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M M Knight

Publications and source records attributed to M M Knight.

39 records · Page 3Linked to original sources

Measurement of the deformation of isolated chondrocytes in agarose subjected to cyclic compression.

Mechanically induced cell deformation is one of a number of possible mechanotransduction pathways by which chondrocytes sense and respond to changes in their mechanical environment. The present study describes a system for measuring the deformation of isolated chondrocytes in agarose during both static and cyclic compression. A test rig mounted on the stage of an inverted microscope was used to apply precise levels of compressive strain to individual cell-agarose constructs bathed in culture medium. Images of the cells were recorded using a CCD video camera attached to the microscope. Cell deformation was quantified in terms of a deformation index (X/Y) representing the ratio of cell diameters measured parallel (X) and perpendicular (Y) to the axis of compression. Cyclic compression between 0 and 15% strain, at 0.3 Hz, resulted in cyclic deformation of the cells at the same frequency. However, during the unstrained phase the cells did not fully recover to their initially spherical morphology (X/Y = 1.0). During the strained phase, the level of deformation (X/Y = 0.59) was initially similar to that observed during static 15% strain. However, this level of cell deformation reduced over a 20 min period of cyclic compression (X/Y = 0.72), although during static compression the cell deformation remained constant. This system may be used to examine cellular events under a range of dynamic mechanical stimuli.

Animals↗

Medication education for children. Is it worthwhile?

Child psychiatric inpatients have little or no control over their treatment. Anxiety and fear are normal reactions to the hospitalization and treatment process. Pharmacotherapy is an intervention that the child often perceives as an indicator of pathology. This article presents various methodologies to teach children about their medication treatment. Puppetry, graphics, art, and discussion were techniques used in a peer group to impart knowledge and decrease anxiety, fear, and negativity.

Adolescent↗

Intracellular mechanics and mechanotransduction associated with chondrocyte deformation during pipette aspiration.

The present study utilised pipette aspiration and simultaneous confocal microscopy to test the hypothesis that chondrocyte deformation is associated with distortion of intracellular organelles and activation of calcium signalling. Aspiration pressure was applied to isolated articular chondrocytes in increments of 2 cm of water every 60 seconds up to a maximum of 10 cm of water. At each pressure increment, confocal microscopy was used to visualise the mitochondria and nucleus labelled with JC-1 and Syto-16, respectively. To investigate intracellular calcium signalling, separate cells were labelled with Fluo 4, rapidly aspirated to 5 cm of water and then imaged for 5 minutes at a tare pressure of 0.1 cm of water. Partial cell aspiration was associated with distortion of the mitochondrial network, elongation of the nucleus and movement towards the pipette mouth. Treatment with cytochalasin D or nocodazole produced an increase in cell aspiration indicating that both the actin microfilaments and microtubules provide mechanical integrity to the cell. When the data was normalised to account for the increased cell deformation, both actin microfilaments and microtubules were shown to be necessary for strain transfer to the intracellular organelles. Mitochondria and nucleus deformation may both be involved in chondrocyte mechanotransduction as well as cellular and intracellular mechanics. In addition, pipette aspiration induced intracellular calcium signalling which may also form part of a mechanotransduction pathway. Alternatively calcium mobilisation may serve to modify actin polymerisation, thereby changing cell mechanics and membrane rigidity in order to facilitate localised cell deformation. These findings have important implications for our understanding of cell mechanics and mechanotransduction as well as interpretation and modelling of pipette aspiration data.

Animals↗