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Biomedical subjects

Kahp Y Suh

Publications and source records attributed to Kahp Y Suh.

At least 19 recordsLinked to original sources

Fabrication of non-biofouling polyethylene glycol micro- and nanochannels by ultraviolet-assisted irreversible sealing.

We present a simple and widely applicable method to fabricate micro- and nanochannels comprised entirely of crosslinked polyethylene glycol (PEG) by using UV-assisted irreversible sealing to bond partially crosslinked PEG surfaces. The method developed here can be used to form channels as small as approximately 50 nm in diameter without using a sophisticated experimental setup. The manufactured channel is a homogeneous conduit made completely from non-biofouling PEG, exhibits robust sealing with minimal swelling and can be used without additional surface modification chemistries, thus significantly enhancing reliability and durability of microfluidic devices. Furthermore, we demonstrate simple analytical assays using PEG microchannels combined with patterned arrays of supported lipid bilayers (SLBs) to detect ligand (biotin)-receptor (streptavidin) interactions.

Acrylates↗

Pumpless, selective docking of yeast cells inside a microfluidic channel induced by receding meniscus.

We present a simple cell docking method induced by receding meniscus to capture non-adherent yeast cells onto microwells inside a microfluidic channel. Microwells were fabricated either by capillary moulding of UV curable polyurethane acrylate (PUA) onto glass substrate or direct replica moulding of poly(dimethyl siloxane) (PDMS). A cell suspension of the budding yeast, Saccharomyces cerevisiae, was introduced into the microfluidic channel by surface tension driven capillary flow and a receding meniscus was subsequently generated by evaporation. As the meniscus progressed, one to multiple yeast cells were spontaneously captured onto microwells by lateral capillary force created at the bottom of the meniscus. Using this cell-based platform, we observed the response of yeast cells upon stimulation by a mating pheromone (alpha-factor) by monitoring the expression of green fluorescent protein (GFP) with time. It was observed that alpha-factor triggered the expression of GFP at 60 min after stimulation and the fluorescence intensity was sustained for an additional 60 min without changes.

Cells, Immobilized↗

Anisotropic rupture of polymer strips driven by Rayleigh instability.

We demonstrate that the separated polymer strips of micro- and sub-micro-length-scales rupture anisotropically along the strip direction, resulting in the formation of distinctly observable, regularly spaced polymer drops. The wavelength of the polymer drops and the surface tension dependence of the rupture behavior are found to be well represented by a relationship derived on the basis of Rayleigh instability. The period is proportional to the square root of the cross-sectional area of the strip and the proportionality constant depends on the contact angle. The rupture of polymer strips into polymer blocks instead of drops, which result when annealed with physically confining walls in place, is found to be well described by the same relationship.

Journal Article↗

Guided three-dimensional growth of functional cardiomyocytes on polyethylene glycol nanostructures.

We introduce well-defined nanopillar arrays of a poly(ethylene glycol) (PEG) hydrogel as a cell culture platform to guide a 3D construct of primary rat cardiomyocytes in vitro for potential tissue engineering applications. Ultraviolet (UV)-assisted capillary lithography was used to fabricate highly uniform approximately 150 nm PEG pillars with approximately 400 nm height. It was found that cell adhesion was significantly enhanced on PEG nanopillars (132 +/- 29 cells/mm2) compared to that on the bare PEG control (39 +/- 17 cells/mm2) (p < 0.05) but substantially reduced compared to that on the glass control (502 +/- 45 cells/mm2) (p < 0.01). Furthermore, in colonizing cardiomyocytes, the nanopillars stimulated self-assembled aggregates among the contacting cells with 3D growth, which is a unique feature for nanopatterned PEG hydrogels as a cell culture substrate. The 3D-grown cardiomyocytes retained their conductive and contractile properties, as evidenced by the observation of beating cardiomyocytes with robust action potential generation.

Animals↗

Nanoengineered multiscale hierarchical structures with tailored wetting properties.

A simple method for fabricating micro/nanoscale hierarchical structures is presented using a two-step temperature-directed capillary molding technique. This lithographic method involves a sequential application of the molding process in which a uniform polymer-coated surface is molded with a patterned mold by means of capillary force above the glass transition temperature of the polymer. Various microstructures and nanostructures were fabricated with minimum resolution down to approximately 50 nm with good reproducibility. Also contact angle measurements of water indicated that two wetting states coexist on a multiscale hierarchical structure where heterogeneous wetting is dominant for the microstructure and homogeneous wetting for the nanostructure. A simple theoretical model combining these two wetting states was presented, which was in good agreement with the experimental data. Using this approach, multiscale hierarchical structures for biomimetic functional surfaces can be fabricated with precise control over geometrical parameters and the wettability of a solid surface can be tailored in a controllable manner.

Journal Article↗

Formation of regular nanoscale undulations on a thin polymer film imprinted by a soft mold.

We observed the formation of regular nanoscale undulations on a polystyrene film when imprinted by a soft poly(dimethylsiloxane) mold above the polymer's glass transition temperature. The shape of the wave was reminiscent of a buckling wave frequently observed for a metal film supported on an elastomeric substrate. We derived a simple theoretical model based on an anisotropic buckling of the polymer film rigidly bound to a substrate, which agrees well with the experiment.

Journal Article↗

Direct confinement of individual viruses within polyethylene glycol (PEG) nanowells.

Individual M13 viruses were spatially confined within wells fabricated from nanomolding of a PEG-based random copolymer. The viruses were selectively adhered to the region pretreated with an antibody against the virus, resulting in individual virus arrays. The polymer surface was found to be highly resistant to the attachment of the virus (approximately 0.02 microm-2), approximately 2 orders of magnitude lower than that on a bare silicon surface. The physical height of the template provided an additional barrier to the attachment of the virus due to entropic penalty in bending of a semi-flexible M13 virus. The effects of pattern size and barrier height were investigated, revealing that a certain critical height is needed to ensure successful confinement within the template for a given pattern size.

Bacteriophage M13↗

Stretched polymer nanohairs by nanodrawing.

A simple, yet innovative, method is presented for fabricating high-aspect-ratio polymer nanohairs (aspect ratio >20) on a solid substrate by sequential application of molding and drawing of a thin polymer film. The polymer film was prepared by spin coating on a rigid or flexible substrate, and the temperature was raised above the polymer's glass transition while in conformal contact with a poly(urethane acrylate) mold having nanocavities. Consequently, capillary forces induced deformation of the polymer melt into the void spaces of the mold and the filled nanostructure was further elongated upon removal of the mold due to tailored adhesive force at the mold/polymer interface. The optimum value of the work of adhesion at the mold/polymer interface ranged from 0.9 to 1.1 times that at the substrate/polymer interface.

Capillary Action↗

Soft lithographic patterning of supported lipid bilayers onto a surface and inside microfluidic channels.

We present simple soft lithographic methods for patterning supported lipid bilayer (SLB) membranes onto a surface and inside microfluidic channels. Micropatterns of polyethylene glycol (PEG)-based polymers were fabricated on glass substrates by microcontact printing or capillary moulding. The patterned PEG surfaces have shown 97 +/- 0.5% reduction in lipid adsorption onto two dimensional surfaces and 95 +/- 1.2% reduction inside microfluidic channels in comparison to glass control. Atomic force microscopy measurements indicated that the deposition of lipid vesicles led to the formation of SLB membranes by vesicle fusion due to hydrophilic interactions with the exposed substrate. Furthermore, the functionality of the patterned SLBs was tested by measuring the binding interactions between biotin (ligand)-labeled lipid bilayer and streptavidin (receptor). SLB arrays were fabricated with spatial resolution down to approximately 500 nm on flat substrate and approximately 1 microm inside microfluidic channels, respectively.

Dimethylpolysiloxanes↗

Control over wettability of polyethylene glycol surfaces using capillary lithography.

We demonstrate that wettability of poly(ethylene glycol) (PEG) surfaces can be controlled using nanostructures with various geometrical features. Capillary lithography was used to fabricate PEG nanostructures using a new ultraviolet (UV) curable mold consisting of functionalized polyurethane with acrylate group (MINS101m, Minuta Tech.). Two distinct wetting states were observed depending of the height of nanostructures. At relatively lower heights (< 300 nm for 150 nm pillars with 500 nm spacing), the initial contact angle of water was less than 80 degrees and the water droplet easily invaded into the surface grooves, leading to a reduced contact angle at equilibrium (Wenzel state). At relatively higher heights (> 400 nm for 150 nm pillars with 500 nm spacing), on the other hand, the nanostructured PEG surface showed hydrophobic nature and no significant change in contact angle was observed with time (Cassie state). The presence of two wetting states was also confirmed by dynamic wetting properties and contact-angle hysteresis. The wetting transition from hydrophilic (bare PEG surface) to hydrophobic (PEG nanostructures) was described by the Cassie-Baxter equation assuming that enhanced hydrophobicity is due to the heterogeneous wetting mediated by an air pocket on the surface. The measured contact angles in the Cassie state were increased with increasing air fraction, in agreement with the theoretical prediction.

Journal Article↗

Characterization of chemisorbed hyaluronic acid directly immobilized on solid substrates.

Hyaluronic acid (HA) has a number of potential biomedical applications in drug delivery and tissue engineering. For these applications, a prerequisite is to understand the characteristic of HA films directly immobilized to solid substrates. Here, we demonstrate that high molecular weight HA can be directly immobilized onto hydrophilic substrates without any chemical manipulation, allowing for the formation of an ultrathin chemisorbed layer. Hyaluronic acid is stabilized on these surfaces through hydrogen bonding between the hydrophilic moieties in HA [such as carboxylic acid (-COOH) or hydroxyl (-OH) groups] with silanol (-SiOH), carboxylic acid or hydroxyl groups on the hydrophilic substrates. Despite the water solubility, the chemisorbed HA layer remained stable on glass or silicon oxide substrates for at least 7 days in phosphate-buffered saline. Furthermore, HA immobilized on silicon and other dioxide surfaces in much higher quantities than other polysaccharides including dextran sulfate, heparin, heparin sulfate, chondroitin sulfate, dermatan sulfate, and alginic acid. This behavior is related to the molecular entanglement and intrinsic stiffness of HA as a result of strong internal and external hydrogen bonding as well as high molecular weight. These results demonstrate that HA can be used to coat surfaces through direct immobilization.

Absorption↗

Molded polyethylene glycol microstructures for capturing cells within microfluidic channels.

The ability to control the deposition and location of adherent and non-adherent cells within microfluidic devices is beneficial for the development of micro-scale bioanalytical tools and high-throughput screening systems. Here, we introduce a simple technique to fabricate poly(ethylene glycol)(PEG) microstructures within microfluidic channels that can be used to dock cells within pre-defined locations. Microstructures of various shapes were used to capture and shear-protect cells despite medium flow in the channel. Using this approach, PEG microwells were fabricated either with exposed or non-exposed substrates. Proteins and cells adhered within microwells with exposed substrates, while non-exposed substrates prevented protein and cell adhesion (although the cells were captured inside the features). Furthermore, immobilized cells remained viable and were stained for cell surface receptors by sequential flow of antibodies and secondary fluorescent probes. With its unique strengths in utility and control, this approach is potentially beneficial for the development of cell-based analytical devices and microreactors that enable the capture and real-time analysis of cells within microchannels, irrespective of cell anchorage properties.

Animals↗

Single nanocrystal arrays on patterned poly(ethylene glycol) copolymer microstructures using selective wetting and drying.

Single nanocrystal arrays were fabricated on sub-microwells of poly(ethylene glycol) (PEG) copolymer using selective wetting on the hydrophilic regions of the exposed substrate surface and subsequent drying. Templates were produced by molding a thin film of a PEG-based random copolymer on hydrophilic substrates such as glass or silicon dioxide. The polymeric microstructures provide a topographical barrier around the well, which makes it possible to create nanocrystal arrays with controlled geometrical features. The size of the nanocrystal was found to decrease with decreasing well size and also decrease with decreasing topological height. A simple empirical equation was derived to predict the size of the crystal as a function of the pattern size and height, which is in good agreement with the experimental data.

Journal Article↗

Polymer elasticity-driven wrinkling and coarsening in high temperature buckling of metal-capped polymer thin films.

We report the critical effects the deformational stress from the elastic nature of a confined polymer layer has during the relaxation process on the buckling of thin metal-polymer bilayer systems (less than 100 nm) even above the temperature at which the polymer is in the liquid flow region. In contrast with what is generally believed, the dispersion force does not play a significant role in the buckling. We also find that the final wrinkled waves take on the shape of wormlike islands. The coarsening leading to the island structure is driven by the growth in amplitude of the dominant wave at the expense of less dominant ones.

Journal Article↗

A soft lithographic approach to fabricate patterned microfluidic channels.

The control of surface properties and spatial presentation of functional molecules within a microfluidic channel is important for the development of diagnostic assays and microreactors and for performing fundamental studies of cell biology and fluid mechanics. Here, we present a simple technique, applicable to many soft lithographic methods, to fabricate robust microchannels with precise control over the spatial properties of the substrate. In this approach, the patterned regions were protected from oxygen plasma by controlling the dimensions of the poly(dimethylsiloxane) (PDMS) stamp and by leaving the stamp in place during the plasma treatment process. The PDMS stamp was then removed, and the microfluidic mold was irreversibly bonded to the substrate. The approach was used to pattern a nonbiofouling poly(ethylene glycol)-based copolymer or the polysaccharide hyaluronic acid within microfluidic channels. These nonbiofouling patterns were then used to fabricate arrays of fibronectin and bovine serum albumin as well as mammalian cells. In addition, further control over the deposition of multiple proteins onto multiple or individual patterns was achieved using laminar flow. Also, cells that were patterned within channels remained viable and capable of performing intracellular reactions and could be potentially lysed for analysis.

Animals↗

Layer-by-layer deposition of hyaluronic acid and poly-L-lysine for patterned cell co-cultures.

A novel method for patterning cellular co-cultures that uses the layer-by-layer deposition of ionic biopolymers is described. Non-biofouling hyaluronic acid (HA) micropatterns were used to immobilize cells and proteins to glass substrates. Subsequent ionic adsorption of poly-L-lysine (PLL) to HA patterns was used to switch the HA surfaces from cell repulsive to adherent thereby facilitating the adhesion of a second cell type. The utility of this approach to pattern co-cultures of hepatocytes or embryonic stem cells with fibroblasts was demonstrated. In addition, the versatility of this approach to generate patterned co-cultures irrespective of the primary cell seeding and relative adhesion of the seeded cells was demonstrated. Thus, the proposed method may be a useful tool for fabricating controlled cellular co-cultures for cell-cell interaction studies and tissue engineering applications.

Adsorption↗

A simple soft lithographic route to fabrication of poly(ethylene glycol) microstructures for protein and cell patterning.

We present a simple, direct soft lithographic method to fabricate poly(ethylene glycol) (PEG) microstructures for protein and cell patterning. This lithographic method involves a molding process in which a uniform PEG film is molded with a patterned polydimethylsiloxane stamp by means of capillary force. The patterned surfaces created by this method provide excellent resistance towards non-specific protein and cell adsorption. The patterned substrates consist of two regions: the molded PEG surface that acts as a resistant layer and the exposed substrate surface that promotes protein or cell adsorption. A notable finding here is that the substrate surface can be directly exposed during the molding process due to the ability to control the wetting properties of the polymer on the stamp, which is a key factor to patterning proteins and cells.

Adsorption↗