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A Novel SERS Substrate Containing Silver Nanoparticle-Deposited Photoresist Microcone Array for Sensitive Biosensing

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IEEE Sensors Journal 2024年第2期24卷 1398-1404页

作者： Lin, Ying-Ting Wang, Sheng-Hsiang Tseng, Yu-Hsin Lin, Ze-Cheng Chen, Jyun-Tu Chang, Cheng-Chung Wang, Gou-Jen National Chung Hsing University Program in Tissue Engineering and Regenerative Medicine Taichung40227 Taiwan National Chung Hsing University Department of Mechanical Engineering Taichung40227 Taiwan Graduate Institute of Biomedical Engineering National Chung Hsing University Taichung40227 Taiwan

In this study, we developed a novel surface-enhanced Raman scattering (SERS) substrate formed by depositing silver nanoparticles (AgNPs) on a silver thin film-coated photoresist (PR) microcone array. The semiconductor photolithographic process was adopted for fabricating the PR microcone array. The hotspot enhancement effect of the SERS substrates was optimized by adjusting the concentration of the trisodium citrate, which was used as a reducing agent and a stabilizing agent for the synthesis of AgNPs. Experimental results showed that the proposed SERS substrate could detect the indicating peaks of Rhodamine 6G (R6G) at a relatively low concentration of 10-12 M. A standard detection curve for HbA1c detection with a linear detection range of 5000-20000 ng/mL with good linearity of 0.994 and a limit of detection (LOD) of 850.2 ng/mL was further established by adding HbA1c agent to artificial blood to simulate a real environmental condition. Recovery rate detection experiments confirmed the reproducibility of the established standard detection curve. © 2001-2012 IEEE.

关键词： Silver nanoparticles

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CHITOSAN-COLLAGEN COMPOSITE SCAFFOLDS FOR GROWTH DIRECTION GUIDANCE OF PC12 AXONS

CHITOSAN-COLLAGEN COMPOSITE SCAFFOLDS FOR GROWTH DIRECTION G...

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ASME 2024 International Design engineering Technical Conferences and Computers and Information in engineering Conference, IDETC-CIE 2024

作者： Tsai, Po-Yu Lin, Ying-Ting Wang, Gou-Jen Graduate Institute of Biomedical Engineering National Chung-Hsing University Taichung Taiwan PhD Program in Tissue Engineering and Regenerative Medicine National Chung-Hsing University Taichung Taiwan Department of Mechanical Engineering Graduate Institute of Biomedical Engineering National Chung-Hsing University Taichung Taiwan

ISBN: (纸本)9780791888421

In this study, nerve conduits of chitosan and collagen composites with biocompatibility and biodegradability were fabricated. The effects of material composition and surface morphology on the axonal growth and directionality of rat epinephric pheochromocytoma PC12 were investigated. The cells were separated by a PDMS structure to simulate nerve injury, and the growth of axons of PC12 cells was observed under the separation of large gaps. At the same time, mitochondria extracted from osteosarcoma cybrid cells (C2-GFP) were added to the culture to investigate the effect of healthy mitochondria on axon differentiation and growth. The experimental results showed that the chitosan-collagen composite film can promote the growth of PC12 axons more effectively than the pure chitosan film. Cell culture results at different groove widths also showed that smaller grooves resulted in more axon numbers and longer axon lengths. Nerve injury simulation experiments showed that the nerve repair could be almost completed after 12 days of culture. Copyright © 2024 by ASME.

关键词： Mitochondria

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An Electrochemical HbA1c Immunosensor With a Simple Electrode of Gold Nanoparticle Monolayer

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IEEE Sensors Journal 2023年第15期23卷 16579-16584页

作者： Tseng, Chun-Chiang Lin, Ying-Ting Wang, Gou-Jen National Chung-Hsing University Department of Mechanical Engineering Taichung40227 Taiwan National Chung-Hsing University Program in Tissue Engineering and Regenerative Medicine Taichung40227 Taiwan National Chung-Hsing University Department of Mechanical Engineering The Graduate Institute of Biomedical Engineering Taichung40227 Taiwan

This study developed a novel electrochemical glycosylated hemoglobin (HbA1c) immunosensor. Polyethylene terephthalate (PET) was used as a substrate. After gold (Au) film sputtering on the PET surface, 1,6-hexanedithol with thiol groups on both terminals was used as an intermediary to attach a monolayer of uniformly distributed Au nanoparticles (AuNPs) to the Au thin film to form a nanostructured electrode. HbA1c antibodies were successfully immobilized on the AuNP monolayer via the self-assembled monolayer (SAM) approach. To mimic a real clinical detection environment, a calibration curve for HbA1c detection was performed in a mixture solution containing heparin sodium, red blood cell (RBC) lysis buffer, artificial human serum, and hemoglobin. The experimental results revealed that the proposed HbA1c immunosensor detected HbA1c in a linear range from 2500 to 10 000 ng/mL with a limit of detection of 2253 ng/mL. The developed HbA1c-sensing chip offers the advantages of simple operation, low cost, short detection time, and provides high feasibility for clinical application. © 2001-2012 IEEE.

关键词： Gold nanoparticles

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Effect of silver/zinc co-dopant on enhancing cytotoxicity and antibacterial property of spray pyrolyzed bioactive glasses

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Journal of Asian Ceramic Societies 2024年第1期12卷 104-116页

作者： Ningsih, Henni Setia Vietanti, Frizka Wong, Chin-Chean Chou, Yu-Jen Department of Mechanical Engineering National Taiwan University of Science and Technology Taipei Taiwan Department of Orthopedics Taipei Medical University Shuang Ho Hospital New Taipei Taiwan Department of Orthopedics School of Medicine College of Medicine Taipei Medical University Taipei Taiwan Research Center of Biomedical Devices Taipei Medical University Taipei Taiwan International PhD Program for Cell Therapy and Regenerative Medicine College of Medicine Taipei Medical University Taipei Taiwan

Owing to the advancement of bioactive materials, bioactive glasses (BGs) are progressively being recognized as promising choices in fields of bone and tissue engineering with their superior properties such as bioactivity and biocompatibility. However, their applications are limited by the fact that the material itself does not possess antibacterial properties. To address this limitation, antibacterial BGs with silver-zinc co-dopant were proposed and successfully synthesized via the one-step spray pyrolysis technique in this work. The phase composition, chemical composition, surface morphology, and nanoscale structures of the obtained BG specimens were characterized using X-ray diffraction, energy dispersive spectroscopy, scanning electron microscopy, and transmission electron microscopy. Moreover, in-vitro bioactivity was assessed through immersion of the SBF solution, while the in-vitro antibacterial property was examined using Escherichia coli bacteria. Meanwhile, the viability of MC3T3-E1 osteoblast cells was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Finally, the results showed positive effects of zinc addition, and the influence of silver and zinc dopants on the in-vitro bioactivity, antibacterial property, and cell viability were discussed. © 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The Korean Ceramic Society and The Ceramic Society of Japan.

关键词： Spray pyrolysis

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Semilicoisoflavone B induces oral cancer cell apoptosis by targeting claspin and ATR-Chk1 signaling pathways

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Environmental Toxicology 2024年第4期39卷 2417-2428页

作者： Hsieh, Ming-Ju Lin, Chia-Chieh Lo, Yu-Sheng Chuang, Yi-Ching Ho, Hsin-Yu Chen, Mu-Kuan Oral Cancer Research Center Changhua Christian Hospital Changhua Taiwan Doctoral Program in Tissue Engineering and Regenerative Medicine College of Medicine National Chung Hsing University Taichung Taiwan Graduate Institute of Biomedical Sciences China Medical University Taichung Taiwan Department of Otorhinolaryngology Head and Neck Surgery Changhua Christian Hospital Changhua Taiwan Department of Post-Baccalaureate Medicine College of Medicine National Chung Hsing University Taichung Taiwan

The prevalence of oral squamous cell carcinoma (OSCC) is increasing worldwide mainly due to poor oral hygiene and unrestricted lifestyle. Advanced-stage OSCC is associated with poor prognosis and a 5-year survival rate of only 30%–50%. The present study was designed to investigate the anticancer effect and mode of action of Glycyrrhiza-derived semilicoisoflavone B (SFB) in 5-fluorourasil (5FU)-resistant human OSCC cell lines. The study findings revealed that SFB significantly reduces OSCC cell viability and colony formation ability by arresting cell cycle at the G2/M and S phases and reducing the expressions of key cell cycle regulators including cyclin A, cyclin B, CDC2, and CDK2. The compound caused a significant induction in the percentage of nuclear condensation and apoptotic cells in OSCC. Regarding pro-apoptotic mode of action, SFB was found to increase Fas-associated death domain and death receptor 5 expressions and reduce decoy receptor 2 expression, indicating involvement of extrinsic pathway. Moreover, SFB was found to increase pro-apoptotic Bim expression and reduce anti-apoptotic Bcl-2 and Bcl-xL expressions, indicating involvement of intrinsic pathway. Moreover, SFB-mediated induction in cleaved caspases 3, 8, and 9 and cleaved poly(ADP-ribose) polymerase confirmed the induction of caspase-mediated apoptotic pathways. Regarding upstream signaling pathway, SFB was found to reduce extracellular signal regulated kinase 1/2 (ERK) phosphorylation to execute its pro-apoptotic activity. The Human Apoptotic Array findings revealed that SFB suppresses claspin expression, which in turn caused reduced phosphorylation of ATR, checkpoint kinase 1 (Chk1), Wee1, and CDC25C, indicating disruption of ATR-Chk1 signaling pathway by SFB. Taken together, these findings indicate that SFB acts as a potent anticancer compound against 5FU-resistant OSCC by modulating mitogen-activated protein kinase (MAPK) and ATR-Chk1 signaling pathways. © 2024 Wiley Periodicals LLC.

关键词： Phosphorylation

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Picrasidine I Regulates Apoptosis in Melanoma Cell Lines by Activating ERK and JNK Pathways and Suppressing AKT Signaling

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Environmental Toxicology 2024年第12期39卷 5309-5320页

作者： Shieu, Mu-Kuei Lin, Chia-Chieh Ho, Hsin-Yu Lo, Yu-Sheng Chuang, Yi-Ching Hsieh, Ming-Ju Department of Dermatology Changhua Christian Hospital Changhua Taiwan Oral Cancer Research Center Changhua Christian Hospital Changhua Taiwan Graduate Institute of Clinical Medicine College of Medicine National Chung Hsing University Taichung Taiwan Doctoral Program in Tissue Engineering and Regenerative Medicine College of Medicine National Chung Hsing University Taichung Taiwan Graduate Institute of Biomedical Sciences China Medical University Taichung Taiwan

World Health Organization data indicate a continuous increase in melanoma incidence, with metastatic melanoma characterized by poor prognosis and drug resistance. The exploration of therapeutics derived from natural products remains an active area of in vitro research. The aim of this study was to determine the antitumor effects of picrasidine I, a natural compound extracted from Picrasma quassioides, against two melanoma cell lines. We selected two metastatic melanoma cell lines, HMY-1 and A2058, for molecular studies, including Western blotting, 4′,6-diamidino-2-phenylindole staining, and flow cytometry. Picrasidine I demonstrated cytotoxic effects against the HMY-1 and A2058 melanoma cell lines. It induced cell cycle arrest in the sub-G1 phase and downregulated cell cycle–related proteins (e.g., cyclin A2, D1, cyclin-dependent kinases 4, and 6). In the intrinsic apoptosis pathway, picrasidine I activated proapoptotic proteins (e.g., Bax, Bak, t-Bid, BimL/S) and suppressed the expression of antiapoptotic proteins (e.g., Bcl-2, Bcl-xL), with an observed increase in the quantity of depolarized cells. In addition, the apoptotic effects of picrasidine I were linked to the activation of the c-Jun N-terminal kinase and extracellular signal-regulated kinase pathways and the inhibition of the protein kinase B signaling pathway. A human apoptosis array indicated claspin inhibition upon picrasidine I treatment, suggesting the potential involvement of picrasidine I in apoptosis and cell cycle regulation. Our findings suggest that picrasidine I has potential as a candidate for treating advanced melanoma, and thus these findings warrant further investigation. The modulation of claspin expression by picrasidine I could be investigated further as a potential biomarker to predict its efficacy in related to advanced stages of melanoma. © 2024 The Author(s). Environmental Toxicology published by Wiley Periodicals LLC.

关键词： Dermatology

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Isokinetic training of lower extremity during the early stage promote functional restoration in elder patients with disability after Total knee replacement (TKR) - a randomized control trial

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BMC Geriatrics 2024年第1期24卷 1页

作者： Cheng, Yuan-Yang Chen, Cheng-Hsu Wang, Shun-Ping Department of Physical Medicine and Rehabilitation Taichung Veterans General Hospital Taichung Taiwan School of Medicine National Yang Ming Chiao Tung University Taipei Taiwan Department of Post-Baccalaureate Medicine College of Medicine National Chung Hsing University Taichung Taiwan Division of Nephrology Department of Internal Medicine Taichung Veterans General Hospital Taichung Taiwan Ph.D. Program in Tissue Engineering and Regenerative Medicine College of Medicine National Chung Hsing University Taichung Taiwan Department of Orthopedics Taichung Veterans General Hospital Taichung Taiwan

Background: Transient progressive weakness and disability of lower limb during the early stage after TKR will increase the risk of fall, but the superior postoperative strength training mode have not been elucidated for functional restoration. This study aimed to compare whether the isokinetic lower limb training is superior to either isotonic or home isometric exercise during early stage after TKR in older people. Methods: A total of 43 recruited old participants (mean age, 68.40 years old) receiving TKR were divided randomly based on the different four-week training modes into three groups including isokinetic, isotonic, and home isometric exercise (control group). The primary outcome was set as functional performance in terms of Timed Up and Go (TUG) test and the secondary outcomes include the peak torque of knee at 60 and 120 degree/ second, Short-Form 36 Health Survey (SF-36), and Western Ontario and McMaster Universities Arthritis index (WOMAC). Results: All of the peak torque measurements of the knee improved significantly in both the isokinetic and the isotonic group, but not in the control group. Although isotonic training resulted in more strength gains, a significant enhancement in TUG test was observed in the isokinetic group only (p = 0.003). However, there were no significantly improvement of TUG test after training in other two groups. SF-36 and WOMAC improved after training in all three groups, with no significant difference in the degree of improvement between groups. Conclusion: Isokinetic training for 4 weeks following TKR effectively improved all the outcome parameters in this study, including the TUG test, lower limb strength, and functional scores. However, both isokinetic and isotonic training modes could be recommended after TKR because of no significant difference in the degree of improvement between these two groups. Trial registration: Clinical trial registration number: NCT02938416. Level of evidence: I © The Author(s) 2024.

关键词： Isokinetic Isometric Isotonic Timed up and go Total knee replacement

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Roles of Neuronal Protein Kinase Cε on Endoplasmic Reticulum Stress and Autophagic Formation in Diabetic Neuropathy

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Molecular Neurobiology 2024年第5期61卷 2481-2495页

作者： Kan, Yu-Yu Chang, Ying-Shuang Liao, Wen-Chieh Chao, Tzu-Ning Hsieh, Yu-Lin School of Medicine College of Medicine National Sun Yat-sen University Kaohsiung 80424 Taiwan Department of Anatomy School of Medicine College of Medicine Kaohsiung Medical University Kaohsiung 80708 Taiwan Doctoral Program in Tissue Engineering and Regenerative Medicine College of Medicine National Chung Hsing University Taichung 40227 Taiwan Department of Post-Baccalaureate Medicine College of Medicine National Chung Hsing University Taichung 40227 Taiwan School of Post-Baccalaureate Medicine College of Medicine Kaohsiung Medical University Kaohsiung 80708 Taiwan Department of Medical Research Kaohsiung Medical University Hospital Kaohsiung 80708 Taiwan

In chronic diabetic neuropathy (DN), the cellular mechanisms of neuropathic pain remain unclear. Protein kinase C epsilon (PKCε) is an intracellular signaling molecule that mediates chronic pain. This paper addresses the long-term upregulated PKCε in DN associated with endoplasmic reticulum (ER) stress and autophagic formation and correlates to chronic neuropathic pain. We found that thermal hyperalgesia and mechanical allodynia course development were associated with PKCε upregulation after DN but not skin denervation. Pathologically, PKCε upregulation was associated with the expression of inositol-requiring enzyme 1α (IRE1α;ER stress–related molecule) and ubiquitin D (UBD), which are involved in the ubiquitin-proteasome system (UPS)-mediated degradation of misfolded proteins under ER stress. Manders coefficient analyses revealed an approximately 50% colocalized ratio for IRE1α(+):PKCε(+) neurons (0.34–0.48 for M1 and 0.40–0.58 for M2 Manders coefficients). The colocalized coefficients of UBD/PKCε increased (M1: 0.33 ± 0.03 vs. 0.77 ± 0.04, p < 0.001;M2: 0.29 ± 0.05 vs. 0.78 ± 0.04;p < 0.001) in the acute DN stage. In addition, the regulatory subunit p85 of phosphoinositide 3-kinase, which is involved in regulating insulin signaling, exhibited similar expression patterns to those of IRE1α and UBD;for example, it had highly colocalized ratios to PKCε. The ultrastructural examination further confirmed that autophagic formation was associated with PKCε upregulation. Furthermore, PKCεv1-2, a PKCε specific inhibitor, reverses neuropathic pain, ER stress, and autophagic formation in DN. This finding suggests PKCε plays an upstream molecule in DN-associated neuropathic pain and neuropathology and could provide a potential therapeutic target. © 2023, The Author(s).

关键词： Autophagy Diabetic Neuropathy ER Stress IRE1α PI3Kp85 PKCε Ubiquitin D

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Synthesis of Methotrexate-Loaded Dumbbell-Shaped Titanium Dioxide/Gold Nanorods Coated with Mesoporous Silica and Decorated with Upconversion Nanoparticles for Near-Infrared-Driven Trimodal Cancer Treatment

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ACS Applied Materials and Interfaces 2023年第28期15卷 33335-33347页

作者： Dash, Pranjyan Thirumurugan, Senthilkumar Tseng, Ching-Li Lin, Yu-Chien Chen, Sung-Lung Dhawan, Udesh Chung, Ren-Jei Taipei10608 Taiwan Graduate Institute of Biomedical Materials and Tissue Engineering College of Biomedical Engineering Taipei Medical University Taipei110 Taiwan International Ph.D. Program in Biomedical Engineering College of Biomedical Engineering Taipei Medical University Taipei110 Taiwan Research Center of Biomedical Device College of Biomedical Engineering Taipei Medical University Taipei110 Taiwan International Ph.D. Program in Cell Therapy and Regenerative Medicine College of Medicine Taipei Medical University Taipei110 Taiwan Centre for the Cellular Microenvironment Division of Biomedical Engineering James Watt School of Engineering Mazumdar-Shaw Advanced Research Centre University of Glasgow GlasgowG116EW United Kingdom Taipei10608 Taiwan

This study prepared dumbbell-shaped titanium dioxide (TiO2)/gold nanorods (AuNRs) coated with mesoporous silica shells (mS) (AuNRs-TiO2@mS). Methotrexate (MTX) was further loaded into the AuNRs-TiO2@mS, and then upconversion nanoparticles (UCNPs) were decorated to form AuNRs-TiO2@mS-MTX: UCNP nanocomposites. TiO2 is used as an intense photosensitizer (PS) to produce cytotoxic reactive oxygen species (ROS), leading to photodynamic therapy (PDT). Concurrently, AuNRs exhibited intense photothermal therapy (PTT) effects and photothermal conversion efficiency. In vitro results suggested that these nanocomposites can kill oral cancer cells (HSC-3) without toxicity through irradiation of NIR laser, owing to the synergistic effect. The in vivo studies indicated that these nanocomposites exhibited excellent antitumor effects through synergistic PDT/PTT/chemotherapy under a near-infrared (NIR) 808 nm laser irradiation. Thus, these AuNRs-TiO2@mS: UCNP nanocomposites have great potential to undergo deep tissue penetration with enhanced synergistic effects through NIR-triggered light for cancer treatment. © 2023 American Chemical Society

关键词： Titanium dioxide

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A novel anode of gold nanoparticle coated Au nanorod arrays for nonenzymatic glucose fuel cells

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Journal of Power Sources 2024年 590卷

作者： Ting, Ya-Chun Lin, Ying-Ting Wang, Gou-Jen Department of Mechanical Engineering National Chung-Hsing University Taichung40227 Taiwan Program in Tissue Engineering and Regenerative Medicine National Chung-Hsing University Taichung40227 Taiwan Graduate Institute of Medical Engineering National Chung-Hsing University Taichung40227 Taiwan Regenerative Medicine and Cell Therapy Research Center Kaohsiung Medical University Kaohsiung80708 Taiwan

In this study, we propose a novel nonenzymatic glucose fuel cell (GFC) with an anode of gold nanoparticle coated Au nanorod arrays (AuNPs@AuNRA). The anode is fabricated through anodic aluminum oxide production, photolithographic patterning, core nickel nanorods electroforming, immersing gold, and gold nanoparticle electrodeposition. A double-chamber GFC, consisting of the proposed anode, a graphene-modified glassy carbon cathode, and a Nafion proton-exchange membrane separating the anode and cathode, is constructed. Experimental results reveal that the proposed GFC can produce a maximum power density of 822 μW/cm2, a Voc of 0.8 V, and an Isc density of 5.15 mA/cm2 at a glucose concentration of 0.2 M. The energy conversion efficiency of the proposed nonenzymatic GFC reaches 51.6%, which can be attributed to the high catalyzation area of the AuNPs@AuNRA anode. Thus, the proposed nonenzymatic GFC exhibit promising feasibility in applications. © 2023 The Authors

关键词： Anodes

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