Sensitivity analysis of a multi-physics model for the vascular microenvironment.

The vascular microenvironment is the scale at which microvascular transport, interstitial tissue properties and cell metabolism interact. The vascular microenvironment has been widely studied by means of quantitative approaches, including multi-physics mathematical models as it is a central system for the pathophysiology of many diseases, such as cancer. The microvascular architecture is a key factor for fluid balance and mass transfer in the vascular microenvironment, together with the physical parameters characterizing the vascular wall and the interstitial tissue. The scientific literature of this field has witnessed a long debate about which factor of this multifaceted system is the most relevant. The purpose of this work is to combine the interpretative power of an advanced multi-physics model of the vascular microenvironment with state of the art and robust sensitivity analysis methods, in order to determine the factors that most significantly impact quantities of interest, related in particular to cancer treatment. We are particularly interested in comparing the factors related to the microvascular architecture with the ones affecting the physics of microvascular transport. Ultimately, this work will provide further insight into how the vascular microenvironment affects cancer therapies, such as chemotherapy, radiotherapy or immunotherapy.

Capsules Rheology in Carreau-Yasuda Fluids.

In this paper, a Multi Relaxation Time Lattice Boltzmann scheme is used to describe the evolution of a non-Newtonian fluid. Such method is coupled with an Immersed-Boundary technique for the transport of arbitrarily shaped objects navigating the flow. The no-slip boundary conditions on immersed bodies are imposed through a convenient forcing term accounting for the hydrodynamic force generated by the presence of immersed geometries added to momentum equation. Moreover, such forcing term accounts also for the force induced by the shear-dependent viscosity model characterizing the non-Newtonian behavior of the considered fluid. Firstly, the present model is validated against well-known benchmarks, namely the parabolic velocity profile obtained for the flow within two infinite laminae for five values of the viscosity model exponent, n = 0.25, 0.50, 0.75, 1.0, and 1.5. Then, the flow within a squared lid-driven cavity for Re = 1000 and 5000 (being Re the Reynolds number) is computed as a function of n for a shear-thinning (n < 1) fluid. Indeed, the local decrements in the viscosity field achieved in high-shear zones implies the increment in the local Reynolds number, thus moving the position of near-walls minima towards lateral walls. Moreover, the revolution under shear of neutrally buoyant plain elliptical capsules with different Aspect Ratio (AR = 2 and 3) is analyzed for shear-thinning (n < 1), Newtonian (n = 1), and shear-thickening (n > 1) surrounding fluids. Interestingly, the power law by Huang et al. describing the revolution period of such capsules as a function of the Reynolds number and the existence of a critical value, Rec, after which the tumbling is inhibited in confirmed also for non-Newtonian fluids. Analogously, the equilibrium lateral position yeq of such neutrally buoyant capsules when transported in a plane-Couette flow is studied detailing the variation of yeq as a function of the Reynolds number as well as of the exponent n.

Vascular journey and adhesion mechanics of micro-sized carriers in narrow capillaries.

In this work a Lattice Boltzmann-Immersed Boundary method is used for predicting the dynamics of rigid and deformable adhesive micro-carriers (1 µm) navigating a capillary by the size of 10 µm with 20% hematocrit. Red cells and particles are modeled as a collection of mass-spring elements responding to a bending potential, an elastic potential and total enclosed area conservation constraint. Furthermore, particle surfaces are uniformly decorated with adhesive molecules (ligands) interacting with receptors disposed on the walls. Particle adhesion is modeled as a short-range ligad-receptor interaction and in term of formation and destruction probability functions that discriminate whether a chemical bond can be formed or destroyed. If a bond is established an attractive elastic force is activated. Particle transport and adhesion are characterized in terms of their ability to reach the capillary peripheries (margination rate) and firmly adhere the vasculature. This analysis is carried out systematically by varying particles' and cells' releasing positions and stiffness (Ca = 0 and 10-2). Moreover, three rigid and soft representative particles are transported on a finer mesh (Δx = 15 nm) and the chemical strength of their adhesive coating is varied (σ = 0.5, 1.0, and 2.0) to precisely analyze the resulting adhesion mechanics. Stiffness is found to weakly influence the margination rate while significantly affect the ability of such constructs to efficiently interact with the endothelium by forming stable chemical bonds.

Quantitative micro-Raman analysis of micro-particles in drug delivery.

Polymeric micro and nanoconstructs are emerging as promising delivery systems for therapeutics and contrast agents in microcirculation. Excellent assets associated with polymeric particulates of tunable shape, size, mechanical and chemical properties may improve the efficiency of delivery and represent the basis of personalized medicine and treatment. Nevertheless, lack of effective techniques of analysis may limit their use in biomedicine and bioengineering. In this paper, we demonstrated Raman Spectroscopy for quantitative characterization of poly lactic-co-glycolic acid (PLGA) micro-plate drug delivery systems. To do so, we (i) acquired bi-dimensional Raman maps of PLGA micro-plates loaded with curcumin at various times of release over multiple particles. We (ii) realized an exploratory analysis of data using the principal component analysis (PCA) technique to find hidden patterns in the data and reduce the dimensionality of the system. Then, we (iii) used an innovative univariate method of analysis of the reduced system to derive quantitative drug release profiles. High performance liquid chromatography (HPLC), the consolidated method of analysis of macro-sized systems, was used for comparison. We found that our system is as efficient as HPLC but, differently from HPLC, it enables quantitative analysis of systems at the single particle level.

A tissue chamber chip for assessing nanoparticle mobility in the extravascular space.

Although a plethora of nanoparticle configurations have been proposed over the past 10 years, the uniform and deep penetration of systemically injected nanomedicines into the diseased tissue stays as a major biological barrier. Here, a 'Tissue Chamber' chip is designed and fabricated to study the extravascular transport of small molecules and nanoparticles. The chamber comprises a collagen slab, deposited within a PDMS mold, and an 800 µm channel for the injection of the working solution. Through fluorescent microscopy, the dynamics of molecules and nanoparticles was estimated within the gel, under different operating conditions. Diffusion coefficients were derived from the analysis of the particle mean square displacements (MSD). For validating the experimental apparatus and the protocol for data analysis, the diffusion D of FITC-Dextran molecules of 4, 40 and 250 kDa was first quantified. As expected, D reduces with the molecular weight of the dextran molecules. The MSD-derived diffusion coefficients were in good agreement with values derived via fluorescence recovery after photobleaching (FRAP), an alternative technique that solely applies to small molecules. Then, the transport of six nanoparticles with similar hydrodynamic diameters (~ 200 nm) and different surface chemistries was quantified. Surface PEGylation was confirmed to favor the diffusion of nanoparticles within the collagen slab, whereas the surface decoration with hyaluronic acid (HA) chains reduced nanoparticle mobility in a way proportional to the HA molecular weight. To assess further the generality of the proposed approach, the diffusion of the six nanoparticles was also tested in freshly excised brain tissue slices. In these ex vivo experiments, the diffusion coefficients were 5-orders of magnitude smaller than for the Tissue Chamber chip. This was mostly ascribed to the lack of a cellular component in the chip. However, the trends documented for PEGylated and HA-coated nanoparticles in vitro were also confirmed ex vivo. This work demonstrates that the Tissue Chamber chip can be employed to effectively and efficiently test the extravascular transport of nanomedicines while minimizing the use of animals.

Deciphering the relative contribution of vascular inflammation and blood rheology in metastatic spreading.

Vascular adhesion of circulating tumor cells (CTCs) is a key step in cancer spreading. If inflammation is recognized to favor the formation of vascular "metastatic niches," little is known about the contribution of blood rheology to CTC deposition. Herein, a microfluidic chip, covered by a confluent monolayer of endothelial cells, is used for analyzing the adhesion and rolling of colorectal (HCT-15) and breast (MDA-MB-231) cancer cells under different biophysical conditions. These include the analysis of cell transport in a physiological solution and whole blood over a healthy and a TNF-α inflamed endothelium with a flow rate of 50 and 100 nl/min. Upon stimulation of the endothelial monolayer with TNF-α (25 ng/ml), CTC adhesion increases from 2 to 4 times whilst cell rolling velocity only slightly reduces. Notably, whole blood also enhances cancer cell deposition from 2 to 3 times, but only on the unstimulated vasculature. For all tested conditions, no statistically significant difference is observed between the two cancer cell types. Finally, a computational model for CTC transport demonstrates that a rigid cell approximation reasonably predicts rolling velocities while cell deformability is needed to model adhesion. These results would suggest that, within microvascular networks, blood rheology and inflammation contribute similarly to CTC deposition, thereby facilitating the formation of metastatic niches along the entire network, including the healthy endothelium. In microfluidic-based assays, neglecting blood rheology would significantly underestimate the metastatic potential of cancer cells.