[Medical engineering].
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Biomedical subjects
Publications and source records attributed to Joji Ando.
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The caveola is a membrane domain that compartmentalizes signal transduction at the cell surface. Normally in endothelial cells, groups of caveolae are found clustered along stress fibers or at the lateral margins in all regions of the cell. Subsets of these clusters appear to contain the signaling machinery for initiating Ca(2+) wave formation. Here we report that induction of cell migration, either by wounding a cell monolayer or by exposing cells to laminar shear stress, causes caveolae to move to the trailing edge of the cell. Concomitant with the relocation of the caveolae, sites of Ca(2+) wave initiation move to the same location. In as much as the relocated caveolae contain elements of the signaling machinery required for ATP-stimulated release of Ca(2+) from the ER, these results suggest that caveolae function as containers that carry this machinery to different cellular locations.
Since ischemia-reperfusion injury continues to be a major problem in reconstructive microsurgery, improvement of experimental models is still desirable. We developed a model that allows direct visualization of flap microcirculation in mice by intravital microscopic techniques. A newly designed skinfold chamber was installed on the dorsum of mice, and microcirculation was inspected with an intravital microscope. An island flap, nourished by the deep circumflex iliac arteries, was elevated after implantation of the chamber, allowing visualization of the microcirculation in the island flap. The island flap was exposed to global ischemia by clamping the pedicles, and the clamps were then released to allow reperfusion. Various microcirculatory responses induced by ischemia-reperfusion were visualized. This model accurately simulated the clinical situation in reconstructive surgery and successfully realized chronic visualization of the flap microcirculation in vivo.
To investigate whether hyperthermic preconditioning can actually protect skin flaps against ischemia/reperfusion injury, the authors first developed a new skin-flap model in 15 mice, a dorsal bipedicle island skin-flap model. Then, another 75 mice were separated into five groups. Mice in Groups 1 to 4 received the same hyperthermic preconditioning, but had different recovery times of 6 hr, 24 hr, 48 hr, and 72 hr, respectively. Mice in Group 5 served as control. Island skin flaps were elevated in all groups, and then were subjected to 8 hr of ischemia and subsequent reperfusion. Flap survival was statistically significantly higher than in controls in animals in Groups 1 and 3, with recovery times of 6 hr and 48 hr, respectively. Mice in Groups 2 and 4 had recovery times of 24 hr and 72 hr, respectively. Hyperthermic preconditioning could thus protect skin flaps against ischemia/reperfusion injury, and there were two optimal periods for such a protective effect.
Mice are popular animals for biomedical studies, but few skin flap models have been reported in them. To investigate the ischaemia/reperfusion phenomenon in skin flaps, we first investigated the vascular anatomy of murine dorsal skin and then designed a suitable murine dorsal skin flap model. In 120 mice, six distinct vascular patterns were identified, one being seen in 111 mice (93%). Based on this finding, in Part 2 of the study, 15 mice had flaps (4 x 4 cm) raised based on the two caudal vascular pedicles of the left and right deep circumflex iliac vessels as a bipedicled flap in which the mean (SD) survival was 96 (5)%. In a further 10 mice, flaps were raised based on a single pedicle, the left deep circumflex iliac vessel, as a monopedicled flap, in which the mean (SD) survival was 71 (12)%. The bipedicled flap model was then used to study ischaemia/reperfusion injury. Twenty flaps were subjected to eight hours of ischaemia and subsequent reperfusion, and their mean (SD) survival was 43 (26)%. Histological assessments were also carried out using neutrophil and leucocyte counts, and significant differences between groups were observed.
The electric control of cellular functions via Ca2+ was formerly suggested. From this viewpoint, the involvement of a Ca2+ channel was studied using bovine fetal arterial endothelial (BFAE) cells in which P2X4, an ATP-operated and fluid shear stress sensitive Ca2+ channel, exists predominantly. An electric stimulus (sine wave, 10 Hz, 10 VPP, 30 s) caused a marked influx of Ca2+ into BFAE cells from an extracellular solution. The magnitude of the [Ca2+]i change increased with a decrease in the frequency in the range from 100 Hz to 5 Hz. Regarding the pathway of this Ca2+ influx, single-cell imaging and an ATP depletion experiment strongly suggested the involvement of a pathway different from P2X4. This pathway was thought to be a non-specific one, because typical Ca2+ channel blockers, such as verapamil, Gd3+, and Co2+, could not inhibit the Ca2+ influx.