An Interface-Particle Interaction Approach for Evaluation of the Co-Encapsulation Efficiency of Cells in a Flow-Focusing Droplet Generator.

Droplet-based microfluidics offers significant advantages, such as high throughput and scalability, making platforms based on this technology ideal candidates for point-of-care (POC) testing and clinical diagnosis. However, the efficiency of co-encapsulation in droplets is suboptimal, limiting the applicability of such platforms for the biosensing applications. The homogeneity of the bioanalytes in the droplets is an unsolved problem. While there is extensive literature on the experimental setups and active methods used to increase the efficiency of such platforms, passive techniques have received less attention, and their fundamentals have not been fully explored. Here, we develop a novel passive technique for investigating cell encapsulation using the finite element method (FEM). The level set method was used to track the interfaces of forming droplets. The effects of walls and the droplet interfaces on relatively large cells were calculated to track them more accurately during encapsulation. The static surface tension force was used to account for the effects of the interfaces on cells. The results revealed that the pairing efficiency is highly sensitive to the standard deviation (SD) of the distance between the cells in the entrance channel. The pairing efficiency prediction error of our model differed by less than 5% from previous experiments. The proposed model can be used to evaluate the performance of droplet-based microfluidic devices to ensure higher precision for co-encapsulation of cells.

Fabrication and characterization of low-cost, bead-free, durable and hydrophobic electrospun membrane for 3D cell culture.

This paper reports the fabrication of electrospun polydimethylsiloxane (PDMS) membranes/scaffolds that are suitable for three-dimensional (3D) cell culture. Through modification the ratio between PDMS and polymethylmethacrylate (PMMA) as carrier polymer, we report the possibility of increasing PDMS weight ratio of up to 6 for electrospinning. Increasing the PDMS content increases the fiber diameter, the pore size, and the hydrophobicity. To our best knowledge, this is the first report describing beads-free, durable and portable electrospun membrane with maximum content of PDMS suitable for cell culture applications. To show the proof-of-concept, we successfully cultured epithelial lung cancer cells on these membranes in a static well plate without surface modification. Surprisingly, due to three-dimensional (3D) and hydrophobic nature of the electrospun fibers, cells aggregated into 3D multicellular spheroids. These easily detachable and cost-effective scaffolds with controllable thicknesses and high tensile strength are good candidates for cell-stretching devices, organ-on-a-chip devices, tissue engineering and studies of non-adherent mammalian cancer stem cells.

Separation of microparticles suspended in a minichannel using laser radiation pressure.

Optical separation, which is a contactless and accurate technique, has been mostly used to manipulate single particles. This work mainly aims to present an effective technique for optical propulsion and separation of a group of microscopic particles that are suspended in liquids. An experimental study is conducted to assess the effect of radiation pressure of a high-power laser on a dilute dispersion of microparticles in water using microscopic image analysis. Results of separation experiments indicate that the manipulation mechanism is capable of sorting the microscopic particles in two size classes. Compared to common optical separators, this configuration has a benefit of separating many particles simultaneously.

Investigation of the different parameters contributing to bubble sticking inside physiological bifurcations.

Gas embolotherapy (GE) is a developing medical method which can be utilized either as an autonomous therapeutic method to treat vascularized solid tumors, or it can be combined with other medical procedures-such as high-intensity focused ultrasound-to improve their efficiency. This paper is dedicated to investigating the different parameters which influence bubble lodging inside human vasculature via 2D-modeling of bubble dynamics in arteries' and arterioles' bifurcations which are potential sticking positions. Values used in the simulations are in accordance with the non-dimensional physiological numbers. It is found out that inlet pressure plays a decisive role in bubble lodging; the lower the value, the higher the possibility of bubble sticking. On the other hand, gravity has a counteracting effect on bubble lodging in arteries, but not on arterioles. The initial length of the bubble is not a determining factor in sticking behavior, even though it affects the flow rate behavior. Surface tension, another critical factor, has a semi-linear impact on bubble resisting power; lowering the surface tension will reduce bubble resistance to the flow, diminishing the possibility of bubble lodging. Finally, it is shown that lower values for the static contact angle impose higher resistance to the flow.

Design of the micropump and mass-transfer compartment of a microfluidic system for regular nonenzymatic glucose measurement.

The aim of this paper is to design and numerically simulate the mass-transfer compartment and piezoelectric micropump of an implantable integrated microfluidic device for regular microdialysis-based nonenzymatic measurement of glucose level in diabetic patients. The device function is based on the process that the piezoelectric micropump pumps the dialysis fluid into the mass-transfer compartment microchannels, where the interstitial fluid (ISF) glucose diffusion into this dialysis fluid gives it a glucose content, then detected and measured in the sensor section. This diffusion takes place through the semipermeable membranes located in the microchannels at the base of the hollow microneedles entering the body skin painlessly. The value of dialysis fluid flow rate (1 µL/min) was chosen so that the best achievable recovery factor can be obtained while the size and time delay of system were being kept at the best minimum possible. In the mass-transfer compartment, the number of microneedles, the dimensions of microchannels and the thickness of membranes were selected so as to achieve the best appropriate recovery factor, minimum possible size as well as considering the fabrication feasibility. Furthermore, in the different parts of micropump, the materials and dimensions were chosen so as to provide the needed flow rate with the best minimum voltage, sufficiently small size and fabrication feasibility.

Atheroprone sites of coronary artery bifurcation: Effect of heart motion on hemodynamics-dependent monocytes deposition.

Atherosclerosis as a common cardiovascular disease is a result of both adverse hemodynamics conditions and monocyte deposition within coronary arteries. It is known that the adhesion of monocytes on the arterial wall and their interaction with the vascular surface are one of the main parameters in the initiation and progression of atherosclerosis. In this work, hemodynamic parameters and monocyte deposition have been investigated in a 3D computational model of the Left Anterior Descending coronary artery (LAD) and its first diagonal branch (D1) under the heart motion. A one-way Lagrangian approach is performed to trace the monocyte particles under different blood flow regimes and heart motion conditions. The hemodynamic results show that the myocardial wall, and also the flow divider wall can be candidates for atheroprone sites. The dynamic movement and pulsatile inlet changed the flow rate between branches about 21% compared to the static case and steady inlet. On the other hand, the calculation of monocytes' depositional behavior illustrates that they settle down downstream the LAD-D1 bifurcation and on the myocardial wall. The deposition rate is closely associated with the inlet type and changing the steady inlet to the sinusoidal and real physiologic profile showed a 150% increase in the deposition rate. These results ensure that the myocardial wall and LAD-D1 bifurcation are the desirable locations for atherosclerosis. These results are in good agreement with the clinical observations.

High-Throughput, Label-Free Isolation of White Blood Cells from Whole Blood Using Parallel Spiral Microchannels with U-Shaped Cross-Section.

Rapid isolation of white blood cells (WBCs) from whole blood is an essential part of any WBC examination platform. However, most conventional cell separation techniques are labor-intensive and low throughput, require large volumes of samples, need extensive cell manipulation, and have low purity. To address these challenges, we report the design and fabrication of a passive, label-free microfluidic device with a unique U-shaped cross-section to separate WBCs from whole blood using hydrodynamic forces that exist in a microchannel with curvilinear geometry. It is shown that the spiral microchannel with a U-shaped cross-section concentrates larger blood cells (e.g., WBCs) in the inner cross-section of the microchannel by moving smaller blood cells (e.g., RBCs and platelets) to the outer microchannel section and preventing them from returning to the inner microchannel section. Therefore, it overcomes the major limitation of a rectangular cross-section where secondary Dean vortices constantly enforce particles throughout the entire cross-section and decrease its isolation efficiency. Under optimal settings, we managed to isolate more than 95% of WBCs from whole blood under high-throughput (6 mL/min), high-purity (88%), and high-capacity (360 mL of sample in 1 h) conditions. High efficiency, fast processing time, and non-invasive WBC isolation from large blood samples without centrifugation, RBC lysis, cell biomarkers, and chemical pre-treatments make this method an ideal choice for downstream cell study platforms.

Endothelial Cells Morphology in Response to Combined WSS and Biaxial CS: Introduction of Effective Strain Ratio.

INTRODUCTION: Endothelial cells (ECs) morphology strongly depends on the imposed mechanical stimuli. These mechanical stimuli include wall shear stress (WSS) and biaxial cyclic stretches (CS). Under combined loading, the effect of CS is not as simple as pure CS. The present study investigates the morphological response of ECs to the realistic mechanical stimuli. METHODS: The cell population is theoretically studied using our previous validated model. The mechanical stimuli on ECs are described using four parameters; WSS magnitude (0 to 2.0 Pa), WSS angle (- 50° to 50°), and biaxial CS in two perpendicular directions (0 to 10%). The morphology of ECs is reported using four parameters; average shape index (SI) and orientation angle (OA) of the cell population as well as the standard deviation (SD) of SI and OA as measures for scattering of cells' SI and OA from these average values. RESULTS: A new effective strain ratio (ESR) is defined as the ratio of the undesirable CS to the desirable one. The obtained results of the model, illustrated that the SI and OA of cells increase with absolute value of ESR. In addition, the scattering in the SI of cells decreases with the absolute value of ESR, which means that the cell shapes become more regular. It is shown that the angular irregularity of cells increases at higher ESR values. CONCLUSIONS: The results indicated that, the defined ESR is a stand-alone parameter for describing the realistic mechanical loading on the ECs and their morphological response.

Three-Dimensional Modeling of Avascular Tumor Growth in Both Static and Dynamic Culture Platforms.

Microfluidic cell culture platforms are ideal candidates for modeling the native tumor microenvironment because they can precisely reconstruct in vivo cellular behavior. Moreover, mathematical modeling of tumor growth can pave the way toward description and prediction of growth pattern as well as improving cancer treatment. In this study, a modified mathematical model based on concentration distribution is applied to tumor growth in both conventional static culture and dynamic microfluidic cell culture systems. Apoptosis and necrosis mechanisms are considered as the main inhibitory factors in the model, while tumor growth rate and nutrient consumption rate are modified in both quiescent and proliferative zones. We show that such modification can better predict the experimental results of tumor growth reported in the literature. Using numerical simulations, the effects of the concentrations of the nutrients as well as the initial tumor radius on the tumor growth are investigated and discussed. Furthermore, tumor growth is simulated by taking into account the dynamic perfusion into the proposed model. Subsequently, tumor growth kinetics in a three-dimensional (3D) microfluidic device containing a U-shaped barrier is numerically studied. For this case, the effect of the flow rate of culture medium on tumor growth is investigated as well. Finally, to evaluate the impact of the trap geometry on the tumor growth, a comparison is made between the tumor growth kinetics in two frequently used traps in microfluidic cell culture systems, i.e., the U-shaped barrier and microwell structures. The proposed model can provide insight into better predicting the growth and development of avascular tumor in both static and dynamic cell culture platforms.

Prediction of Necrotic Core and Hypoxic Zone of Multicellular Spheroids in a Microbioreactor with a U-Shaped Barrier.

Microfluidic devices have been widely used for biological and cellular studies. Microbioreactors for three-dimensional (3D) multicellular spheroid culture are now considered as the next generation in in vitro diagnostic tools. The feasibility of using 3D cell aggregates to form multicellular spheroids in a microbioreactor with U-shaped barriers has been demonstrated experimentally. A barrier array is an alternative to commonly used microwell traps. The present study investigates oxygen and glucose concentration distributions as key parameters in a U-shaped array microbioreactor using finite element simulation. The effect of spheroid diameter, inlet concentration and flow rate of the medium are systematically studied. In all cases, the channel walls are considered to be permeable to oxygen. Necrotic and hypoxic or quiescent regions corresponding to both oxygen and glucose concentration distributions are identified for various conditions. The results show that the entire quiescent and necrotic regions become larger with increasing spheroid diameter and decreasing inlet and wall concentration. The shear stress (0.5⁻9 mPa) imposed on the spheroid surface by the fluid flow was compared with the critical values to predict possible damage to the cells. Finally, optimum range of medium inlet concentration (0.13⁻0.2 mM for oxygen and 3⁻11 mM for glucose) and flow rate (5⁻20 µL/min) are found to form the largest possible multicellular spheroid (500 µm), without any quiescent and necrotic regions with an acceptable shear stress. The effect of cell-trap types on the oxygen and glucose concentration inside the spheroid was also investigated. The levels of oxygen and glucose concentration for the microwell are much lower than those for the other two traps. The U-shaped barrier created with microposts allows for a continuous flow of culture medium, and so improves the glucose concentration compared to that in the integrated U-shaped barrier. Oxygen concentration for both types of U-shaped barriers is nearly the same. Due to the advantage of using U-shaped barriers to culture multicellular spheroids, the results of this paper can help to choose the experimental and design parameters of the microbioreactor.

Challenge in particle delivery to cells in a microfluidic device.

Micro and nanotechnology can potentially revolutionize drug delivery systems. Novel microfluidic systems have been employed for the cell culture applications and drug delivery by micro and nanocarriers. Cells in the microchannels are under static and dynamic flow perfusion of culture media that provides nutrition and removes waste from the cells. This exerts hydrostatic and hydrodynamic forces on the cells. These forces can considerably affect the functions of the living cells. In this paper, we simulated the flow of air, culture medium, and the particle transport and deposition in the microchannels under different angles of connection inlet. It was found that the shear stress induced by the medium culture flow is not so high to damage the cells and that it is roughly uniform in the cell culture section (CCS). However, the local shear stresses in the other parts of the microchip differ by changing the angles of the connection inlet. The results showed that the particle deposition was a function of the particle size, the properties of the fluid, and the flow rate. At a lower air flow rate, both small and large particles deposited in the entrance region and none of them reached the CCS. Once the airflow rate increased, the drag of the flow could overcome the diffusion of the small particles and deliver them to the CCS so that more than 88% of the 100 nm and 98% of the 200 nm particles deposited in the CCS. However, larger particles with average diameters in micrometers could not reach the CCS by the airflow even at high flow rate. In contrast, our findings indicated that both small and large particles could be delivered to the CCS by liquid flow. Our experimental data confirm that microparticles (with diameters of 5 and 20 µm) suspended in a liquid can reach the CCS at a well-adjusted flow rate. Consequently, a liquid carrier is suggested to transport large particles through microchannels. As a powerful tool, these numerical simulations provide a nearly complete understanding of the flow field and particle patterns in microchips which can significantly lower the trial and error in the experiment tests and accordingly save researchers considerable cost and time for drug delivery to the cell in the microchip by micro/nanocarriers.

A microfabricated platform for the study of chondrogenesis under different compressive loads.

Microfluidic devices are beneficial in miniaturizing and multiplexing various cellular assays in a single platform. Chondrogenesis is known to pertain to chemical, topographical, and mechanical cues in the microenvironment. Mechanical cues themselves have numerous parameters such as strain magnitude, frequency, and stimulation time. Effects of different strain magnitudes on the chondrogenic differentiation of adult stem cells have not been explored thoroughly. Here, a new multilayer microdevice is presented for the unidirectional compressive stimulation of cells in a three-dimensional cell culture. Numerical simulations were performed to evaluate and optimize the design. Results showed a favorable highly uniform axial strain distribution and negligible radial and circumferential strain for the optimized design. Moreover, an experimental study was performed on rabbit adipose-derived stem cells encapsulated in-situ in alginate hydrogel. Strain levels of 20%, 15%, 10%, 5%, and 0% were studied simultaneously on a microfluidic platform. Dynamic mechanical compression positively influenced cellular viability and upregulated collagen II, Sox-9, and aggrecan expression in the absence of exogenous growth factors. The expression of collagen type II as specific marker for articular chondrocytes was further confirmed by immunofluorescence staining of collagen type II. Taking together, 10% strain can be considered as optimal stimulation factor for chondrogenic differentiation of adipose derived stem cells.

A high-performance polydimethylsiloxane electrospun membrane for cell culture in lab-on-a-chip.

Thin porous membranes are important components in a microfluidic device, serving as separators, filters, and scaffolds for cell culture. However, the fabrication and the integration of these membranes possess many challenges, which restrict their widespread applications. This paper reports a facile technique to fabricate robust membrane-embedded microfluidic devices. We integrated an electrospun membrane into a polydimethylsiloxane (PDMS) device using the simple plasma-activated bonding technique. To increase the flexibility of the membrane and to address the leakage problem, the electrospun membrane was fabricated with the highest weight ratio of PDMS to polymethylmethacrylate (i.e., 6:1 w/w). The membrane-integrated microfluidic device could withstand a flow rate of up to 50 µl/min. As a proof of concept, we demonstrated that such a compartmentalized microfluidic platform could be successfully used for cell culture with the capability of providing a more realistic in vivo-like condition. Human lung cancer epithelial cells (A549) were seeded on the membrane from the top microchannel, while the continuous flow of the culture medium through the bottom microchannel provided a shear-free cell culture condition. The tortuous micro-/nanofibers of the membrane immobilized the cells within the hydrophobic micropores and with no need of extracellular matrix for cell adhesion and cell growth. The hydrophobic surface conditions of the membrane were suitable for anchorage-independent cell types. To further extend the application of the device, we qualitatively showed that rinsing the membrane with ethanol prior to cell seeding could temporarily render the membrane hydrophilic and the platform could also be used for anchorage-dependent cells. Due to the three-dimensional (3D) topography of the membranes, three different configurations were observed, including individual single cells, monolayer cells, and 3D cell clusters. This cost-effective and robust compartmentalized microfluidic device may open up new avenues in translational medicine and pharmacodynamics research.

A multiscale approach for determining the morphology of endothelial cells at a coronary artery.

The morphology of endothelial cells (ECs) may be an indication for determining atheroprone sites. Until now, there has been no clinical imaging technique to visualize the morphology of ECs in the arteries. The present study introduces a computational technique for determining the morphology of ECs. This technique is a multiscale simulation consisting of the artery scale and the cell scale. The artery scale is a fluid-structure interaction simulation. The input for the artery scale is the geometry of the coronary artery, that is, the dynamic curvature of the artery due to the cardiac motion, blood flow, blood pressure, heart rate, and the mechanical properties of the blood and the arterial wall, the measurements of which can be obtained for a specific patient. The results of the artery scale are wall shear stress (WSS) and cyclic strains as the mechanical stimuli of ECs. The cell scale is an inventive mass-and-spring model that is able to determine the morphological response of ECs to any combination of mechanical stimuli. The results of the multiscale simulation show the morphology of ECs at different locations of the coronary artery. The results indicate that the atheroprone sites have at least 1 of 3 factors: low time-averaged WSS, high angle of WSS, and high longitudinal strain. The most probable sites for atherosclerosis are located at the bifurcation region and lie on the myocardial side of the artery. The results also indicated that a higher dynamic curvature is a negative factor and a higher pulse pressure is a positive factor for protection against atherosclerosis.

Numerical Simulation of the Behavior of Toroidal and Spheroidal Multicellular Aggregates in Microfluidic Devices with Microwell and U-Shaped Barrier.

A microfluidic system provides an excellent platform for cellular studies. Most importantly, a three-dimensional (3D) cell culture model reconstructs more accurately the in vivo microenvironment of tissue. Accordingly, microfluidic 3D cell culture devices could be ideal candidates for in vitro cell culture platforms. In this paper, two types of 3D cellular aggregates, i.e., toroid and spheroid, are numerically studied. The studies are carried out for microfluidic systems containing U-shaped barrier as well as microwell structure. For the first time, we obtain oxygen and glucose concentration distributions inside a toroid aggregate as well as the shear stress on its surface and compare its performance with a spheroid aggregate of the same volume. In particular, we obtain the oxygen concentration distributions in three areas, namely, oxygen-permeable layer, multicellular aggregates and culture medium. Further, glucose concentration distributions in two regions of multicellular aggregates and culture medium are investigated. The results show that the levels of oxygen and glucose in the system containing U-shaped barriers are far more than those in the system containing microwells. Therefore, to achieve high levels of oxygen and nutrients, a system with U-shaped barriers is more suited than the conventional traps, but the choice between toroid and spheroid depends on their volume and orientation. The results indicate that higher oxygen and glucose concentrations can be achieved in spheroid with a small volume as well as in horizontal toroid with a large volume. The vertical toroid has the highest levels of oxygen and glucose concentration while the surface shear stress on its surface is also maximum. These findings can be used as guidelines for designing an optimum 3D microfluidic bioreactor based on the desired levels of oxygen, glucose and shear stress distributions.

Pulsatile blood flow in total cavopulmonary connection: a comparison between Y-shaped and T-shaped geometry.

Single-ventricle anomaly is a hereditary heart disease that is characterized by anatomical malformations. The main consequence of this malformation is desaturated blood flow, which without proper treatment increases the risk of death. The classical treatment is based on a three-stage palliative procedure which should begin from the first few days of patient's life. The final stage is known as Fontan procedure, in which inferior vena cava is directly connected to pulmonary arteries without going through the ventricle. This connection is called total cavopulmonary connection (TCPC). After surgery, the single ventricle supplies adequate and saturated systemic blood flow to the body; however, TCPC contains low pressure and low flow pulsatility. To overcome this problem, a new method is proposed wherein pulsatile blood will be directed to the TCPC through the stenosed main pulmonary artery. In this study, through the use of Computational Fluid Dynamics, T-shaped (MRI-based) and Y-shaped (computer-generated) geometries are compared in order to determine the influence of this modification on pulsation of blood flow as well as energy loss in pulmonary arteries. The results indicate that energy loss in Y-shaped geometry is far less than T-shaped geometry, while the difference in flow pulsatility is insignificant.

Organ-Tumor-on-a-Chip for Chemosensitivity Assay: A Critical Review.

With a mortality rate over 580,000 per year, cancer is still one of the leading causes of death worldwide. However, the emerging field of microfluidics can potentially shed light on this puzzling disease. Unique characteristics of microfluidic chips (also known as micro-total analysis system) make them excellent candidates for biological applications. The ex vivo approach of tumor-on-a-chip is becoming an indispensable part of personalized medicine and can replace in vivo animal testing as well as conventional in vitro methods. In tumor-on-a-chip, the complex three-dimensional (3D) nature of malignant tumor is co-cultured on a microfluidic chip and high throughput screening tools to evaluate the efficacy of anticancer drugs are integrated on the same chip. In this article, we critically review the cutting edge advances in this field and mainly categorize each tumor-on-a-chip work based on its primary organ. Specifically, design, fabrication and characterization of tumor microenvironment; cell culture technique; transferring mechanism of cultured cells into the microchip; concentration gradient generators for drug delivery; in vitro screening assays of drug efficacy; and pros and cons of each microfluidic platform used in the recent literature will be discussed separately for the tumor of following organs: (1) Lung; (2) Bone marrow; (3) Brain; (4) Breast; (5) Urinary system (kidney, bladder and prostate); (6) Intestine; and (7) Liver. By comparing these microchips, we intend to demonstrate the unique design considerations of each tumor-on-a-chip based on primary organ, e.g., how microfluidic platform of lung-tumor-on-a-chip may differ from liver-tumor-on-a-chip. In addition, the importance of heart⁻liver⁻intestine co-culture with microvasculature in tumor-on-a-chip devices for in vitro chemosensitivity assay will be discussed. Such system would be able to completely evaluate the absorption, distribution, metabolism, excretion and toxicity (ADMET) of anticancer drugs and more realistically recapitulate tumor in vivo-like microenvironment.

Three-dimensional simulation of urine concentrating mechanism in a functional unit of rat outer medulla. I. Model structure and base case results.

The urine formation and excretion system have long been of interest for mathematicians and physiologists to elucidate the obscurities within the process happens in renal tissue. In this study, a novel three-dimensional approach is utilized for modeling the urine concentrating mechanism in rat renal outer medulla which is essentially focused on demonstrating the significance of tubule's architecture revealed in anatomic studies and physiological literature. Since nephrons and vasculatures work interdependently through a highly structured arrangement in outer medulla which is dominated by vascular bundles, a detailed functional unit is proposed based on this specific configuration. Furthermore, due to relatively lesser influence of vasa recta on interstitial medullary osmolality and osmotic gradients as well as model structure simplicity, central core assumption is employed. The model equations are based on three spatial dimensional mass, momentum and species transport equations as well as standard expressions for solutes and water transmural transport. Our model can simulate preferential interactions between different tubules and it is shown that such interactions promote solute cycling and subsequently, enhance urine-concentrating capability. The numerical results are well consistent with tissue slice experiments and moreover, our model predicts more corticomedullary osmolality gradient in outer medulla than previous influential 1-D simulations.