Asst.Prof. Panwong Kuntanawat

Year	Title	Journal
2022	A Bench-Top Approach for Isolation of Single Antibody Producing Chinese Hamster Ovary (CHO) Cells Using a Microwell-Based Microfluidic Device Fuadiyah S., Chotchindakun K., Phatthanakun R., Kuntanawat P., Yamabhai M.	Micromachines
Abstract: Genetically-modified monoclonal cell lines are currently used for monoclonal antibody (mAbs) production and drug development. The isolation of single transformed cells is the main hindrance in the generation of monoclonal lines. Although the conventional limiting dilution method is time-consuming, laborious, and skill-intensive, high-end approaches such as fluorescence-activated cell sorting (FACS) are less accessible to general laboratories. Here, we report a bench-top approach for isolating single Chinese hamster ovary (CHO) cells using an adapted version of a simple microwell-based microfluidic (MBM) device previously reported by our group. After loading the cell suspension to the device, the electrostatically trapped cells can be viewed under a microscope and transferred using a micropipette for further clone establishment. Compared to the conventional method, the invented approach provided a 4.7-fold increase in the number of single cells isolated per round of cell loading and demonstrated a 1.9-fold decrease in total performing time. Additionally, the percentage of correct single-cell identifications was significantly improved, especially in novice testers, suggesting a reduced skill barrier in performing the task. This novel approach could serve as a simple, affordable, efficient, and less skill-intensive alternative to the conventional single-cell isolation for monoclonal cell line establishment. © 2022 by the authors. Keyword: cell line generation; limiting dilution; microfluidic device; monoclonal cell lines; single-cell cloning Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85141820257&doi=10.3390%2fmi13111939&partnerID=40&md5=0c40f3203b742f3555b0c412f35d56b7 DOI: 10.3390/mi13111939
2022	In Vitro Preparation and Evaluation of Chitosan/Pluronic F-127 Hydrogel as a Local Delivery of Crude Extract ofPhycocyanin for Treating Gingivitis Li M., Kamdenlek P., Kuntanawat P., Eawsakul K., Porntaveetus T., Osathanon T., Manaspon C.	Chiang Mai University Journal of Natural Sciences
Abstract: The phycocyanin (PC) in the crude extract is one of the main active compounds that has significant anti-inflammatory properties and the potential to treat gingival inflammation, the common oral disease. This work reports the preparation and characterization of chitosan/pluronic F-127 hydrogels entrapping the crude extracts of PC from Arthrospira platensis (C005H and C005L) as a local drug delivery system aiming to prolong PC release for the treatment of gingivitis. The results showed that the total phenolic content (TPC) values of the crude extracts were in the range of 2 to 5 μg GAE/100g and presented above 85% inhibition in the protein denaturation test and over 65% using a lipoxygenase (LOX) inhibition test. The hydrogels incorporating the crude extracts from C005H and C005L were perfectly prepared via the electrostatic interaction between chitosan and pluronic F-127 with very high encapsulation efficiency. The crude extracts of PC C005H and C005L were released over 70% from the loaded hydrogel within 6 hours under artificial saliva conditions. The anti-inflammatory activity of the released supernatant from the hydrogels after 6 hours was around 32-47% by LOX inhibition. The hydrogel vehicles and loaded-hydrogels did notshow any cytotoxic effects against mouse fibroblast cell lines (L929) and humangingival fibroblast cells. Our current work shows that the crude extract PC-loadedchitosan/pluronic F-127 hydrogel is biocompatibility with human cells and showsprolonged crude extract release properties, suggesting it as an alternativetreatment approach for gingivitis. © 2022 Author (s). This is an open access article distributed under the term of the Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author (s) and the source. Keyword: Chitosan; Gingivitis; Hydrogel; Phycocyanin; Pluronic f-127 Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85142233008&doi=10.12982%2fCMUJNS.2022.052&partnerID=40&md5=d3864079cf6ae1c445e27b04b3a1df00 DOI: 10.12982/CMUJNS.2022.052
2021	THE DEVELOPMENT OF AN ALTERNATIVE ALGAL LIPID QUANTIFICATION METHOD BASED ON RESISTANCE OF SCREEN PRINTED CARBON PAPER ELECTRODES Kuntanawat P., Kunhorm P., Siangdung W., Bunnag B., Phatthanakun R., Surareungchai W.	Suranaree Journal of Science and Technology
Abstract: Quantification of algal lipids may require large sample sizes and specialized equipment. Here we present an alternative principle using screen-printed carbon electrodes. Based on a set of experiments using soy bean oil, glyceryl trioleate and crude lipids extracted from Spirulina (Arthrospira platensis), Ankistrodesmus sp. and Chlorella vulgaris, the normalized resistance of the electrodes changes with concentration of the lipid deposited. The lipid concentration- normalized resistance relationships are of bell patterns with a linear range started from concentration = 0–20 mg/ ml and ended around 30–100 mg/ml, depending on lipid types. The sensitivity of the measurement is improved by about an order when fatty acid methyl esters obtained from Ankistrodesmus was used instead of the crude extract of the same algae. The technique consumes a dry algal biomass of 0.1–4 mg for an analysis. The amount is much less than that conventionally required for the gravitational analysis (80–100 mg) and smaller than that demanded in the sample preparation for analysis with GC (7–10 mg). The technique has a potential to develop into a cost- effective and less equipment- dependent lipid quantification for microfarms. © 2021. All Rights Reserved. Keyword: Algal lipid; biodiesel; lipid quantification; screen-printed carbon electrodes Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85126351576&partnerID=40&md5=68624dc9cc8113b62f0b6a5360ac45da DOI:
2020	New method for arbuscular mycorrhizal fungus spore separation using a microfluidic device based on manual temporary flow diversion Srisom K., Tittabutr P., Teaumroong N., Lapwong Y., Phatthanakun R., Sirivisoot S., Kuntanawat P.	Mycorrhiza
Abstract: Arbuscular mycorrhizal fungi are beneficial components often included in biofertilizers. Studies of the biology and utilization of these fungi are key to their successful use in the biofertilizer industry. The acquisition of isolated spores is a required step in these studies; however, spore quality control and spore separation are bottlenecks. Filtered and centrifuged spores have to be hand-picked under a microscope. The conventional procedure is skill-demanding, labor-intensive, and time-consuming. Here, we developed a microfluidic device to aid manual separation of spores from a filtered and centrifuged suspension. The device is a single spore streamer equipped with a manual temporary flow diversion (MTFD) mechanism to select single spores. Users can press a switch to generate MTFD when the spore arrives at the selection site. The targeted spore flows in a stream to the collection chamber via temporary cross flow. Using the device, spore purity, the percentage of spore numbers against the total number of particles counted in the collecting chamber reached 96.62% (median, n = 10) which is greater than the spore purity obtained from the conventional method (88.89% (median, n = 10)). © 2020, Springer-Verlag GmbH Germany, part of Springer Nature. Keyword: Arbuscular Mycorrhiza; Manual temporary flow diversion mechanism; Microfluidic device; Spore separation Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85090764937&doi=10.1007%2fs00572-020-00986-4&partnerID=40&md5=e52cd00fd53b8ae40b6efa6668f801dd DOI: 10.1007/s00572-020-00986-4
2018	The biomechanical role of overall-shape transformation in a primitive multicellular organism: A case study of dimorphism in the filamentous cyanobacterium Arthrospira platensis Chaiyasitdhi A., Miphonpanyatawichok W., Riehle M.O., Phatthanakun R., Surareungchai W., Kundhikanjana W., Kuntanawat P.	PLoS ONE
Abstract: Morphological transformations in primitive organisms have long been observed; however, its biomechanical roles are largely unexplored. In this study, we investigate the structural advantages of dimorphism in Arthrospira platensis, a filamentous multicellular cyanobacterium. We report that helical trichomes, the default shape, have a higher persistence length (Lp), indicating a higher resistance to bending or a large value of flexural rigidity (kf), the product of the local cell stiffness (E) and the moment of inertia of the trichomes’ cross-section (I). Through Atomic Force Microscopy (AFM), we determined that the E of straight and helical trichomes were the same. In contrast, our computational model shows that I is greatly dependent on helical radii, implying that trichome morphology is the major contributor to kf variation. According to our estimation, increasing the helical radii alone can increase kf by 2 orders of magnitude. We also observe that straight trichomes have improved gliding ability, due to its structure and lower kf. Our study shows that dimorphism provides mechanical adjustability to the organism and may allow it to thrive in different environmental conditions. The higher kf provides helical trichomes a better nutrient uptake through advection in aquatic environments. On the other hand, the lower kf improves the gliding ability of straight trichomes in aquatic environments, enabling it to chemotactically relocate to more favorable territories when it encounters certain environmental stresses. When more optimal conditions are encountered, straight trichomes can revert to their original helical form. Our study is one of the first to highlight the biomechanical role of an overall-shape transformation in cyanobacteria. © 2018 Chaiyasitdhi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85046861178&doi=10.1371%2fjournal.pone.0196383&partnerID=40&md5=f9f0dfd2d5c408c2bce462850548c4a0 DOI: 10.1371/journal.pone.0196383
2017	Increased osteoblast growth with graphene oxide in a microwell-based biochip Ramdongbang A., Kunhorm P., Kuntanawat P., Laksanasopin T., Sirivisoot S.	BMEiCON 2016 - 9th Biomedical Engineering International Conference
Abstract: This study investigated a biocompatibility of graphene oxide (GO) nanoflakes in culture medium with bone-forming cells or pre-osteoblasts using a microwell-based microfluidic chip made from polydimethylsiloxane (PDMS) and glass slide. The osteoblast growth was studied with GO in three concentrations (0, 25 and 50 μg/ml) at three time points (2, 4 and 6 hours) by a light microscopy. A percentage of relative cell numbers per microwell was analyzed using ImageJ program. The results suggested that with GO suspension at the concentration of 25 μg/ml the relative cell-survival percentages after 4 and 6 hours of culture in microwell-based biochips were highest when compared with other conditions. Whereas, at the concentration of 50 μg/ml GO caused adverse effect to pre-osteoblast cells at every time point. The microfluidic biochip provides an easy platform to track cells and determine cell growth with nanomaterials in a small volume of suspension. © 2016 IEEE. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85015952061&doi=10.1109%2fBMEiCON.2016.7859625&partnerID=40&md5=86d3ee8eb6e91f1bea4acbe4dd218b7a DOI: 10.1109/BMEiCON.2016.7859625
2014	An electrostatic microwell-based biochip for phytoplanktonic cell trapping Kuntanawat P., Ruenin J., Phatthanakun R., Kunhorm P., Surareungchai W., Sukprasong S., Chomnawang N.	Biomicrofluidics
Abstract: A simple microwell-based microfluidic chip for microalgal cells trapping was fabricated. An electrostatic cell trapping mechanism, enabled by a positively charged glass surface, was used. The chip was capable of capturing multiple algal cell types. In the case of filamentous Spirulina platensis, we observed single filament occupancy of up to ~30% available wells, as high as some previously proposed methods. Captured filaments were not of any preferential size, suggesting well randomized cell trapping. It was found that the electrostatic attraction did not affect the cell growth. Total replacement of liquid inside the wells could be achieved by pumping new solutions via the inlet, making single cell experiments in controlled chemical conditions possible. After the top layer of the chip was removed, cells in the wells could be simply transferred using a micropipette, turning the chip into a platform for strain selection. © 2014 AIP Publishing LLC. Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84929587429&doi=10.1063%2f1.4882196&partnerID=40&md5=a9b01cd7d0d31c882511d3c0f7618ab7 DOI: 10.1063/1.4882196
2009	Does the thickness of surface-bound hydrogel substrate affect adherent cell behaviours? Kuntanawat P., Wilkinson C.D.W., Riehle M.O.	European Cells and Materials
Abstract: [No abstract available] Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84860448161&partnerID=40&md5=65b89e3f5facb8739646bb5500e7f406 DOI: