Jin-Ho Park

Electrically conductive graphene/polyacrylamide hydrogels produced by mild chemical reduction for enhanced myoblast growth and differentiation

Graphene and graphene derivatives, such as graphene oxide (GO) and reduced GO (rGO), have been extensively employed as novel components of biomaterials because of their unique electrical and mechanical properties. These materials have also been used to fabricate electrically conductive biomaterials that can effectively deliver electrical signals to biological systems. Recently, increasing attention has been paid to electrically conductive hydrogels that have both electrical activity and a tissue-like softness. In this study, we synthesized conductive graphene hydrogels by mild chemical reduction of graphene oxide/polyacrylamide (GO/PAAm) composite hydrogels to obtain conductive hydrogels. The reduced hydrogel, r(GO/PAAm), exhibited muscle tissue-like stiffness with a Youngs modulus of approximately 50kPa. The electrochemical impedance of r(GO/PAAm) could be decreased by more than ten times compared to that of PAAm and unreduced GO/PAAm. In vitro studies with C2C12 myoblasts revealed that r(GO/PAAm) significantly enhanced proliferation and myogenic differentiation compared with unreduced GO/PAAm and PAAm. Moreover, electrical stimulation of myoblasts growing on r(GO/PAAm) graphene hydrogels for 7days significantly enhanced the myogenic gene expression compared to unstimulated controls. As results, our graphene-based conductive and soft hydrogels will be useful as skeletal muscle tissue scaffolds and can serve as a multifunctional platform that can simultaneously deliver electrical and mechanical cues to biological systems.nnnSTATEMENT OF SIGNIFICANCEnGraphene-based conductive hydrogels presenting electrical conductance and a soft tissue-like modulus were successfully fabricated via mild reduction of graphene oxide/polyacrylamide composite hydrogels to study their potential to skeletal tissue scaffold applications. Significantly promoted myoblast proliferation and differentiation were obtained on our hydrogels. Additionally, electrical stimulation of myoblasts via the graphene hydrogels could further upregulate myogenic gene expressions. Our graphene-incorporated conductive hydrogels will impact on the development of new materials for skeletal muscle tissue engineering scaffolds and bioelectronics devices, and also serve as novel platforms to study cellular interactions with electrical and mechanical signals.

An optical fiber-based LSPR aptasensor for simple and rapid in-situ detection of ochratoxin A

Label-free biosensing methods that rely on the use of localized surface plasmon resonance (LSPR) have attracted great attention as a result of their simplicity, high sensitivity, and relatively low cost. However, in-situ analysis of real samples using these techniques has remained challenging because colloidal nanoparticles (NPs) can be unstable at certain levels of pH and salt concentration. Even in the case of a chip-type LSPR sensor that can resolve the instability problem by employing NPs immobilized on the substrate, loading of a sample to sensor chip with exact volume control can be difficult for unskilled users. Herein, we report an optical-fiber-based LSPR aptasensor that can avoid these problems and serve as a portable and simple system for sensitive detection of a small mycotoxin, ochratoxin A (OTA), in real samples. The optical fiber coated with aptamer-modified gold nanorods (GNRs) is simply dipped into a solution containing OTA and subjected to LSPR analysis. Quantitative analysis of OTA is performed by measuring the spectral red shift of the LSPR peak of GNRs. Under optimized conditions, the LSPR peak shift displays a linear response (R2 = 0.9887) to OTA in the concentration range from 10pM to 100nM, with a limit of detection of 12.0pM (3S). The developed sensor shows a high selectivity for OTA over other mycotoxins such as zearalenone (ZEN) and ochratoxin B (OTB), and shows an accurate detection capability for OTA in real grape juice samples.

A Localized Surface Plasmon Resonance (LSPR)-based, simple, receptor-free and regeneratable Hg2+ detection system

A simple, receptor-free and regeneratable Hg(2+) sensor, which utilizes localized surface plasmon resonance (LSPR) shifts of a gold nanorod (GNR), has been developed. Precipitation induced by coordination of Hg(2+) to citrate alters the local refractive index (RI) around the GNR surface on glass slide, promoting a red-shift in its LSPR absorption peak. This phenomenon is used to design a sensor that enables quantitative detection of Hg(2+) in the 1nM to 1mM concentration range with good linearity (0.9507 correlation coefficient) and limit of detection (LOD) is reached to 0.38nM. A high selectivity of this sensor for Hg(2+) is demonstrated by the specific LSPR red-shift of 27.67nm promoted by Hg(2+) in comparison to those caused by other metal ions. In addition, the reusability of the new sensor chip is shown by its successful reuse eight-times following successive washing/precipitation steps. Lastly, the sensor displays excellent recoveries in spiking test with real water samples, such as tap water, lake and river. The simple combination of precipitation of Hg(2+)-citrate complex and the LSPR red-shift has led to the design of a novel sensing strategy for Hg(2+) detection.

A highly sensitive and widely adaptable plasmonic aptasensor using berberine for small-molecule detection

Localized surface plasmon resonance (LSPR) biosensors allow label-free detection of small molecules in molecular binding events; however, they are limited by a relatively low sensitivity and narrow dynamic range. Here, we report highly sensitive small-molecule detection by LSPR peak shift exploiting the G-quadruplex (GQx) structure-binding characteristic of known GQx binders to enhance the LSPR signal of a plasmonic aptasensor. Six known GQx binders (thiazole orange, malachite green, crystal violet, zinc protoporphyrin IX, thioflavin T, and berberine) were tested for their ability to enhance the LSPR signal. Among these, berberine (BER) induced the largest LSPR peak shift by interacting with the GQx structure formed by the aptamer/target binding event on a gold nanorod surface. This specific binding performance was confirmed by the fluorescence signal of BER and through repeated cycles of BER addition and washing on the plasmonic sensing chip. The proposed plasmonic aptasensor respectively showed limit of detection (LOD) of 0.56, 0.63, 0.87 and 1.05 pM for ochratoxin A, aflatoxin B1, adenosine triphosphate and potassium ions, which was 1000-fold higher than that in BER-free condition, and a wide dynamic range from 10 pM to 10μM. In addition, the proposed LSPR aptasensor could effectively be used to quantitatively analyze small molecules in real samples.

Quantitative analysis of lard in animal fat mixture using visible Raman spectroscopy

Food adulteration is a serious issue that requires verification and strict management due to healthcare, morality, and social value problems. In the context of fat, food manufacturers blend lard with vegetable oils or animal fats for convenience and gaining economic benefits. Thus, we herein report the classification of 4 animal fats, e.g., beef tallow, pork lard, chicken fat, duck oil, using Raman spectroscopy combined with simple calculation of intensity ratios of Raman signal at vibrational modes corresponding to unsaturated fatty acids and total fatty acids. Various calculated values of the species were compared to find a feature that is able to classify each fats using Raman peak ratio. As a result, we suggested Oil gauge (OG) as a standard feature for classification of the fats in Raman analysis field. Furthermore, a quantification of the lard in other fat was accomplished with good linear correlation using the OG values.

Dark-field spectral imaging microscope for localized surface plasmon resonance-based biosensing

Localized surface plasmon resonance (LSPR) of metal nanoparticles makes red-shift of extinction wavelength with an increase in the refractive index at the surface of the metal nanoparticles. Since biomolecules bound to the metal nanoparticle’s surface induce refractive index change, biosensing based on LSPR effect can be possible by monitoring scattering or absorption spectrum changes. Generally, however, conventional method detects ensemble averaged LSPR signal of a huge number of metal nanoparticles. Here, we have constructed a dark-field spectral imaging microscope in order to monitor the scattering spectra of individual metal nanoparticles, simultaneously. Gold nanorod (GNR) and aptamer are employed to detect ochratoxin A (OTA) related to a carcinogenic illness. An aptamer-target binding mechanism promotes wavelength shift of extinction spectra due to refractive index change within sensing volume of GNR by structural change of aptamer. A number of GNRs can be identified in a dark-field LSPR image, simultaneously. A typical spectrum of a GNR exhibits red-shift after target binding of molecules and OTA detection is extended to the very low concentration of 1 pM level.

A localized surface plasmon resonance (LSPR) immunosensor for CRP detection using 4-chloro-1-naphtol (4-CN) precipitation

In this study, C-reactive protein (CRP) was detected by monitoring of LSPR shift promoted by precipitation of 4-chloro-1-naphthol (4-CN). The precipitation occurred by horseradish peroxide (HRP) catalyst which is modified at CRP-detection antibody utilized in sandwich enzyme-linked immunosorbent assay (ELISA) on gold nano bipyramid (GNBP) substrate. Due to 4-CN precipitates which are located nearby the surface of GNBP, local refractive index (RI) and molecular density were greatly increased. This phenomenon eventually induced strong spectral red-shift of absorption band of GNBP. An excellent linear relationship (R2=0.9895) between the LSPR shift and CRP concentration was obtained in the range from 100 pg/mL to 100 ng/mL and limit of detection (LOD) was reached to 87 pg/mL.

Note: Automatic laser-to-optical-fiber coupling system based on monitoring of Raman scattering signal

We developed an automatic laser-to-optical-fiber coupling (ALOC) system that is based on the difference in the Raman scattering signals of the core and cladding of the optical fiber. This system can be easily applied to all fields of fiber optics since it can perform automatic optical coupling within a few seconds regardless of the core size or the condition of the output end of the optical fiber. The coupling time for a commercial single-mode fiber for a wavelength of 632.8 nm (core diameter: 9 μm, cladding diameter: 125 μm) is ~1.5 s. The ALOC system was successfully applied to single-mode-fiber Raman endoscopy for the measurement of the Raman spectrum of carbon nanotubes.