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.

Extensor indicis proprius tendon transfer using shear wave elastography.

The means for judging optimal tension during tendon transfers are approximate and not very quantifiable. The purpose of this study was to demonstrate the feasibility of quantitatively assessing muscular mechanical properties intraoperatively using ultrasound elastography (shear wave elastography [SWE]) during extensor indicis proprius (EIP) transfer. We report two cases of EIP transfer for post-traumatic rupture of the extensor pollicis longus muscle. Ultrasound acquisitions measured the elasticity modulus of the EIP muscle at different stages: rest, active extension, active extension against resistance, EIP section, distal passive traction of the tendon, after tendon transfer at rest and then during active extension. A preliminary analysis was conducted of the distribution of values for this modulus at the various transfer steps. Different shear wave velocity and elasticity modulus values were observed at the various transfer steps. The tension applied during the transfer seemed close to the resting tension if a traditional protocol were followed. The elasticity modulus varied by a factor of 37 between the active extension against resistance step (565.1 kPa) and after the tendon section (15.3 kPa). The elasticity modulus values were distributed in the same way for each patient. The therapeutic benefit of SWE elastography was studied for the first time in tendon transfers. Quantitative data on the elasticity modulus during this test may make it an effective means of improving intraoperative adjustments.

TIEG-null mice display an osteopenic gender-specific phenotype.

TGFbeta inducible early gene-1 (TIEG) was originally cloned from human osteoblasts (OB) and has been shown to play an important role in TGFbeta/Smad signaling, regulation of gene expression and OB growth and differentiation. To better understand the biological role of TIEG in the skeleton, we have generated congenic TIEG-null (TIEG(-/-)) mice in a pure C57BL/6 background. Through the use of DXA and pQCT analysis, we have demonstrated that the femurs and tibias of two-month-old female TIEG(-/-) mice display significant decreases in total bone mineral content, density, and area relative to wild-type (WT) littermates. However, no differences were observed for any of these bone parameters in male mice. Further characterization of the bone phenotype of female TIEG(-/-) mice involved mechanical 3-point bending tests, micro-CT, and histomorphometric analyses of bone. The 3-point bending tests revealed that the femurs of female TIEG(-/-) mice have reduced strength with increased flexibility compared to WT littermates. Micro-CT analysis of femurs of two-month-old female TIEG(-/-) mice revealed significant decreases in cortical bone parameters compared to WT littermates. Histomorphometric evaluation of the distal femur revealed that female TIEG(-/-) mice also display a 31% decrease in cancellous bone area, which is primarily due to a decrease in trabecular number. At the cellular level, female TIEG(-/-) mice exhibit a 42% reduction in bone formation rate which is almost entirely due to a reduction in double labeled perimeter. Differences in mineral apposition rate were not detected between WT and TIEG(-/-) mice. Taken together, these findings suggest that female TIEG(-/-) mice are osteopenic mainly due to a decrease in the total number of functional/mature OBs.

Macroscopic-microscopic characterization of the passive mechanical properties in rat soleus muscle.

The purpose of the study was to investigate changes in passive mechanical properties of the soleus muscle of the rat during the first year of life. These mechanical changes were quantified at a macroscopic (whole muscle) and a microscopic level (fiber) and were correlated with biochemical and morphological properties. Three passive mechanical tests (a relaxation test, a ramp stretch test and a stretch release cycle test) with different amplitudes and velocities were performed on isolated soleus muscles and fibers in rats at ages 1 (R1), 4 (R4) and 12 (R12) months. Mechanical parameters (dynamic and static forces, stresses and normalized stiffness) were recorded and measured. The morphological properties (size of fibers and muscles) for the three groups of rats were assessed by light microscopy which allowed us to observe the evolution of the fiber type (I, IIc and IIa) in the belly region and along the longitudinal axis of the muscle. In addition, biochemical analyses were performed at the level of the whole muscle in order to determine the collagen content. The results of the passive mechanical properties between the macroscopic (muscle) and microscopic (fiber) levels showed a similar evolution. Thus, an increase of the dynamic and static forces appeared between 1 and 4 months while a decrease of the passive tension occurred between 4 and 12 months. These mechanical changes were correlated to the morphological properties. In addition, the size of the three fibers type which grew with age could explain the increase of forces between 1 and 4 months. Furthermore, the biochemical analysis showed an increase of the collagen content during the same period which could also be associated with the increase of the passive forces. After 4 months, the passive tension decreased while the size of the fiber continued to increase. The biochemical analysis showed a decrease of the collagen content after 4 months, which could explain the loss of passive tension in the whole muscle. Concerning the similar loss at the fiber level, other assumptions are required such as a myofibril loss process and an increase of intermyofibrillar spaces. The originality of this present study was to compare the passive mechanical properties between two different levels of anatomical organization within the soleus muscle of the rat and to explain these mechanical changes in terms of biochemical and morphological properties.