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The viscoelastic properties of single, attached C2C12 myoblasts were measured using a recently developed cell loading device. The device allows global compression of an attached cell, while simultaneously measuring the associated forces. The viscoelastic properties were examined by performing a series of dynamic experiments over two frequency decades (0.1-10 Hz) and at a range of axial strains (approximately 10-40%). Confocal laser scanning microscopy was used to visualize the cell during these experiments. To analyze the experimentally obtained force-deformation curves, a nonlinear viscoelastic model was developed. The nonlinear viscoelastic model was able to describe the complete series of dynamic experiments using only a single set of parameters, yielding an elastic modulus of 2120 +/- 900 Pa for the elastic spring, an elastic modulus of 1960 +/- 1350 for the nonlinear spring, and a relaxation time constant of 0.3 +/- 0.12 s. To our knowledge, it is the first time that the global viscoelastic properties of attached cells have been quantified over such a wide range of strains. Furthermore, the experiments were performed under optimal environmental conditions and the results are, therefore, believed to reflect the viscoelastic mechanical behavior of cells, such as would be present in vivo.

Skeletal muscle cells are sensitive to sustained compression, which can lead to the development of pressure sores. Although it is known that this type of tissue breakdown depends on the magnitude and duration of the applied load, the exact relationship between cell deformation and damage remains unclear. To gain more insight into this process, a method has been developed, that incorporates the use of a new loading device and confocal microscopy. The loading device is able to compress individual cells, either statically or dynamically, while measuring the resulting forces. Experiments can be performed under ideal environmental conditions, comparable with those of a CO2 incubator. First compression experiments on C2C12 mouse myoblasts showed the shape changes that cells undergo during static compression by the loading device. Calculations using the three-dimensional confocal images showed no change in volume and an increase in the surface area of the cell as a result of compression. The device presented here provides a useful way to monitor the biomechanical response of skeletal muscle cells during long-term compression experiments. Therefore it will contribute to the knowledge about strain-induced cell damage, as seen in pressure sores and other mechanically induced clinical conditions.

Abstract-Quantifying three-dimensional deformation of cells under mechanical load is relevant when studying cell deformation in relation to cellular functioning. Because most cells are anchorage dependent for normal functioning, it is desired to study cells in their attached configuration. This study reports new threedimensional morphometric measurements of cell deformation during stepwise compression experiments with a recently developed cell loading device. The device allows global, unconfined compression of individual, attached cells under optimal environmental conditions. Three-dimensional images of fluorescently stained myoblasts were recorded with confocal microscopy and analyzed with image restoration and three-dimensional image reconstruction software to quantify cell deformation. In response to compression, cell width, cross-sectional area, and surface area increased significantly with applied strain, whereas cell volume remained constant. Interestingly, the cell and the nucleus deformed perpendicular to the direction of actin filaments present along the long axis of the cell. This strongly suggests that this anisotropic deformation can be attributed to the preferred orientation of actin filaments. A shape factor was introduced to quantify the global shape of attached cells. The increase of this factor during compression reflected the anisotropic deformation of the cell.