Microphysiological endothelial models to characterize subcutaneous drug absorption.

The high variability in subcutaneous bioavailability of protein therapeutics is poorly understood, contributing to critical delays in patient access to new therapies. Preclinical animal and in vitro models fail to provide a physiologically relevant testbed to parse potential contributors to human bioavailability, therefore new strategies are necessary. Here, we present a microphysiological model of the human hypodermal vasculature at the injection site to study the interactions of administered protein therapeutics within the microenvironment that influence subcutaneous bioavailability. Our model combines human dermal endothelial cells, fibroblasts, and adipocytes, self-assembled into three-dimensional, perfusable microvessels that express relevant extracellular matrix. We demonstrate the utility of the model for measurement of biophysical parameters within the hypodermal microenvironment that putatively impact protein kinetics and distribution at the injection site. We propose that microphysiological models of the subcutaneous space have applications in preclinical development of protein therapeutics intended for subcutaneous administration with optimal bioavailability.

Pre- and post-industrial levels of polycyclic aromatic hydrocarbons in sediments from the Estuary and Gulf of St. Lawrence (eastern Canada).

The concentrations of 23 polycyclic aromatic hydrocarbons (PAHs; 16 parent PAHs and 7 alkyl-PAHs) were determined in 45 surface sediment and 7 basal sediment box core samples retrieved from the Estuary and Gulf of St. Lawrence in eastern Canada. The concentration sums of 16 priority PAHs (Σ16PAHs) in the surface sediments (representing modern times or at least younger than the last decade) ranged from 71 to 5672 ng g-1. Σ16PAHs in the basal sediments ranged from 93 to 172 ng g-1 among the pre-industrial samples (pre-1900 common era or CE) and from 1216 to 1621 ng g-1 among the early post-industrial samples (~1930s and ~1940s CE). The highest Σ16PAH values occurred in samples retrieved from the Baie-Comeau-Matane area, an area affected by intense industrial anthropogenic activities. Source-diagnostic PAH ratios suggest a predominance of pyrogenic sources via atmospheric deposition, with a minor contribution of petrogenic seabed pockmark sources. The PAH concentrations in the sediments from the study areas reveal low ecological risks to benthic or other organisms living near the water-sediment interface.

Spatial and temporal distributions of polycyclic aromatic hydrocarbons in sediments from the Canadian Arctic Archipelago.

The concentrations of 23 polycyclic aromatic hydrocarbons (PAHs; 16 parent and 7 alkylated PAHs) were determined in 113 surface marine sediment samples, 13 on-land sediment samples and 8 subsampled push cores retrieved from the Canadian Arctic Archipelago (CAA). PAHs were extracted via accelerated solvent extraction and quantified via gas chromatography-mass spectrometry. The sums of the concentrations of 16 priority PAHs in the surface sediments ranged from 7.8 to 247.7 ng g-1 (dry weight basis, dw). The PAH inputs to the sediments have remained constant during the last century. Source-diagnostic ratios and statistical analysis suggest that the PAHs in the CAA mainly originate from natural petrogenic sources, with some pyrogenic sources. Temporal trends did not indicate major source shifts and largely indicated petrogenic inputs. Overall, the sediments retrieved from the CAA have low PAH concentrations, which indicates a low ecological risk for benthic or other organisms living near the water-sediment interface.

The CCL2-CCR2 astrocyte-cancer cell axis in tumor extravasation at the brain.

Although brain metastases are common in cancer patients, little is known about the mechanisms of cancer extravasation across the blood-brain barrier (BBB), a key step in the metastatic cascade that regulates the entry of cancer cells into the brain parenchyma. Here, we show, in a three-dimensional in vitro BBB microvascular model, that astrocytes promote cancer cell transmigration via their secretion of C-C motif chemokine ligand 2 (CCL2). We found that this chemokine, produced primarily by astrocytes, promoted the chemotaxis and chemokinesis of cancer cells via their C-C chemokine receptor type 2 (CCR2), with no notable changes in vascular permeability. These findings were validated in vivo, where CCR2-deficient cancer cells exhibited significantly reduced rates of arrest and transmigration in mouse brain capillaries. Our results reveal that the CCL2-CCR2 astrocyte-cancer cell axis plays a fundamental role in extravasation and, consequently, metastasis to the brain.

Microheart: A microfluidic pump for functional vascular culture in microphysiological systems.

Advances in microphysiological systems have prompted the need for long-term cell culture under physiological flow conditions. Conventional laboratory pumps typically lack the ability to deliver cell culture media at the low flow rates required to meet the physiological ranges of fluid flow, and are often pulsatile or require flow reversal. Here, a microfluidic-based pump is presented, which allows for the controlled delivery of media for vascular microphysiological applications. The performance of the pump was characterized in a range of microfluidic systems, including straight channels of varying dimensions and self-assembled microvascular networks. A theoretical framework was developed based on lumped element analysis to predict the performance of the pump for different fluidic configurations and a finite element model of the included check-valves. The use of the pump for microvascular physiological studies demonstrated the utility of this system to recapitulate vascular fluid transport phenomena in microphysiological systems, which may find applications in disease models and drug screening.

The effects of luminal and trans-endothelial fluid flows on the extravasation and tissue invasion of tumor cells in a 3D in vitro microvascular platform.

Throughout the process of metastatic dissemination, tumor cells are continuously subjected to mechanical forces resulting from complex fluid flows due to changes in pressures in their local microenvironments. While these forces have been associated with invasive phenotypes in 3D matrices, their role in key steps of the metastatic cascade, namely extravasation and subsequent interstitial migration, remains poorly understood. In this study, an in vitro model of the human microvasculature was employed to subject tumor cells to physiological luminal, trans-endothelial, and interstitial flows to evaluate their effects on those key steps of metastasis. Luminal flow promoted the extravasation potential of tumor cells, possibly as a result of their increased intravascular migration speed. Trans-endothelial flow increased the speed with which tumor cells transmigrated across the endothelium as well as their migration speed in the matrix following extravasation. In addition, tumor cells possessed a greater propensity to migrate in close proximity to the endothelium when subjected to physiological flows, which may promote the successful formation of metastatic foci. These results show important roles of fluid flow during extravasation and invasion, which could determine the local metastatic potential of tumor cells.

Interstitial flow promotes macrophage polarization toward an M2 phenotype.

Tumor tissues are characterized by an elevated interstitial fluid flow from the tumor to the surrounding stroma. Macrophages in the tumor microenvironment are key contributors to tumor progression. While it is well established that chemical stimuli within the tumor tissues can alter macrophage behaviors, the effects of mechanical stimuli, especially the flow of interstitial fluid in the tumor microenvironment, on macrophage phenotypes have not been explored. Here, we used three-dimensional biomimetic models to reveal that macrophages can sense and respond to pathophysiological levels of interstitial fluid flow reported in tumors (∼3 µm/s). Specifically, interstitial flow (IF) polarizes macrophages toward an M2-like phenotype via integrin/Src-mediated mechanotransduction pathways involving STAT3/6. Consistent with this flow-induced M2 polarization, macrophages treated with IF migrate faster and have an enhanced ability to promote cancer cell migration. Moreover, IF directs macrophages to migrate against the flow. Since IF emanates from the tumor to the surrounding stromal tissues, our results suggest that IF could not only induce M2 polarization of macrophages but also recruit these M2 macrophages toward the tumor masses, contributing to cancer cell invasion and tumor progression. Collectively, our study reveals that IF could be a critical regulator of tumor immune environment.