Browsing by Author "Hung, Yao-Ching"

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A bio-inspired computing model for ovarian carcinoma classification and oncogene detection
(2015-04-01)
Tsai, Meng-Hsiun
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Chen, Mu-Yen
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Huang, Steve G.
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Hung, Yao-Ching
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Wang, Hsin-Chieh
Motivation: Ovarian cancer is the fifth leading cause of cancer deaths in women in the western world for 2013. In ovarian cancer, benign tumors turn malignant, but the point of transition is difficult to predict and diagnose. The 5-year survival rate of all types of ovarian cancer is 44%, but this can be improved to 92% if the cancer is found and treated before it spreads beyond the ovary. However, only 15% of all ovarian cancers are found at this early stage. Therefore, the ability to automatically identify and diagnose ovarian cancer precisely and efficiently as the tissue changes from benign to invasive is important for clinical treatment and for increasing the cure rate. This study proposes a new ovarian carcinoma classification model using two algorithms: a novel discretization of food sources for an artificial bee colony (DfABC), and a support vector machine (SVM). For the first time in the literature, oncogene detection using this method is also investigated. Results: A novel bio-inspired computing model and hybrid algorithms combining DfABC and SVM was applied to ovarian carcinoma and oncogene classification. This study used the human ovarian cDNA expression database to collect 41 patient samples and 9600 genes in each pathological stage. Feature selection methods were used to detect and extract 15 notable oncogenes. We then used the DfABC-SVM model to examine these 15 oncogenes, dividing them into eight different classifications according to their gene expressions of various pathological stages. The average accuracyof the eight classification experiments was 94.76%. This research also found some oncogenes that had not been discovered or indicated in previous scientific studies. The main contribution of this research is the proof that these newly discovered oncogenes are highly related to ovarian or other cancers.
A nanodevice for rapid modulation of proliferation, apoptosis, invasive ability, and cytoskeletal reorganization in cultured cells
(2010)
Hung, Yao-Ching
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Pan, Hsu-An
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Tai, Shih-Ming
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Huang, G. Steve
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材料科學與工程學系奈米科技碩博班
We have fabricated a nanodevice composed of a matrix of nine nanodot arrays with various dot sizes, ranging from a flat surface to 10 nm, 50 nm, 100 nm, and 200 nm arrays. HELA, C33A, ES2, PA-1, TOV-112D, TOV-21G, MG63, and NIH-3T3 cells were seeded onto the device and cultured for three days. To evaluate the size-dependent effect of nanodot arrays on cell growth, indices corresponding to cell proliferation, apoptosis, cell adhesion, and cytoskeletal organization were defined. VD(50) is defined as the diameter of nanodots on which 50% of the cell population remains viable. AD(50) is defined as the diameter of nanodots on which 50% of the cell population appears to have an apoptosis-like morphology. FD(50) is the diameter of nanodots that promotes the formation of 50% of the focal adhesions compared to cells grown on a flat surface. CD(50) is defined as the diameter of nanodots on which cells have half the amount of microfilament bundles compared to cells grown on a flat surface. We were able to distinguish between the invasive ability of HELA versus later-staged C33A cells. Ovarian cancer cell lines (ES2, PA-1, TOV-112D, and TOV-21G) also exhibited differential growth parameters that are associated with cell type, grade, and stage. Modulation of the growth of MG63 cells was also achieved. More broadly, we have established a platform that can be used to assess basic parameters of cell growth. A simplified fabrication process ensures mass production and lowers cost. According to our results, the device is capable of distinguishing among cancer cell lines at various stages and also provides basic design parameters for artificial implants. Our device will serve as a convenient and fast tool for tissue engineering and cancer treatment.
A Nanodot Array Modulates Cell Adhesion and Induces an Apoptosis-Like Abnormality in NIH-3T3 Cells
(2009-08-01)
Pan, Hsu-An
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Hung, Yao-Ching
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Su, Chia-Wei
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Tai, Shih-Ming
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Chen, Chiun-Hsun
Micro-structures that mimic the extracellular substratum promote cell growth and differentiation, while the cellular reaction to a nanostructure is poorly defined. To evaluate the cellular response to a nanoscaled surface, NIH 3T3 cells were grown on nanodot arrays with dot diameters ranging from 10 to 200 nm. The nanodot arrays were fabricated by AAO processing on TaN-coated wafers. A thin layer of platinum, 5 nm in thickness, was sputtered onto the structure to improve biocompatibility. The cells grew normally on the 10-nm array and on flat surfaces. However, 50-nm, 100-nm, and 200-nm nanodot arrays induced apoptosis-like events. Abnormality was triggered after as few as 24 h of incubation on a 200-nm dot array. For cells grown on the 50-nm array, the abnormality started after 72 h of incubation. The number of filopodia extended from the cell bodies was lower for the abnormal cells. Immunostaining using antibodies against vinculin and actin filament was performed. Both the number of focal adhesions and the amount of cytoskeleton were decreased in cells grown on the 100-nm and 200-nm arrays. Pre-coatings of fibronectin (FN) or type I collagen promoted cellular anchorage and prevented the nanotopography-induced programed cell death. In summary, nanotopography, in the form of nanodot arrays, induced an apoptosis-like abnormality for cultured NIH 3T3 cells. The occurrence of the abnormality was mediated by the formation of focal adhesions.
Assessment of gold nanoparticles as a size-dependent vaccine carrier for enhancing the antibody response against synthetic foot-and-mouth disease virus peptide
(2010-05-14)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Lin, Wei-Hsu
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Huang, Guewha Steven
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材料科學與工程學系
To assess the ability of gold nanoparticles (GNPs) to act as a size-dependent carrier, a synthetic peptide resembling foot-and-mouth disease virus (FMDV) protein was conjugated to GNPs ranging from 2 to 50 nm in diameter (2, 5, 8, 12, 17, 37, and 50 nm). An extra cysteine was added to the C-terminus of the FMDV peptide (pFMDV) to ensure maximal conjugation to the GNPs, which have a high affinity for sulfhydryl groups. The resultant pFMDV-GNP conjugates were then injected into BALB/c mice. Immunization with pFMDV-keyhole limpet hemocyanin (pFMDV-KLH) conjugate was also performed as a control. Blood was obtained from the mice after 4, 6, 8, and 10 weeks and antibody titers against both pFMDV and the carriers were measured. For the pFMDV-GNP immunization, specific antibodies against the synthetic peptide were detected in the sera of mice injected with 2, 5, 8, 12, and 17 nm pFMDV-GNP conjugates. Maximal antibody binding was noted for GNPs of diameter 8-17 nm. The pFMDV-GNPs induced a three-fold increase in the antibody response compared to the response to pFMDV-KLH. However, sera from either immunized mouse group did not exhibit an antibody response to GNPs, while the sera from pFMDV-KLH-immunized mice presented high levels of binding activity against KLH. Additionally, the uptake of pFMDV-GNP in the spleen was examined by inductively coupled plasma mass spectroscopy (ICP-MS) and transmission electron microscopy (TEM). The quantity of GNPs that accumulated in the spleen correlated to the magnitude of the immune response induced by pFMDV-GNP. In conclusion, we demonstrated the size-dependent immunogenic properties of pFMDV-GNP conjugates. Furthermore, we established that GNPs ranging from 8 to 17 nm in diameter may be ideal for eliciting a focused antibody response against a synthetic pFMDV peptide.
Assessment of the In Vivo Toxicity of Gold Nanoparticles
(2009-08-01)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Liau, Ian
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Huang, G. Steve
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材料科學與工程學系
The environmental impact of nanoparticles is evident; however, their toxicity due to their nanosize is rarely discussed. Gold nanoparticles (GNPs) may serve as a promising model to address the size-dependent biological response to nanoparticles because they show good biocompatibility and their size can be controlled with great precision during their chemical synthesis. Naked GNPs ranging from 3 to 100 nm were injected intraperitoneally into BALB/C mice at a dose of 8 mg/kg/week. GNPs of 3, 5, 50, and 100 nm did not show harmful effects; however, GNPs ranging from 8 to 37 nm induced severe sickness in mice. Mice injected with GNPs in this range showed fatigue, loss of appetite, change of fur color, and weight loss. Starting from day 14, mice in this group exhibited a camel-like back and crooked spine. The majority of mice in these groups died within 21 days. Injection of 5 and 3 nm GNPs, however, did not induce sickness or lethality in mice. Pathological examination of the major organs of the mice in the diseased groups indicated an increase of Kupffer cells in the liver, loss of structural integrity in the lungs, and diffusion of white pulp in the spleen. The pathological abnormality was associated with the presence of gold particles at the diseased sites, which were verified by ex vivo Coherent anti-Stoke Raman scattering microscopy. Modifying the surface of the GNPs by incorporating immunogenic peptides ameliorated their toxicity. This reduction in the toxicity is associated with an increase in the ability to induce antibody response. The toxicity of GNPs may be a fundamental determinant of the environmental toxicity of nanoparticles.
Camellia sinensis tea melanin suppresses transformation of the aryl hydrocarbon receptor and prevents against dioxin-induced toxicity in mice
(2008-02-01)
Hung, Yao-Ching
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Huang, Guewha Steven
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Sava, Vasyl M.
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Makan, Svetlana Y.
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Hong, Meng-Yen
The suppressive effects of Camellia sinensis tea melanin (CSTM) on transformation of aryl hydrocarbon receptor (AhR) induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were disclosed for the first time. CSTM strongly inhibited TCDD-induced toxicity with IC50 equalling 20.4 mu g mL(-1). Daily administration of CSTM (40 mg kg(-1), p.o.) prevented TCDD-induced body weight loss, ameliorated TCDD-induced mortality and prevented TCDD-induced hepatomegaly and thymic atrophy. Co-administration of CSTM significantly inhibited TCDD-induced hepatic CYP1 A1 activity. CSTM retarded transformation of AhR in vitro. In animals treated with CSTM, the antibody-secreting cells produced significantly (P < 0.05) more antibodies (32-34%) than the antigen control. Administration of TCDD caused a suppression of antibody-forming cells of 29-33% against the antigen control level. Co-administration of CSTM restored immunity to the control level. We demonstrated that CSTM directly competed with TCDD during the transformation of AhR and suppressed the downstream activation of genes associated with TCDD toxicity.
CLINICAL GENOMICS IDENTIFIES THE EXPRESSION OF HUMAN OVARIAN CARCINOMA
(2009-09-01)
Tsai, Meng-Hsiun
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Huang, Steve G.
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Tsaur, Shyh-Chang
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Hung, Yao-Ching
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Chiu, Sheng-Hsiung
Background: Human ovarian carcinoma is a silent killer. Most patients with human ovarian carcinoma are diagnosed in the advanced stages. Currently, no effective means of early detection are available. identifying genes that distinguish early cancers from normal tissue may provide insight to the initiation and progression of ovarian cancer. Methods: A large-scale gene expression profiling using 9,600 genes-complementary DNA array to study ovarian tumors was conducted. We analyzed 13 benign tumors, 22 invasive cancers (seven stage I and 15 stage III), and 16 normal ovaries. Cluster analysis revealed two major groups: the early-stage cluster that included 13 benign tumors and seven stage I cancers; and the late-stage cluster that included all stage III cancers. Results: There were 433 upregulated and 60 downregulated genes from the early-stage cluster, and 395 upregulated and 101 downregulated genes from the late-stage cluster. These genes showed altered functions for tumor/cancers, functions such as cell growth, cell cycle control, differentiation, signaling, apoptosis, ion transport, and morphogenesis. of most interest is the downregulation of nuclear receptor proteins. Differentially expressed genes between early-stage cluster and late-stage clusters were identified. We found 90 genes preferentially overly expressed and 165 genes relatively downregulated at late-stage cancers. The advanced cancer appears differently from early-stage cancer in cell growth, cell adhesion, hormone activity, morphology, and cell-cell signaling compared with benign or early-stage cancer. of particular interest are enzymes corresponding to pteridine biosynthesis, reported for the first time in this study. Real-time reverse transcription polymerase chain reaction showed that GCH1, KRT18, H3F3A, and MYBL2 were overly expressed in all cancer tissues, while MDFI, ARP3BETA, COL16A1, and MLN discriminated late-stage from early-stage cancers. Conclusion: The microarray provides valuable resources for comparing expression profiles of cancers at various stages and helps research the diagnosis and mechanism of ovarian cancer. [International journal of Gerontology 2009; 3(3): 163-169]
Control of growth and inflammatory response of macrophages and foam cells with nanotopography
(2012-07-16)
Mohiuddin, Mohammed
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Pan, Hsu-An
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Hung, Yao-Ching
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Huang, Guewha Steven
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材料科學與工程學系
Macrophages play an important role in modulating the immune function of the human body, while foam cells differentiated from macrophages with subsequent fatty streak formation play a key role in atherosclerosis. We hypothesized that nanotopography modulates the behavior and function of macrophages and foam cells without bioactive agent. In the present study, nanodot arrays ranging from 10aEuro to 200aEuronm were used to evaluate the growth and function of macrophages and foam cells. In the quantitative analysis, the cell adhesion area in macrophages increased with 10- to 50-nm nanodot arrays compared to the flat surface, while it decreased with 100- and 200-nm nanodot arrays. A similar trend of adhesion was observed in foam cells. Immunostaining, specific to vinculin and actin filaments, indicated that a 50-nm surface promoted cell adhesion and cytoskeleton organization. On the contrary, 200-nm surfaces hindered cell adhesion and cytoskeleton organization. Further, based on quantitative real-time polymerase chain reaction data, expression of inflammatory genes was upregulated for the 100- and 200-nm surfaces in macrophages and foam cells. This suggests that nanodots of 100aEuroaEuro parts per thousand and 200aEuronm triggered immune inflammatory stress response. In summary, nanotopography controls cell morphology, adhesions, and proliferation. By adjusting the nanodot diameter, we could modulate the growth and expression of function-related genes in the macrophages and foam cell system. The nanotopography-mediated control of cell growth and morphology provides potential insight for designing cardiovascular implants.
Control of In Vivo Transport and Toxicity of Nanoparticles by Tea Melanin
(2012)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Hong, Meng-Yeng
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Onischuk, Andrei Aleksandrovich
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Chiou, Jin Chern
"Nanoparticles are unfamiliar to researchers in toxicology. Toxicity may be generated simply due to the reduction in size. Compounds that prevent or cure toxic materials may not work on nanoparticles. Furthermore, as there are more and more applications of nanoparticles in drug delivery and in vivo imaging, controlling the transport and toxicity will be primary concerns for medical application of nanoparticles. Gold nanoparticles (GNPs) if injected intraperitoneally into mice can enter hippocampus and induce cognitive impairment. GNPs caused a global imbalance of monoamine levels, specifically affecting the dopaminergic and serotonergic neurons. Pretreatment of tea melanin significantly prevented the deposition of GNPs in mouse brains, especially in the hippocampus. Pretreatment of melanin completely alleviated GNP-induced impairment of cognition. Pre-administration of melanin stably maintained monoamines at normal profiles. Melanin completely prevented the invasion of GNPs into the Cornu Ammonis region of the hippocampus shown by coherent anti-Stoke Raman scattering microscopy. Here we show that the administration of tea melanin prevented the accumulation of Au in brain, the imbalance of monoamines, and the impairment of cognition in mice. The current study provides a therapeutic approach to toxicity of nanoparticles and a novel strategy to control the transport of GNP in mouse brain."
Detection of gold nanoparticles using an immunoglobulin-coated piezoelectric sensor
(2008-12-10)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Chen, Kaochao
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Huang, Guewha Steven
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材料科學與工程學系
Since the existence of nanoparticles in our environment has already attracted considerable attention due to their possible toxic impact on biological systems, the field detection of nanoparticles is becoming a technology that will be much in need. We have constructed a piezoelectric sensor with an antibody-coated electrode. The antiserum can bind gold nanoparticles with a high degree of selectivity and sensitivity. The biosensor thus constructed can detect 4, 5, or 6 nm gold nanoparticles (GNPs) depending on the coated antiserum. The sensitivity for the detection of 5 nm GNPs was 10.3 +/- 0.9 ng Hz(-1), with the low limit of detection at 5.5 ng. A quartz crystal microbalance (QCM) sensor was capable of detecting GNPs and other types of nanoparticle, such as ZnO, or Fe(3)O(4). The current study provides, for the first time, a platform for detecting nanoparticles in a convenient, economical manner.
Effects of static magnetic fields on the development and aging of Caenorhabditis elegans
(2010-06-15)
Hung, Yao-Ching
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Lee, Jia-Huey
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Chen, Huang-Meng
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Huang, Guewha Steven
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材料科學與工程學系奈米科技碩博班
The current study investigated the possible effects of static magnetic fields (SMFs) on the developmental and aging processes of Caenorhabditis elegans. Nematodes were grown in the presence of SMFs of strengths varying from 0 to 200 mT. The rate of development and the lifespan were recorded. Treatment with a 200 mT SMF reduced the development time from the L2 to the L3 stage by 20%, from L3 to L4 by 23%, and from L4 to young adult by 31%. After SMF treatment, the average lifespan was reduced from 31 days to 24 days for wild-type nematodes. The up-regulation of clk-1, lim-7, daf-2, unc-3 and age-1 after SMF treatment was verified by quantitative real-time RT-PCR. Apparently, induction of gene expression is selective and dose dependent. The total developmental time was significantly reduced for the lin-4, lin-14, lin-41 and lim-7 mutants, but not for the let-7, clk-1, unc-3 and age-1 mutants. Lifespan analyses revealed that the let-7, unc-3 and age-1 mutants were not affected by SMF treatment. Here we show that SMFs accelerate nematode development and shorten nematode lifespan through pathways associated with let-7, clk-1, unc-3 and age-1.
Gold nanoparticles caused learning impairment in mice
(2009)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Liau, Ian
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Lin, Li-Wei
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Hong, Meng-Yeng
Aims : To investigate the toxic effect of gold nanoparticles (GNPs) to the learning and memory of mice. Main methods: Naked GNPs of 17 nm and 37 nm were synthesized, purified, and injected intraperitoneally into BALB/C mice at a dose of 8 mg/kg/week. Passive avoidance test was performed to detect the possible leaning impairment. Biochemical analysis to examine levels of monoamine and acetylcholine in mouse brain was also performed. ICP-MS was performed to examine the presence of GNP in brain. Localization of GNP at hippocampus was surveyed by ex vivo Coherent anti-Stoke Raman scattering (CARS) microscopy. Transmission electron microscopy (TEM) was also performed to verify the cellular location of GNPs at hippocampus. Key findings: Both 17 nm and 37 nm GNPs induced severe sickness in rnice. However, only 17 rim GNP impaired the learning and memory of mice. GNP treatment elevated levels of dopamine from 114.5 ng/mouse brain to 143.6 ng/mouse brain for 17 nm (p<0.01) and to 138.2 for 37 nm GNP (p<0.05). Serotinin was significantly reduced by 17 nm GNP treatment from 57.2 ng/mouse brain to 44.3 ng/mouse brain (p<0.05). ICP-MS indicated the presence of GNPs in every part of the brain. CARS microscopy showed that 17 nm GNP was located at the Cornu Amonis regions of hippocampus where neuronal cells clustered, while 37 nm GNP was excluded from the cell clustered region. TEM and EDAX verified the presence of 17 nm GNP in the cytoplasm and in the dendrite of pyramidal cell, while 37 rim GNP was found in the extracellular region of neuronal cells. TEM image also indicated that both endocytosis and free diffusion coexisted for the invasion of 17 nm GNP. Significance: The current study provided evidence that nanoparticles were capable of entering into brain and affecting normal brain function. The size-dependent invading ability of GNPs provides an extra dimension for drug delivery.
Nanosurface design of dental implants for improved cell growth and function
(2012-08-24)
Pan, Hsu-An
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Hung, Yao-Ching
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Chiou, Jin-Chern
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Tai, Shih-Ming
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Chen, Hsin-Hung
A strategy was proposed for the topological design of dental implants based on an in vitro survey of optimized nanodot structures. An in vitro survey was performed using nanodot arrays with dot diameters ranging from 10 to 200 nm. MG63 osteoblasts were seeded on nanodot arrays and cultured for 3 days. Cell number, percentage undergoing apoptotic-like cell death, cell adhesion and cytoskeletal organization were evaluated. Nanodots with a diameter of approximately 50 nm enhanced cell number by 44%, minimized apoptotic-like cell death to 2.7%, promoted a 30% increase in microfilament bundles and maximized cell adhesion with a 73% increase in focal adhesions. An enhancement of about 50% in mineralization was observed, determined by von Kossa staining and by Alizarin Red S staining. Therefore, we provide a complete range of nanosurfaces for growing osteoblasts to discriminate their nanoscale environment. Nanodot arrays present an opportunity to positively and negatively modulate cell behavior and maturation. Our results suggest a topological approach which is beneficial for the design of dental implants.
Psoralea corylifolia extract ameliorates experimental osteoporosis in ovariectomized rats
(2007)
Tsai, Meng-Hsiun
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Huang, G. Steven
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Hung, Yao-Ching
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Bin, Liu
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Liao, Lung-Te
We evaluated the protective effect of Psoralea corylifolia L. (PCL) extract on the ovariectomized (OVX) rat model. The biochemical markers of bone turnover, calcium metabolism, and calcium balance were examined. PCL extract (25 mg or 50 mg/kg body weight/day) was orally administrated to OVX rats for 3 months. PCL extract did not alter weight gain or uterus weight in OVX rats. PCL extract significantly increased serum Ca (calcium) levels (p < 0.05, vs. OVX group) as well as decreased urinary Ca excretion (p < 0.05, vs. OVX group) in OVX rats. The upregulation of serum osteocalcin level by ovariectomy was suppressed by treatment with PCL extract in rats (p < 0.05, vs. OVX group). PCL extract increased bone mineral density at 50 mg/kg body weight/day in OVX rats (p < 0.05, vs. OVX group). Our results indicate that orally administrated PCL extract can decrease urinary calcium excretion and decrease serum osteocalcin in OVX rats, resulting in positive effects on bone mineral density as well as bone formation. In conclusion, our studies showed that PCL might be a potential candidate for treatment of postmenopausal osteoporosis.
Size-dependent impairment of cognition in mice caused by the injection of gold nanoparticles
(2010-12-03)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Lin, Li-Wei
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Liau, Ian
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Hong, Meng-Yeng
We explored the size-dependent impairment of cognition in mice caused by the injection of gold nanoparticles (GNPs). GNPs of 17 and 37 nm in diameter were injected intraperitoneally into BALB/c mice at doses ranging from 0.5 to 14.6 mg kg(-1). ICP-MS was performed on brain tissue collected 1, 14 and 21 days after the injection. A passive-avoidance test was performed on day 21. Monoamine levels were determined on day 21. The microscopic distribution of GNPs in the hippocampus was examined using coherent anti-Stokes Raman scattering (CARS) microscopy and transmission electron microscopy (TEM). The results indicated that 17 nm GNPs passed through the blood-brain barrier more rapidly than 37 nm GNPs. Treatment with 17 nm GNPs decreased the latency time, which was comparable to the effect of scopolamine treatment, while 37 nm GNPs showed no significant effect. Dopamine levels and serotonin levels in the brain were significantly altered by the injection of 17 and 37 nm GNPs. GNPs affected dopaminergic and serotonergic neurons. CARS microscopy indicated that 17 nm GNPs entered the Cornu Ammonis (CA) region of the hippocampus, while 37 nm GNPs were excluded from the CA region. TEM verified the presence of 17 nm GNPs in the cytoplasm of pyramidal cells. In this study, we showed that the ability of GNPs to damage cognition in mice was size-dependent and associated with the ability of the particles to invade the hippocampus. The dosage and duration of the treatment should be taken into account if GNPs are used in the future as vehicles to carry therapeutic agents into the brain.
The spatial and temporal control of cell migration by nanoporous surfaces through the regulation of ERK and integrins in fibroblasts
(2013-01-01)
Pan, Hsu-An
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Liang, Jia-You
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Hung, Yao-Ching
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Lee, Chia-Hui
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Chiou, Jin-Chern
Nanotopography controls cell behaviours, such as cell adhesion and migration. However, the mechanisms responsible for topology-mediated cellular functions are not fully understood. A variety of nanopores was fabricated on 316L stainless steel to investigate the effects of spatial control on the growth and function of fibroblasts, the temporal regulation of integrins, and their effects on migration. The NIH-3T3 fibroblast cell line was cultured on the nanopore surfaces, whose pore diameters ranged from 40 to 210 nm. The 40 and 75 nm nanopores enhanced cell proliferation, focal adhesion formation and protein expression of vinculin and beta-tubulin after 24 h of incubation. Integrin expression was analysed by qPCR, which showed the extent of spatial and temporal regulation achieved by the nanopores. The protein expression of pERK1/2 was greatly attenuated in cells grown on 185 and 210 nm nanopore surfaces at 12 and 24 h. In summary, the 40 and 75 nm nanopore surfaces promoted cell adhesion and migration in fibroblasts by controlling the temporal expression of integrins and ERK1/2. The current study provides insight into the improvement of the design of stainless steel implants and parameters that affect biocompatibility. The ability to regulate the expression of integrin and ERK1/2 using nanopore surfaces could lead to further applications of surface modification in the fields of biomaterials science and tissue engineering. (C) 2012 Elsevier Ltd. All rights reserved.
Thea sinensis melanin prevents cisplatin-induced nephrotoxicity in mice
(2007-07-01)
Hung, Yao-Ching
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Huang, G. Steven
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Lin, Li-Wei
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Hong, Meng-Yen
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Se, Pei-San
The preventive effect of Thea sinensis melanin (TSM) against cisplatin-induced nephrotoxicity was studied on ICR mice. Animals were given 20 mg/kg i.p. of cisplatin, and TSM was injected i.p. in doses 10-40 mg/kg 2 h before intoxication. The protective effects were evidenced by a complete inhibition of the cisplatin-induced elevation of serum Blood Urea nitrogen (BUN), prevention of oxidative stress, and complete blockade of cisplatin-induced elevation of serum creatinine. TSM by itself, however, did not affect the renal functional parameters, including serum BUN and creatinine. Real-time RT-PCR was applied to quantify mRNA levels of cisplatin-treated mouse kidney compared to normal mouse kidney for selected marker genes. Cisplatin treatment increases mRNA levels 40-fold for glutathione-S-transferases (Gstp2), 15-fold for soluble epoxide hydrolase (Ephx1), 15-fold for lipocalin 2 (Lcn2), 9-fold for lysozyme (Lyz), 5-fold for UDP glycosyltransferase 2 (Utg2b), 30-fold for survival motor neuron (Smn1), 30-fold for guanidinoacetate methyltransferase (Gamt), 80-fold for urine retinol binding protein (Rbp4), 60-fold for aminopeptidase N (Apn), 60-fold for cytochrome P450 (Cyp2d18), and 100-fold for ornithine aminotransferase (Oat). Pre-administration of TSM restored normal expression of marker genes for cisplatin-treated mouse kidneys. TSM by itself, however, did not affect the transcription for marker genes. Results obtained demonstrate that TSM pre-administration can prevent the renal toxic effects of cisplatin. (c) 2007 Elsevier Ltd. All rights reserved.
Topographic control of the growth and function of cardiomyoblast H9c2 cells using nanodot arrays
(2012-01-01)
Pan, Hsu-An
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Hung, Yao-Ching
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Sui, Yu-Ping
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Huang, G. Steve
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材料科學與工程學系
Cardiovascular stents require optimised control for the enhancement or inhibition epithelial and smooth muscle cell growth in close contact with the implant. Here we propose that the surface topology in contact with the living cells could be designed to control and optimise the growth and function of such cells. The cardiomyoblast H9c2 was cultured on nanodot arrays with dot diameters ranging between 10 and 200 nm. On the 50-nm nanodot arrays H9c2 showed maximum attachment and proliferation with largest cell area and extended lamellipodia. In contrast, 53.7% and 72.6% reductions of growth were observed on the 100- and 200-nm nanodot arrays after 3 days. Immunostaining indicated that nanodots smaller than 50-nm induced cell adhesion and cytoskeleton organization. Expression of genes associated with fibrosis and hypertrophy was up-regulated in cells grown on 100-nm nanodots. Western blot data showed high levels of expression for vinculin and plasminogen activator inhibitor-1 for cells cultured on 50-nm nanodots. Nanotopography controls cell adhesion, morphology and proliferation. By adjusting the diameter of the nanodots, we could modulate the growth and expression of function-related genes and proteins of H9c2 cardiomyoblasts. The current study provides insights for improved design of artificial implants and parameters that affect biocompatibility. (C) 2011 Elsevier Ltd. All rights reserved.
Ultrasensitive Detection of Cymbidium Mosaic Potexvirus Using a Single-Wall Carbon Nanotube-Functionalized Quartz Crystal Microbalance
(2010)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Chiou, Jin-Chern
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Wang, Hui-Liang
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Huang, Hung-Shu
We have developed an ultrasensitive, convenient, real-time platform for detecting Cymbidium mosaic potexvirus (CymMV) based on single-wall carbon nanotube (SWNT)-functionalized quartz crystal microbalance (QCM) sensors. Functionalization was achieved by coating the QCM electrode with SWNTs, followed by 1,1'-carbonyldiimidazole-activated Tween 20 (CDI-Tween 20) modification and conjugation of antibodies. Sensitivity was enhanced from 2.18 to 11.5 Hz ng(-1) when 0.1 mu g mL(-1) CymMV was applied. The low limit of detection of SWNT-functionalized QCM sensors was improved from 2.08 to 0.502 ng. The SWNT-functionalized QCM sensor was successfully used to quantify the amount of CymMV contained in infected orchid leaves. Compared to enzyme-linked immunosorbent assay (ELISA), SWNT-functionalized QCM sensors are fast, economical, and ultra-sensitive, with comparable sensitivities. The current study demonstrates the application of QCM sensors as a convenient platform to detect and quantify CymMV. (C) 2010 The Japan Society of Applied Physics