The malleus and incus in the human middle ear are linked by the incudo-malleolar joint (IMJ). The mobility of the human IMJ under physiologically relevant acoustic stimulation and its functional role in middle-ear sound transmission are still debated. In this study, spatial stapes motions were measured during acoustic stimulation (0.25-8 kHz) in six fresh human temporal bones for two conditions of the IMJ: (1) normal IMJ and (2) IMJ with experimentally-reduced mobility. Stapes velocity was measured at multiple points on the footplate using a scanning laser Doppler vibrometry (SLDV) system, and the 3D motion components were calculated under both conditions of the IMJ. The artificial reduction of the IMJ mobility was confirmed by measuring the relative motion between the malleus and the incus. The magnitudes of the piston-like motion of the stapes increased with the reduced IMJ mobility above 2 kHz. The increase was frequency dependent and was prominent from 2 to 4 kHz and at 5.5 kHz. The magnitude ratios of the rocking-like motions to the piston-like motion were similar for both IMJ conditions. The frequency-dependent change of the piston-like motion after the reduction of the IMJ mobility suggests that the IMJ is mobile under physiologically relevant levels of acoustic stimulation, especially at frequencies above 2 kHz. The malleus and incus in the human middle ear are linked by the incudo-malleolar joint 10 (IMJ). The mobility of the human IMJ under physiologically relevant acoustic stimulation and 11 its functional role in middle-ear sound transmission are still debated. In this study, spatial 12 stapes motions were measured during acoustic stimulation (0.25-8 kHz) in six fresh human 13 temporal bones for two conditions of the IMJ: (1) normal IMJ and (2) IMJ with 14 experimentally-reduced mobility. Stapes velocity was measured at multiple points on the 15 footplate using a scanning laser Doppler vibrometry (SLDV) system, and the 3D motion 16 components were calculated under both conditions of the IMJ. The artificial reduction of the 17 IMJ mobility was confirmed by measuring the relative motion between the malleus and the 18 incus. The magnitudes of the piston-like motion of the stapes increased with the reduced IMJ 19 mobility above 2 kHz. The increase was frequency dependent and was prominent from 2-4 20 kHz and at 5.5 kHz. The magnitude ratios of the rocking-like motions to the piston-like 21 motion were similar for both IMJ conditions. The frequency-dependent change of the piston-22 like motion after the reduction of the IMJ mobility suggests that the IMJ is mobile under 23 physiologically relevant levels of acoustic stimulation, especially at frequencies above 2 kHz. 24 2

The annular ligament provides a compliant connection of the stapes to the oval window. To estimate the stiffness characteristics of the annular ligament, human temporal bone measurements were conducted. A force was applied sequentially at several points on the stapes footplate leading to different patterns of displacement with different amounts of translational and rotational components. The spatial displacement of the stapes footplate was measured using a laser vibrometer. The experiments were performed on several stapes with dissected chain and the force was increased stepwise, resulting in load-deflection curves for each force application point. The annular ligament exhibited a progressive stiffening characteristic in combination with an inhomogeneous stiffness distribution. When a centric force, orientated in the lateral direction, was applied to the stapes footplate, the stapes head moved laterally and in the posterior-inferior direction. Based on the load-deflection curves, a mechanical model of the annular ligament was derived. The mathematical representation of the compliance of the annular ligament results in a stiffness matrix with a nonlinear dependence on stapes displacement. This description of the nonlinear stiffness allows simulations of the sound transfer behavior of the middle ear for different preloads.

Both SLDV and 3D LDV data indicate that sound transmission in the skull bone causes rigid-body-like motion at low frequencies whereas transverse deformations and travelling waves were observed above 2 kHz, with propagation speeds of approximately of 450 m/s at 8 kHz.

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BACKGROUND: The malleus-incus complex (MIC) plays a crucial role in the hearing process as it transforms and transmits acoustically-induced motion of the tympanic membrane, through the stapes, into the inner-ear. However, the transfer function of the MIC under physiologically-relevant acoustic stimulation is still under debate, especially due to insufficient quantitative data of the vibrational behavior of the MIC. This study focuses on the investigation of the sound transformation through the MIC, based on measurements of three-dimensional motions of the malleus and incus with a full six degrees of freedom (6 DOF). METHODS: The motion of the MIC was measured in two cadaveric human temporal bones with intact middle-ear structures excited via a loudspeaker embedded in an artificial ear canal, in the frequency range of 0.5-5 kHz. Three-dimensional (3D) shapes of the middle-ear ossicles were obtained by sequent micro-CT imaging, and an intrinsic frame based on the middle-ear anatomy was defined. All data were registered into the intrinsic frame, and rigid body motions of the malleus and incus were calculated with full six degrees of freedom. Then, the transfer function of the MIC, defined as velocity of the incus lenticular process relative to velocity of the malleus umbo, was obtained and analyzed. RESULTS: Based on the transfer function of the MIC, the motion of the lenticularis relative to the umbo reduces with frequency, particularly in the 2-5 kHz range. Analysis of the individual motion components of the transfer function indicates a predominant medial-lateral component at frequencies below 1 kHz, with low but considerable anterior-posterior and superior-inferior components that become prominent in the 2-5 kHz range. CONCLUSION: The transfer function of the human MIC, based on motion of the umbo and lenticularis, has been visualized and analyzed. While the magnitude of the transfer function decreases with frequency, its spatio-temporal complexity increases significantly. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. M A N U S C R I P T A C C E P T E D ACCEPTED MANUSCRIPT

The piston-like (translation normal to the footplate) and rocking-like (rotation along the long and short axes of the footplate) are generally accepted as motion components of the human stapes. It has been of issue whether in-plane motions, i.e., transversal movements of the footplate in the oval window, are comparable to these motion components. In order to quantify the in-plane motions the motion at nine points on the medial footplate was measured in five temporal bones with the cochlea drained using a three-dimensional (3D) laser Doppler vibrometer. It was found that the stapes shows in-plane movements up to 19.1 ± 8.7% of the piston-like motion. By considering possible methodological errors, i.e., the effects of the applied reflective glass beads and of alignment of the 3D laser Doppler system, such value was reduced to be about 7.4 ± 3.1%. Further, the in-plane motions became minimal (≈ 4.2 ± 1.4% of the piston-like motion) in another plane, which was anatomically within the footplate. That plane was shifted to the lateral direction by 118 μm, which was near the middle of the footplate, and rotated by 4.7° with respect to the medial footplate plane.

Under large quasi-static loads, the incudo-malleolar joint (IMJ), connecting the malleus and the incus, is highly mobile. It can be classified as a mechanical filter decoupling large quasi-static motions while transferring small dynamic excitations. This is presumed to be due to the complex geometry of the joint inducing a spatial decoupling between the malleus and incus under large quasi-static loads. Spatial Laser Doppler Vibrometer (LDV) displacement measurements on isolated malleus-incus-complexes (MICs) were performed. With the malleus firmly attached to a probe holder, the incus was excited by applying quasi-static forces at different points. For each force application point the resulting displacement was measured subsequently at different points on the incus. The location of the force application point and the LDV measurement points were calculated in a post-processing step combining the position of the LDV points with geometric data of the MIC. The rigid body motion of the incus was then calculated from the multiple displacement measurements for each force application point. The contact regions of the articular surfaces for different load configurations were calculated by applying the reconstructed motion to the geometry model of the MIC and calculate the minimal distance of the articular surfaces. The reconstructed motion has a complex spatial characteristic and varies for different force application points. The motion changed with increasing load caused by the kinematic guidance of the articular surfaces of the joint. The IMJ permits a relative large rotation around the anterior-posterior axis through the joint when a force is applied at the lenticularis in lateral direction before impeding the motion. This is part of the decoupling of the malleus motion from the incus motion in case of large quasi-static loads. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractUnder large quasi-static loads, the incudo-malleolar joint (IMJ), connecting the malleus and the incus, is highly mobile. It can be classified as a mechanical filter decoupling large quasi-static motions while transferring small dynamic excitations. This is presumed to be due to the complex geometry of the joint inducing a spatial decoupling between the malleus and incus under large quasi-static loads.Spatial Laser Doppler Vibrometer (LDV) displacement measurements on isolated malleus-incus-complexes (MICs) were performed. With the malleus firmly attached to a probe holder, the incus was excited by applying quasi-static forces at different points. For each force application point the resulting displacement was measured ...

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