi-segment foot model, assessing robustness by progressively reducing the number of skin markers. ResultsAverage errors were about 1{degrees} and 0.5 mm when using subject-specific synergies and below 7{degrees} and 4 mm when scaling the generic model, matching or exceeding the accuracy of existing models. Reliable reconstruction was obtained using only four foot markers. In clinical gait analysis, the model showed superior discrimination between populations and enabled assessment of transverse arch deformation, not accessible with conventional models. ConclusionThe proposed synergy-based model provides an accurate, low-complexity framework for reconstructing bone-level foot and ankle kinematics, substantially simplifying gait analysis while improving biomechanical interpretability. This framework supports future integration with dynamic models aimed at studying load transmission in the foot.

Background and ObjectivesLung cancer is one of the most frequently diagnosed cancers worldwide. While non-surgical treatment options have increased in number and efficacy, lung resection for primary cancers is still a mainstay of treatment. Lung resection has been shown to impair right ventricular function, although the mechanism for the impairment remains unclear. Wave intensity is increasingly used as a metric for increased post-operative afterload. Here, we develop a computational framework to assess the impact of simulated lung resection on wave intensity to establish that post-operative changes in wave intensity are attributable to the change in pulmonary artery morphometry. MethodsWe analyse a 48 pulmonary arterial surfaces segmented from CT images in patients with no evidence of lung disease to obtain 1D representations of the pulmonary vasculature. For each pulmonary vasculature we sequentially remove vessel branches to mimic post-operative morphometric changes to the arterial network. Using an established 1D computational flow model, we simulate pulsate blood flow in 44 pre-operative cases and 1596 post-operative cases. We compute wave intensity in the main, right, and left pulmonary arteries for all simulations. ResultsWe compare the change in computed wave intensities pre-versus post-operatively to the results of an experimental clinical study comparing pre- and post-operative wave intensity in a 27 patient cohort. We see good agreement between the changes in the parameters of wave intensity between this study and those reported in the clinical study. Further, we capture flow distribution the changes pre-versus post-operatively which indicates that the computational model behaves as expected. ConclusionsIn this preliminary study on a computational framework to capture changes in pulmonary arterial haemodynamics following lung resection, we have shown that our model and analysis pipeline is capable of capturing post-operative changes to wave intensity and flow redistribution between the pulmonary arteries following lung resection. These results motivate further research to develop and validate a patient specific model which is an area of active research for us.

ObjectivePulsed field ablation (PFA) relies on irreversible electroporation to create nonthermal cardiac lesions, yet real-time indicators of electroporation progression and validated lethal electric field thresholds remain limited. This study aimed to develop a bioimpedance-based metric for real-time monitoring of cardiac electroporation, evaluate the impact of myocardial anisotropy under electroporation conditions, and derive waveform-specific lethal electric field thresholds. IntroductionCurrent PFA procedures lack direct intraoperative feedback on lesion formation, and uncertainty remains regarding the role of myocardial fiber orientation in shaping electric field distributions. Because electroporation dynamically alters tissue electrical properties, monitoring these changes during treatment may improve prediction of ablation outcomes. MethodsPFA was delivered to fresh ex vivo porcine ventricular tissue using clinically relevant and energy-matched waveforms with pulse widths from 1 to 100 {micro}s. Inter-burst broadband electrical impedance spectroscopy was performed using a low-voltage diagnostic waveform to quantify burst-resolved impedance changes. Lesions were visualized using metabolic staining, then finite element models incorporating nonlinear electroporation-dependent conductivity were used to compare anisotropic and homogenized electric field distributions. Lethal electric field thresholds were estimated by fitting simulated contours to measured lesion areas and validated using uniform electric fields generated by a parallel electrode array. ResultsAcross all waveforms, impedance measurements showed a rapid initial decrease followed by stabilization, indicating early electroporation saturation. Burst-to-burst percent change in impedance slope provided a consistent, waveform-agnostic metric of electroporation progression. Lesion morphology was not systematically influenced by fiber orientation, and modeling demonstrated that electroporation-induced conductivity increases homogenized tissue anisotropy. Lethal electric field thresholds increased with decreasing pulse width, ranging from 517 {+/-} 46 V/cm (100 {micro}s) to 1405 {+/-} 55 V/cm (1 {micro}s), and were validated under uniform field conditions. ConclusionBioimpedance-assisted monitoring enables real-time assessment of cardiac electroporation, while electroporation-induced homogenization supports simplified modeling and standardized PFA treatment design.

The effects of specific footwear features on biomechanical parameters are often confounded by simultaneous changes in other shoe conditions, making it difficult to identify the isolated effect of material and design properties on relevant biomechanical outcomes. This study aimed to propose a tool, namely the Modular Footwear Setup (MFS), to assess the effects of midsole modifications on lower limb joint kinematics and in-shoe plantar pressure. The MFS uses a micro-hook-and-loop fastening system and a custom alignment device to enable fast, strong, and reliable midsole attachment/detachment to/from the upper. Accuracy and repeatability of the MFS in replicating the biomechanical outcomes of a control shoe featuring the same upper and midsole were tested in 10 healthy participants (5M,5F; age=33.2{+/-}9.2 yrs; BMI=21.5{+/-}2.8 kg/m2). Participants were asked to walk wearing both the MFS and the standard control shoe in three sessions. Kinematics of lower limb joints were measured via inertial measurement units, while capacitive pressure insoles were used to measure in-shoe plantar pressure. Intraclass correlation coefficient (ICC) was used to assess the repeatability of kinematic and pressure measurements between sessions. Statistical Parametric Mapping analysis did not identify significant differences in joint kinematics between conditions. While the MFS exhibited slightly lower peak pressure at the rearfoot, pressure parameters were not statistically different in the other foot regions. The MFS demonstrated good-to-excellent inter-session repeatability (ICC 0.84-0.97) for peak and mean pressure. Participants reported similar levels of comfort and stability in both shoes. The findings of the present study suggest the MFS has the potential to be a reliable and accurate tool for evaluating the effect of midsole features on relevant biomechanical parameters. This modular approach may improve data-driven footwear design by providing a consistent platform for testing the effects of midsole designs and materials across various applications, including therapeutic, safety, and athletic shoes.

It is increasingly necessary to both study biology in 3D and obtain quantitative measurements. Not all 3D-reconstructions are created equal, particularly when using the anatomical model as a basis for force calculations, i.e. computational modeling. Here, we compare 3D anatomical reconstructions from two emerging imaging modalities: 4D ultrasound (4DUS) and light sheet fluorescent microscopy (LSFM) against our previous nano-computed tomography (nanoCT) cohort data, using the tortuous highly intricate pharyngeal arch artery system of the chick embryo as a test bed. We highlight modality-specific morphological image acquisition discrepancies and their influence on subsequent computational fluid dynamics results. Overall, LSFM accurately captured quantitative volumetric measurements of small rapidly-changing vascular morphologies while 4DUS systematically inflated small tortuous vessels. Differences in image-based morphology changes led to significant changes in computationally-obtained force magnitudes and flow patterns linked to vessel angle and tortuosity. This validates LSFM as a comparative preclinical vascular quantitative imaging tool and suggests that 4DUS needs extensive 3D anatomical validation for non cardiac chamber vessels.

BackgroundPersonalized computational neuromusculoskeletal models have great potential for optimizing the design of clinical treatments for movement impairments. While many software tools address specific parts of the model personalization and treatment optimization processes, they typically require significant programming experience to use and do not cover the full breadth of these two processes. Furthermore, published neuromusculoskeletal modeling studies typically do not provide all of the minute methodological details needed for others to reproduce the work. Consequently, researchers seeking to develop skills in the model personalization and treatment optimization processes face a steep learning curve due to the lack of detailed training materials that demonstrate both processes for real-life clinical problems using real-life subject movement data. MethodsThis article presents detailed training tutorials for the model personalization and treatment optimization processes using two real-life clinical problems and the Neuromusculoskeletal Modeling (NMSM) Pipeline. The first clinical problem involves the design of personalized gait modifications and high tibial osteotomy surgery for an individual with bilateral medial knee osteoarthritis, where the goal is to reduce the peak adduction moment in both knees to a specified target level. The second clinical problem involves the design of a synergy-based functional electrical stimulation prescription for an individual post-stroke with impaired walking function, where the goal is to equalize the propulsive and braking impulses between the two legs. Both tutorials were evaluated as course projects given to novice users in a combined undergraduate/graduate mechanical engineering course. ResultsBoth tutorials produced personalized neuromusculoskeletal models and associated dynamically consistent tracking optimizations that closely reproduced subject-specific experimental joint angles, joint moments, ground reaction forces and moments, and (if applicable) muscle activations measured during walking. Subsequent design optimizations predicted personalized treatments that achieved target values of peak knee adduction moments or propulsive and braking impulses. ConclusionsThe detailed step-by-step tutorials presented with this article are the first to walk users step-by-step through the entire process of creating personalized neuromusculoskeletal models and then using them to design personalized treatments for clinical problems. These tutorials can be used to introduce new users to the NMSM Pipeline and as projects in neuromusculoskeletal modeling courses.

Advanced footwear technology (AFT) shoes incorporate increased sole thickness and compliant midsole materials that may alter running biomechanics. While these effects have been widely studied during level running, little is known about how sole thickness influences running style and stability during uphill running. This study examined the effects of two AFT shoes differing in sole thickness (35 mm-AFT35; 50 mm-AFT50) and a traditional control shoe (27 mm-CON27) on running style and stability during uphill running. Seventeen experienced male runners performed treadmill running at a 10% incline at 6.5 and 10 km/h in three shoe conditions. Running style was assessed using duty factor, normalized step frequency, center-of-mass oscillation, vertical and leg stiffness, and lower-limb joint kinematics. Running stability was evaluated using local dynamic stability via the maximum Lyapunov exponent and detrended fluctuation analysis of stride time. Duty factor and normalized step frequency did not differ between shoes. However, AFT shoes showed greater center-of-mass oscillation (p = 0.004), lower vertical stiffness (p = 0.022) compared to CON27. Joint kinematics revealed significant shoe effects at the ankle (p = 0.001), particularly increased dorsiflexion and eversion in AFT conditions. Running stability showed only minor changes. Local dynamic stability differed at the trunk (p = 0.027), with reduced stability in AFT50 compared with CON27 (p = 0.006), while global stability remained unchanged. No shoe x speed interactions were observed for any variable. Overall, uphill running style and stability remained largely preserved across shoe conditions, suggesting that sole thickness alone had limited influence.

BackgroundIn aortic arch surgery, bilateral selective antegrade cerebral perfusion (bSACP) maintains cerebral blood flow during circulatory arrest. bSACP is often delivered using a single pump with a Y-connector, dividing the flow. Current practice has veered towards perfusion of the left common carotid artery by cannula and the right subclavian artery or axillary artery by a vascular graft. Under this configuration, inflow distribution may be sensitive to left-sided cannula resistance, particularly in patients with limited collateral circulation, potentially reducing left-hemispheric pressure and flow despite bSACP. We investigated how cannula design influences perfusion pressure and arterial inflow distribution during bSACP. MethodsFour perfusion cannulas with different flow resistances were characterized using bench measurements (40-200 ml/min) and computational fluid dynamics (CFD). The CFD cannula models were then integrated into patient-specific CFD models of the cerebral circulation from three patients with varying collateral circulation/capacity. Both flow- and pressure-controlled pump strategies were simulated. ResultsBench measurements showed substantial variation in flow resistance between the cannulas, which was accurately reproduced by CFD. For the patient-specific analysis, cannula choice affected perfusion through roughly doubled pressure laterality and halved left-side inflow between the most extreme cannulas. Still, perfusion pressure was kept within recommended levels in two subjects but was low in one. Left-side arterial inflow varied between 70-150 ml/min. ConclusionsWe isolated the effects of cannula design on cerebral pressure and blood inflow distribution during bSACP, highlighting potential pitfalls in patients with limited collateral circulation.

Dysphagia, difficulty swallowing due to irritation or damage to the esophagus, is one of the most common complications following anterior cervical discectomy and fusion (ACDF), the most frequently performed cervical spine procedure in the United States. Surgical retraction hardware imposes sustained compression on the esophagus during surgery, generating nonuniform stress and strain fields that may contribute to temporary postoperative soft tissue damage. Current intraoperative assessment relies on visual inspection and manual inspection by the surgical team and does not provide quantitative measures of esophageal deformation, strain, or retraction displacement. Here, we present a comprehensive mechanics analysis of esophageal compression during ACDF that integrates experiments on esophageal phantoms, nonlinear finite element modeling, and theoretical thick-wall scaling relationships. Modeling results quantify peak contact pressures and corresponding stress distributions, identifying conditions under which circumferential strain in the compressed esophageal wall increases sharply as localized pressures approach the upper physiological range ([~]6-17 kPa). Parametric investigation of retractor blade width, placement depth, and polymeric biocompatible coating properties demonstrates that targeted, yet mechanically simple, design modifications can help to attenuate strain concentrations. In particular, the introduction of compliant polymeric coatings redistributes contact loads and reduces peak wall stress by up to 20% relative to unbuffered blades (17 kPa to 13.5 kPa). Increasing blade width from 20 mm to 50 mm further decreases peak interface stress from 2.48 kPa to 0.45 kPa, corresponding to an 82% reduction. Reducing these stresses may help limit mechanically induced complications such as postoperative dysphagia. Experiments performed on esophageal phantoms with embedded pressure sensors replicate surgical ACDF retraction protocols under displacement-controlled conditions. This setup establishes physiologically relevant loading and enables quantitative validation of computational predictions by correlating measured voltage output with contact pressure and esophageal deformation. Measured relationships between applied retraction displacement, contact pressure, and tissue deformation govern stress amplification during ACDF retraction. Together, these results establish a predictive mechanics framework that links retractor blade design variables to esophageal stress fields, providing quantitative criteria to mitigate soft tissue damage during ACDF. HIGHLIGHTSO_LI2D and 3D finite element models quantify esophageal wall stress during anterior cervical discectomy and fusion (ACDF) retraction. C_LIO_LIRetractor blade geometry influences stress distribution, with wider blades reducing localized tissue loading by up to 82% likely associated with post-surgical dysphagia. C_LIO_LICompliant polymeric buffer layers attenuate pressure and smoothen stress gradients to reduce peak tissue loading by up to 20% during retraction. C_LI

ObjectiveAchilles tendon ruptures lead to long-term structural and functional deficits. Prior research that sought to identify optimal rehabilitation techniques was fundamentally limited by the inability to continuously monitor Achilles tendon loading during rehabilitation. Our objective was to develop a data-driven model that predicts per-step peak Achilles tendon loading from only a single, boot-mounted accelerometer. MethodsNineteen patients recovering from an acute Achilles tendon rupture completed in-lab walking trials while wearing an instrumented immobilizing boot. A boot-mounted inertial measurement unit provided acceleration signals used for prediction, while a force-sensing insole provided ground truth tendon-loading data through a validated ankle moment balance. We developed a stance-detection algorithm, as well as a personalized one-dimensional convolutional neural network (1D-CNN) to estimate per-step peak Achilles tendon load. Our training framework incorporated a small patient-specific personalization sample and was evaluated on held-out steps. ResultsThe stance detection algorithm identified stance phases with 99.8% precision and mean timing errors of 27.3 ms for heel strike and 61.9 ms for toe-off. The CNN estimated per-step peak Achilles tendon load with a mean absolute error of 0.14 bodyweights (R2=0.68) across rupture patients. ConclusionContinuous, objective estimation of Achilles tendon loading during early rehabilitation is feasible using a single, boot-mounted accelerometer. Model errors were small (9%) relative to the wide range of tendon loading exhibited during immobilizing boot walking. Our proposed approach enables clinicians to continuously monitor mechanical loading during a previously unobservable rehabilitation period and provides a foundation for personalized rehabilitation guidance after Achilles rupture.

Thoracic Endovascular Aortic Repair (TEVAR) is a minimally invasive procedure for the treatment of thoracic aortic pathologies, such as Thoracic Aortic Aneurysm (TAA). Computational simulations can provide valuable insights into TEVAR outcomes and complications prior to surgery, making them a useful tool in the procedural planning. In this work, Fluid-Structure Interaction (FSI) computational simulations are carried out in ten pre-TEVAR patient-specific TAA cases, for which post-TEVAR outcomes are known, to quantify the hemodynamic drag forces acting on the aortic wall. Based on these results, this study proposes a new risk factor R to predict the occurrence of type I and III endoleaks. The patient cohort is divided in a calibration set, used to associate specific R values with three different risk levels, and a validation set, to test the risk factor efficacy. Based on the risk factor values obtained for the calibration set, R[&le;] 0.33 is associated with low risk of endoleak formation, 0.33 < R[&le;] 0.67 with moderate risk, and R > 0.67 with high risk. Once it is applied to the validation set,the risk factor is able to predict the formation of a type Ia endoleak. The risk factor proposed in this work is capable of identifying all the endoleak cases analysed, as well as conditions known to increase the risk of TEVAR complications. This study represents a preliminary attempt to determine whether pre-TEVAR hemodynamics can effectively predict post-TEVAR complications and thereby aid clinicians in the pre-operative planning.

IntroductionArteries, like other soft tissues, exhibit viscoelastic mechanical behavior, meaning their response to stress and strain is time dependent. This implies that the way arteries deform depends not only on the amount of force applied but also on the rate at which the force is applied. This study investigates the effects of different loading rates on the mechanical behavior of human femoropopliteal arteries (FPAs) to understand their rate-dependent characteristics. MethodsHuman FPA specimens were collected from 14 donors, including 7 males and 7 females, aged 45-55 years. A 10x10 mm segment was isolated, mounted onto a biaxial testing device, and subjected to varying loading rates (10 to 50 mN/s). Mechanical responses were recorded, and stress-stretch curves were analyzed. Statistical analyses, including mixed-design ANOVA, assessed the impact of sex and loading rates on tissue stiffness. ResultsResults indicated significant loading-rate dependency, particularly in the circumferential direction. Stretch values decreased with increasing loading rates, more prominently in the circumferential than in the longitudinal direction (p-value<0.01). Statistical analyses revealed no significant interaction between sex and loading rate, though male arteries exhibited slightly higher compliance than female arteries. DiscussionThe findings demonstrate that the mechanical response of FPAs is highly dependent on the loading rate, with more pronounced effects observed in the circumferential direction. At higher loading rates, the human FPAs demonstrated a stiffer response in the circumferential direction. DedicationWe dedicate this work to the memory of our late student, Ali Zolfaghari Sichani, who passed away tragically during his doctoral studies. Ali performed the majority of the experiments and the initial analysis reported in this paper. His passion, dedication, and hard work were the foundation of this research, and he is deeply missed.

The development of large-scale, three-dimensional human tissues is crucial for various applications in therapeutic tissue engineering, disease modeling, and drug testing. However, due to the diffusion limit of oxygen, the lack of functional vascular networks is a significant limitation in maintaining these engineered tissues in the laboratory. To address this challenge, we present a systematic, model-based design process for artificial supply networks that can ensure a sufficient supply of oxygen and nutrients to engineered human tissue. Our approach combines mathematical models of fluid dynamics, cell metabolism, and network properties to identify key parameters influencing the supply performance. We demonstrate the applicability and possibilities of this design process by simulating different network structures, including cuboid and rhombic do-decahedral honeycombs, under various conditions. Our results show that the structure of the artificial supply network, oxygen concentration, and solute flow within the network strongly influence cellular metabolic activity and viability. We also examine the effects of non-uniform cell density, channel blockage, and long channel length on the oxygen distribution inside the cell-containing tissue compartment. Our findings highlight the importance of considering these factors in the design of artificial supply networks for large-scale engineered human tissues. This study provides a promising approach for quickly exploring the vast design space of possible network structures under different conditions for desired cell and tissue states, ultimately contributing to the development of more efficient and effective tissue engineering strategies.

Various ankle-foot conditions (e.g., fractures, diabetic foot ulcers, and post-surgical recovery) require periods of complete non-weightbearing followed by gradually increasing partial loadings. However, existing assistive devices often provide inconsistent or uncomfortable offloading during gait. Additionally, prolonged proximal leg offloading can contribute to muscle atrophy, reduced bone density, and overuse of other body segments. We present a novel offloading ankle-foot orthosis (OLAFO) designed to overcome these limitations. The OLAFO features a patient-specific load-bearing shank brace, designed through a digital workflow and fabricated from a 3D-printed core reinforced with carbon-fiber composite lamination. Interlocking serrated side struts, adjustable in 2 mm increments, modulate load sharing between the shank and plantar surfaces. Furthermore, the OLAFO incorporates contact plates with a rocker profile informed by roll-over-shape measurements to support forward progression and gait symmetry. Proof-of-concept biomechanical verification in one able-bodied participant evaluated complete offloading, five partial-loading levels, and normal gait using a pressure walkway to compute vertical ground reaction forces and impulses. In complete offloading, the affected foot generated no contact pressures. Across partial-loading levels, the foot impulse increased from 14% to 53% of the total load and scaled linearly with strut height adjustments, supporting clinician-prescribed loading increments. Contralateral stance duration increased only modestly compared to commonly used assistive devices, indicating reduced compensatory loading on the intact limb. These findings demonstrate the proof-of-concept feasibility of the OLAFO, highlighting its potential for verifying full offloading and prescribing partial-loading targets during rehabilitation. Future research will evaluate performance across patient populations and clinical rehabilitation tasks.

Research questionCan tissue clearing, combined with volumetric imaging, enable reliable, quantitative three-dimensional analysis of follicles and vasculature in intact human ovarian tissue? DesignA CUBIC-based clearing protocol was adapted for human ovarian medulla and cryopreserved cortex. Tissue from reproductive-aged donors was cleared, fluorescently labeled, and imaged using confocal and light sheet microscopy. Tissue expansion, imaging depth, and vascular morphometrics were quantified and follicle density was compared to conventional histology. ResultsClearing produced optically transparent tissue with a linear expansion factor of 1.2 across cortex and medulla. Imaging depth increased 6.5-11-fold in cortex and 6-8-fold in medulla. Follicle density measurements in immunolabeled cleared cortex were comparable to histology, supporting the validity of volumetric follicle quantification. Light sheet microscopy of lectin-labeled cortex revealed no significant donor-to-donor differences in vascular morphometrics, including mean vessel diameters of 12-14 {micro}m, branch point densities of 632-965 points/mm3, vessel length densities of 117-175 mm/mm3, and volume fractions of 1.9-2.3%. Volumetric imaging further illustrated heterogeneous spatial relationships between follicles and surrounding vessels. ConclusionTissue clearing and volumetric imaging complement routine histology and enable quantitative three-dimensional investigation of follicle-vascular interactions in intact human ovarian tissue, providing a framework for advancing fertility preservation and ovarian tissue transplantation research.

PurposeFibrosis is the pathological remodeling of the extracellular matrix (ECM) that is largely orchestrated by activated fibroblasts. The mechanical properties of the ECM change drastically during fibrosis, and fibroblasts become increasingly activated by mechanical environments that mimic the properties of fibrotic tissues. While the effects of increased elastic modulus (stiffness) on fibroblast activation have been well-studied, the impact of changes in viscoelasticity are less clear. Here, we sought to determine how fibroblast activation is altered by changes in viscoelasticity in a three-dimensional, fibrillar microenvironment. MethodsWe employed 3D alginate collagen I hydrogels with independently tunable stiffness and stress relaxation rates. Dermal fibroblasts were encapsulated in hydrogels with four distinct mechanical profiles (soft: 3 kPa or stiff: 10 kPa, fast stress relaxing: {tau}1/2 {approx} 160 s or slow stress relaxing: {tau}1/2 {approx} 1600 s). We assessed fibroblast activation by changes in cell morphology, expression of key activation markers, and evidence of ECM remodeling. ResultsFibrillar alginate collagen networks enhanced fibroblast spreading, -smooth muscle actin stress fiber formation, and fibroblast activation protein- expression in matrices that were slow relaxing or stiff. The presence of the fibrillar network further enhanced fibroblast activation, independent of the changes driven by matrix viscoelasticity. ECM remodeling was also promoted by slow relaxing matrices, with increased fibronectin deposition and more remodeling of the local collagen fiber network. ConclusionsOur results demonstrate that fibroblast activation is highly responsive to matrix stress relaxation rate, and that models incorporating fibrillar, viscoelastic networks can provide new insights into the role of ECM mechanics driving fibroblast activation.

There is more to musculotendon path modeling than aligning a cable to reflect the geometric features of a muscle-tendon unit. From the perspective of simulation accuracy, the key is to replicate the length- and moment arm-joint angle relations of the target muscle. In this study, we propose an effect-oriented approach of automated path modeling, via the hybrid calibration based on muscle surface mesh and moment arm. The task is formulated as an optimization problem with a threefold objective for the path to: 1) pass through multiple ellipses representing muscle cross-sections, 2) yield moment arms that match experimental measurements, and 3) yield moment arms with the designated signs. The performance of our optimization framework is demonstrated with the musculoskeletal surface mesh from the Visible Human Male and moment arm datasets from literature--producing 42 paths that are anatomically realistic and biomechanically accurate in 20.1 min. Our optimization framework is gradient-specified, which is faster and more accurate than using the default numerical gradient, making it applicable for large-scale subject-specific uses.