Seung Yun Yang

Cell-laden Microengineered and Mechanically Tunable Hybrid Hydrogels of Gelatin and Graphene Oxide

Incorporating graphene oxide inside GelMA hydrogels enhances their mechanical properties and reduces UV-induced cell damage while preserving their favorable characteristics for 3D cell encapsulation. NIH-3T3 fibroblasts encapsulated in GO-GelMA microgels demonstrate excellent cellular viability, proliferation, spreading, and alignment. GO reinforcement combined with a multi-stacking approach offers a facile engineering strategy for the construction of complex artificial tissues.

A bio-inspired swellable microneedle adhesive for mechanical interlocking with tissue

Achieving significant adhesion to soft tissues while minimizing tissue damage poses a considerable clinical challenge. Chemical-based adhesives require tissue-specific reactive chemistry, typically inducing a significant inflammatory response. Staples are fraught with limitations including high-localized tissue stress and increased risk of infection, and nerve and blood vessel damage. Here, inspired by the endoparasite Pomphorhynchus laevis which swells its proboscis to attach to its host’s intestinal wall, we have developed a biphasic microneedle array that mechanically interlocks with tissue through swellable microneedle tips, achieving ~ 3.5 fold increase in adhesion strength compared to staples in skin graft fixation, and removal force of ~ 4.5 N/cm2 from intestinal mucosal tissue. Comprising a poly(styrene)-block-poly(acrylic acid) swellable tip and non-swellable polystyrene core, conical microneedles penetrate tissue with minimal insertion force and depth, yet high adhesion strength in their swollen state. Uniquely, this design provides universal soft tissue adhesion with minimal damage, less traumatic removal, reduced risk of infection and delivery of bioactive therapeutics.

Microstructured barbs on the North American porcupine quill enable easy tissue penetration and difficult removal

North American porcupines are well known for their specialized hairs, or quills that feature microscopic backward-facing deployable barbs that are used in self-defense. Herein we show that the natural quill’s geometry enables easy penetration and high tissue adhesion where the barbs specifically contribute to adhesion and unexpectedly, dramatically reduce the force required to penetrate tissue. Reduced penetration force is achieved by topography that appears to create stress concentrations along regions of the quill where the cross sectional diameter grows rapidly, facilitating cutting of the tissue. Barbs located near the first geometrical transition zone exhibit the most substantial impact on minimizing the force required for penetration. Barbs at the tip of the quill independently exhibit the greatest impact on tissue adhesion force and the cooperation between barbs in the 0–2 mm and 2–4 mm regions appears critical to enhance tissue adhesion force. The dual functions of barbs were reproduced with replica molded synthetic polyurethane quills. These findings should serve as the basis for the development of bio-inspired devices such as tissue adhesives or needles, trocars, and vascular tunnelers where minimizing the penetration force is important to prevent collateral damage.

Visible Light-Triggered On-Demand Drug Release from Hybrid Hydrogels and its Application in Transdermal Patches

On-demand release from stimuli-responsive hydrogels has received great attention due to an increasing clinical need. Here, we have prepared spherical hydrogel beads showing visible light-induced volume change at body temper-ature. By spray injection of the monomer solution using the alginate templ-ating method, hybrid beads of several hundred micrometers, consisting of temperature-responsive poly(N-isopropylacrylamide-co-vinyl-2-pyrrolidinone) hydrogel and magnetite nanoparticles (MNP), are produced. MNP dispersed in the hydrogel matrix absorbed visible light and generated heat, increasing the temperature of the matrix and resulting in shrinkage of the beads proportional to light intensity. It is demonstrated that light-induced volume change of dexamethasone-loaded hybrid beads result in on-demand and localized release of the drug by exposure to moderate visible light. As a potential application of the light-sensitive hybrid hydrogel beads, a transdermal patch is developed that incorporates drug-loaded hydrogel beads in multiple drug reservoirs, achieving enhanced release of a model drug when exposed to visible light. This platform should be applicable to on-demand, sequential, and long-term release of drugs via light exposure.

A self-adherent, bullet-shaped microneedle patch for controlled transdermal delivery of insulin

Abstract Proteins are important biologic therapeutics used for the treatment of various diseases. However, owing to low bioavailability and poor skin permeability, transdermal delivery of protein therapeutics poses a significant challenge. Here, we present a new approach for transdermal protein delivery using bullet‐shaped double‐layered microneedle (MN) arrays with water‐swellable tips. This design enabled the MNs to mechanically interlock with soft tissues by selective distal swelling after skin insertion. Additionally, prolonged release of loaded proteins by passive diffusion through the swollen tips was obtained. The bullet‐shaped MNs provided an optimal geometry for mechanical interlocking, thereby achieving significant adhesion strength (˜ 1.6 N cm− 2) with rat skin. By harnessing the MNs reversible swelling/deswelling property, insulin, a model protein drug, was loaded in the swellable tips using a mild drop/dry procedure. The insulin‐loaded MN patch released 60% of insulin when immersed in saline over the course of 12 h and approximately 70% of the released insulin appeared to have preserved structural integrity. An in vivo pilot study showed a prolonged release of insulin from swellable MN patches, leading to a gradual decrease in blood glucose levels. This self‐adherent transdermal MN platform can be applied to a variety of protein drugs requiring sustained release kinetics. Graphical abstract A bio‐inspired, self‐adherent microneedle (MN) patch is designed for effective transdermal protein drug delivery. The dual‐functional MN patch achieved not only a firm adhesion to live animal skin tissue, but also a prolonged insulin drug delivery following a mild loading process into swellable tips with minimal loss of biofunctionality. This transdermal delivery platform using self‐adherent MN patches can be applied for a variety of protein drugs requiring sustained release kinetics. Figure. No Caption available.

A Portable Chemotaxis Platform for Short and Long Term Analysis

Flow-based microfluidic systems have been widely utilized for cell migration studies given their ability to generate versatile and precisely defined chemical gradients and to permit direct visualization of migrating cells. Nonetheless, the general need for bulky peripherals such as mechanical pumps and tubing and the complicated setup procedures significantly limit the widespread use of these microfluidic systems for cell migration studies. Here we present a simple method to power microfluidic devices for chemotaxis assays using the commercially available ALZET® osmotic pumps. Specifically, we developed a standalone chemotaxis platform that has the same footprint as a multiwell plate and can generate well-defined, stable chemical gradients continuously for up to 7 days. Using this platform, we validated the short-term (24 hours) and long-term (72 hours) concentration dependent PDGF-BB chemotaxis response of human bone marrow derived mesenchymal stem cells.

Sample-free quantification of blood biomarkers via laser-treated skin

Surface modified microneedle (MN) arrays are being developed to capture circulating biomarkers from the skin, but inefficiency and unreliability of the current method limit its clinical applications. We describe here that illumination of a tiny area of the skin with hemoglobin-preferably absorbent laser increased the amount of circulating biomarkers in the upper dermis by more than 1000-fold. The hemoglobin-specific light altered the permeability of capillaries leading to extravasation of molecules but not blood cells beneath the skin involved. When specific probe-coated MN arrays were applied into the laser-treated skin, the biomarkers accumulated in the upper dermis were reliably, accurately, and sufficiently captured as early as 15 min of the assay. The maximal binding occurred in 1 h in a manner independent of penetration depth or a molecular mass of the biomarker. With anti-fluorescein isothiocyanate (FITC)-MNs, we were able to measure blood concentrations of FITC in mice receiving FITC intravenously. The sensitivity and accuracy were comparable to those attained by fluorescence spectrophotometer. Likewise, MNs containing influenza hemagglutinin (HA) could detect anti-HA antibody in mice or swine receiving influenza vaccines as effectively as standard immunoassays. The novel, minimally invasive approach holds great promise for measurement of multiple biomarkers by a single array for point-of-care diagnosis.

Sub-100 nm gold nanohole-enhanced Raman scattering on flexible PDMS sheets.

Surface-enhanced Raman spectroscopy (SERS) is a highly sensitive vibrational spectroscopy technique enabling detection of multiple analytes at the molecular level in a nondestructive and rapid manner. In this work, we introduce a new approach to fabricate deep subwavelength-scaled (sub-100 nm) metallic nanohole arrays (quasi-3D metallic nanoholes) on flexible and highly efficient SERS substrates. Target structures have been fabricated using a two-step process consisting of (i) direct pattern transfer of spin-coated polymer films onto polydimethylsiloxane (PDMS) substrates by plasma etching with transferred anodic aluminum oxide masks, and (ii) producing SERS-active substrates by functionalization of the etched polymeric films followed by Au deposition. Such an all-dry, top-down lithographic approach enables on-demand patterning of SERS-active metallic nanoholes with high structural fidelity even onto flexible and stretchable substrates, thus making possible multiple sensing modes in a versatile fashion. For example, metallic nanoholes on flexible PDMS substrates are highly amenable to their integration with curved glass sticks, which can be used in optical fiber-integrated SERS systems. Au surfaces immobilized by probe DNA molecules show a selective enhancement of Raman scattering with Cy5-labeled complementary DNA (as compared to flat Au surfaces), demonstrating the potential of using the quasi-3D Au nanohole arrays for bio-sensing applications.

Photo-crosslinkable comb-type copolymers bearing a benzophenone moiety for the enhanced swelling kinetics of hydrogels

Photo-polymerization has been intensively studied for the preparation of hydrogels because of several attractive features, including independent spatial control with photomasks, mild reaction conditions, and a simple procedure. Benzophenone (BP) and its derivatives are representative photoactive materials for the crosslinking of polymer chains; however, the incorporation of BP in a polymer chain results in low solubility in water, low swelling ratio, and slow swelling kinetics of the hydrogels. In this work, we have synthesized photo-crosslinkable copolymers of grafted PNIPAm and a benzophenone moiety. We could fabricate the hydrogels by UV illumination of the synthesized copolymers, and found that the fabricated hydrogels showed enhanced swelling kinetics through free chains of comb-type copolymers. We also demonstrated volume changes of the 3D hydrogel triggered by temperature, suggesting that the prepared copolymers can be applied to the fabrication of stimuli-responsive and fast responding origami patterns, actuators, and so on.

Biocompatibility of a PLA-based composite containing hydroxyapatite derived from waste bones of dolphin Neophocaena asiaeorientalis

Natural hydroxyapatite (HA), derived from waste bones of several animal species, has received much attention as a material for bone grafts and fillers and has a role as a coating for metal implants because of its biocompatibility and non-toxicity. To investigate the applicability of HA derived from waste bones of novel animal sources, the biocompatibility and toxicity of a poly-l-lactic acid (PLA)-based composite containing HA derived from the backbone of the dolphin Neophocaena asiaeorientalis (HANA) were examined in Sprague-Dawley (SD) rats. HANA powder showed X-ray diffraction peak patterns that corresponded to those of standard HA. Among five composites prepared from different combinations of PLA and HANA (7:3, 6:4, 5:5, 4:6, and 3:7), a PLA/HANA composite manufactured with a 6:4 PLA:HANA ratio had high surface roughness (453xa0nm), 10.3xa0N of maximum load, and 451.9xa0MPa of module elasticity. After implantation in the subcutaneous region of SD rats for 8xa0weeks, the amount of confluent, aggregated structures of multilayered cells on the PLA/HANA implant surface was greater than that on the PLA surface, although both implants were completely covered with adhesive cells. During the implant period, the initial intact form of the PLA/HANA composite broke into small fragments with few inflammatory cells in the contact region and no indication of significant toxicity. Taken together, the results suggest that HANA may have good biocompatibility and be non-toxic as it did not induce an immune response in SD rats.