Leveraging avidin/biotin interaction to quantify permeability of microvessels-on-a-chip

Microvessels-on-a-chip have enabled in vitro studies to closely simulate in vivo microvessel environment. However, assessing microvessel permeability, a functional measure of microvascular exchange, has not been attainable in nonpermeable microfluidic platforms. This study developed a new approach that enables permeability coefficients (Ps) to be quantified in microvessels developed in nonpermeable chip platforms by integrating avidin/biotin technology. Microvessels were developed on biotinylated fibronectin-coated microfluidic channels. Solute transport was assessed by perfusing microvessels with fluorescence-labeled avidin. Avidin molecules that crossed endothelium were captured by substrate biotin and recorded with real-time confocal images. The Ps was derived from the rate of avidin/biotin accumulation at the substrate relative to solute concentration difference across microvessel wall. Avidin tracers with different physiochemical properties were used to characterize the barrier properties of the microvessel wall. The measured baseline Ps and inflammatory mediator-induced increases in Ps and EC [Ca2+]i resembled those observed in intact microvessels. Importantly, the spatial accumulation of avidin/biotin at substrate defines the transport pathways. Glycocalyx layer is well-formed on endothelium and its degradation increased transcellular transport without affecting EC junctions. This study demonstrated that in vitro microvessels developed in this simply designed microfluidics structurally possess in vivo-like glycocalyx layer and EC junctions and functionally recapitulate basal barrier properties and stimuli-induced responses observed in intact microvessels. This new approach overcomes the limitations of nonpermeable microfluidics and provides an easily executed highly reproducible in vitro microvessel model with in vivo microvessel functionality, suitable for a wide range of applications in blood and vascular research and drug development.

Numerical analysis of the biomechanical effects on micro-vessels by ultrasound-driven cavitation

Purpose: The goal of this study was to evaluate the biomechanical effects such as sonoporation or permeability, produced by ultrasound- driven microbubbles (UDM) within microvessels with various parameters. Methods: In this study, a bubble-fluid-solid coupling system was established through combination of finite element method. The stress, strain and permeability of the vessel wall were theoretically simulated for different ultrasound frequencies, vessel radius and vessel thickness. Results: the bubble oscillation induces the vessel wall dilation and invagination under a pressure of 0.1 MPa. The stress distribution over the microvessel wall was heterogeneous and the maximum value of the midpoint on the inner vessel wall could reach 0.7 MPa as a frequency ranges from 1 to 3 MHz, and a vessel radius and an initial microbubble radius fall within the range of 3.5–13 μm and 1–4 μm, respectively. With the same conditions, the maximum shear stress was equal to 1.2 kPa and occurred at a distance of ±5 μm from the midpoint of 10 μm and the maximum value of permeability was 3.033 × 10–13. Conclusions: Results of the study revealed a strong dependence of biomechanical effects on the excitation frequency, initial bubble radius, and vessel radius. Numerical simulations could provide insight into understanding the mechanism behind bubble-vessel interactions by UDM, which may explore the potential for further improvements to medical applications.

A time-dependent multi-layered mathematical model of filtration and solute exchange, the revised Starling principle and the Landis experiments

Cell oxygenation and nutrition is vitally important for human and animal life. Oxygen and nutrients are transported by the blood stream and cross microvessel walls to penetrate the cell’s membrane. Pathological alterations in the transport of oxygen, and other nutrition elements, across microvessel walls may have serious consequences to cell life, possibly leading to localized cell necrosis. We present a transient model of plasma filtration and solute transport across microvessel walls by coupling flow and transport equations, the latter being non-linear in solute concentration. The microvessel wall is modeled through the superimposition of two or more membranes with different physical properties, representing key structural elements. With this model, the combined effect of the endothelial cells, the glycocalyx and other coating membranes specific of certain microvessels, can be analyzed. We investigate the role of transient external pressures in the study of trans-vascular filtration and solute exchange during the drop of blood capillary pressure due to the pathological decrease of blood volume called hypovolaemia, as well as hemorrhage. We discuss the advantage of using a multi-layered model, rather than a model considering the microvessel wall as a single and homogeneous membrane.

Influence of Starling's Hypothesis and Joule Heating on Peristaltic Flow of an Electrically Conducting Casson Fluid in a Permeable Microvessel

Peristaltic transport of electrically conducting blood through a permeable microvessel is investigated by considering the Casson model in the presence of an external magnetic field. The reabsorption process across the permeable microvessel wall is regarded to govern by Starling's hypothesis. Under the long wavelength approximation and low-Reynolds number assumption, the nonlinear governing equations along with the boundary conditions are solved using a perturbation technique. Starling's hypothesis at the microvessel wall provides a second-order ordinary differential equation to be solved numerically for pressure distribution which in turn gives the stream function and temperature field. Also, the location of the interface between the plug and core regions is obtained from the axial velocity. Due to an increasing reabsorption process, the axial velocity is found to increase initially but decreases near the outlet. The temperature is appreciably intensified by virtue of the Joule heating produced due to the electrical conductivity of blood.

Static Stress Distribution in Microvessel Wall with a Layered Model

Based on the vascular membrane stress model and the pseudo-elastic vessel model, the combination constitutive model with a layered structure in microvessel is presented in this paper. By using obtained constitutive equations of the current model, the circumferential stress of the membrane intimal (inner) layer and the three-dimensional stress distribution of the structural outer layer are analyzed. Under the initial blood pressure state, the vascular static stress changes with the inner stiffness increase are also discussed. The results show that with inner stiffness increasing, the stress of outer layer is less affected but the circumferential stress of the intimal layer is increased significantly, which may be one potential risk factor for the vascular injury. These analysis methods and its conclusions have some theoretical significance for studying the problems of arteriosclerosis and other diseases, and preventing the occurrence of related diseases.

Adhesion of malignant mammary tumor cells MDA-MB-231 to microvessel wall increases microvascular permeability via degradation of endothelial surface glycocalyx

To investigate the effect of tumor cell adhesion on microvascular permeability ( P) in intact microvessels, we measured the adhesion rate of human mammary carcinoma MDA-MB-231, the hydraulic conductivity (Lp), the P, and reflection coefficient (σ) to albumin of the microvessels at the initial tumor cell adhesion and after ∼45 min cell perfusion in the postcapillary venules of rat mesentery in vivo. Rats (Sprague-Dawley, 250–300 g) were anesthetized with pentobarbital sodium given subcutaneously. A midline incision was made in the abdominal wall, and the mesentery was gently taken out and arranged on the surface of a glass coverslip for the measurement. An individual postcapillary venule was perfused with cells at a rate of ∼1 mm/s, which is the mean blood flow velocity in this type of microvessels. At the initial tumor cell adhesion, which was defined as one adherent cell in ∼100- to 145-μm vessel segment, Lp was 1.5-fold and P was 2.3-fold of their controls, and σ decreased from 0.92 to 0.64; after ∼45-min perfusion, the adhesion increased to ∼5 adherent cells in ∼100- to 145-μm vessel segment, while Lp increased to 2.8-fold, P to 5.7-fold of their controls, and σ decreased from 0.92 to 0.42. Combining these measured data with the predictions from a mathematical model for the interendothelial transport suggests that tumor cell adhesion to the microvessel wall degrades the endothelial surface glycocalyx (ESG) layer. This suggestion was confirmed by immunostaining of heparan sulfate of the ESG on the microvessel wall. Preserving of the ESG by a plasma glycoprotein orosomucoid decreased the P to albumin and reduced the tumor cell adhesion.

TNF-α activation of arterioles and venules alters distribution and levels of ICAM-1 and affects leukocyte-endothelial cell interactions

The observation that leukocyte-endothelial cell (EC) interactions are localized to specific regions on the microvessel wall suggests that adhesion molecule distribution is not uniform. We investigated ICAM-1 distribution and leukocyte-EC interactions in blood-perfused microvessels (<80 μm) in cremaster muscle of anesthetized mice, using intravital confocal microscopy and immunofluorescent labeling. Variability of ICAM-1 expression directly determines leukocyte adhesion distribution within the venular microcirculation and contributes to leukocyte rolling in arterioles during inflammation. The number of rolling interactions increased with ICAM-1 intensity ( r2 = 0.69, P < 0.05), and rolling velocity was lower in regions of higher ICAM-1 intensity. In controls, venular ICAM-1 expression was approximately twofold higher than in arterioles. After TNF-α treatment, ICAM-1 expression was significantly increased, 2.8 ± 0.2-fold in arterioles and 1.7 ± 0.2-fold in venules ( P < 0.05). ICAM-1 expression on activated arteriolar ECs only reached the level of control venular ICAM-1. Arteriolar but not venular ECs underwent redistribution of ICAM-1 among cells; some cells increased and some decreased ICAM-1 expression, magnifying the variability of ICAM-1. TNF-α treatment increased the length of bright fluorescent regions per unit vessel length (42%, control; 70%, TNF-α) along the arteriolar wall, whereas no significant change was observed in venules (60%, control; 63%, TNF-α). The spatial distribution and expression levels of adhesion molecules in the microcirculation determine the timing and placement of leukocyte interactions and hence significantly impact the inflammatory response. That arteriolar ECs respond to TNF-α by upregulation of ICAM-1, although in a different way compared with venules, suggests an explicit role for arterioles in inflammatory responses.