On the role of the actin cytoskeleton and nucleus in the biomechanical response of spread cells
Reynolds, Noel H.
Dowling, Enda P.
McMeeking, Robert M.
McGarry, J. Patrick
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Reynolds, NH,Ronan, W,Dowling, EP,Owens, P,McMeeking, RM,McGarry, JP (2014) 'On the role of the actin cytoskeleton and nucleus in the biomechanical response of spread cells'. Biomaterials, 35 :4015-4025.
Micropipette aspiration (MA) has been used extensively in biomechanical investigations of un-adhered cells suspended in media. In the current study, a custom MA system is developed to aspirate substrate adhered spread cells. Additionally, the system facilitates immuno-fluorescent staining of aspirated cells to investigate stress fibre redistribution and nucleus deformation during MA. In response to an applied pressure, significantly lower aspiration length is observed for untreated contractile cells compared to cells in which actin polymerisation is chemically inhibited, demonstrating the important contribution of stress fibres in the biomechanical behaviour of spread cells. Additional experiments are performed in which untreated contractile cells are subjected to a range of applied pressures. Computational finite element simulations reveal that a viscoelastic material model for the cell cytoplasm is incapable of accurately predicting the observed aspiration length over the range of applied pressures. It is demonstrated that an active computational framework that incorporates stress fibre remodelling and contractility must be used in order to accurately simulate MA of untreated spread cells. Additionally, the stress fibre distribution observed in immuno-fluorescent experimental images of aspirated cells is accurately predicted using the active stress fibre modelling framework. Finally, a detailed experimental computational investigation of the nucleus mechanical behaviour demonstrates that the nucleus is highly deformable in cyto, reaching strain levels in excess of 100% during MA. The characterisation of stress fibres and nucleus biomechanics in spread cells presented in the current study can potentially be used to guide tissue engineering strategies to control cell behaviour and gene expression. (c) 2014 Elsevier Ltd. All rights reserved.
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