Increase In Number Of Capillaries Research Articles

Regional changes in the masseter muscle of rats after reduction of blood supply

The masticatory musculature is an integral functional part of the stomatognathic system and influences craniofacial morphogenesis and morphology. This animal study aimed to investigate the morphological consequences of restricted regional blood supply to the m. masseter. A total of 20 adult male Wistar rats (Rattus norvegicus Berkenhout) were divided into an experimental group and a control group comprised of 10 animals respectively and kept under standardized conditions. The experimental group underwent a dextrolateral complete surgical ligation of the a. carotis communis and, after 5 weeks, specimens were taken from the masseters. The muscle samples were analyzed immunohistochemically for fiber distribution and capillary density. Analysis revealed a discrete increase in the proportion of type I fibers with a significant increase of capillary number per area. Although no agreement exists on the alterations occurring in chronically ischemic muscles, it may be assumed that chronic ischemia evokes histomorphological adaptation processes similar to endurance training effects.

Nitric oxide and p38 MAP kinase mediate shear stress‐dependent inhibition of MMP‐2 production in microvascular endothelial cells

Chronic exposure of the skeletal muscle microcirculation to elevated shear stress-induces angiogenesis. Previous studies observed that shear stress-induced capillary growth involves luminal sprouting, or internal division, of the capillaries, which is characterized by a minimal proliferative response and the retention of an intact basement membrane. Matrix metalloproteinases (MMPs) are associated with the process of abluminal sprouting angiogenesis, but may not be required for the process of luminal division during capillary growth. We analyzed the production of MMP-2, using both the in vivo model of prazosin-induced angiogenesis in rat skeletal muscle, and cultured microvascular endothelial cells exposed to laminar shear stress. We found that MMP-2 was not elevated in capillaries of shear stress-stimulated skeletal muscle, despite a significant increase in capillary number in response to a shear stress stimulus. In cultured microvascular endothelial cells, MMP-2 mRNA and protein levels were attenuated significantly in response to shear stress exposure. This effect on MMP-2 was reversed by nitric oxide (NO) synthase inhibition using LNNA. In contrast, exposure of static cultures of endothelial cells to NO donors significantly reduced MMP-2 production. Shear stress exposure and NO donors both modified phosphorylation levels of several members of the MAPK family. Treatment of shear stress-exposed cells with the p38 MAPK inhibitor, SB203580, abolished the shear stress-mediated reduction in MMP-2 mRNA. Thus, our data provide strong evidence that elevated shear stress inhibits MMP-2 production in microvascular endothelial cells, an effect that is mediated by signal pathways involving both production of NO and activation of p38 MAPK.

Erythroid cells play essential roles in angiogenesis by bone marrow cell implantation

Bone marrow cell implantation (BMI) has been utilized to treat patients with limb and heart ischemia. BMI provides angiogenic precursors and angiogenic cytokine-producing cells, especially erythroid cells. In this study, we induced in vitro angiogenesis cultures and in vivo BMI simulation using a murine limb ischemia model to examine the role of erythroid cells and the effect of erythropoietin (EPO). Human erythroid colonies (BFU-e) induced capillary networks around the colonies in vitro. Erythroid cells in human bone marrow produced vascular endothelial growth factor and placental growth factor. The angiogenic effects of erythroid cells were further amplified in the presence of EPO. Limb-ischemic mice were treated with BMI +/- EPO, and limb survival, blood flow recovery, and muscle histology were analyzed. Treatment with whole bone marrow cells + EPO significantly improved limb survival and blood flow. The cumulative effects of EPO on BMI induced and increase in capillary number and artery enlargement. Erythroid cells were essential for the in vivo effects of BMI, and CD14-positive cells supported the biological effects. In addition to the direct effect of EPO on angiogenesis, EPO showed indirect effect on angiogenesis through amplifying the angiogenic effects by erythroid cells supported by CD14-positive cells.

Sample Pulse Broadening in Taylor Flow Microchannels for Screening Applications

AbstractThe effect of different parameters on the residence time distribution of a Taylor flow microreactor was investigated. The reactor residence time distribution was calculated from the tracer residence time distribution of a single unit cell. It was found that generally an increase in Bodenstein number, capillary number or slug length increased the spread of the residence times. A case study comparing the effect of two different sets of design and operating parameters on the distribution spread showed that spreading can be minimized by careful choice of parameters, in particular slug length.

Platelet-Derived Microparticles Induce Angiogenesis and Stimulate Post-Ischemic Revascularization.

Platelet attachment to the subcellular matrix at injured sites of the vasculature is followed by their activation and release of microparticles. Platelet-derived microparticles (PMP) have been shown to be involved in the regulation of hemostasis. However, little is known about the role of PMP in the regulation of angiogenesis and related clinical conditions. We have recently demonstrated that platelets as a cellular system induce angiogenic responses both in vitro and in vivo. In the present study, we investigated the potential role of PMP in angiogenesis. A strong dose-dependent pro-angiogenic effect of PMP in the rat aortic ring model (5.3±2.1 mm2 surface covered with sprouting vessels versus 0.24±0.2 mm2 in the control, p<0.001) was observed. This effect was reversed by selective inhibition of VEGF, bFGF and PDGF (surface covered with vessels 0.7±0.5 mm2, 1.7±1.5 mm2, and 2.4±1.2 mm2, respectively, p<0.02 versus control), but not by inhibition of heparanase (5.1±0.8 mm2, p>0.5 versus control). PMP exert their stimulatory effect via PI3-kinase, Src kinase and ERK, whereas protein kinase C seems not to be involved, as judged by the aortic ring sprouting model. Using confocal and electron microscopy, we also demonstrate that PMP bind to non-activated endothelial cells. In addition, PMP markedly increased invasion of human endothelial cells through a layer of matrigel. This effect was abolished by an inhibitor of VEGF receptor tyrosine phosphorylation or laminaran sulfate (heparanase inhibitor). It was also partially reduced by PDGF blocking mAb, whereas blocking of bFGF had no effect. Furthermore, we have demonstrated that PMP induce angiogenesis in an in vivo model, in which beads (30 μl) of 4% agarose gel containing the substances under study were transplanted subcutaneously into mice. Image analysis of the capillary area revealed the following: control beads − 0.2±0.05 mm2, VEGF + bFGF containing beads − 4.8±1.1 mm2, PMP (100 μg/ml) containing beads − 5.1±1.3 mm2, p<0.001 versus control. The latter finding was further supported by immunohistochemical staining of the skin in the vicinity of the beads for von Willebrand factor, a marker of endothelial cells (control − 4.0±3.2, VEGF+bFGF − 12±4.4, PMP − 17±6.5 capillaries per view field, p<0.05 versus control). Finally, we explored the potential effect of PMP in a rat myocardial infarction model. Ischemia was induced by LAD ligation followed by injection of either PMP or PBS into the ischemic region. Preliminary evaluation of the LAD myocardial territory in sham-operated animals revealed 157±42.0 capillaries per view field. In contrast, number of capillaries observed 3 weeks after induction of ischemia was reduced to 34±21.5. When PMP were injected into the ischemic region, there was an increase in capillary number up to 97±27.3. In conclusion, PMP induce angiogenesis in both in vitro and in vivo models. Local injection of PMP into the ischemic myocardium may improve revascularization.

Angiogenesis induced by endothelial nitric oxide synthase gene through vascular endothelial growth factor expression in a rat hindlimb ischemia model.

Because the mechanism of the angiogenic property of nitric oxide (NO) was not fully understood in vivo, we focused on the role of vascular endothelial growth factor (VEGF) in angiogenesis induced by endothelial NO synthase (eNOS) gene transfer. After intramuscular injection of eNOS DNA into a rat ischemic hindlimb, transfection of eNOS vector resulted in a significant increase in eNOS protein 1 week after transfection. In addition, tissue concentrations of nitrite and nitrate were significantly increased in rats transfected with the eNOS gene up to 2 weeks after transfection. The increase in tissue nitrite and nitrate concentrations was completely inhibited by NG-nitro-L-arginine methyl ester (L-NAME). In contrast, serum concentrations of nitrite and nitrate and blood pressure were not changed by eNOS gene transfer. Importantly, overexpression of the eNOS gene resulted in a significant increase in peripheral blood flow, whereas L-NAME inhibited the increase in blood flow. Interestingly, basal blood flow was significantly lower in rats treated with L-NAME than in control rats. A significant increase in capillary number was consistently detected in rats transfected with the eNOS gene at 4 weeks after transfection, accompanied by a significant increase in VEGF. Moreover, administration of neutralizing anti-VEGF antibody abolished the increase in blood flow and capillary density induced by eNOS plasmid injection. Overall, intramuscular injection of bovine eNOS plasmid induced therapeutic angiogenesis in a rat ischemic hindlimb model, a potential therapy for peripheral arterial disease. The stimulation of angiogenesis by NO might be due to upregulation of local VEGF expression.

Nonequilibrium capillary pressure and relative permeability curves of porous media

AbstractHistory matching of unsteady immiscible displacement experiments performed on model porous media reveals that nonequilibrium capillary pressure and relative permeability functions are sensitive to the dominating flow pattern. As the capillary number increases, the network displacement pattern changes gradually from invasion percolation cluster to stable displacement. For primary drainage, the capillary pressure and relative permeability curves are increasing functions of the capillary number. The dependence of the relevant macroscopic parameters on the capillary number is consistent with power laws derived from scaling arguments of gradient percolation theory, which describes the growth of the displacement front. The material coefficient of the thermodynamic theory of capillary pressure changes mildly with fluid saturation and is a decreasing function of the capillary number.

Pore-level modeling of drainage: crossover from invasion percolation fingering to compact flow.

A pore-level model of drainage, which has been quantitatively validated, is used to study the effect of increased injection rate (i.e., increased capillary number) upon the flow, with matched-viscosity fluids. For small enough capillary number, the flows from the model correctly reproduce the flows from the invasion percolation with trapping (IPWT) model. As the capillary number is increased, the early-time flows mimic those of the IPWT-model, but then deviate towards compact flow at a characteristic time that decreases as the capillary number increases. That is, the larger the capillary number, the sooner the flow crosses over from IPWT flows towards compact (linear) flows.

Hydrodynamic Interaction between Spheres Coated with Deformable Thin Liquid Films

In this article, we considered the hydrodynamic interaction between two unequal spheres coated with thin deformable liquids in the asymptotic lubrication regime. This problem is a prototype model for drop coalescence through the so-called “film drainage” mechanism, in which the hydrodynamic contribution comes dominantly from the lubrication region apart from the van der Waals interaction force. First, a general formulation was derived for two unequal coated spheres that experienced a head-to-head collision at a very close proximity. The resulting set of the evolution equations for the deforming film shapes and stress distributions was solved numerically. The film shapes and hydrodynamic interaction forces were determined as functions of the separation distance, film thickness, viscosity ratios, and capillary numbers. The results show that as the two spheres approach each other, the films begin to flatten and eventually to form negative curvature (or a broad dimple) at their forehead areas in which high lubrication pressure is formed. The dimple formation occurs earlier as the capillary number increases. For large capillary numbers, the film liquids are drained out from their forehead areas and the coated liquid films rupture before the two films “touch” each other. Meanwhile, for small capillary numbers, the gap liquid is drained out first and the two liquid films eventually coalesce.

Effect of compatibilization on the breakup of polymeric drops in shear flow

A block copolymer may be added as a compatibilizer during polymer processing in order to promote intimate mixing of thermodynamically immiscible homopolymers. The action of this compatibilizer can only partially be attributed to its effect on the interfacial tension between the immiscible homopolymers. Here the additional contributions of the compatibilizer are directly probed by measuring the capillary number during coalescence experiments. Model blends consisting of polyisobutylene (PIB) and polydimethylsiloxane (PDMS), compatibilized with various amounts of a PIB–PDMS diblock copolymer, are used for this purpose. The mean capillary number of the droplets is determined from the mechanical frequency response of the blends. With increasing amounts of compatibilizer, a systematic increase in steady shear capillary number is seen, to values well above the critical capillary number for droplet breakup of uncompatibilized systems. This indicates that a simple decrease in interfacial tension is not the only effect of adding the compatibilizer to these immiscible blends. Past simulations suggest that these results are associated with gradients in interfacial tension (Marangoni stresses) induced by the gradients of compatibilizer concentration due to flow. Direct evidence of the presence of such interfacial tension gradients along the surface of compatibilized drops was obtained by optical microscopy.

Exercise adaptation attenuates VEGF gene expression in human skeletal muscle.

Angiogenesis is a component of the multifactoral adaptation to exercise training, and vascular endothelial growth factor (VEGF) is involved in extracellular matrix changes and endothelial cell proliferation. However, there is limited evidence supporting the role of VEGF in the exercise training response. Thus we studied mRNA levels of VEGF, using quantitative Northern analysis, in untrained and trained human skeletal muscle at rest and after a single bout of exercise. Single leg knee-extension provided the acute exercise stimulus and the training modality. Four biopsies were collected from the vastus lateralis muscle at rest in the untrained and trained conditions before and after exercise. Training resulted in a 35% increase in muscle oxygen consumption and an 18% increase in number of capillaries per muscle fiber. At rest, VEGF/18S mRNA levels were similar before (0.38 +/- 0.04) and after (1.2 +/- 0.4) training. When muscle was untrained, acute exercise greatly elevated VEGF/18S mRNA levels (16.9 +/- 6.7). The VEGF/18S mRNA response to acute exercise in the trained state was markedly attenuated (5.4 +/- 1.3). These data support the concept that VEGF is involved in exercise-induced skeletal muscle angiogenesis and appears to be subject to a negative feedback mechanism as exercise adaptations occur.

Numerical analysis of the deformation of an adherent drop under shear flow.

The adhesion of leukocytes to substrates is an important biomedical problem and has drawn extensive research. In this study, employing both single and compound drop models, we investigate how hydrodynamics interacts with an adherent liquid drop in a shear flow. These liquid drop models have recently been used to describe the rheological behavior of leukocytes. Numerical simulation confirms that the drop becomes more elongated when either capillary number or initial contact angle increases. Our results show that there exists a thin region between the drop and the wall as the drop undergoes large stretching, which allows high pressure to build up and provides a lift force. In the literature, existing models regard the leukocyte as a rigid body to calculate the force and torque acting on the drop in order to characterize the binding between cell receptors and endothelial ligands. The present study indicates that such a rigid body model is inadequate and the force magnitude obtained from it is less than half of that obtained using the deformable drop models. Furthermore, because of its much higher viscosity, the cell nucleus introduces a hydrodynamic time scale orders of magnitude slower than the cytoplasm. Hence the single and compound drops experience different dynamics during stretching, but exhibit very comparable steady-state shapes. The present work offers a framework to facilitate the development of a comprehensive dynamic model for blood cells.

Dynamics of drops in branched tubes

The flow of two-dimensional deformable drops through branching (bifurcating) tubes is studied numerically using a boundary integral formulation. The undeformed drop diameter is assumed to be less than the tube diameter. Capillary numbers between 10−2 and 1 are considered. Flow in the branching tube is characterized by the fraction of fluid which enters each of the two downstream branches. The likelihood of drops entering the high-flow-rate branch increases as (i) the viscosity ratio between the drops and suspending fluid decreases, (ii) the capillary number increases, and (iii) the drop size increases. Hydrodynamic interactions between the suspended drops increase the number of drops which enter the low-flow-rate branch. The implications of these results for dispersion processes and local transport are explored. The disturbance flow created by drops passing over ‘dead-end’ pores or cavities results in fluid transfer between the pore and the free stream; suspensions may then be effective in improving the ‘cleaning’ of porous materials.

Water/Oil Relative Permeability Endpoints of Intermediate-Wet, Low-Permeability Rocks

Summary The relative permeability endpoints of low-permeability, intermediate-wet mudstones increased with flow rate. This was because of both a decrease in capillary end effect and an increase in capillary number. Hence, conventional analysis will incorrectly describe field behavior. We recommend an improved methodology. We imaged steady-state saturation profiles at the end of different rate floods on one sample. We analyzed these profiles, which were axially nonuniform, to obtain capillary-end-effect-corrected data for each flow rate. The saturation distributions were also radially nonuniform. We characterized core-scale heterogeneity using a miscible flow test and examined the influence of heterogeneity. We conducted long-term tests on three short samples to study the higher-capillary-number regime. These tests displayed the existence of a critical capillary number, where the water relative permeability endpoint increased dramatically. The corresponding rate was over a magnitude smaller than flow rates traditionally recommended to eliminate capillary end effects.

In Vivo Fluorescence Microscopy of Microcirculation in the Renal Cortex of Mice

In vivo fluorescence microscopy was used for experimental studies of the renal cortical microcirculation in mice. The effects of i.v. infusions of mannitol and iohexol were studied in untreated mice as well as in mice after temporary renal ischemia. All infusions produced marked effects on the distribution and velocity of cortical blood flow with no differences between iohexol and mannitol. The renal cortical blood flow was inhomogeneous with respect to different capillaries and there was an initial rise in the number of capillaries with increased blood flow after infusions of both mannitol and iohexol. There was an initial increase in the number of capillaries with a decreased blood flow following 30 min of ischemia. This increase was reduced by both types of infusions, suggesting a beneficial effect on renal cortical blood flow. Neither mannitol nor contrast medium exhibited any adverse effects in ischemic kidneys that would be likely to affect renal function. However, 5 min after the infusions there was an increasing number of capillaries with decreased blood flow in all groups.

The Quantitative Development of Glomerular Capillaries in Rats with Special Reference to Unbiased Stereological Estimates of Their Number and Sizes

The kidneys of 14 female Wistar rats from 5 days to 18 months postnatally have been perfusion-fixed to study the development of glomerular capillaries using design-based stereological techniques. This report uses a topological definition of a capillary structural unit which forms the basis for the estimation of the number of glomerular capillaries using histological sections. The number of mature glomeruli is about 27,500 for all animals older than 5 days, whereas the 5-day-old animals have 21,100 mature glomeruli. Postnatally, the mean glomerular volume increases rapidly for the first 2 months after which it increases at a much slower rate. At first there is a rapid increase in total number, length, and surface area of glomerular capillaries with age; after 2 months the increase is rather moderate. The rapid growth of glomerular capillaries is anisomorphic, and the restrained growth is roughly isomorphic. The mean length of glomerular capillaries is nearly constant at 56 μm in mature and immature rats. The cross-sectional area rises only twofold from the most immature to the oldest rats. Using an approximation of the geometric factor in the law of Poiseuille, a decreasing resistance of glomerular capillaries with age is obtained. This is in agreement with the falling resistance of maturing renal vasculature and the increasing single-nephron glomerular filtration rate. When taking into account the constant mean capillary length and moderate increase in cross-sectional area of glomerular capillaries in growing rats, it appears that the branching of capillaries, forming a complicated network of serial and parallel connections, seems to be the structural basis for normal glomerular capillary growth.

In Vivo Fluorescence Microscopy of Microcirculation in the Renal Cortex of Mice

In vivo fluorescence microscopy was used for experimental studies of renal cortical microcirculation in 46 mice. The cortical circulation was studied after i.v. infusions of mannitol of various osmolalities as well as nonionic low osmolar (iohexol) and ionic high osmolar (metrizoate) contrast media. All infusions produced an increase in the number of capillaries with increased blood flow, significantly more pronounced after the infusion of iohexol, metrizoate and mannitol 1.46 mol/l than after the infusion of mannitol 0.3 mol/l. However, the renal cortical blood flow was inhomogeneous with respect to different capillaries. While the blood flow was increased in some capillaries it was to some extent simultaneously decreased in others, significantly more after infusion of metrizoate than after infusion of mannitol 0.3 mol/l.

Minimum wet thickness in extrusion slot coating

An experimental study has been carried out to determine the minimum wet thickness of extrusion slot coating. It was found that there exists a critical capillary number Ca* for Newtonian solutions within the range of experimental conditions: below Ca*, the minimum wet thickness depends strongly on the coating speed and the fluid viscosity, or the capillary number Ca. Above Ca*, the minimum wet thickness is independent of the capillary number and is a function of the coating gap only. Comparing with slide coating, we have found that the minimum wet thickness for both coating methods are very close if Ca < Ca*. As Ca > Ca*, the minimum wet thickness for extrusion slot coating will become constant as the capillary number increases and the dimensionless minimum wet thickness will reach a constant value, but the minimum wet thickness for slide coating will increase as the capillary number increases. The experimentally determined minimum wet thickness for extrusion slot coating has been compared with the theoretical predictions of Ruschak, and of Higgins and Scriven. The effect of bead vacuum was also investigated, it was found that increasing bead vacuum can effectively reduce the minimum wet thickness particularly if the fluid viscosity is low and the coating gap is narrow.

Enhancement of incisional wound healing and neovascularization in normal rats by thrombin and synthetic thrombin receptor-activating peptides.

To better define thrombin-receptor interactions, we synthesized human thrombin peptides and identified binding-domain peptides that bind thrombin receptors and activate mitogenic signals (Glenn, K.C., G.H. Frost, J.S. Bergmann, and D.H. Carney. 1988. Pept. Res. 1:65-73). Treatment of full dermal dorsal incisions with a single topical application of thrombin receptor-activating peptide (TRAP-508) or human alpha-thrombin in saline enhances 7-d incisional breaking strength in normal rats up to 82% or 55% over saline-treated controls, respectively. Control wounds require approximately 11.5 d to achieve breaking strength equivalent to TRAP-treated wounds at day 7. Thus, a single application of TRAP accelerates healing, shifting the time course forward by up to 4.5 d. Histological comparisons at day 7 show more type I collagen, less evidence of prolonged inflammation, and an increase in number and maturity of capillaries in TRAP- and thrombin-treated incisions. Angiograms also show 50-65% more functional vascularization going across thrombin- and TRAP-treated surgical incisions. Thus, alpha-thrombin and thrombin peptides, such as those released following injury, appear to initiate or enhance signals required for neovascularization and wound healing. The ability to accelerate normal wound healing events with synthetic peptides representing receptor binding domains of human thrombin may offer new options for management of wound healing in man.

Red muscle fibre and capillary dimensions in different sizes of a tilapia, Oreochromis niloticus (Trewavas)

Measurements of muscle dimensions that affect respiration in relation to body weight were carried out in a tilapia, Oreochromis niloticus. The fish used in all measurements weighed 0.65–812.3 g. The data were analysed with respect to body weight using logarithmic transformations (log Y=log a+blog W). The slopes (b) of the log/log regression lines for weight of body trunk red muscle, average cross-sectional area of muscle fibre, average number of capillaries in direct contact with a muscle fibre, average capillary contact length with a fibre as a fraction of average fibre circumference and number of capillaries mm2 of fibre cross-sectional area [NA(cƒ)] were 1.16, 0.221, 0.084, 0.015, and −0.137 respectively. These results show that there is an increase in muscle cross-sectional fibre area and number of capillaries in contact with muscle fibres whereas number of capillaries supplying a unit area of muscle fibre decreases during development. There is development of new capillaries with increase in cross-sectional area of red muscle fibres.