Alexander A. Spector Mohammed Ameen Department of Biomedical Engineering and Center for Computational Medicine and Biology, Johns Hopkins University, 720 Rutland Ave., Baltimore, MD 21205 Panos G. Charalambides Department of Mechanical Engineering, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 Aleksander S. Popel Department of Biomedical Engineering and Center for Computational Medicine and Biology, Johns Hopkins University, 720 Rutland Ave., Baltimore, MD 21205 Nanostructure, Effective Properties, and Deformation Pattern of the Cochlear Outer Hair Cell Cytoskeleton We consider the mechanical properties of the outer hair cell cytoskeleton. The cytoskel- eton is represented as a set of microdomains of different sizes and orientations composed of actin filaments and spectrin crosslinks. An intermediate material between domains is also introduced. The domain characteristics are randomly generated and the histograms of the cytoskeleton stiffness moduli are obtained. We solve an inverse problem and esti- mate the stiffness of the crosslink and connective molecule in the intermediate material. We discovered a pattern of highly inhomogeneous deformation of the cytoskeleton where the circumferential strain is primarily determined by the deformation of the intermediate material. DOI: 10.1115/1.1448521 Introduction Outer hair cells, receptor/effector cells in the cochlea of the mammalian ear, play a critical role in the active amplification, sharp frequency selectivity, and characteristic nonlinearities of the cochlea 1–3. The outer hair cell exhibits a unique form of mo- tility, named electromotility 4, changing its length in response to changes of the cell membrane wallelectric potential. The outer hair cells produce the active force 5,6, affect vibration of the basilar membrane and other cochlear components 7–9, and pro- vide energy for the active processes in the cochlea. The outer hair cell is an elongated cylinder with a pressurized liquid core bounded by a three-layer composite wall 1. The cell cytoskel- eton is represented by the intermediate layer between the outer- most plasma membrane and the innermost subsurface cisternae. The cytoskeleton, which supports the cell’s cylindrical shape, is important for normal turgor pressure in the cell. The cytoskeleton is also critical for the active force production, a mechanism that includes generation of the active strains in the plasma membrane and their transmission through the composite cell wall 10. In first experimental studies with demembranated cells 11, the molecular structure of the cytoskeleton was analyzed and its lon- gitudinal stiffness was measured. As a result of such analysis, a model of the cytoskeleton as a regular two-dimensional network of coiled helical actin filaments connected by longitudinal spectrin crosslinks was proposed. The connection of the cytoskeleton to the plasma membrane was studied, and the geometric parameters of a system of radial pillars between the two components of the cell wall were estimated 12. Later, it was discovered 13that the outer hair cell cytoskeleton has a more complicated, less regu- lar, nanostructure. It was shown that the cytoskeleton is composed of small domains of different sizes 200–1000 nm longand ori- entations, where each domain is composed of actin filaments and spectrin crosslinks. The nanostructure of the cytoskeleton is such that its mechanical properties in the longitudinal along the cell and circumferential around the celldirections are expected to be different. Indeed, significant anisotropy was shown by directly measuring the longitudinal and circumferential stiffness of an iso- lated cytoskeleton 14. The cytoskeleton determines effective an- isotropy of the whole outer hair cell wall because other compo- nents of the wall are practically isotropic. Anisotropy of the whole wall was estimated and the ratio of the circumferential stiffness modulus over that for the longitudinal direction was found to be in the range 3.5–4 15–17. In another study 18, anisotropy of the outer hair cell wall was also confirmed, but the ratio of the stiff- ness moduli was close to 1.5. The cytoskeleton in other cells has been a topic of intensive studies because of its importance for the cell structural integrity, deformability, and signal transduction. The behavior and properties of the red blood cell cytoskeleton 19–23and effective moduli of the endothelial cell cytoskeleton 24were analyzed on the basis of these cells’ nanostructure. Modeling of the rearrangement of the endothelial cell cytoskel- eton in response to external forces stretchesor concentration of the filaments was also developed 25,26. In the present paper, we consider the multiple-domain nano- structure of the outer hair cell cytoskeleton and introduce an in- termediate material connecting adjacent domains Fig. 1. We con- sider random variation of the major intradomain geometric parameters together with domain orientation and size. We assume that these characteristics are distributed within ranges known from experimental measurements. By using the finite element method and generating the random parameters of the model, we obtain histograms of the elements of the 33 symmetric matrix of the anisotropic stiffness moduli of the cytoskeleton. We find that the cytoskeleton exhibits a form of anisotropy more general than orthotropy. By applying our model to experimental values of the longitudinal and circumferential stiffness of the cytoskeleton, we estimate the stiffness of the molecules entering our model. We show that the stiffness of the hypothetical connective molecule in the intermediate material is close to that of the spectrin crosslink. Contributed by the Bioengineering Division for publication in the JOURNAL OF BIOMECHANICAL ENGINEERING. Manuscript received by the Bioengineering Divi- sion April 2001; revised manuscript received November 2001. Associate Editor: C. Dong. 180 Õ Vol. 124, APRIL 2002 Copyright © 2002 by ASME Transactions of the ASME