In this study, we investigated the effects of the diameter of microfibers on the orientation (angle between cells' major axis and the substrate fiber long axis) of adhered cells. For this purpose, mouse fibroblast L929 cells were cultured on the surface of PLGA fibers of defined diameters ranging from 10 to 242 mum, and their adhesion and alignment was quantitatively analyzed. It was found that the mean orientation of cells and the spatial variation of cell alignment angle directly related to the microfiber diameter. Cells that were cultured on microfibrous scaffolds oriented along the long axis of the microfiber and the orientation increased as the fiber diameter decreased. For the fiber diameter of 10 microm, the mean orientation was 3.0 +/- 0.2 degrees (mean +/- SE), whereas for a diameter of 242 microm, it decreased to 37.7 +/- 2.1 degrees . Using these studies we demonstrate that fibroblasts have a characteristic alignment on microscale fibers and that the microscale fiber diameter plays a critical role in cellular orientation. The ability to control cellular alignment on engineered tissue scaffold can be a potentially powerful approach to recreate the microscale architecture of engineered tissues. This may be important for engineering a variety of human tissues such as tendon, muscle and nerves as well as applications in 3D tissue culture and drug screening.

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