Arthur B. Ritter

Microvascular transport and endothelial cell alterations preceding skeletal muscle damage in ischemia and reperfusion injury

We determined the leakage of macromolecules using FITC-dextran-150 as a tracer and measured the extent of no-reflow phenomenon by video field analysis. The cremaster muscle of anesthetized rats was fashioned as a single layer, splayed on a lucite chamber and suffused with bicarbonate solution at 35 degrees C. After a 1 hour period of baseline data collection, ischemia was produced by cross-clamping the cremasteric vascular pedicle for periods of 30 minutes and 2 hours in separate experiments. Macromolecular leakage was visualized after reinstitution of perfusion. Leakage occurred at postcapillary venules 15 to 50 micron in diameter and quickly spread to the interstitium. The magnitude of leakage decreased as a function of time with continuous buffer suffusion, but remained higher than in the control period. No reflow occurred in approximately 30 percent of the muscle microvasculature upon reperfusion. The no-reflow values at 30 minute and 2 hour periods of ischemia were significantly different from the control values but were not from each other. Electron micrographs demonstrated endothelial cell swelling and migration of leukocytes and normal myocytes after 1 hour of reperfusion following 2 hours of ischemia. Our results demonstrate that permeability changes, occurrence of no reflow, and leukocyte migration precede the onset of damage to skeletal muscle in ischemia and reperfusion injury.

Analysis of microvascular permeability to macromolecules by video-image digital processing

The dynamics of macromolecular transport across microvascular walls were studied in the hamster cheek pouch by intravital fluorescence microscopy. A graded series of fluorescein isothiocyanate-labeled dextrans (FITC-Dx) of 20,000-70,000 MW was used as macromolecular tracers. The time-dependent extravasation of FITC-Dxs was videotaped for about 2.5 hr to allow for tracer equilibration in the interstitial space. Permeation of macromolecules from individual microvessels was quantified by digital video-image processing. Histograms of the light intensity distributions for selected fields at various times were measured and used to construct integral optical density-time profiles of the extravasated fluorochromes for particular leaky sites. A nonlinear regression algorithm was employed to determine the effective microvascular permeability (P) for the macromolecules studied using a one-dimensional two-compartmental diffusion model and a step change in macromolecular concentration at the boundary. The calculated Ps (X 10(-8) cm/sec) were 47.8 +/- 8.7 for FITC-Dx 20; 31.7 +/- 5.9 for FITC-Dx 40, and 17.5 +/- 4.1 for FITC-Dx 70. Our values are comparable to those obtained by whole organ techniques. The observed differences can be explained by explicit consideration of interstitial resistance in the calculations.

RESIDUAL STRESSES IN OSCILLATING THORACIC ARTERIES REDUCE CIRCUMFERENTIAL STRESSES AND STRESS GRADIENTS

The purpose of this paper is to examine the effects of residual stresses and strains in the oscillating arteries on the stress distribution in the vascular wall. We employ a static theory of large elastic deformations for orthotropic material (Chuong and Fung, 1986, J. biomech. Engng 108, 189-192) with the acceleration term added to make the theory dynamic. We use the static elastic parameters of residual stresses in our analysis because the dynamic parameters are not available in the literature. Our analysis reveals that the effect of considering the residual stresses is to decrease the very large circumferential stresses at the inner wall by 62% and reduces the stress gradient through the arterial wall by 94% compared to the case when residual stresses are ignored. Thus, because the arteries do contain residual stresses, the consequent lower stresses at the inner wall and the reduced stress gradient may reduce the progression of atheroma. Our computations show that the stress gradients do not depend on the heart rate.

Effect of platelet-activating factor on leukocyte adhesion to microvascular endothelium. Time course and dose-response relationships

The hamster cheek pouch microcirculation was used to investigate the effects of platelet-activating factor (PAF) on leukocyte adhesion to microvascular walls by means of intravital microscopy. PAF was applied topically at concentrations ranging from 10−11 to 10−5 M. An inverse relationship between PAF concentration and number of adhering white cells per 100-Μm length was found in venules ranging in diameter from 10 to 60 Μm (grouped into 10-Μm intervals). Importantly, the PAF-induced adhesion of leukocytes lasted for the 3-h experimental period. We postulate that induction of leukocyte adhesion to venular endothelium is an important role of PAF in inflammatory processes.

Assessment of ischemia reperfusion injury in skeletal muscle by macromolecular clearance.

Qualitative changes in skeletal muscle injury after ischemia are well known; however, quantitative assessments have not been well documented. We have determined microvascular permeability changes by measuring the clearance of fluorescein-labeled dextran of MW 150,000 (FITC-Dextran-150). The cremaster muscle of anesthetized rats was fashioned as a single layer, splayed on a lucite chamber and suffused with bicarbonate buffer solution at 35 degrees C. Clearance is the product of suffusion rate times the ratio of suffusate to plasma concentrations of FITC-Dx 150. After a 1-hr period of baseline data collection, ischemia was produced by cross-clamping the cremasteric vascular pedicle for periods of 30 min and 2 hr in separate experiments. Clearance of FITC-Dx 150 increased from a control value (mean +/- SE) of 8.3 +/- 2.7 to 29.9 +/- 8.1 microliters/min/g after reperfusion following a 30-min period of ischemia, and from a control value of 36.2 +/- 13.6 to 274 +/- 94.5 after 2 hr of ischemia. The differences were statistically significant (P less than 0.05). Our results show a significant increase in microvascular permeability occurring after only 30 min of ischemia. They also demonstrate a direct relationship between the extent of the permeability change and the duration of the ischemic period.

Biomedical engineering principles

Introduction: Modeling of Physiological Processes Cell Physiology and Transport Principles and Biomedical Applications of Hemodynamics A Systems Approach to Physiology The Cardiovascular System Biomedical Signal Processing Signal Acquisition and Processing Techniques for Physiological Signal Processing Examples of Physiological Signal Processing Principles of Biomechanics Practical Applications of Biomechanics Biomaterials Principles of Biomedical Capstone Design Unmet Clinical Needs Entrepreneurship: Reasons why Most Good Designs Never Get to Market An Engineering Solution in Search of a Biomedical Problem to Solve Tissue Engineering Future Trends in BME

Optimal patterns for suturing wounds

A mathematical model for computing stresses in sutured human skin wounds is presented. The model uses the incremental law of elasticity and elastic constants valid for in vivo orthotropic skin. The model is applied to compute the principal stress and displacements resulting from suturing small elliptical and circular wounds in a large flat sheet of skin, in order to determine the optimal suturing patterns. It is observed that the average stress index for a circular wound sutured toward the center is almost double that of a wound sutured transverse to the diameter. Thus, the latter type of suturing pattern is preferable. Similarly, suturing an elliptical wound transversely produces a lower average stress index than a circular wound of the same area. It is also found that the optimal ratio of semi-major to semi-minor axis of an elliptical wound is near 3 (for abdominal wounds), i.e., this ratio produces the most uniform stresses along the wound edges, where wound healing is slowest. Since high stresses have adverse effects on healing and blood flow, this work, depicting regions of high stresses, may be used along with other biological factors to help predict regions of slower healing in sutured wounds.

Reproducibility of microvascular permeability responses to successive topical applications of bradykinin in the hamster cheek pouch

Abstract Bradykinin increases leakage of macromolecules at postcapillary venules. Whether the same sites respond to repeated applications of bradykinin is not known. Knowledge of this phenomenon is of importance in ascertaining the mechanisms of vascular wall repair in inflammatory processes. We used the hamster cheek pouch preparation, suffused with buffer bicarbonate (pH 7.4). Fluorescein isothiocyanate-dextran 150 injected iv was used as a tracer of macromolecular microvascular transport. An area of cheek pouch representing 0.8 cm 2 was scanned using epi- and transillumination light microscopy before, during and after bradykinin application. Video recordings of individual fields representing 1 mm 2 were examined to determine the number of leakage sites present as well as their position in the field. Usually, 20–40 min of suffusion with buffer was sufficient to wash away the effects of any given dose of bradykinin. For fields observed, 85% of the leakage sites induced by the first dose were the same sites induced by the second application of the same dose of bradykinin. Whether or not the same site responds to a subsequent application may depend upon the type of microvascular wall repair which ensues. We propose that (a) relaxation of bradykinin-contracted endothelial cells is a mechanism of microvascular wall repair consistent with those leakage sites which repeatedly respond to bradykinin stimulation, and (b) platelet plugging of interendothelial gaps may have occurred in those leakage sites refractory to subsequent applications of bradykinin.

Measurement of skin stretch via light reflection

A noninvasive technique for measuring the stretch of skin is described. The technique utilizes changes in the reflectivity of polarized light intensity as a monitor of skin stretch. Measurements of in vitro pigskin and in vivo human skin show that the reflectivity of polarized light intensity increases linearly with stretch. The changes in diffusive reflectivity properties of skin result from the alterations that take place in the roughness across the thickness of the skin layers due to stretch. Conceptually, as the roughness of a layer decreases with stretch, a smoother reflecting media is produced, resulting in a proportional increase in the specular reflection. Results can be easily extended to a real-time stretch analysis of large tissue areas that would be applicable for mapping the stretch of skin.

STRESSES AND STRAINS IN THE PASSIVE LEFT VENTRICLE

In this paper, we estimate the stresses and strains from the equatorial region down to the apex of the heart by modeling the passive left ventricle as a frustrum of a thick hollow cone. Large deformation theory has been employed in this analysis. Furthermore, the effects of residual stresses and the anisotropy due to muscle fiber orientation have been included. It is observed that circumferential stress, which is the most important physiologically, decreases considerably at the endocardium and is more evenly distributed through the wall when residual stresses are taken into account. The stresses also decrease as we go from the equatorial region to the apex. Because heart muscles physically have residual stresses, the consequent lower stress gradient through the wall enhances the diastolic function of the left ventricle.