Multimodal imaging and functional analysis of the chick NMDA retinal damage model.

OBJECTIVES: The chick is rapidly becoming a standardized preclinical model in vision research to study mechanisms of ocular disease. We seek to comprehensively evaluate the N-methyl-D-aspartate (NMDA) model of excitotoxic retinal damage using multimodal imaging, functional, and histologic approaches in NMDA-damaged, vehicle-treated, and undamaged chicks. METHODS: Chicks were either left undamaged in both eyes or were injected with NMDA in the left eye and saline (vehicle) in the right eye. TUNEL assay was performed on chicks to assess levels of retinal cell death one day post-injection of NMDA or saline and on age-matched untreated chicks. Spectral domain optical coherence tomography (SD-OCT) was performed weekly on chicks and age-matched controls day 1 (D1) up to D28 post-injection. Light adapted electroretinograms (ERG) were performed alongside SD-OCT measurements on post-injection chicks along with age-matched untreated controls. RESULTS: Untreated and vehicle-treated eyes had no TUNEL positive cells while NMDA-treated eyes accumulated large numbers of TUNEL positive cells in the Inner Nuclear Layer (INL), but not other layers, at D1 post injection. Significant inner retina swelling or edema was found on SD-OCT imaging at D1 post-injection which resolved at subsequent timepoints. Both the INL and the inner plexiform layer significantly thinned by one-week post-injection and did not recover for the duration of the measurements. On ERG, NMDA-treated eyes had significantly reduced amplitudes of all parameters at D1 with all metrics improving over time. The b-wave, oscillatory potentials, and ON/OFF bipolar responses were the most affected with at least 70% reduction immediately after damage compared to the fellow eye control. CONCLUSION: This study establishes a normative baseline on the retinal health and gross functional ability as well as intraocular pressures of undamaged, vehicle-treated, and NMDA-damaged chicks to provide a standard for comparing therapeutic treatment studies in this important animal model.

Quasistatic kinetic avalanches and self-organized criticality in deviatorically loaded granular media.

The behavior of granular media under quasistatic loading has recently been shown to attain a stable evolution state corresponding to a manifold in the space of micromechanical variables. This state is characterized by sudden transitions between metastable jammed states, involving the partial micromechanical rearrangement of the granular medium. Using numerical simulations of two-dimensional granular media under quasistatic biaxial compression, we show that the dynamics in the stable evolution state is characterized by scale-free avalanches well before the macromechanical stationary flow regime traditionally linked to a self-organized critical state. This, together with the nonuniqueness and the nonmonotony of macroscopic deformation curves, suggests that the statistical avalanche properties and the susceptibilities of the system cannot be reduced to a function of the macromechanical state. The associated scaling exponents are nonuniversal and depend on the interactions between particles. For stiffer particles (or samples at low confining pressure) we find distributions of avalanche properties compatible with the predictions of mean-field theory. The scaling exponents decrease below the mean-field values for softer interactions between particles. These lower exponents are consistent with observations for amorphous solids at their critical point. We specifically discuss the relationship between microscopic and macroscopic variables, including the relation between the external stress drop and the internal potential energy released during kinetic avalanches.

Correction: Effects of collagen matrix and bioreactor cultivation on cartilage regeneration of a full-thickness critical-size knee joint cartilage defects with subchondral bone damage in a rabbit model.

[This corrects the article DOI: 10.1371/journal.pone.0196779.].

Effects of collagen matrix and bioreactor cultivation on cartilage regeneration of a full-thickness critical-size knee joint cartilage defects with subchondral bone damage in a rabbit model.

Cartilage has limited self-repair ability. The purpose of this study was to investigate the effects of different species of collagen-engineered neocartilage for the treatment of critical-size defects in the articular joint in a rabbit model. Type II and I collagen obtained from rabbits and rats was mixed to form a scaffold. The type II/I collagen scaffold was then mixed with rabbit chondrocytes to biofabricate neocartilage constructs using a rotating cell culture system [three-dimensional (3D)-bioreactor]. The rabbit chondrocytes were mixed with rabbit collagen scaffold and rat collagen scaffold to form neoRBT (neo-rabbit cartilage) and neoRAT (neo-rat cartilage) constructs, respectively. The neocartilage matrix constructs were implanted into surgically created defects in rabbit knee chondyles, and histological examinations were performed after 2 and 3 months. Cartilage-like lacunae formation surrounding the chondrocytes was noted in the cell cultures. After 3 months, both the neoRBT and neoRAT groups showed cartilage-like repair tissue covering the 5-mm circular, 4-mm-deep defects that were created in the rabbit condyle and filled with neocartilage plugs. Reparative chondrocytes were aligned as apparent clusters in both the neoRAT and neoRBT groups. Both neoRBT and neoRAT cartilage repair demonstrated integration with healthy adjacent tissue; however, more integration was obtained using the neoRAT cartilage. Our data indicate that different species of type II/I collagen matrix and 3D bioreactor cultivation can facilitate cartilage engineering in vitro for the repair of critical-size defect.

The possible influence of osmotic poration on cell membrane water permeability.

It has been hypothesized that pores in the plasma membrane form under conditions of rapid water efflux, allowing extracellular ice to grow into the cytoplasm under conditions of rapid freezing. When cells with intracellular ice are thawed slowly, the transmembrane ice crystal expands through recrystallization causing the cell to lyse. One of the implications of this hypothesis is that osmotic pores will provide an alternative route for water movement under conditions of osmotically induced flow. We show that the plasma membrane water permeability of a fibroblast cell changes as a function of the osmotic pressure gradient that is used to drive water movement. It is further shown that cell volume is more important than the magnitude of water flux in causing this departure from a uniform water permeability. We suggest that these data provide evidence of a transient route for water movement across cell membranes.

Retardation of ice growth in glass capillaries: measurement of the critical capillary radius.

An experiment was designed to compare the freezing of an aqueous solution in glass microcapillaries and in thin films. The velocity dependence of the ice front propagation in glass capillaries with radii of 87.5 microm-1.5 microm was observed. A critical capillary radius r(0), corresponding to certain thermal conditions, was obtained, below which the ice growth inside the capillaries was retarded. This critical capillary radius is further related to lambda(0), the smallest wavelength used in the Mullins-Sekerka criterion for the instability analysis of bulk solidifications [Mullins and Sekerka, J. Appl. Phys. 35, 444 (1964)]. It was found that for the present hypothesis, r(0)=lambda(0)/4 gives good predictions. The relation between the propagation velocity (or cooling rate) and the critical radius (or pore size) is summarized in a chart for applications in capillary-porous media, such as in the freezing of biological tissues.

A finite element model for ice ball evolution in a multi-probe cryosurgery.

The ice formation in a water body is examined for the computation of temperature field, phase change and a moving ice-water interface whose location is not known á priori. This is classically referred to as the Stefan problem [Rubinstein, L.I. (1971) The Stefan Problem (American Mathematical Society, Providence, Rhode Island 02904]. Based on the Duvaut [Duvaut, G. (1973) "Résolution d'un probléme Stefan" C.R. Acad Sci. Paris 276, 1461-1463] transformation, the governing equations for heat conduction are formulated within a variational principle that is readily amenable to a standard finite element solution without remeshing. Numerical simulation results pertaining to the freezing of tumour tissue in a multi-cryoprobe cryosurgery are presented. These results lend both quantitative and graphical support to the current empirical standards of "effective therapy" in view of refining clinical applications.

Measurement of freezing point depression of water in glass capillaries and the associated ice front shape.

Variations of the Kelvin equation [W. Thomson, Philos. Mag. 42, 448 (1871)] to describe the freezing point depression of water in capillaries exist in the literature. The differing equations, coupled with the uncertainty in input parameters, lead to various predictions. The difference between the predictions may become substantial when the capillary size decreases much below micron dimensions. An experiment was designed to investigate the predicted values using a customized directional solidification stage. The capillary freezing point depression for glass tubes with radii of 87 microm-3 microm was successfully measured. The image of the ice-water interface at equilibrium was also digitally captured and analyzed to examine the contact angle and the interface shape as well. Both are important for examining the hemispherical interface assumption that was exclusively used in the theoretical derivations. Finally, an equilibrium analysis of the thermodynamic system leads to a theoretical discussion of the problem. The effect of the temperature gradient on the interface shape is addressed, and an engineering criterion for the critical temperature gradient above which the effect must be considered for the interface shape calculation is derived.