Multiscale analysis of Klf10's impact on the passive mechanical properties of murine skeletal muscle.

Skeletal muscle is a hierarchical structure composed of multiple organizational scales. A major challenge in the biomechanical evaluation of muscle relates to the difficulty in evaluating the experimental mechanical properties at the different organizational levels of the same tissue. Indeed, the ability to integrate mechanical properties evaluated at various levels will allow for improved assessment of the entire tissue, leading to a better understanding of how changes at each level evolve over time and/or impact tissue function, especially in the case of muscle diseases. Therefore, the purpose of this study was to analyze a genetically engineered mouse model (Klf10 KO: Krüppel-Like Factor 10 knockout) with known skeletal muscle defects to compare the mechanical properties with wild-type (WT) controls at the three main muscle scales: the macroscopic (whole muscle), microscopic (fiber) and submicron (myofibril) levels. Passive mechanical tests (ramp, relaxation) were performed on two types of skeletal muscle (soleus and extensor digitorum longus (EDL)). Results of the present study revealed muscle-type specific behaviors in both genotypes only at the microscopic scale. Interestingly, loss of Klf10 expression resulted in increased passive properties in the soleus but decreased passive properties in the EDL compared to WT controls. At the submicron scale, no changes were observed between WT and Klf10 KO myofibrils for either muscle; these results demonstrate that the passive property differences observed at the microscopic scale (fiber) are not caused by sarcomere intrinsic alterations but instead must originate outside the sarcomeres, likely in the collagen-based extracellular matrix. The macroscopic scale revealed similar passive mechanical properties between WT and Klf10 KO hindlimb muscles. The present study has allowed for a better understanding of the role of Klf10 on the passive mechanical properties of skeletal muscle and has provided reference data to the literature which could be used by the community for muscle multiscale modeling.

The Effect of Freezing Time on Muscle Fiber Mechanical Properties.

The purpose of this study was to investigate the effect of freezing time on the functional behavior of mouse muscle fibers. Passive mechanical tests were performed on single soleus muscle fibers from fresh (0 month) and preserved (stored at -20°C for 3, 6, 9 and 12 months) 3 month old mice. The Young's modulus and the dynamic and the static stresses were measured. A viscoelastic Hill model of 3rd order was used to fit the experimental relaxation test data. The statistical analysis corresponding to the elastic modulus of single muscle fibers did not differ when comparing fresh and stored samples for 3 and 6 months at -20 °C. From 9 months, fibers were less resistant and the mechanical properties were damaged. The primary goal of this study was to complete the gold standard process of muscle fiber preservation for subsequent mechanical property studies. We have demonstrated that muscle fibers can be stored at -20°C for up to 6 months without altering their mechanical properties.

Estimation of accuracy of patient-specific musculoskeletal modelling: case study on a post polio residual paralysis subject.

For patients with patterns ranging out of anthropometric standard values, patient-specific musculoskeletal modelling becomes crucial for clinical diagnosis and follow-up. However, patient-specific modelling using imaging techniques and motion capture systems is mainly subject to experimental errors. The aim of this study was to quantify these experimental errors when performing a patient-specific musculoskeletal model. CT scan data were used to personalise the geometrical model and its inertial properties for a post polio residual paralysis subject. After having performed a gait-based experimental protocol, kinematics data were measured using a VICON motion capture system with six infrared cameras. The musculoskeletal model was computed using a direct/inverse algorithm (LifeMod software). A first source of errors was identified in the segmentation procedure in relation to the calculation of personalised inertial parameters. The second source of errors was subject related, as it depended on the reproducibility of performing the same type of gait. The impact of kinematics, kinetics and muscle forces resulting from the musculoskeletal modelling was quantified using relative errors and the absolute root mean square error. Concerning the segmentation procedure, we found that the kinematics results were not sensitive to the errors (relative error<1%). However, a strong influence was noted on the kinetics results (deviation up to 71%). Furthermore, the reproducibility error showed a significant influence (relative mean error varying from 5 to 30%). The present paper demonstrates that in patient-specific musculoskeletal modelling variations due to experimental errors derived from imaging techniques and motion capture need to be both identified and quantified. Therefore, the paper can be used as a guideline.

Automated analysis of MR image of hip: geometrical evaluation of the Legg-Calve-Perthes disease.

This study proposes semi-automatic determination of geometrical features in hip magnetic resonance (MR) images in order to evaluate the Legg-Calvé-Perthes disease (LCPD). Nine anatomical points on a hip image are selected by a clinician; then eight geometrical indexes of the hip joint are calculated: acetabulum head index (AHI), Wiberg angle (VCE), inner acetabular coverage angle (VCI), acetabular inclination angle (HTE), femoral shaft-neck angle (CC'D), circularity (C), convex deficiency factor (CDF) and pillar height deficiency factor (HDF) for the head region. The geometrical parameters are evaluated on 46 hip images of young patients with unilateral LCPD: 23 images concern the affected hip and 23 the unaffected hip. The extraction of the region of interest is done with a seeded region growing method. All the data were centered and reduced, and were subjected to principal component analysis. Supervised classification is applied with discriminant analysis and k-nearest neighbours classification. The AHI appears to be the best discriminant attribute (maximum between-class variance ratio). Cross-validation tests indicate that we can at most reduce the parameters to five (AHI, CC'D, DHF, DCF and VCE). The classification error rate for the linear discriminant method is 12.5%.

Comparative particle-induced cytotoxicity toward macrophages and fibroblasts.

The cytotoxicity caused by the debris resulting from wear of prostheses can produce major damage to tissues around the implant. We have compared particle internalization by macrophages and fibroblasts in vitro and analyzed cell death. J774.2 macrophages and L929 fibroblasts were incubated with 0.43 and 2.81 microm alumina particles or 0.45 and 3.53 microm polystyrene (PS) beads. Incubation of J774.2 cells with alumina particles of both sizes and 0.5 and 1.0 mg/ml PS beads significantly decreased cell numbers in a particle concentration-dependent manner. L929 cells were not affected by lower concentrations of 0.43 microm alumina particles (which aggregate at high concentrations) and they internalized 0.45 microm PS beads without any decrease in cell numbers. Particles were more cytotoxic for macrophages than for fibroblasts. Particles caused the size of both types of cells to increase in correlation with cytotoxicity. Trypan blue exclusion and lactate dehydrogenase release showed cell membrane leakage for both types of cells incubated with PS beads for 24 h. Apoptosis was assessed by annexin V-FITC, propidium iodide staining and assay of caspase 3 activity. Macrophage death appeared to depend on both necrosis, caused mainly by 3.53 microm PS beads, and apoptosis, mainly due to 0.45 microm PS beads. The release of the inflammatory cytokine IL-6 appears to be nonlinearly correlated with cytotoxicity. Thus, the size of the internalized particles affects macrophages and fibroblasts differently, and the increase in cell size can be used as a preliminary criterion of particle cytotoxicity in vitro.