Detection of mitoses in embryonic epithelia using motion field analysis.

Although computer simulations indicate that mitosis may be important to the mechanics of morphogenetic movements, algorithms to identify mitoses in bright field images of embryonic epithelia have not previously been available. Here, the authors present an algorithm that identifies mitoses and their orientations based on the motion field between successive images. Within this motion field, the algorithm seeks 'mitosis motion field prototypes' characterised by convergent motion in one direction and divergent motion in the orthogonal direction, the local motions produced by the division process. The algorithm uses image processing, vector field analyses and pattern recognition to identify occurrences of this prototype and to determine its orientation. When applied to time-lapse images of gastrulation and neurulation-stage amphibian (Ambystoma mexicanum) embryos, the algorithm achieves identification accuracies of 68 and 67%, respectively and angular accuracies of the order of 30 degrees , values sufficient to assess the role of mitosis in these developmental processes.

Digital tracking algorithm reveals the influence of structural irregularities on joint movements in the human cervical spine.

BACKGROUND: Disc height loss and osteophytes change the local mechanical environment in the spine; while previous research has examined kinematic dysfunction under degenerative change, none has looked at the influence of disc height loss and osteophytes throughout movement. METHODS: Twenty patients with pain related to the head, neck or shoulders were imaged via videofluoroscopy as they underwent sagittal-plane flexion and extension. A clinician graded disc height loss and osteophytes as "severe/moderate", "mild", or "none". A novel tracking algorithm quantified motions of each vertebra. This information was used to calculate intervertebral angular and shear displacements. The digital algorithm made it practical to track individual vertebrae in multiple patients through hundreds of images without bias. FINDINGS: Cases without height loss/osteophytes had a consistent increase in intervertebral angular displacement from C2/C3 to C5/C6, like that of healthy individuals, and mild height losses did not produce aberrations that were systematic or necessarily discernable. However, joints with moderate to severe disc height loss and osteophytes exhibited reduced range of motion compared to adjacent unaffected joints in that patient and corresponding joints in patients without structural irregularities. INTERPRETATION: Digitally-obtained motion histories of individual joints allowed anatomical joint changes to be linked with changes in joint movement patterns. Specifically, disc height loss and osteophytes were found to influence cervical spine movement in the sagittal plane, reducing angular motions at affected joints by approximately 10% between those with and without height loss and osteophytes. Further, these joint changes were associated with perturbed intervertebral angular and shear movements.

High strain rate compressive properties of bovine muscle tissue determined using a split Hopkinson bar apparatus.

The polymeric split Hopkinson pressure bar (PSHPB) apparatus is introduced as a means for measuring the high strain rate (1,000-2,500 s(-1)) compressive properties of soft tissues. Issues related to specimen design are discussed, and protocols are presented for specimen preparation. Proposed specimen geometries were validated using high-speed photography. Stress-strain data were obtained for high strain rate compression of bovine muscle tissue to strains as high as 80%. The stress-strain curves were found to be strain rate-sensitive and concave upward, as is typical of soft tissues. Rigor had a significant impact on the material properties between 5 and 24 h post mortem, while at longer times, properties returned essentially to their pre-rigor values. This study presents some of the first published high rate properties of muscle tissue, data that are urgently for advanced modeling of the human body and for evaluation of safety systems for the human body.

The mechanics of cell sorting and envelopment.

Aggregates of embryonic cells undergo a variety of intriguing processes including sorting by histological type and envelopment of cell masses of one type by another. It has long been held that these processes were driven by differential adhesions, as embodied in the famous differential adhesion hypothesis (DAH). Here, we use analytical mechanics to investigate the forces that are generated by various sub-cellular structures including microfilaments, cell membranes and their associated proteins, and by sources of cell-cell adhesions. We consider how these forces cause the triple junctions between cells to move, and how these motions ultimately give rise to phenomena such as cell sorting and tissue envelopment. The analyses show that, contrary to the widely accepted DAH, differential adhesions alone are unable to drive sorting and envelopment. They show, instead, that these phenomena are driven by the combined effect of several force generators, as embodied in an equivalent surface or interfacial tension. These unconventional findings follow directly from the relevant surface physics and mechanics, and are consistent with well-known cell sorting and envelopment experiments, and with recent computer simulations.

Mechanical effects of cell anisotropy on epithelia.

Theoretical, numerical and experimental methods are used to develop a comprehensive understanding of how cell shape affects the mechanical characteristics of two-dimensional aggregates such as epithelia. This is an important step in relating the mechanical properties of tissues to those of the cells of which they are composed. Statistical mechanics is used to derive formulas for the in-plane stresses generated by tensions gamma along cell-cell interfaces in sheets with anisotropic cellular fabric characterized by average cell aspect ratio kappa. These formulas are then used to investigate self-deformation (strain relaxation) of an anisotropic sheet composed of cells of thickness h and having effective viscosity mu. Finite element simulations of epithelia and of isolated cells and novel relaxation studies of specimens of embryonic epithelia reported herein are consistent with the predictions of the theory. In all cases, geometric factors cause the relaxation responses to be more complex than a single decaying exponential.

A computer model for reshaping of cells in epithelia due to in-plane deformation and annealing.

Although cell reshaping is fundamental to the mechanics of epithelia, technical barriers have prevented the methods of mechanics from being used to investigate it. These barriers have recently been overcome by the cell-based finite element formulation of Chen and Brodland. Here, parameters to describe the fabric of an epithelium in terms of cell shape and orientation and cell edge density are defined. Then, rectangular "patches" of model epithelia having various initial fabric parameters are generated and are either allowed to anneal or are subjected to one of several patterns of in-plane deformation. The simulations show that cell reshaping lags the deformation history, that it is allayed by cell rearrangement and that it causes the epithelium as a whole to exhibit viscoelastic mechanical properties. Equations to describe changes in cell shape due to annealing and in-plane deformation are presented.

Novel lap test determines the mechanics of delamination between annular lamellae of the intervertebral disc.

Delamination between lamellae of the annulus fibrosus is a crucial stage of intervertebral disc herniation, and to better understand the mechanics of the delamination process, a novel lap test was devised. Specimens consisting of two adjacent, naturally bonded lamellae were obtained from the cervical region of frozen porcine spines. They were cut into specimens nominally 3.5mm wide by 7 mm long and tabs of the deep and superficial layers were removed from opposite ends of the specimens so that a 4.5-5.0mm long intact interface remained between the lamellae. Specimens were mounted in a BioTester tensile instrument using BioRake attachments having 5 sharpened points side-by-side, and they were strained at 2%/s. Force-time curves were obtained and, using tracking software, a detailed map was made of the time course of the displacements within the specimens. Extensibility of the lamellae themselves was found to substantially complicate interpretation of the data. The experiments, together with mathematical analyses and finite element models, show that much of the shear load is transferred between lamellae at the ends of the bonded region, a finding of clinical importance. The inter-lamellae bond was found to have a peak strength of 0.30 ± 0.05 N/mm of specimen width (not to be confused with lap length), and the remarkable ability to carry substantial load even when lamellae had displaced up to 10mm relative to each other.

Biaxial mechanical testing of human sclera.

The biomechanical environment of the optic nerve head (ONH), of interest in glaucoma, is strongly affected by the biomechanical properties of sclera. However, there is a paucity of information about the variation of scleral mechanical properties within eyes and between individuals. We thus used biaxial testing to measure scleral stiffness in human eyes. Ten eyes from 5 human donors (age 55.4+/-3.5 years; mean+/-SD) were obtained within 24h of death. Square scleral samples (6mm on a side) were cut from each ocular quadrant 3-9 mm from the ONH centre and were mechanically tested using a biaxial extensional tissue tester (BioTester 5000, CellScale Biomaterials Testing, Waterloo). Stress-strain data in the latitudinal (toward the poles) and longitudinal (circumferential) directions, here referred to as directions 1 and 2, were fit to the four-parameter Fung constitutive equation W=c(e(Q)-1), where Q=c(1)E(11)(2)+c(2)E(22)(2)+2c(3)E(11)E(22) and W, c's and E(ij) are the strain energy function, material parameters and Green strains, respectively. Fitted material parameters were compared between samples. The parameter c(3) ranged from 10(-7) to 10(-8), but did not contribute significantly to the accuracy of the fitting and was thus fixed at 10(-7). The products cc(1) and cc(2), measures of stiffness in the 1 and 2 directions, were 2.9+/-2.0 and 2.8+/-1.9 MPa, respectively, and were not significantly different (two-sided t-test; p=0.795). The level of anisotropy (ratio of stiffness in orthogonal directions) was 1.065+/-0.33. No statistically significant correlations between sample thickness and stiffness were found (correlation coefficients=-0.026 and -0.058 in directions 1 and 2, respectively). Human sclera showed heterogeneous, near-isotropic, nonlinear mechanical properties over the scale of our samples.