Sungsook Ahn

Gold nanoparticle-incorporated human red blood cells (RBCs) for X-ray dynamic imaging

Time-resolved dynamic imaging of bio-fluids can provide valuable information for clinical diagnosis and treatment of circulatory disorders. Quantitative information on non-transparent blood flows can be directly obtained by particle-tracing dynamic X-ray imaging, which needs better spatial resolution and enhanced image contrast compared to static imaging. For that use, tracer particles tagging along the flow streams are critically required. In this study, taking the advantage of high X-ray absorption, gold nanoparticles (AuNPs) are incorporated into human red blood cells (RBC) to produce contrast-enhanced tracers designed for dynamic X-ray imaging of blood flows. RBCs are advantageous tracers for blood flow measurements since they are natural and primary components of blood. The loading efficiency of AuNPs into RBCs is investigated in terms of the surface properties of the AuNPs. The AuNP-incorporated RBC provides a potential in the dynamic X-ray imaging of blood flows which can be used for clinical applications.

Gold Nanoparticle Contrast Agents in Advanced X-ray Imaging Technologies

Recently, there has been significant progress in the field of soft- and hard-X-ray imaging for a wide range of applications, both technically and scientifically, via developments in sources, optics and imaging methodologies. While one community is pursuing extensive applications of available X-ray tools, others are investigating improvements in techniques, including new optics, higher spatial resolutions and brighter compact sources. For increased image quality and more exquisite investigation on characteristic biological phenomena, contrast agents have been employed extensively in imaging technologies. Heavy metal nanoparticles are excellent absorbers of X-rays and can offer excellent improvements in medical diagnosis and X-ray imaging. In this context, the role of gold (Au) is important for advanced X-ray imaging applications. Au has a long-history in a wide range of medical applications and exhibits characteristic interactions with X-rays. Therefore, Au can offer a particular advantage as a tracer and a contrast enhancer in X-ray imaging technologies by sensing the variation in X-ray attenuation in a given sample volume. This review summarizes basic understanding on X-ray imaging from device set-up to technologies. Then this review covers recent studies in the development of X-ray imaging techniques utilizing gold nanoparticles (AuNPs) and their relevant applications, including two- and three-dimensional biological imaging, dynamical processes in a living system, single cell-based imaging and quantitative analysis of circulatory systems and so on. In addition to conventional medical applications, various novel research areas have been developed and are expected to be further developed through AuNP-based X-ray imaging technologies.

Controlled cellular uptake and drug efficacy of nanotherapeutics

Cellular uptake pathway of nanoparticle (NP) is different from that of free drugs. Therefore, NP-mediated nanotherapeutics can be designed to overcome the adverse effects of free drugs. However, synthetic NPs are typically trapped in the endosome and have difficulty to reach the cytosol because of the characteristic endocytosis, where the endosomal membranes wrap-up the introduced NPs. In this study, the Spacer molecules linking the apoptotic anticancer drug and the gold NP (AuNP) are designed and cellular uptake procedure and drug deployment in the cancer cells are controlled. X-ray nanoscopy and two-photon microscopy are employed to observe the AuNPs in a cell in-situ without additional dye molecule or imaging agent introduction on an AuNP. We confirm that the effective design of the Spacer molecules importantly control the cellular interaction of the AuNPs. This technology can be generalized to broad biomedical applications utilizing nanotherapeutics-mediated diagnosis and new-concepted disease treatment technologies.

Gold Nanoparticle Flow Sensors Designed for Dynamic X-ray Imaging in Biofluids

X-ray-based imaging is one of the most powerful and convenient methods in terms of versatility in applicable energy and high performance in use. Different from conventional nuclear medicine imaging, contrast agents are required in X-ray imaging especially for effectively targeted and molecularly specific functions. Here, in contrast to much reported static accumulation of the contrast agents in targeted organs, dynamic visualization in a living organism is successfully accomplished by the particle-traced X-ray imaging for the first time. Flow phenomena across perforated end walls of xylem vessels in rice are monitored by a gold nanoparticle (AuNP) (approximately 20 nm in diameter) as a flow tracing sensor working in nontransparent biofluids. AuNPs are surface-modified to control the hydrodynamic properties such as hydrodynamic size (DH), zeta-potential, and surface plasmonic properties in aqueous conditions. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray nanoscopy (XN), and X-ray microscopy (XM) are used to correlate the interparticle interactions with X-ray absorption ability. Cluster formation and X-ray contrast ability of the AuNPs are successfully modulated by controlling the interparticle interactions evaluated as flow-tracing sensors.

Flow tracing microparticle sensors designed for enhanced X-ray contrast.

In applying the X-ray particle image velocimetry (PIV) technique to biofluid flows, the most pivotal prerequisite is suitable flow tracing sensors which should be detected effectively by the X-ray imaging system. In this study, to design those flow tracing sensors, X-ray contrast agent Iopamidol was encapsulated into the poly(vinyl alcohol) (PVA) microparticles crosslinked by glutaraldehyde (GA). The characteristics of the fabricated particle sensors were determined by optical microscopy, scanning electron microscopy, dynamic light scattering, laser Doppler electrophoresis and nuclear magnetic resonance spectroscopy ((1)H NMR). The amount of Iopamidol in the microparticles was measured using the energy dispersive X-ray spectroscopy (EDS) and (1)H NMR. The physical properties of the PVA microparticles are effectively controlled in terms of the average particle size, degree of crosslinking, degree of swelling and encapsulation efficiency of Iopamidol. By changing the amount of crosslinker, the degree of crosslinking and the efficiency of the Iopamidol encapsulation reached to the optimal. To some extent, the zeta-potential of the PVA microparticles is increased in less ionic media where the particles can effectively repel each other prohibiting aggregation. The X-ray absorption ability of the designed tracing sensors was examined by a synchrotron X-ray imaging technique. The X-ray absorption coefficients of the particle sensors were expressed by an exponential law assuming the spherical shape of the microparticles. The X-ray contrast agent, Iopamidol, was successfully encapsulated into the bio-compatible and bio-degradable PVA. With the controlled physical properties of the flow tracing sensors designed in this study, the particle sensors exhibit excellent X-ray absorption contrast fairly applicable in biological systems.

In vivo measurements of blood flow in a rat using X-ray imaging technique.

To measure instantaneous velocity fields of venous blood flow in a rat using X-ray particle tracking method. Gold nanoparticles (AuNPs) incorporated chitosan microparticles were applied as biocompatible flow tracers. After intravenous injection of the AuNP-chitosan particles into 7- to 9-week-old male rat vein, X-ray images of particle movement inside the cranial vena cava were consecutively captured. Individual AuNP-chitosan particles in the venous blood flow were clearly observed, and the corresponding velocity vectors were successfully extracted. The measured velocity vectors are in good agreement with the theoretical velocity profile suggested by Casson. This is the first trial to measure blood flow in animals under in vivo conditions with X-ray imaging technique. The results show that X-ray particle tracking technique has a great potential for in vivo measurements of blood flow, which can extend to various biomedical applications related with the diagnosis of circulatory vascular diseases.

Chitosan microparticles incorporating gold as an enhanced contrast flow tracer in dynamic X-ray imaging

In situ monitoring of a biofluid can provide important information on circulatory disorders and a basic understanding on the metabolic mechanisms of living organisms. X-ray imaging has significant advantages as one of the most popular diagnostic tools to seethrough various biological systems. Particle traced velocity field measurement is one of the most popular methods for quantitative analysis of dynamic flow motion. In this study we have developed chitosan microparticles incorporating gold nanoparticles (AuNP) as a new enhanced contrast flow tracer for dynamic X-ray imaging. Gold is a useful material possessing high X-ray absorption ability and also biocompatibility. We chose chitosan as an AuNP delivery system because it can effectively trap AuNPs at high yield. In particular, the unique gold ion reduction ability of and compatibility with surface-modified chitosans are effectively utilized. The physical properties of the Au-chitosan microparticles can be controlled by varying the molecular weight of the chitosan employed and the AuNP incorporation methodology. The environment of the particles and the type of applied X-ray essentially determine the imaging efficiency. The designed chitosan microparticles incorporating Au have been successfully applied to track the digestive mechanisms occurring in delicate insects such as live mosquitoes.

Properties of iopamidol-incorporated poly(vinyl alcohol) microparticle as an X-ray imaging flow tracer.

We have recently reported on poly(vinyl alcohol) microparticles containing X-ray contrast agent, iopamidol, designed as a flow tracer working in synchrotron X-ray imaging ( Biosens. Bioelectron. 2010 , 25 , 1571 ). Although iopamidol is physically encapsulated in the microparticles, it displays a great contrast enhancement and stable feasibility in in vitro human blood pool. Nonetheless, a direct relation between the absolute amount of incorporated iopamidol and the enhancement in imaging efficiency was not observed. In this study, physical properties of the designed microparticle are systematically investigated experimentally with theoretical interpretation to correlate an enhancement in X-ray imaging efficiency. The compositional ratio of X-ray contrast agent in polymeric microparticle is controlled as 1/1 and 10/1 [contrast agent/polymer microparticle (w/w)] with changed degree of cross-linkings. Flory-Huggins interaction parameter (χ), retractive force (τ) and degree of swelling of the designed polymeric microparticles are investigated. In addition, the hydrodynamic size (D(H)) and ζ-potential are evaluated in terms of environment responsiveness. The physical properties of the designed flow tracer microparticles under a given condition are observed to be strongly related with the X-ray absorption efficiency, which are also supported by the Beer-Lambert-Bouguer law. The designed microparticles are almost nontoxic with a reasonable concentration and time period, enough to be utilized as a flow tracer in various biomedical applications. This study would contribute to the basic understanding on the physical property connected with the imaging efficiency of contrast agents.

Interactive Ion-Mediated Sap Flow Regulation in Olive and Laurel Stems: Physicochemical Characteristics of Water Transport via the Pit Structure

Sap water is distributed and utilized through xylem conduits, which are vascular networks of inert pipes important for plant survival. Interestingly, plants can actively regulate water transport using ion-mediated responses and adapt to environmental changes. However, ionic effects on active water transport in vascular plants remain unclear. In this report, the interactive ionic effects on sap transport were systematically investigated for the first time by visualizing the uptake process of ionic solutions of different ion compositions (K+/Ca2+) using synchrotron X-ray and neutron imaging techniques. Ionic solutions with lower K+/Ca2+ ratios induced an increased sap flow rate in stems of Olea europaea L. and Laurus nobilis L. The different ascent rates of ionic solutions depending on K+/Ca2+ ratios at a fixed total concentration increases our understanding of ion-responsiveness in plants from a physicochemical standpoint. Based on these results, effective structural changes in the pit membrane were observed using varying ionic ratios of K+/Ca2+. The formation of electrostatically induced hydrodynamic layers and the ion-responsiveness of hydrogel structures based on Hofmeister series increase our understanding of the mechanism of ion-mediated sap flow control in plants.

Detection of circulating tumor cells via an X-ray imaging technique

Detailed information on the location and the size of tumor cells circulating through lymphatic and blood vessels is useful to cancer diagnosis. Metastasis of cancers to other non-adjacent organs is reported to cause 90% of deaths not from the primary tumors. Therefore, effective detection of circulating tumors cells (CTCs) related to metastasis is emphasized in cancer treatments. With the use of synchrotron X-ray micro-imaging techniques, high-resolution images of individual flowing tumor cells were obtained. Positively charged gold nanoparticles (AuNPs) which were inappropriate for incorporation into human red blood cells were selectively incorporated into tumor cells to enhance the image contrast. This approach enables images of individual cancer cells and temporal movements of CTCs to be captured by the high X-ray absorption efficiency of selectively incorporated AuNPs. This new technology for in vivo imaging of CTCs would contribute to improve cancer diagnosis and cancer therapy prognosis.