Isolating the influences of fluid dynamics on selectin-mediated particle rolling at venular junctional regions.

The objective of this study was to isolate the impact of hydrodynamics on selectin-mediated cell rolling in branched microvessels. Significant advancements have been made in furthering the understanding of complex interactions between biochemical and physical factors in the inflammatory cascade in simplified planar geometries. However, few studies have sought to quantify the effects of branched configurations and to isolate the effects of associated fluid forces. Experimental techniques were developed to perform in vitro adhesion experiments in Y-shaped micro-slides. The micro-slides were coated with P-selectin and microspheres coated with Sialyl-Lewisx were observed as they rolled in the chambers at different wall shear stresses. Study results revealed that microsphere rolling velocities and rolling flux were lowest in regions closest to the apex of a junctional region and were dependent on both branch angle and wall shear stress. The regions closest to the junctional region were shown to have low bulk flow velocities and shear stresses using computational fluid dynamics (CFD) modeling. Collectively, the study demonstrates that despite the presence of a uniform coating of P-selectin, hydrodynamic factors associated with the chamber geometry yield non-uniform effects on particle behavior. These findings could explain why cells have been observed to preferentially adhere or transmigrate near junctional regions. Future characterization of inflammatory processes in microvascular network configurations is therefore crucial for furthering our fundamental understanding of inflammation.

Reverse engineering life: physical and chemical mimetics for controlled stem cell differentiation into cardiomyocytes.

Our ability to manipulate stem cells in order to induce differentiation along a desired developmental pathway has improved immeasurably in recent years. That is in part because we have a better understanding of the intracellular and extracellular signals that regulate differentiation. However, there has also been a realization that stem cell differentiation is not regulated only by chemical signals but also by the physical milieu in which a particular stem cell exists. In this regard we are challenged to mimic both chemical and physical environments. Herein we describe a method to induce stem cell differentiation into cardiomyocytes using a combination of chemical and physical cues. This method can be applied to produce differentiated cells for research and potentially for cell-based therapy of cardiomyopathies.

A separate role for ICAM-1 and fluid shear in regulating leukocyte interactions with straight regions of venular wall and venular convergences.

OBJECTIVE: Variation in expression of adhesion molecules plays a key role in regulating leukocyte behavior, but the contribution of fluid shear to these interactions cannot be ignored. Here, we dissected the effects of each of these factors on leukocyte behavior in different venular regions. MATERIALS AND METHODS: Leukocyte behavior was quantified in blood-perfused microvascular networks in anesthetized mouse cremaster muscle, using intravital confocal microscopy. ICAM-1 expression and fluid shear rate were quantified by using ICAM-1 fluorescent labeling, fluorescent particle tracking, and computational fluid dynamics. RESULTS: Tumor necrosis factor alpha induced an increase in ICAM-1 expression and abolished the differences observed among control venules of different sizes. Consequently, leukocyte adhesion was increased to a similar level across all vessel sizes [5.1+/-0.46 leukocytes/100 microm vs. 2.1+/-0.47 (control)], but remained significantly higher in venular convergences (7.8+/-0.4). Leukocyte transmigration occurred primarily in the smallest venules and venular convergences (23.9+/-5.1 and 31.9+/-2.7 leukocytes/10,000 microm(2) tissue, respectively). In venular convergences, the two inlet vessels are predicted to create a region of low velocity, increasing leukocyte adhesion probability. CONCLUSIONS: In straight regions of different-sized venules, the variability in ICAM-1 expression accounts for the differences in leukocyte behavior; in converging regions, fluid shear potentially has a greater effect on leukocyte endothelial cell interactions.

The distribution of rolling neutrophils in venular convergences.

The aim of this study was to characterize the distribution of adherent leukocytes in branched venular convergences in vivo. Intravital microscopy was used to obtain video images of leukocyte adhesion in multiple branched sites in mouse cremaster muscle, during the mild inflammatory response induced by surgical preparation. The average number of cells/vessel length was obtained over several minutes for seven venular convergences with varying geometrical configurations. Results from this study demonstrate a strong tendency of leukocytes to adhere at junctional points between converging vessels. Different vessel configurations were studied and results were shown to be insensitive to precise vessel geometry. Thus, in post-capillary venules, leukocytes are most likely to adhere at points between converging vessels, regardless of the precise geometrical properties or configuration of the vessels. Hydrodynamic mechanisms due to flow behavior through convergences likely play a significant role in determining locations of cellular adhesion. Future work should concentrate on quantifying the relative contributions of hydrodynamic and biochemical mechanisms to aid in understanding disease processes and development of treatments or therapeutics.

Impact of the Fåhraeus effect on NO and O2 biotransport: a computer model.

Nitric oxide (NO) and oxygen (O2) transport in the microcirculation are coupled in a complex manner, since enzymatic production of NO depends on O2 availability, NO modulates vascular tone and O2 delivery, and tissue O2 consumption is reversibly inhibited by NO. The authors investigated whether NO bioavailability is influenced by the well-known Fåhraeus effect, which has been observed for over 70 years. This phenomenon occurs in small-diameter blood vessels, where the tube hematocrit is reduced below systemic hematocrit as a plasma boundary layer forms near the vascular wall when flowing red blood cells (rbcs) migrate toward the center of the bloodstream. Since hemoglobin in the bloodstream is thought to be the primary scavenger of NO in vivo, this might have a significant impact on NO transport. To investigate this possibility, the authors developed a multilayered mathematical model for mass transport in arterioles using finite element numerical methods to simulate coupled NO and O2 transport in the blood vessel lumen, plasma layer, endothelium, vascular wall, and surrounding tissue. The Fåhraeus effect was modeled by varying plasma layer thickness while increasing core hematocrit based on conservation of mass. Key findings from this study are that (1) despite an increase in the NO scavenging rate in the core with higher hematocrit, the model predicts enhanced vascular wall and tissue NO bioavailability due to the relatively greater resistance for NO diffusion through the plasma layer; (2) increasing the plasma layer thickness also increases the resistance for O2 diffusion, causing a larger P(O2) gradient near the vascular wall and decreasing tissue O2 availability, although this can be partially offset with inhibition of O2 consumption by higher tissue NO levels; (3) the Fåhraeus effect can become very significant in smaller blood vessels (diameters <30 microm); and (4) models that ignore the Fåhraeus effect may underestimate NO concentrations in blood and tissue.

Interactions between NO and O2 in the microcirculation: a mathematical analysis.

Biotransport of nitric oxide (NO) and of oxygen (O(2)) in the microcirculation are inherently interdependent, since all nitric oxide synthase (NOS) isoforms (eNOS, nNOS, and iNOS) require O(2) to produce NO. Furthermore, tissue O(2) consumption is reversibly inhibited by NO. To investigate these complex interactions, a mathematical model was developed for coupled mass transport of NO and O(2) around a cylindrical arteriole using finite element computational methods. Steady-state radial NO and O(2) gradients in the bloodstream, plasma layer, endothelium, vascular wall, and surrounding tissue were simulated for different conditions. Special cases of the model were solved, including O(2)-dependent NO production from eNOS alone, and with additional NO production from either nNOS or iNOS at specified locations. The model predicts that (a) concentration changes in one species can have significant effects on transport of the other species with the degree of interaction dependent on spatial gradients; (b) eNOS NO production rates required to maintain the concentration of NO in the vascular wall are more dependent on NO scavenging in blood than in tissue; (c) relatively low rates of NO production in tissue from either nNOS or iNOS can elevate vascular wall NO, compensating for possible reductions in NO production from eNOS; (d) depending on their physical location, nNOS and iNOS can be very sensitive to O(2); and (e) increased tissue NO can increase O(2) delivery to more distal regions by inhibiting O(2) consumption in other regions.