Wentao Shi

Characterization of vascular permeability using a biomimetic microfluidic blood vessel model

The inflammatory response in endothelial cells (ECs) leads to an increase in vascular permeability through the formation of gaps. However, the dynamic nature of vascular permeability and external factors involved is still elusive. In this work, we use a biomimetic blood vessel (BBV) microfluidic model to measure in real-time the change in permeability of the EC layer under culture in physiologically relevant flow conditions. This platform studies the dynamics and characterizes vascular permeability when the EC layer is triggered with an inflammatory agent using tracer molecules of three different sizes, and the results are compared to a transwell insert study. We also apply an analytical model to compare the permeability data from the different tracer molecules to understand the physiological and bio-transport significance of endothelial permeability based on the molecule of interest. A computational model of the BBV model is also built to understand the factors influencing transport of molecules of different sizes under flow. The endothelial monolayer cultured under flow in the BBV model was treated with thrombin, a serine protease that induces a rapid and reversible increase in endothelium permeability. On analysis of permeability data, it is found that the transport characteristics for fluorescein isothiocyanate (FITC) dye and FITC Dextran 4k Da molecules are similar in both BBV and transwell models, but FITC Dextran 70k Da molecules show increased permeability in the BBV model as convection flow (Peclet number > 1) influences the molecule transport in the BBV model. We also calculated from permeability data the relative increase in intercellular gap area during thrombin treatment for ECs in the BBV and transwell insert models to be between 12% and 15%. This relative increase was found to be within range of what we quantified from F-actin stained EC layer images. The work highlights the importance of incorporating flow in in vitro vascular models, especially in studies involving transport of large size objects such as antibodies, proteins, nano/micro particles, and cells.

Generation of Customizable Micro-wavy Pattern through Grayscale Direct Image Lithography.

With the increasing amount of research work in surface studies, a more effective method of producing patterned microstructures is highly desired due to the geometric limitations and complex fabricating process of current techniques. This paper presents an efficient and cost-effective method to generate customizable micro-wavy pattern using direct image lithography. This method utilizes a grayscale Gaussian distribution effect to model inaccuracies inherent in the polymerization process, which are normally regarded as trivial matters or errors. The measured surface profiles and the mathematical prediction show a good agreement, demonstrating the ability of this method to generate wavy patterns with precisely controlled features. An accurate pattern can be generated with customizable parameters (wavelength, amplitude, wave shape, pattern profile, and overall dimension). This mask-free photolithography approach provides a rapid fabrication method that is capable of generating complex and non-uniform 3D wavy patterns with the wavelength ranging from 12 μm to 2100 μm and an amplitude-to-wavelength ratio as large as 300%. Microfluidic devices with pure wavy and wavy-herringbone patterns suitable for capture of circulating tumor cells are made as a demonstrative application. A completely customized microfluidic device with wavy patterns can be created within a few hours without access to clean room or commercial photolithography equipment.

Shear induced alignment of short nanofibers in 3D printed polymer composites

3D printing of composite materials offers an opportunity to combine the desired properties of composite materials with the flexibility of additive manufacturing in geometric shape and complexity. In this paper, the shear-induced alignment of aluminum oxide nanowires during stereolithography printing was utilized to fabricate a nanowire reinforced polymer composite. To align the fibers, a lateral oscillation mechanism was implemented and combined with wall pattern printing technique to generate shear flow in both vertical and horizontal directions. A series of specimens were fabricated for testing the composite materials tensile strength. The results showed that mechanical properties of the composite were improved by reinforcement of nanofibers through shear induced alignment. The improvement of tensile strength was approximately ∼28% by aligning the nanowires at 5 wt% (∼1.5% volume fraction) loading of aluminum oxide nanowires.

Short fiber reinforced 3d printed ceramic composite with shear induced alignment

This paper accounts for utilization of shear induced alignment method during ceramic stereolithography. Lateral oscillation mechanism, combined with 3d printed wall pattern, was employed to generate necessary shear to align fiber in desired direction. First, semicircular channel pattern was printed to assess the effect of difference between wall direction and oscillation direction on the fiber alignment. Then, flexural strength of ceramic matrix was tested with nickel coated carbon fiber and ceramic fiber reinforcements. The results demonstrated that the shear induced alignment further improves the flexural strength compare to randomly distributed samples. Flexural strength of aligned samples with 1.0 wt% carbon fiber loading was improved by ~90% compared to randomly orientated samples and by ~333% compared to unreinforced samples. Finally, fracture surface morphology of the flexural strength test specimens was evaluated. The main fracture mechanism was observed as fiber pull-out.

Facile Tumor Spheroids Formation in Large Quantity with Controllable Size and High Uniformity

A facile method for generation of tumor spheroids in large quantity with controllable size and high uniformity is presented. HCT-116 cells are used as a model cell line. Individual tumor cells are sparsely seeded onto petri-dishes. After a few days of growth, separated cellular islets are formed and then detached by dispase while maintaining their sheet shape. These detached cell sheets are transferred to dispase-doped media under orbital shaking conditions. Assisted by the shear flow under shaking and inhibition of cell-to-extracellular matrix junctions by dispase, the cell sheets curl up and eventually tumor spheroids are formed. The average size of the spheroids can be controlled by tuning the cell sheet culturing period and spheroid shaking period. The uniformity can be controlled by a set of sieves which were home-made using stainless steel meshes. Since this method is based on simple petri-dish cell culturing and shaking, it is rather facile for forming tumor spheroids with no theoretical quantity limit. This method has been used to form HeLa, A431 and U87 MG tumor spheroids and application of the formed tumor spheroids in drug screening is also demonstrated. The viability, 3D structure, and necrosis of the spheroids are characterized.

A Facile Way to Fabricate Transparent Superhydrophobic Surfaces

A fast, easy, and low-cost way to fabricate transparent superhydrophobic (SHP) surfaces is developed. By simply mixing silica nanoparticles (SiNPs), polydimethylsiloxane (PDMS) and heptane to form a suspension, dip- or drop-coating the suspension onto different surfaces, transparent SHP surfaces can be obtained. By tuning the ratio of the three components above, transparency of the coating can reach more than 90% transmittance in the visible region, while static water contact angle of the coating can reach as high as 162°. Dynamic contact angle study shows the advancing contact angle and receding contact angle of water can be as high as 168° and 161°, and the resulting contact angle hysteresis can be as low as 7°. The reported facile way of fabricating transparent superhydrophobic (SHP) surfaces is potential for applications which need both optical transparency and self-cleaning capability, such as solar cells, optical equipment, and visible microfluidic chips.

Antibody-coated nanoparticles are promising molecular probes for microscopic analysis of cell behavior

Cells are complex systems that respond to local environment and interact with their neighbors through various receptors expressed on the surface. Because of the presence of artifacts associated with microscope images during the fixing process, microscopy assays of fixed and stained cells cannot provide enough information to characterize cellular behaviors and cell–cell interactions [1]. Therefore, development of novel technologies capable of monitoring cellular scale events in real time is of significant importance for providing valuable kinetic and spatial information of cells [2]. Highly sensitive fluorescence microscopy has been widely used for real-time investigation and monitoring of spatiotemporal cellular events such as cell adhesion, cell migration as well as cell–cell, viral–cell, protein–cell and protein–protein interactions [3–5]. For instance, genetically labeled cells with green fluorescent protein (GFP) are being used to visualize cancer cells and metastatic process in live tissue or in the intact animal by whole-body imaging. The development of functional fluorophores and the discovery of GFP-modified versions such as yellow fluorescent protein and cyan fluorescent protein have provided the platform to image several events simultaneously at the single-cell level [1]. One example would be usage of multicolor fluorescence microscopy to study the entry of nanoparticles (NPs) and viruses into living or fixed cells. Dualcolor fluorescent imaging technique is also utilized for subcellular real-time imaging of cancer cell trafficking. By labeling cancer cells with GFP in the nucleus and red fluorescent protein in the cytoplasm, cancer cell trafficking in lymphatic vessels of nude live mice are studied using whole-body imaging. Fluorescent microscopy has further been combined with other microscopy techniques such as atomic force microscopy or electron microscopy to help broaden the understanding of subtle biological interactions and processes [6]. However, the labeling procedures are usually complex and the chemical modification introduced by the fluorescent probe can affect normal cell behavior [7].