Browsing by Author "Cheng, Chia-Liang"

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Alpha-bungarotoxin binding to target cell in a developing visual system by carboxylated nanodiamond
(2008-05-21)
Liu, Kuang-Kai
;
Mei-Fang Chen
;
Chen, Po-Yi
;
Lee, Tony J. F.
;
Cheng, Chia-Liang
Biological molecules conjugating with nanoparticles are valuable for applications including bio-imaging, bio-detection, and bio-sensing. Nanometer-sized diamond particles have excellent electronic and chemical properties for bio-conjugation. In this study, we manipulated the carboxyl group produced on the surface of nanodiamond (carboxylated nanodiamond, cND) for conjugating with alpha-bungarotoxin (alpha-BTX), a neurotoxin derived from Bungarus multicinctus with specific blockade of alpha7-nicotinic acetylcholine receptor (alpha 7-nAChR). The electrostatic binding of cND-alpha-BTX was mediated by the negative charge of the cND and the positive charge of the alpha-BTX in physiological pH conditions. Sodium dodecyl sulfate-polyacrylamide gel analysis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF-MS) spectra displayed that alpha-BTX proteins were conjugated with cND particles via non-covalent bindings. The green fluorescence of the cND particles combining with the red fluorescence of tetramethylrhodamine-labeled a-BTX presented a yellow color at the same location, which indicated that alpha-BTX proteins were conjugated with cND particles. Xenopus laevis's oocytes expressed the human alpha 7-nAChR proteins by microinjection with alpha 7-nAChR mRNA. The cND-alpha-BTX complexes were bound to alpha 7-nAChR locating on the cell membrane of oocytes and human lung A549 cancer cells analyzed by laser scanning confocal microscopy. The choline-evoked alpha 7-nAChR-mediated inward currents of the oocytes were blocked by cND-alpha-BTX complexes in a concentration-dependent manner using two-electrode voltage-clamp recording. Furthermore, the fluorescence intensity of cND-alpha-BTX binding on A549 cells could be quantified by flow cytometry. These results indicate that cND-conjugated alpha-BTX still preserves its biological activity in blocking the function of alpha 7-nAChR, and provide a visual system showing the binding of alpha-BTX to alpha 7-nAChR.
Bio-imaging of alpha-bungarotoxin binding to target cell by carboxylated nanodiamond
(2008-04-01)
Chao, Jui-I
;
Liu, Kuang-Kai
;
Chen, Mei-Fang
;
Chen, Po-Yi
;
Lee, Tony J. F.
Biocompatible and detectable carboxylated nanodiamond on human cell
(2007-08-15)
Liu, Kuang-Kai
;
Cheng, Chia-Liang
;
Chang, Chia-Ching
;
Chao, Jui-I
;
生物科技學系
Surface-modified carboxylated nanometre-sized diamond (cND) has been applied for the conjugation of biological molecules such as DNA and protein. In this study, we evaluated the biocompatibility and detection of cNDs and carbon nanotubes on human lung A549 epithelial cells and HFL-1 normal fibroblasts. Treatment with 5 or 100 nm cND particles, 0.1 - 100 mu g ml(-1), did not reduce the cell viability and alter the protein expression profile in lung cells; however, carbon nanotubes induced cytotoxicity in these cells. The cNDs particles were accumulated in A549 cells, which were observed by atomic force microscopy and laser scanning confocal microscopy. Both 5 and 100 nm cNDs particles exhibited the green fluorescence and were ingested into cells. Moreover, the fluorescence intensities were increased in cells via a concentration-dependent manner after treatment with 5 and 100 nm cNDs, which can be detected by flow cytometer analysis. The fluorescence intensities of 5 nm cNDs were relative higher than 100 nm cNDs in cells at equal concentration treatment. The observation demonstrated that cND-interacting with cell is detectable by a confocal microscope, flow cytometer and atomic force microscope. These nanoparticles may be useful for further biomedical applications based on the properties of uptake ability, detectability and little cytotoxicity in human cells.
CARRIER COMPRISING NANODIAMOND
(2011-01-13)
Chao, Jui-I
;
Chen, Chin-Piao
;
Cheng, Chia-Liang
The present invention provides a carrier including a nanodiamond (ND) particle and a linker covalently bound to the ND particle, in which the linker is presented by the formula: —R1—O(R2)m-Q-. In addition, the present invention further provides a carrier having an active unit covalently bound to the linker, in which the active unit is a drug, a vitamin or a biological molecule.
Covalent linkage of nanodiamond-paclitaxel for drug delivery and cancer therapy
(2010-08-06)
Liu, Kuang-Kai
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Zheng, Wen-Wei
;
Wang, Chi-Ching
;
Chiu, Yu-Chung
;
Cheng, Chia-Liang
A nanoparticle-conjugated cancer drug provides a novel strategy for cancer therapy. In this study, we manipulated nanodiamond (ND), a carbon nanomaterial, to covalently link paclitaxel for cancer drug delivery and therapy. Paclitaxel was bound to the surface of 3-5 nm sized ND through a succession of chemical modifications. The ND-paclitaxel conjugation was measured by atomic force microscope and nuclear magnetic resonance spectroscopy, and confirmed with infrared spectroscopy by the detection of deuterated paclitaxel. Treatment with 0.1-50 mu g ml(-1) ND-paclitaxel for 48 h significantly reduced the cell viability in the A549 human lung carcinoma cells. ND-paclitaxel induced both mitotic arrest and apoptosis in A549 cells. However, ND alone or denatured ND-paclitaxel (after treatment with strong alkaline solution, 1 M NaOH) did not induce the damage effects on A549 cells. ND-paclitaxel was taken into lung cancer cells in a concentration-dependent manner using flow cytometer analysis. The ND-paclitaxel particles were located in the microtubules and cytoplasm of A549 cells observed by confocal microscopy. Furthermore, ND-paclitaxel markedly blocked the tumor growth and formation of lung cancer cells in xenograft SCID mice. Together, we provide a functional covalent conjugation of ND-paclitaxel, which can be delivered into lung carcinoma cells and preserves the anticancer activities on the induction of mitotic blockage, apoptosis and anti-tumorigenesis.
Endocytic carboxylated nanodiamond for the labeling and tracking of cell division and differentiation in cancer and stem cells
(2009-09-01)
Liu, Kuang-Kai
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Wang, Chi-Ching
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Cheng, Chia-Liang
;
Chao, Jui-I
;
生物科技學系
Nanodiamond (ND) is carbon nanomaterial developing for biological applications in recent years. In this study, we investigated the location and distribution of 100 nm carboxylated ND particles in cell division and differentiation. ND particles were taken into cells by macropinocytosis and clathrin-mediated endocytosis pathways. However, the cell growth ability was not altered by endocytic ND particles after long-term cell culture for 10 days in both A549 lung cancer cells and 3T3-L1 embryonic fibroblasts. ND particles were equal separating into two daughter cells of cell division approximately. Finally, the cell retained a single ND's cluster in cytoplasm after sub-cultured for several generations. Interestingly, ND's clusters were carried inside of cell but without inducing damages after long-term cell culture. Moreover, ND particles did not interfere with the gene or protein expressions on the regulation of cell cycle progression and adipogenic differentiation. Together, these findings provide that endocytic ND particles are non-cytotoxic in cell division and differentiation, which can be applied for the labeling and tracking of cancer and stem cells. (c) 2009 Elsevier Ltd. All rights reserved.
Laser induced popcornlike conformational transition of nanodiamond as a nanoknife
(2008-07-21)
Chang, Chia-Ching
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Chen, Pei-Hsin
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Chu, Hsueh-Liang
;
Lee, Tzu-Cheng
;
Chou, Ching-Chung
Nanodiamond (ND) is surrounded by layers of graphite on its surface. This unique structure feature creates unusual fluorescence spectra, which can be used as an indicator to monitor its surface modification. Meanwhile, the impurity, nitroso (C - N = O) inside the ND can be photolyzed by two-photon absorption, releasing NO to facilitate the formation of a sp(3) diamond structure in the core of ND and transforming it into a sp(2) graphite structure. Such a conformational transition enlarges the size of ND from 8 to 90 nm, resulting in a popcornlike structure. This transition reaction may be useful as nanoknives in biomedical application. (c) 2008 American Institute of Physics.
Nano-manipulation on cancer cells by protein-nanodiamond complex.
(2007-01-01)
Chang, Chia-Ching
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Chao, Jui-I
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Ho, Yen-Peng
;
Sun, Ken-Wen
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Cheng, Chia-Liang
Nanometer-sized diamond particle as a probe for biolabeling
(2007-09-01)
Chao, Jui-I.
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Perevedentseva, Elena
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Chung, Pei-Hua
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Liu, Kuang-Kai
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Cheng, Chih-Yuan
A novel method is proposed using nanometer- sized diamond particles as detection probes for biolabeling. The advantages of nanodiamond's unique properties were demonstrated in its biocompatibility, nontoxicity, easily detected Raman signal, and intrinsic fluorescence from its natural defects without complicated pretreatments. Carboxylated nanodiamond's ( cND's) penetration ability, noncytotoxicity, and visualization of cND- cell interactions are demonstrated on A549 human lung epithelial cells. Protein- targeted cell interaction visualization was demonstrated with cND- lysozyme complex interaction with bacteria Escherichia coli. It is shown that the developed biomolecule- cND complex preserves the original functions of the test protein. The easily detected natural fluorescent and Raman intrinsic signals, penetration ability, and low cytotoxicity of cNDs render them promising agents in multiple medical applications.
Toxicity and detection of carboxylated nanodiamonds on human lung epithelial cells
(2007-04-01)
Chao, Jui-I
;
Liu, Kuang-Kai
;
Cheng, Chia-Liang
;
Chang, Chia-Ching
;
生物科技學系