Kai-Hsiang Chuang

Neuronal Specificity of Acupuncture Response: A fMRI Study with Electroacupuncture

Recently, neuronal correlates of acupuncture stimulation in human brain have been investigated by functional neuroimaging. The preliminary findings suggest that acupuncture at analgesic points involves the pain-related neuromatrix and may have acupoint-brain correlation. Although multiple models of control stimulations have been applied to address the specificity of the needling effect clinically, their impacts have not been evaluated by functional neuroimaging. With the advantage of objective parameter setting, electroacupuncture (EA) was used in this study to devise three distinct controls for real EA, i.e., mock EA (no stimulation), minimal EA (superficial and light stimulation), and sham EA (same stimulation as real EA) applied at nonmeridian points. Fifteen healthy volunteers received real EA at analgesic point Gallbladder 34 (Yanglinquan), sham EA, and one of either mock EA or minimal EA over the left leg in counter-balanced orders. Multisubject analysis showed that sham EA and real EA both activated the reported distributed pain neuromatrix. However, real EA elicited significantly higher activation than sham EA over the hypothalamus and primary somatosensory-motor cortex and deactivation over the rostral segment of anterior cingulate cortex. In the comparison of minimal EA versus mock EA, minimal EA elicited significantly higher activation over the medial occipital cortex. Single-subject analysis showed that superior temporal gyrus (encompassing the auditory cortex) and medial occipital cortex (encompassing the visual cortex) frequently respond to minimal EA, sham EA, or real EA. We concluded that the hypothalamus-limbic system was significantly modulated by EA at acupoints rather than at nonmeridian points, while visual and auditory cortical activation was not a specific effect of treatment-relevant acupoints and required further investigation of the underlying neurophysiological mechanisms.

Superparamagnetic iron oxide – Loaded poly (lactic acid)-d-α-tocopherol polyethylene glycol 1000 succinate copolymer nanoparticles as MRI contrast agent

We developed a strategy to formulate supraparamagnetic iron oxides (SPIOs) in nanoparticles (NPs) of biodegradable copolymer made up of poly(lactic acid) (PLA) and d-alpha-tocopherol polyethylene glycol 1000 succinate (TPGS) for medical imaging by magnetic resonance imaging (MRI) of high contrast and low side effects. The IOs-loaded PLA-TPGS NPs (IOs-PNPs) were prepared by the single emulsion method and the nanoprecipitation method. Effects of the process parameters such as the emulsifier concentration, IOs loading in the nanoparticles, and the solvent to non-solvent ratio on the IOs distribution within the polymeric matrix were investigated and the formulation was then optimized. The transmission electron microscopy (TEM) showed direct visual evidence for the well dispersed distribution of the IOs within the NPs. We further investigated the biocompatibility and cellular uptake of the IOs-PNPs in vitro with MCF-7 breast cancer cells and NIH-3T3 mouse fibroblast in close comparison with the commercial IOs imaging agent Resovist. MRI imaging was further carried out to investigate the biodistribution of the IOs formulated in the IOs-PNPs, especially in the liver to understand the liver clearance process, which was also made in close comparison with Resovist. We found that the PLA-TPGS NPs formulation at the clinically approved dose of 0.8 mg Fe/kg could be cleared within 24 h in comparison with several weeks for Resovist. Xenograft tumor model MRI confirmed the advantages of the IOs-PNPs formulation versus Resovist through the enhanced permeation and retention (EPR) effect of the tumor vasculature.

Mapping resting-state functional connectivity using perfusion MRI

Resting-state, low-frequency (<0.08 Hz) fluctuations of blood oxygenation level-dependent (BOLD) magnetic resonance signal have been shown to exhibit high correlation among functionally connected regions. However, correlations of cerebral blood flow (CBF) fluctuations during the resting state have not been extensively studied. The main challenges of using arterial spin labeling perfusion magnetic resonance imaging to detect CBF fluctuations are low sensitivity, low temporal resolution, and contamination from BOLD. This work demonstrates CBF-based quantitative functional connectivity mapping by combining continuous arterial spin labeling (CASL) with a neck labeling coil and a multi-channel receiver coil to achieve high perfusion sensitivity. In order to reduce BOLD contamination, the CBF signal was extracted from the CASL signal time course by high frequency filtering. This processing strategy is compatible with sinc interpolation for reducing the timing mismatch between control and label images and has the flexibility of choosing an optimal filter cutoff frequency to minimize BOLD fluctuations. Most subjects studied showed high CBF correlation in bilateral sensorimotor areas with good suppression of BOLD contamination. Root-mean-square CBF fluctuation contributing to bilateral correlation was estimated to be 29+/-19% (N=13) of the baseline perfusion, while BOLD fluctuation was 0.26+/-0.14% of the mean intensity (at 3 T and 12.5 ms echo time).

Imaging brain deoxyglucose uptake and metabolism by glucoCEST MRI

2-Deoxy-D-glucose (2DG) is a known surrogate molecule that is useful for inferring glucose uptake and metabolism. Although 13C-labeled 2DG can be detected by nuclear magnetic resonance (NMR), its low sensitivity for detection prohibits imaging to be performed. Using chemical exchange saturation transfer (CEST) as a signal-amplification mechanism, 2DG and the phosphorylated 2DG-6-phosphate (2DG6P) can be indirectly detected in 1H magnetic resonance imaging (MRI). We showed that the CEST signal changed with 2DG concentration, and was reduced by suppressing cerebral metabolism with increased general anesthetic. The signal changes were not affected by cerebral or plasma pH, and were not correlated with altered cerebral blood flow as demonstrated by hypercapnia; neither were they related to the extracellular glucose amounts as compared with injection of D- and L-glucose. In vivo 31P NMR revealed similar changes in 2DG6P concentration, suggesting that the CEST signal reflected the rate of glucose assimilation. This method provides a new way to use widely available MRI techniques to image deoxyglucose/glucose uptake and metabolism in vivo without the need for isotopic labeling of the molecules.

Improved neuronal tract tracing using manganese enhanced magnetic resonance imaging with fast T1 mapping

There has been growing interest in using manganese‐enhanced MRI (MEMRI) to detect neuronal activation, neural architecture, and neuronal connections. Usually Mn2+ produces a very wide range of T1 change. In particular, in neuronal tract tracing experiments the site of Mn2+ injection can have very short T1 while distant regions have small T1 reductions, primarily due to dilution of Mn2+. Most MEMRI studies use T1‐weighted sequences, which can only give optimal contrast for a narrow range of T1 changes. To improve sensitivity to the full extent of Mn2+ concentrations and to optimize detection of low concentrations of Mn2+, a fast T1 mapping sequence based on the Look and Locker technique was implemented. Phantom studies demonstrated less than 6.5% error in T1 compared to more conventional T1 measurements. Using center‐out segmented EPI, whole‐brain 3D T1 maps with 200‐μm isotropic resolution were obtained in 2 h from rat brain. Mn2+ transport from the rat olfactory bulb through appropriate brain structures could be detected to the amygdala in individual animals. The method reliably detected less than 7% reductions in T1. With this quantitative imaging it should be possible to study more extensive pathways using MEMRI and decrease the dose of Mn2+ used. Magn Reson Med, 2006. Published 2006 Wiley‐Liss, Inc.

Multimodal tumor imaging by iron oxides and quantum dots formulated in poly (lactic acid)-D-alpha-tocopheryl polyethylene glycol 1000 succinate nanoparticles.

This work developed a multimodal imaging system by co-encapsulating superparamagnetic iron oxides (IOs) and quantum dots (QDs) in the nanoparticles of poly (lactic acid) - d-α-tocopheryl polyethylene glycol 1000 succinate (PLA-TPGS) for concurrent imaging of the magnetic resonance imaging (MRI) and the fluorescence imaging to combine their advantages and to overcome their disadvantages as well as to promote a sustained and controlled imaging with passive targeting effects to the diseased cells. The QDs and IOs-loaded PLA-TPGS NPs were prepared by a modified nanoprecipitation method, which were then characterized for their size and size distribution, zeta potential and the imaging agent encapsulation efficiency. The transmission electron microscopy (TEM) images showed direct evidence for the well-dispersed distribution of the QDs and IOs within the PLA-TPGS NPs. The cellular uptake and the cytotoxicity of the PLA-TPGS NPs formulation of QDs and IOs were investigated in vitro with MCF-7 breast cancer cells, which were conducted in close comparison with the free QDs and IOs at the same agent dose. The Xenograft model was also conducted for biodistribution of the QDs and IOs-loaded PLA-TPGS NPs among the various organs, which showed greatly enhanced tumor imaging due to the passively targeting effects of the NPs to the tumor. Images of tumors were acquired in vivo by a 7T MRI scanner. Further ex vivo images of the tumors were obtained by confocal laser scanning microscopy. Such a multimodal imaging system shows great advantages of both contrast agents making the resultant probe highly sensitive with good depth penetration, which confirms the diagnosis obtained from each individual imaging. With therapeutics co-encapsulation and ligand conjugation, such nanoparticles system can realize a multi-functional system for medical diagnosis and treatment.

Synthesis of Manganese Ferrite/Graphene Oxide Nanocomposites for Biomedical Applications

In this study, MnFe(2)O(4) nanoparticle (MFNP)-decorated graphene oxide nanocomposites (MGONCs) are prepared through a simple mini-emulsion and solvent evaporation process. It is demonstrated that the loading of magnetic nanocrystals can be tuned by varying the ratio of graphene oxide/magnetic nanoparticles. On top of that, the hydrodynamic size range of the obtained nanocomposites can be optimized by varying the sonication time during the emulsion process. By fine-tuning the sonication time, MGONCs as small as 56.8 ± 1.1 nm, 55.0 ± 0.6 nm and 56.2 ± 0.4 nm loaded with 6 nm, 11 nm, and 14 nm MFNPs, respectively, are successfully fabricated. In order to improve the colloidal stability of MGONCs in physiological solutions (e.g., phosphate buffered saline or PBS solution), MGONCs are further conjugated with polyethylene glycol (PEG). Heating by exposing MGONCs samples to an alternating magnetic field (AMF) show that the obtained nanocomposites are efficient hyperthermia agents. At concentrations as low as 0.1 mg Fe mL(-1) and under an 59.99 kA m(-1) field, the highest specific absorption rate (SAR) recorded is 1588.83 W g(-1) for MGONCs loaded with 14 nm MFNPs. It is also demonstrated that MGONCs are promising as magnetic resonance imaging (MRI) T(2) contrast agents. A T(2) relaxivity value (r(2) ) as high as 256.2 (mM Fe)(-1) s(-1) could be achieved with MGONCs loaded with 14 nm MFNPs. The cytotoxicity results show that PEGylated MGONCs exhibit an excellent biocompatibility that is suitable for biomedical applications.

IMPACT: Image-based physiological artifacts estimation and correction technique for functional MRI

Functional MRI (fMRI) signal variation induced by respiratory and cardiac motion affects the activation signal and limits the accuracy of analysis. Current physiological motion correction methods require either synchronization with external monitoring of respiration and heartbeat, specialized pulse sequence design, or k‐space data. The IMage‐based Physiological Artifacts estimation and Correction Technique (IMPACT), which is free from these constraints, is described. When images are acquired fast enough to sample physiological motion without aliasing, respiratory and cardiac signals can be directly estimated from magnitude images. Physiological artifacts are removed by reordering images according to the estimated respiratory and cardiac phases and then subtracting the Fourier‐fitted variation from magnitude images. Compared with the k‐space–based method, this method can efficiently and effectively reduce the overall signal fluctuation in the brain and increase the activated area. With this new technique, physiological artifacts can be reduced using traditional fMRI pulse sequences, and existing data can be corrected and reanalyzed without additional experiments. Magn Reson Med 46:344–353, 2001.

Vitamin E (d-alpha-tocopheryl-co-poly(ethylene glycol) 1000 succinate) micelles-superparamagnetic iron oxide nanoparticles for enhanced thermotherapy and MRI

We synthesized vitamin E TPGS (d-α-Tocopheryl-co-poly(ethylene glycol) 1000 succinate) micelles for superparamagnetic iron oxides formulation for nanothermotherapy and magnetic resonance imaging (MRI), which showed better thermal and magnetic properties, and in vitro cellular uptake and lower cytotoxicity as well as better in vivo therapeutic and imaging effects in comparison with the commercial Resovist and the Pluronic F127 micelles reported in the recent literature. The superparamagnetic iron oxides originally coated with oleic acid and oleylamine were formulated in the core of the TPGS micelles using a simple solvent-exchange method. The IOs-loaded TPGS showed greatest colloidal stability due to the critical micelle concentration (CMC) of vitamin E TPGS. Highly monodisperse and water soluble suspension was obtained which were stable in 0.9% normal saline for a period of 12 days. The micelles were characterized for their size and size distribution. Their morphology was examined through transmission electron microscopy (TEM). The enhanced thermal and superparamagnetic properties of the IOs-loaded TPGS micelles were assessed. Cellular uptake and cytotoxicity were investigated in vitro with MCF-7 cancer cells. Relaxivity study showed that the IOs-loaded TPGS micelles can have better effects for T2-weighted imaging using MRI. T2 mapped images of xenograft grown on SCID mice showed that the TPGS micelle formulation of IOs had ∼1.7 times and ∼1.05 times T2 decrease at the tumor site compared to Resovist and the F127 micelle formulation, respectively.

Brain redox imaging using blood–brain barrier-permeable nitroxide MRI contrast agent

Reactive oxygen species (ROS) and compromised antioxidant defense may contribute to brain disorders such as stroke, amyotrophic lateral sclerosis, etc. Nitroxides are redox-sensitive paramagnetic contrast agents and antioxidants. The ability of a blood—brain barrier (BBB)-permeable nitroxide, methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (MC-P), as a magnetic resonance-imaging (MRI) contrast agent for brain tissue redox imaging was tested. MC-P relaxation in rodent brain was quantified by MRI using a fast Look-Locker T1-mapping sequence. In the cerebral cortex and thalamus, the MRI signal intensity increased up to 50% after MC-P injection, but increased only by 2.7% when a BBB-impermeable nitroxide, 3CxP (3-carboxy-2,2,5,5,5-tetramethylpyrrolidine-1-oxyl) was used. The maximum concentrations in the thalamus and cerebral cortex after MC-P injection were calculated to be 1.9±0.35 and 3.0±0.50 mmol/L, respectively. These values were consistent with the ex vivo data of brain tissue and blood concentration obtained by electron paramagnetic resonance (EPR) spectroscopy. Also, reduction rates of MC-P were significantly decreased after reperfusion following transient MCAO (middle cerebral artery occlusion), a condition associated with changes in redox status resulting from oxidative damage. These results show the use of BBB-permeable nitroxides as MRI contrast agents and antioxidants to evaluate the role of ROS in neurologic diseases.