Visitsattapongse, Sarinporn

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Visitsattapongse, Sarinporn
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Visitsattapongse, S.
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Email
sarinporn.vi@kmitl.ac.th

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    A Novel Approach for Optimizing Molecularly Imprinted Polymer Composition in Electrochemical Detection of Collagen Peptides
    (2025-11-01)
    Vongmanee, Naphatsawan
    ;
    Nampeng, Jindapa
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    Rattanapithan, Katesirin
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    Sriwichai, Phuritasinee
    ;
    Pintavirooj, Chuchart
    Collagen peptides are key structural proteins that play an important role in maintaining the integrity and proper function of multiple tissues in the human body. Their breakdown is recognized as an important biomarker for various degenerative conditions, including the loss of muscle mass, joint and bone disorders, and compromised skin health. Current analytical approaches for collagen detection, such as ultraviolet spectrometry, enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), and histochemical staining, are widely used but often expensive, time-consuming, and reliant on specific laboratory instrumentation, limiting their practicality for routine or rapid diagnostics. This study reports a novel biosensor for collagen peptide detection based on molecularly imprinted polymers (MIPs) integrated with screen-printed electrodes (SPEs). Electrochemical measurements revealed a clear correlation between collagen concentration and current response, confirming effective molecular binding within the imprinted matrix. The optimized MIP-modified electrode exhibited a detection range of 0.1–1000 µg/mL with a limit of detection (LOD) of 1.0106 µg/mL, limit of quantification (LOQ) of 4.46 µg/mL, sensitivity of 8.3816, and correlation coefficient (R<sup>2</sup> = 0.9436). These results highlight strong selectivity and sensitivity toward collagen peptides. The proposed MIP-based biosensor provides a rapid, low-cost platform for detecting collagen degradation products and holds potential for early diagnosis and future clinical applications in degenerative disease monitoring.
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    Store-operated calcium entry facilitates LPS-induced superoxide anion-dependent macrophage extracellular traps
    (2025-07-09)
    Nguyen, Thang Ngoc
    ;
    Lin, Tzu Chien
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    Chimphlee, Waratchaya
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    Siew, Kon Xuen
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    Vongmanee, Naphatsawan
    Macrophage extracellular traps (METs) represent a recently discovered complex defence mechanism that is distinct from phagocytosis and involves the release of DNA and antibacterial proteins. They play an important role in pathogen removal, and calcium ions (Ca 2+) have also been reported to be involved. In the present study, we identified METotic cells using digitonin as an alternative to Triton X-100, coupled with immunofluorescence staining using lamin antibodies. The limited permeability of digitonin ensures exclusive intranuclear antibody labelling of MET cells, therefore providing a straightforward and intuitive differentiation method. We found that under lipopolysaccharide stimulation, macrophages undergo store-operated Ca 2+ entry (SOCE) to facilitate Ca 2+ influx. Elevation of cytoplasmic Ca 2+ levels by SOCE promotes the generation of superoxide anions by NADPH oxidase (NOX), ultimately leading to METosis. In summary, our study strengthens the role of Ca 2+ in NOX-dependent METosis, which differs from previous studies focusing on Ca 2+ in the NOX-independent pathway. Our research reveals that Ca 2+ -mediated regulation of NOX plays a crucial role in METosis, especially in SOCE, and provides novel ideas for future research.
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    Ultrasensitive Label-Free Electrochemical Detection of Pseudomonas aeruginosa Using a Surface Molecularly Imprinted Polymer-Modified Screen-Printed Electrode
    (2026-06-01)
    Vongmanee, Naphatsawan
    ;
    Nampeng, Jindapa
    ;
    Pintavirooj, Chuchart
    ;
    Visitsattapongse, Sarinporn
    Pseudomonas aeruginosa is a major opportunistic pathogen frequently associated with nosocomial infections, such as pneumonia, urinary tract infections, and wound infections, particularly in immunocompromised or hospitalized patients. These infections are often difficult to treat due to the pathogen’s intrinsic antibiotic resistance and biofilm-forming ability. Therefore, rapid and selective detection of P. aeruginosa is essential for early diagnosis and effective infection control. In this study, a novel surface-imprinted MIP design uniquely combines methacrylamide (MAM), acrylamide (AAM), and vinylpyrrolidone (VP) monomers to generate recognition cavities that are complementary to the surface morphology and physicochemical properties of Pseudomonas aeruginosa cells. Unlike traditional MIP approaches, this surface imprinting strategy provides improved stability and reproducibility, without relying on biological recognition elements like antibodies or aptamers. This novel approach enabled us to achieve an ultralow LOD of 1 CFU/mL over a linear range of 1–10<sup>4</sup> CFU/mL, demonstrating excellent analytical performance. In addition, the sensor exhibited good reproducibility with an RSD of 5–12%. The novelty of this work lies in the use of a surface-imprinted MIP strategy combined with a multi-monomer system to enhance bacterial recognition and sensing performance. Overall, the proposed MIP-based electrochemical biomimetic sensor offers a rapid, cost-effective, and portable platform with strong potential for the detection of P. aeruginosa in clinical and environmental applications.
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    Electrochemical Characterization of a Molecularly Imprinted Polymer Sensor for the Selective Recognition of Type II Collagen in Joint Degeneration Monitoring
    (2026-02-01)
    Nampeng, Jindapa
    ;
    Vongmanee, Naphatsawan
    ;
    Pintavirooj, Chuchart
    ;
    Visitsattapongse, Sarinporn
    Type II collagen is a primary fibrillar component of articular cartilage, and its early degradation is a key biomarker of joint-degenerative disorders such as osteoarthritis, rheumatoid arthritis, gout, etc. Reliable detection at low concentrations remains challenging due to limited assay accessibility, complex analytical procedures, and nonspecific responses in multicomponent biological matrices. This research reports the development of a Molecularly Imprinted Polymer (MIP)–based electrochemical sensor engineered for the selective recognition of type II collagen. A series of monomer formulations were evaluated, and the 1AAM:2VP composition produced a well-defined imprinted layer on screen-printed carbon electrodes, yielding the highest electrochemical sensitivity and linearity. The optimized sensor exhibited strong anodic and cathodic responses proportional to increasing collagen concentrations, with a calibration slope corresponding to an R<sup>2</sup> value of 0.9394. Minimal signal interference was observed, confirming high molecular selectivity. The limit of detection (LOD) was calculated to be approximately 0.065 µg/mL. These characteristics demonstrate that the proposed MIP sensor provides a low-cost, accessible, and highly selective analytical platform suitable for early-stage cartilage degeneration monitoring.
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    Electrochemical Biosensors by Means of Molecularly Imprinted Polymers (MIPs) Cortisol Recognition
    (2025-02-01)
    Nampeng, Jindapa
    ;
    Vongmanee, Naphatsawan
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    Pintavirooj, Chuchart
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    Chiu, Wen Tai
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    Visitsattapongse, Sarinporn
    Depression and anxiety are two common mental health issues that require serious attention, as they have significant impacts on human well-being, with both being emotionally and physically reflected in the increasing number of suicide cases globally. The World Health Organization (WHO) estimated that about 322 million people around the world experienced mental illnesses in 2017, and this number continues to increase. Cortisol is a major stress-controlled hormone that is regulated by the hypothalamic–pituitary–adrenal (HPA) axis. The HPA axis has three main components, including the hypothalamus, pituitary gland, and adrenal gland, where cortisol, the primary stress hormone, is released. It plays crucial roles in responding to stress, energy balance, and the immune system. The cortisol level in the bloodstream usually increases when stress develops. Molecularly imprinted polymers (MIPs) have been highlighted in terms of creating artificial bioreceptors by mimicking the shape of detected biomolecules, making natural bioreceptor molecules no longer required. MIPs can overcome the limitations of chemicals and physical properties reducing over time and the short-time shelf life of natural bioreceptors. MIPs’ benefits are reflected in their ease of use, high sensitivity, high specificity, reusability, durability, and the lack of requirement for complicated sample preparation before use. Moreover, MIPs incur low costs in manufacturing, giving them a favorable budget for the market with simple utilization. MIPs can be formulated by only three key steps, including formation, the polymerization of functional monomers, and the creation of three-dimensional cavities mimicking the shape and size of targeting molecules. MIPs have a high potential as biosensors, especially working as bioanalytics for protein, anti-body, antigen, or bacteria detection. Herein, this research proposes an MIP-based cortisol biosensor in which cortisol is imprinted on methyl methacrylate (MMA) and methacrylic acid (MAA) produced by UV polymerization. This MIP-based biosensor may be an alternative method with which to detect and monitor the levels of hormones in biological samples such as serum, saliva, or urine due to its rapid detection ability, which would be of benefit for diagnosing depression and anxiety and prescribing treatment. In this study, quantitative detection was performed using an electrochemical technique to measure the changes in electrical signals in different concentrations of a cortisol solution ranging from 0.1 to 1000 pg/mL. The MIP-based biosensor, as derived by calculation, achieved its best detection limit of 1.035 pg/mL with a gold electrode. Tests were also performed on molecules with a similar molecular structure, including Medroxyprogesterone acetate and drospirenone, to ensure the sensitivity and accuracy of the sensors, demonstrating a low sensitivity and low linear response.
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    Enhanced deep learning model for prediction of diabetic mellitus on optical coherence tomography angiography images
    (2026-01-01)
    Visitsattapongse, Sarinporn
    ;
    Rithcharung, Preeyarat
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    Santiprabhob, Jeerunda
    ;
    Lertbannaphong, Ornsuda
    ;
    Sermsripong, Wasawat
    Background: Diabetes mellitus (DM) is a chronic metabolic disease characterized by dysregulated blood glucose. Prolonged DM can lead to diabetic retinopathy (DR), in which retinal capillaries are damaged by sustained hyperglycemia. Optical coherence tomography angiography (OCTA) is a non-invasive imaging modality for visualizing retinal microvasculature and can detect early changes in both DM patients with and without DR. However, it requires expert evaluation, making early detection costly and time-consuming. This study aimed to develop a high-performance deep learning framework that can classify OCTA images into three groups of DM, such as normal, good glycemic control, and poor glycemic control. Methods: OCTA datasets of horizontal B-scans and en face scans from 300 participants aged 8–18 years were analyzed, including normal controls, DM patients with good glycemic control, and DM patients with poor control (HbA1c ≥8%). For each participant, a 3 mm × 3 mm foveal-centered en face image of the deep capillary plexus (DCP) and a horizontal B-scan through the foveal center of the right eye were selected. Several convolutional and transformer-based models were evaluated, with ConvNeXt (a ConvNet for the 2020s) chosen as the baseline for its superior performance. To enhance generalization and convergence, progressive resizing and the Lookahead optimization strategy were applied, while class-wise augmentation was used to balance the training set without altering the test distribution. Results: The baseline ConvNeXt achieved F1 scores of 0.7877 (B-scans) and 0.7424 (en face). After doing enhancement using progressive resizing and Lookahead optimization, performance improved to 0.8319 and 0.8567 (Wilcoxon signed-rank tests, P<0.05). Conclusions: Our proposed method for DM classification from OCTA images provided promising results while ensuring resource efficiency and rapid evaluation. Clinically, accurate classification of DM status is valuable for assessing the risk of DR progression. Thus, it can be served as an assistive tool for clinical decision support in DR management.
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    Molecularly Imprinted Polymer-Based Electrochemical BioSensors for Haemophilus influenzae Rapid Detection
    (2026-03-01)
    Vongmanee, Naphatsawan
    ;
    Nampeng, Jindapa
    ;
    Pintavirooj, Chuchart
    ;
    Visitsattapongse, Sarinporn
    Haemophilus influenzae (H. influenzae) is an important respiratory pathogen that can cause various invasive and non-invasive bacterial infections requiring rapid and sensitive detection. In recent years, electrochemical biosensors have emerged as a practical alternative for pathogen detection due to their high sensitivity, portability and short analysis time. Molecularly imprinted polymers (MIPs) are a class of synthetic receptors designed to mimic biological recognition through template-directed polymerization. In this study, an electrochemical biosensor based on MIPs was developed for the selective detection of H. influenzae. The polymeric film composed of methacrylamide (MAM), acrylamide (AAM), and vinylpyrrolidone (VP) monomers was fabricated on a gold screen-printed electrode (gold-SPE). The results of cyclic voltammetry (CV) revealed a strong redox current shift corresponding to bacteria concentrations within an analytical range of 1–10,000 CFU/mL with LOD 1.03 CFU/mL, with relative standard deviation (RSD) values below 9% across the tested concentration range. The optimized composition yielded and exhibited excellent selectivity when tested against non-target bacteria such as Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus.
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    C. albicans Detection with Electrochemical Sensors by Using Molecular Imprinted Polymer Technique
    (2026-03-01)
    Vongmanee, Naphatsawan
    ;
    Nampeng, Jindapa
    ;
    Pintavirooj, Chuchart
    ;
    Visitsattapongse, Sarinporn
    Candida albicans (C. albicans) is an opportunistic fungal pathogen and a major cause of nosocomial infections, especially in immunocompromised patients. Conventional diagnostic approaches such as blood culture and biochemical assays are accurate but require multi-step sample processing and prolonged turnaround times, limiting their applicability for rapid clinical screening. In the present study, we developed an electrochemical biosensor based on molecularly imprinted polymer (MIP) technology for the rapid and selective detection of intact C. albicans cells. The MIP layer was electropolymerized onto a screen-printed carbon electrode (SPCE), forming selective recognition cavities complementary to the fungal morphology. Electrochemical characterization and detection were performed using cyclic voltammetry in phosphate-buffered saline (PBS). The system demonstrated a wide linear detection range, enabling reliable quantification of C. albicans across concentrations spanning from 1 to 10<sup>4</sup> CFU/mL and achieved an ultralow limit of detection (LOD) of 1.30 CFU/mL, demonstrating high sensitivity. High selectivity was confirmed against E. coli, S. aureus, and P. aeruginosa, demonstrating that the imprinted cavities effectively distinguish fungal cells from bacterial contaminants. These findings highlight the promise of MIP-based electrochemical biosensors as a simple, low-cost, and portable alternative for early fungal diagnostics.
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    YAP inactivation-mediated autophagy inhibition contributes to cisplatin resistance in ovarian cancer cells
    (2026-12-01)
    Nguyen, Ngoc Thang
    ;
    Hsieh, Meng Ru
    ;
    Nguyen, Hieu Dac Hanh
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    Chimphlee, Waratchaya
    ;
    Visitsattapongse, Sarinporn
    Background: Cisplatin resistance remains a critical barrier in the treatment of high-grade serous ovarian cancer (HGSOC). Although Hippo−YAP signaling regulates cancer progression, its contribution to cisplatin resistance is still poorly understood. Here, we demonstrate that YAP is inactivated and sequestered in the cytoplasm of cisplatin-resistant ovarian cancer cells. Results: This cytosolic retention is mediated by Hippo kinases MST1/2 and LATS1/2, as well as ERK signaling, resulting in increased phosphorylation of YAP at its inhibitory site (Ser397) and reduced phosphorylation at its activating site (Tyr357). Restoration of YAP activity through genetic overexpression or pharmacological induction of nuclear YAP accumulation significantly reversed cisplatin resistance. Mechanistically, YAP inactivation impaired cisplatin-induced autophagy. Cisplatin robustly triggered autophagy in parental cells, as evidenced by LC3 puncta formation; however, this autophagic response was blunted in resistant cells. Overexpression of YAP further suppressed LC3 puncta formation and Beclin-1 expression, and increased p62 accumulation in cisplatin-resistant ovarian cancer cells. Autophagy inhibition using 3-methyladenine (3-MA) resensitized resistant cells to cisplatin. Conclusion: Collectively, these findings reveal that YAP inactivation contributes to cisplatin resistance by abrogating autophagy formation and identify YAP reactivation as a potential strategy to overcome chemoresistance in ovarian cancer.
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    Bcl-2 upregulates calcium efflux through PMCA and NCX1 to preserve intracellular calcium homeostasis and confer resistance to apoptosis
    (2026-07-01)
    Lin, Tzu Chien
    ;
    Lee, Ming Jyun
    ;
    Nguyen, Ngoc Thang
    ;
    Hsiao, Shih Chuan
    ;
    Chen, Ying Chi
    Bcl-2 has been shown to regulate intracellular calcium (Ca²⁺) homeostasis, thereby influencing cellular metabolism, survival, and apoptosis. While previous studies have primarily focused on Bcl-2′s role in modulating Ca²⁺ levels within the endoplasmic reticulum and mitochondria, extracellular Ca²⁺ is a major determinant of cellular activation and Ca²⁺ homeostasis. Therefore, it is important to investigate whether Bcl-2 also contributes to the regulation of Ca²⁺ flux across the plasma membrane. A vector control, wild-type Bcl-2, and a Bcl-2 mutant were stably expressed in MDCK (Madin-Darby Canine Kidney) cells that lacked endogenous Bcl-2 expression. Real-time intracellular Ca²⁺ measurements by a single-cell fluorimeter were performed to measure Ca²⁺ release and influx with the ratiometric Ca<sup>2+</sup> indicator Fura-2 AM. Our results demonstrate that Bcl-2 enhances store-operated Ca²⁺ entry (SOCE)-mediated Ca²⁺ influx, a key mechanism underlying spontaneous Ca²⁺ oscillations. Furthermore, Bcl-2 upregulates the expression of plasma membrane Ca²⁺ ATPase (PMCA) and Na<sup>+</sup>-Ca²⁺ exchanger 1 (NCX1), which mediate Ca²⁺ extrusion from the cytosol. Pharmacological inhibition of PMCA with resveratrol (RES) and of NCX1 with ORM-10103 suppressed spontaneous Ca²⁺ oscillations, with PMCA playing a more dominant role than NCX1. Additionally, both RES and ORM-10103 exacerbated thapsigargin-induced Ca²⁺ cytotoxicity. Collectively, our findings reveal that Bcl-2 promotes Ca²⁺ influx to sustain oscillatory signaling and facilitates Ca²⁺ efflux to prevent cytotoxic accumulation, thereby maintaining Ca²⁺ homeostasis. This dual regulatory function provides new insights into Bcl-2–mediated Ca²⁺ signaling and its biological significance in apoptosis inhibition, independent of its interactions with other Bcl-2 family proteins.