Stalled cerebral capillary blood flow in mouse models of essential thrombocythemia and polycythemia vera revealed by in vivo two-photon imaging.

BACKGROUND: Essential thrombocythemia (ET) and polycythemia vera (PV) are myeloproliferative neoplasms (MPNs) that share the JAK2(V617F) mutation in hematopoietic stem cells, leading to excessive production of predominantly platelets in ET, and predominantly red blood cells (RBCs) in PV. The major cause of morbidity and mortality in PV and ET is thrombosis, including cerebrovascular occlusive disease. OBJECTIVES: To identify the effect of excessive blood cells on cerebral microcirculation in ET and PV. METHODS: We used two-photon excited fluorescence microscopy to examine cerebral blood flow in transgenic mouse models that mimic MPNs. RESULTS AND CONCLUSIONS: We found that flow was 'stalled' in an elevated fraction of brain capillaries in ET (18%), PV (27%), mixed MPN (14%) and secondary (non-MPN) erythrocytosis (27%) mice, as compared with controls (3%). The fraction of capillaries with stalled flow increased when the hematocrit value exceeded 55% in PV mice, and the majority of stalled vessels contained only stationary RBCs. In contrast, the majority of stalls in ET mice were caused by platelet aggregates. Stalls had a median persistence time of 0.5 and 1 h in ET and PV mice, respectively. Our findings shed new light on potential mechanisms of neurological problems in patients with MPNs.

Novel microcatheters for selective intra-arterial injection of fluid in the rat brain.

BACKGROUND AND PURPOSE: The internal carotid artery (ICA) in the rat has a single extracranial branch, which supplies the muscles of mastication. The rat ICA also has multiple intracranial branches including (from proximal to distal): multiple small perforating arteries which supply the hypothalamus and the anterior choroidal artery which supplies the choroid plexus and part of the basal ganglia. At the ICA terminus, the vessel bifurcates into the anterior and middle cerebral arteries. The purpose of this study was to demonstrate selective injection of ICA branches in the rat. MATERIALS AND METHODS: Microcatheters (mucath1 and mucath2) were fabricated by plugging the tip of 169-mum outer diameter polyimide tubing and perforating the sidewalls. A 450-mum polydimethyl-siloxane cylinder was affixed to the distal tip of mucath2 but not mucath1. We evaluated the territory of mucath1 injection ex vivo using magnetization-prepared rapid acquisition of gradient echo MR imaging of brain specimens injected at necropsy. Territories of mucath1 and mucath2 injection were evaluated in vivo with dynamic susceptibility-weighted contrast-enhanced MR imaging. The territory of mucath2 also was evaluated in vivo with fused static microPET/T1 MR images performed after [(18)F] fluorodeoxyglucose ((18)FDG) injection. We evaluated additional catheterized and injected animals at 48 hours using physical examination, T2 MR images, and postmortem brain histologic specimens. RESULTS: Gadolinium-diethylene-triamine pentaacetic acid (Gd-DTPA) and (18)FDG injected through mucath1 selectively opacified the ipsilateral cerebral hemisphere, with no contralateral opacification. Gd-DTPA injected through mucath2 selectively opacified the territories of the hypothalamic perforating arteries, and anterior choroidal artery. There was no iatrogenic complication 48 hours after 20- to 25-minute injections performed with mucath1 or mucath2. CONCLUSIONS: We have developed 2 microcatheters which can be placed in the ICA for selective injection of its branches. One microcatheter selectively injects the ipsilateral cerebral hemisphere. The other selectively injects only the hypothalamus and lateral thalamus.

Fabrication and characterization of microfluidic probes for convection enhanced drug delivery.

Convection enhanced drug delivery (CED) is a promising therapeutic method for treating diseases of the brain by enhancing the penetration of drugs. Most controlled release delivery methods rely on diffusion from a source to transport drugs throughout tissue. CED relies on direct infusion of drugs into tissue at a sufficiently high rate so that convective transport of drug is at least as important as diffusive transport. This work describes the fabrication and characterization of microfluidic probes for CED protocols and the role diffusion plays in determining penetration. Microfluidic channels were formed on silicon substrates by employing a sacrificial photoresist layer encased in a parylene structural layer. Flow in the microchannels was characterized by applying constant upstream pressures from 35 to 310 kPa, which resulted in flow rates of 0.5-4.5 microL/min. The devices were used to infuse Evans Blue and albumin in hydrogel brain phantoms. The results of these infusions were compared to a simple convection-diffusion model for infusions into porous media. In vivo infusions of albumin were performed in the gray matter of rats at upstream pressures of 35, 70, and 140 kPa. The microfabricated probes show reduced evidence of backflow along the device-tissue interface when compared with conventional needles used for CED.

Effects of particle concentration on the partitioning of suspensions at small divergent bifurcations.

Experimental results are reported for the low Reynolds number flow of a suspension of spherical particles through a divergent capillary bifurcation consisting of a straight tube of circular cross-section that splits to form two tubes of equal diameter. The partitioning of particles between the downstream branches of the bifurcation is measured as a fraction of the partitioning of total volume (particles + suspending fluid) between the branches. Two bifurcation geometries are examined: a symmetric Y-shaped bifurcation and a nonsymmetric T-shaped bifurcation. This experiment focuses on the role of hydrodynamic interactions between particles on the partitioning of particles at the bifurcation. The particle diameter, made dimensionless with respect to the diameter of the branch tubes, ranges from 0.4 to 0.8. Results show that hydrodynamic interactions among the particles are significant at the bifurcation, even for conditions where interactions are unimportant in the straight branches away from the bifurcation. As a result of hydrodynamic interactions among particles at the bifurcation, the partitioning of particles between the branches is affected for particle volume fractions as small as 2 percent. The experimental results show that the effect of particle volume fraction is to diminish the inhomogeneity of particle partitioning at the bifurcation. However, the magnitude of this effect depends strongly on the overall shape of the bifurcation geometry, and, in particular on the angles between the branches.

The motion of model cells at capillary bifurcations.

The motion of a rigid particle suspended in a two-dimensional flow through a bifurcating channel is calculated numerically. The calculated particle trajectories depend on the partitioning of total volume at the bifurcation, the size of the particle relative to channel dimensions, and the upstream lateral position of the particle. Results are used to deduce the flux of particles in a dilute suspension entering each branch of a bifurcation as a function of the total volumetric flow entering each branch. The calculated results exhibit several qualitative features observed in experimental data for red cell flux at capillary bifurcations.

Unsteady cell distributions in capillary networks.

A numerical simulation is used to study the distribution and flux of red blood cells in capillary networks. The relationship between the motion of an individual cell in a single capillary and the overall transport of cells throughout a network is investigated.Interpretation of results is greatly facilitated by the use of a computer graphics representation of the simulation. Steady-state and certain time-dependent cell distribution problems are studied, with a special focus on the sensitivity of the results to the presence of a small number of white cells.