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Contractility of cells in wound site is important to understand pathological wound healing and develop therapeutic strategies. In particular, contractile force generated by cells is a basic element for designing artificial three-dimensional cell culture scaffolds. Direct assessment of deformation of three-dimensional structured materials has been used to calculate contractile forces by averaging total forces with respect to the cell population number. However, macroscopic methods have offered only lower bounds of contractility due to experimental assumptions and the large variance of the spatial and temporal cell response. In the present study, cell contractility was examined microscopically in order to measure contractile forces generated by individual cells on self-standing fiber scaffolds that were fabricated via femtosecond laser-induced two-photon polymerization. Experimental assumptions and calculation errors that arose in previous studies of macroscopic and microscopic contractile force measurements could be reduced by adopting a columnar buckling model on individual, standing fiber scaffolds. Via quantifying eccentric critical loads for the buckling of fibers with various diameters, contractile forces of single cells were calculated in the range between 30-116 nN. In the present study, a force magnitude of approximately 200 nN is suggested as upper bound of the contractile force exerted by single cells. In addition, contractile forces by multiple cells on a single fiber were calculated in the range between 241-709 nN.

Development of materials and fabrication techniques lead the growth of three-dimensional cell culture matrices in biomedical engineering. In this work, we present a method for fabricating self-standing fiber scaffolds by two-photon polymerization induced by a femtosecond laser. The aligned fibers are 330 μm long with a diameter of 6–9 μm. Depending on the pitch of the aligned fibers, various cell morphologies are distinguished via three-dimensional images. Furthermore, the morphologies of fibroblast cells (NIH-3T3) and epithelial cells (MDCK) on the fiber scaffolds are studied to show the effect of high curvature (3–4.5 μm radii) on cell morphology. NIH-3T3 cells that contain straight pattern of actin microfilament bundles are extended and partly wrap single fibers or tend to reside between fibers. On the other hand, MDCK cells that contain circular pattern of actin microfilament bundles cover the fiber peripheral surface exhibiting high aspect ratio elongation. These results indicate that cell morphology on fiber scaffolds is influenced by the pattern of actin microfilament bundles.