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Morphology and Adhesion Strength of Myoblast Cells on Photocurable Gelatin under Native and Non-native Micromechanical Environments

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Physical Chemistry of Biosystems, Institute of Physical Chemistry, University of Heidelberg, Heidelberg 69120, Germany
Department of Chemistry, Saitama University, Saitama 338-8570, Japan
§ Division of Biomolecular Chemistry, Institute for Materials Chemistry and Engineering, Kyushu University, Fukuoka 819-0395, Japan
Genome Biotechnology Laboratory, Kanazawa Institute of Technology, Ishikawa 924-0838, Japan
Institute for Integrated Cell-Material Sciences (WPI iCeMS), Kyoto University, 606-8501, Kyoto, Japan
*Tel +81-(0)-92-802-2507, Fax +81-(0)-92-802-2509, e-mail [email protected] (S.K.); Tel +49-(0)-6221-544916, Fax +49-(0)-6221-544918, e-mail [email protected] (M.T.).
Cite this: J. Phys. Chem. B 2013, 117, 15, 4081–4088
Publication Date (Web):March 26, 2013
https://doi.org/10.1021/jp4008224
Copyright © 2013 American Chemical Society

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    Abstract

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    We have quantitatively determined how the morphology and adhesion strength of myoblast cells can be regulated by photocurable gelatin gels, whose mechanical properties can be fine-tuned by a factor of 103 (0.1 kPa ≤ E ≤ 140 kPa). The use of such gels allows for the investigation of mechanosensing of cells not only near the natural mechanical microenvironments (E ∼ 10 kPa) but also far below and beyond of the natural condition. Optical microscopy and statistical image analysis revealed that myoblast cells sensitively adopt their morphology in response to the substrate elasticity at E ∼ 1–20 kPa, which can be characterized by the significant changes in the contact area and order parameters of actin cytoskeletons. In contrast, the cells in contact with the gels with lower elastic moduli remained almost round, and the increase in the elasticity beyond E ∼ 20 kPa caused no distinct change in morphology. In addition to the morphological analysis, the adhesion strength was quantitatively evaluated by measuring the critical detachment pressure with an aid of intensive pressure waves induced by picosecond laser pulses. This noninvasive technique utilizing extremely short pressure waves (pulse time width ∼100 ns) enables one to determine the critical pressure for cell detachment with reliable statistics while minimizing the artifacts arising from the inelastic deformation of cells. The adhesion strength also exhibited a transition from weak adhesion to strong adhesion within the same elasticity range (E ∼ 1–20 kPa). A clear correlation between the cell morphology and adhesion strength suggests the coupling of the strain of the substrate and the mechanosensors near focal adhesion sites.

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    Representative results of image analysis for the actin fiber orientation and a log–log pot of projected area of C2C12 cells as a function of gel elasticity. This material is available free of charge via the Internet at http://pubs.acs.org.

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