One of the great challenges in science and engineering today is to develop technologies to improve the health of people in the poorest regions of the world. Here we integrated new procedures for manufacturing, fluid handling and signal detection in microfluidics into a single, easy-to-use point-of-care (POC) assay that faithfully replicates all steps of ELISA, at a lower total material cost. We performed this 'mChip' assay in Rwanda on hundreds of locally collected human samples. The chip had excellent performance in the diagnosis of HIV using only 1 μl of unprocessed whole blood and an ability to simultaneously diagnose HIV and syphilis with sensitivities and specificities that rival those of reference benchtop assays. Unlike most current rapid tests, the mChip test does not require user interpretation of the signal. Overall, we demonstrate an integrated strategy for miniaturizing complex laboratory assays using microfluidics and nanoparticles to enable POC diagnostics and early detection of infectious diseases in remote settings.

There is a growing interest in the pivotal role of exosomes in cancer and in their use as biomarkers. However, despite the importance of the microenvironment for cancer initiation and progression, monolayer cultures of tumor cells still represent the main in vitro source of exosomes. As a result, their environmental regulation remains largely unknown. Here, we report a three-dimensional tumor model for studying exosomes, using Ewing's sarcoma type 1 as a clinically relevant example. The bioengineered model was designed based on the hypothesis that the 3-dimensionality, composition and stiffness of the tumor matrix are the critical determinants of the size and cargo of exosomes released by the cancer cells. We analyzed the effects of the tumor microenvironment on exosomes, and the effects of exosomes on the non-cancer cells from the bone niche. Exosomes from the tissue-engineered tumor had similar size distribution as those in the patients' plasma, and were markedly smaller than those in monolayer cultures. Bioengineered tumors and the patients' plasma contained high levels of the Polycomb histone methyltransferase EZH2 mRNA relatively to their monolayer counterparts. Notably, EZH2 mRNA, a potential tumor biomarker detectable in blood plasma, could be transferred to the surrounding mesenchymal stem cells. This study provides the first evidence that an in vitro culture environment can recapitulate some properties of tumor exosomes.

Cellular communities in living tissues act in concert to establish intricate microenvironments, with complexity difficult to recapitulate in vitro. We report a method for docking numerous cellularized hydrogel shapes (100-1,000 μm in size) into hydrogel templates to construct 3D cellular microenvironments. Each shape can be uniquely designed to contain customizable concentrations of cells and molecular species, and can be placed into any spatial configuration, providing extensive compositional and geometric tunability of shapecoded patterns using a highly biocompatible hydrogel material. Using precisely arranged hydrogel shapes, we investigated migratory patterns of human mesenchymal stem cells and endothelial cells. We then developed a finite element gradient model predicting chemotactic directions of cell migration in micropatterned cocultures that were validated by tracking ∼2,500 individual cell trajectories. This simple yet robust hydrogel platform provides a comprehensive approach to the assembly of 3D cell environments.microtechnologies | tissue assembly | angiogenesis | modeling | diffusion B iological tissues are composed of cellular "building blocks" that cooperate to provide tissue-specific functions (1-6). Specific cells, molecules, and their geometric assembly establish a biological system, whether it is a vascular network surrounded by parenchymal cells (7), a developing tissue (8-11), or a metastatic tumor (12-16). In vitro culture systems designed to control the 3D presentation of multiple cells and molecular species in a biologically relevant matrix are needed to faithfully recapitulate intricate biological niches (17-23). Previous attempts to assemble complex tissue structures in vitro lacked the specificity and yield to control large numbers of cells and soluble factors (22-24), had limited resolution at the microscale (25, 26), and used matrices without adequate biological function (27-32). Therefore, spatial microenvironmental control of biological systems has been difficult to achieve. Such control is important in many processes, including the migratory formation of vasculature, where gradient patterns dictate growth of vascular sprouts. For example, endothelial cell (EC) stabilization via mesenchymal stem cell (MSC) interactions is known to facilitate the maturation of blood vessels impacting many physiologic systems, from tumors to engineered tissues (33-35). Recent studies clearly showed the importance of microscale gradients for vasculogenesis (36-38).We report a method by which microsized 3D hydrogels are shape-coded for their biological and physical properties and docked by iterative sedimentation into shape-matching hydrogel templates. Microenvironmental niches were fabricated using gelatin methacrylate (GelMA), a modified native protein with excellent biocompatibility, tunable mechanical properties, and micrometer-scale patterning resolution (39, 40). GelMA was molded into diverse geometric shapes with dimensions of 100-1,000 μm, after encapsulating cells or labeled molecular specie...

We describe a microfluidic system that can control, in real time, the microenvironments of mammalian cells in naturally derived 3D extracellular matrix (ECM). This chip combines pneumatically actuated valves with an individually addressable array of 3D cell-laden ECM; actuation of valves determines the pathways for delivering reagents through the chip and for exchanging diffusible factors between cell chambers. To promote rapid perfusion of reagents through 3D gels (with complete exchange of reagents within the gel in seconds), we created conduits above the gels for fluid flow, and microposts to stabilize the gels under high perfusion rates. As a biological demonstration, we studied spatially segregated mouse embryonic stem cells and mouse embryonic fibroblasts embedded in 3D Matrigel over days of culture. Overall, this system may be useful for high-throughput screening, single-cell analysis and studies of cell-cell communication, where rapid control of 3D cellular microenvironments is desired.

The challenging task of heart regeneration is being pursued in three related directions: derivation of cardiomyocytes from human stem cells, in vitro engineering and maturation of cardiac tissues, and development of methods for controllable cell delivery into the heart. In this review, we focus on tissue engineering methods that recapitulate biophysical signaling found during normal heart development and maturation. We discuss the use of scaffold-bioreactor systems for engineering functional human cardiac tissues, and the methods for delivering stem cells, cardiomyocytes and engineered tissues into the heart.

Despite the prevalence of microfluidic-based heterogeneous immunoassays (where analytes in solution are captured on a solid surface functionalized with a capture molecule), there is incomplete understanding of how assay parameters influence the amount of captured analytes. This study presents computational results and corresponding experimental binding assays in which the capture of analytes is studied under variations in both mass transfer and surface binding, constrained by real-world assay conditions of finite sample volume, assay time, and capture area. Our results identify: 1) a "reagent-limited" regime which exists only under the constraints of finite sample volume and assay time; 2) a critical flow rate (e.g. 0.5 microL min(-1) under our assay conditions) to gain the maximum signal with the fastest assay time; 3) an increase in signal by using a short concentrated plug (e.g. 5 microL, 100 nM) rather than a long dilute plug (e.g. 50 microL, 10 nM) of sample; 4) the possibility of spending a considerable fraction of the assay time out of the reaction-limited regime. Overall, an improved understanding of fundamental physical processes may be particularly beneficial for the design of point-of-care assays, where volumes of reagents and available samples are limited, and the desired time-to-result short.

Study Design Controlled laboratory study, case-control design. Objective To evaluate spine kinematics and gait characteristics in people with nonspecific chronic neck pain. Background People with chronic neck pain present with a number of sensorimotor and biomechanical alterations, yet little is known about the influence of neck pain on gait and motions of the spine during gait. Methods People with chronic nonspecific neck pain and age- and sex-matched asymptomatic controls walked on a treadmill at 3 different speeds (self-selected, 3 km/h, and 5 km/h), either with their head in a neutral position or rotated 30°. Tridimensional motion capture was employed to quantify body kinematics. Neck and trunk rotations were derived from the difference between the transverse plane component of the head and thorax and thorax and pelvis angles to provide an indication of neck and trunk rotation during gait. Results Overall, the patient group showed shorter stride length compared to the control group (P<.001). Moreover, the patients with neck pain showed smaller trunk rotations (P<.001), regardless of the condition or speed. The difference in the amount of trunk rotation between groups became larger for the conditions of walking with the head rotated. Conclusion People with chronic neck pain walk with reduced trunk rotation, especially when challenged by walking with their head positioned in rotation. Reduced rotation of the trunk during gait may have long-term consequences on spinal health. J Orthop Sports Phys Ther 2017;47(4):268-277. Epub 3 Feb 2017. doi:10.2519/jospt.2017.6768.

Current in vitro models fall short in deciphering the mechanisms of cardiac hypertrophy induced by volume overload. We developed a pneumatic microfluidic platform for high-throughput studies of cardiac hypertrophy that enables repetitive (hundreds of thousands of times) and robust (over several weeks) manipulation of cardiac μtissues. The platform is reusable for stable and reproducible mechanical stimulation of cardiac μtissues (each containing only 5000 cells). Heterotypic and homotypic μtissues produced in the device were pneumatically loaded in a range of regimes, with real-time on-chip analysis of tissue phenotypes. Concentrated loading of the three-dimensional cardiac tissue faithfully recapitulated the pathology of volume overload seen in native heart tissue. Sustained volume overload of μtissues was sufficient to induce pathological cardiac remodeling associated with upregulation of the fetal gene program, in a dose-dependent manner.