Assoc.Prof. Parinya Noisa

Year	Title	Journal
2023	Encapsulation of HaCaT Secretome for Enhanced Wound Healing Capacity on Human Dermal Fibroblasts Heebkaew N., Promjantuek W., Chaicharoenaudomrung N., Phonchai R., Kunhorm P., Soraksa N., Noisa P.	Molecular Biotechnology
Abstract: In the epidermal and dermal layers of the skin, diverse cell types are reconstituted during the wound healing process. Delays or failures in wound healing are a major issue in skin therapy because they prevent the normal structure and function of wounded tissue from being restored, resulting in ulceration or other skin abnormalities. Human immortalized keratinocytes (HaCAT) cells are a spontaneously immortalized human keratinocyte cell line capable of secreting many bioactive chemicals (a secretome) that stimulate skin cell proliferation, rejuvenation, and regeneration. In this study, the HaCaT secretome was encapsulated with polyesters such as poly (lactic-co-glycolic acid) (PLGA) and cassava starch in an effort to maximize its potential. According to the estimated mechanism of the HaCaT secretome, all treatments were conducted on immortalized dermal fibroblast cell lines, a model of wound healing. Encapsulation of HaCaT secretome and cassava starch enhanced the effectiveness of cell proliferation, migration, and anti-aging. On the other hand, the levels of reactive oxygen species (ROS) were lowered, activating antioxidants in immortalized dermal fibroblast cells. The HaCaT secretome induced in a dose-dependent manner the expression of antioxidant-associated genes, including SOD, CAT, and GPX. Six cytokines, including CCL2 and MCP-1, influenced immunoregulatory and inflammatory processes in cultured HaCAT cells. HaCaT secretome encapsulated in cassava starch can reduce ROS buildup by boosting antioxidant to stimulate wound healing. Hence, the HaCaT secretome may have a new chance in the cosmetics business to develop components for wound prevention and healing. © 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature. Keyword: Antioxidants; Cassava starch; Encapsulation; Fibroblasts; HaCaT secretome; Keratinocytes; Skin regeneration; Wound healing Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85151538267&doi=10.1007%2fs12033-023-00732-z&partnerID=40&md5=caf788ec88a21f8af624e823112e5731 DOI: 10.1007/s12033-023-00732-z
2022	Effect of morpholine and charge distribution of cyanine dyes on cell internalization and cytotoxicity Wangngae S., Chansaenpak K., Weeranantanapan O., Piyanuch P., Sumphanapai T., Yamabhai M., Noisa P., Lai R.-Y., Kamkaew A.	Scientific Reports
Abstract: To improve the potency of Heptamethine cyanines (Hcyanines) in cancer research, we designed and synthesized two novel Hcyanines based theranostic probes, IR794-Morph and IR794-Morph-Mpip, to enhance cancer cell internalization and targeting. In acidic conditions that resemble to tumour environment, both IR794 derivatives exhibited broad NIR absorption band (704‒794 nm) and fluorescence emission (798‒828 nm) that is suitable for deep seated tumour imaging. Moreover, in vitro study revealed that IR794-Morph-Mpip exhibited better cancer targetability towards various cancer cell lines under physiological and slightly acidic conditions compared to normal cells. IR794-Morph-Mpip was fast internalized into the cancer cells within the first 5 min and mostly localized in lysosomes and mitochondria. In addition, the internalized signal was brighter when the cells were in the hypoxic environment. Furthermore, cellular uptake mechanism of both IR794 dyes, investigated via flow cytometry, revealed that endocytosis through OATPs receptors and clathrin-mediated endocytosis were the main routes. Moreover, IR794-Morph-Mpip, displayed anti-cancer activity towards all tested cancer cell types with IC50 below 7 μM (at 6 h incubation), which is approximately three times lower than that of the normal cells. Therefore, increasing protonated cites in tumour environment of Hcyanines together with incorporating morpholine in the molecule can enhance structure-inherent targeting of these dyes. © 2022, The Author(s). Keyword: Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85126075936&doi=10.1038%2fs41598-022-07533-5&partnerID=40&md5=5512595c89ffa6073ab58454dd9b0299 DOI: 10.1038/s41598-022-07533-5
2022	Cordycepin attenuates migration and invasion of HSC-4 oral squamous carcinoma cells through autophagy-dependent FAK/Akt and MMP2/MMP9 suppression Binlateh T., Uppatcha N., Thepchai J., Pleungtuk Y., Noisa P., Hutamekalin P., Jitprasertwong P.	Journal of Dental Sciences
Abstract: Background/purpose: Cordycepin has been proposed anti-cancer effects, however, it is unclear whether and how cordycepin affects oral squamous carcinoma cell (OSCC) migration and invasion. This study aimed to investigate the effect of cordycepin on migration and invasion of OSCC (HSC-4 cells), and its underlying mechanism. Materials and methods: Cell viability was measured with MTT assay. Migrative and invasive abilities were determined by scratch wound healing, agarose spot and transwell invasion assays, respectively. Monodasylcadaverine (MDC) staining, immunofluorescence staining of LC3 and RT-PCR evaluated the gene expression of LC3 and p62 were applied to investigate autophagy. MMP2 and MMP9 gene expression and activity were examined by RT-PCR and gelatin zymography. Expression of caspase 3, cleaved caspase 3, FAK, p-FAK, Akt and p-Akt was determined by Western blot. Results: Cordycepin significantly inhibited HSC-4 cell migration and invasion in a concentration-dependent manner. Cordycepin treatment caused an induction of autophagy, as evidenced by increased MDC fluorescence intensity and MDC positive cells, and upregulated expression level of LC3 gene. In addition, inhibition of autophagy by chloroquine (CQ) significantly abolished cordycepin-inhibited HSC-4 cell migration and invasion, demonstrating that cordycepin-inhibited migration and invasion was mediated by autophagy. Mechanistic studies showed that cordycepin significantly suppressed FAK and Akt phosphorylation, and MMP2 and MMP9 activities. Conversely, CQ pre-incubation significantly restored its expression and activity in cordycepin-treated cells. Conclusion: Cordycepin induces autophagy to suppress FAK and Akt phosphorylation, and MMP2 and MMP9 activity, which responsible for the attenuation of HSC-4 cell migration and invasion. © 2022 Association for Dental Sciences of the Republic of China Keyword: Autophagy; Cordycepin; Invasion; LC3; Migration; MMP2 Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85126911482&doi=10.1016%2fj.jds.2022.03.002&partnerID=40&md5=c2d64f2111d2d9cfae8f71132aad58a1 DOI: 10.1016/j.jds.2022.03.002
2022	Combination of Melatonin and Small Molecules Improved Reprogramming Neural Cell Fates via Autophagy Activation Sotthibundhu A., Nopparat C., Natphopsuk S., Phuthong S., Noisa P., Govitrapong P.	Neurochemical Research
Abstract: Reprogramming cell fates towards mature cell types are a promising cell supply for treating degenerative diseases. Recently, transcription factors and some small molecules have turned into impressive modulating elements for reprogramming cell fates. Melatonin, a pineal hormone, has neuroprotective functions including neural stem cell (NSC) proliferative and differentiative modulation in both embryonic and adult brain. We developed a protocol that could be implemented in the direct reprogramming of human skin fibroblast towards neural cells by using histone deacetylase (HDAC) inhibitor, glycogen synthase kinase-3 (GSK3) inhibitor (CHIR99021), c-Jun N-terminal kinase (JNK) inhibitor, rho-associated protein kinase inhibitor (Y-27632), cAMP activator, and melatonin treatment. We found that melatonin enhanced neural-transcription factor genes expressions, including brain-specific homeobox/POU domain protein 2 (BRN2), Achaete-Scute Family BHLH transcription Factor 1 (ASCL1), and Myelin Transcription Factor 1 Like (MYT1L). Melatonin also increased the expression of different neural-specific proteins such as doublecortin (DCX), Sex determining region Y-box 2 (Sox2), and neuronal nuclei (NeuN) compared with other five small molecules (valproic acid (VPA), CHIR99021, Forskolin, 1,9 pyrazoloanthrone (SP600125), and Y-27632) combination in the presence and absence of melatonin. A noticeable upregulation of autophagy proteins (microtubule-associated protein 1A/1B-light chain 3 (LC3) and Beclin-1) were seen in the melatonin treatment during the induction period while these were reverted in the presence of L-leucine, an autophagy inhibitor. In addition, the expression of NeuN was also significantly reduced by L-leucine. Collectively, our findings revealed an activation of autophagy during neural induction; melatonin enhanced reprogramming efficiency for neuron induction through the modulation of autophagy activation. © 2021, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature. Keyword: Autophagy; Direct reprogramming; Fibroblast; Melatonin Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85137958173&doi=10.1007%2fs11064-021-03382-2&partnerID=40&md5=2eff3027d8011b50b1f4dddfab9c089d DOI: 10.1007/s11064-021-03382-2
2022	Indomethacin-based near-infrared photosensitizer for targeted photodynamic cancer therapy Siriwibool S., Wangngae S., Chansaenpak K., Wet-osot S., Lai R.-Y., Noisa P., Sukwattanasinitt M., Kamkaew A.	Bioorganic Chemistry
Abstract: Near-IR fluorescent sensitizers based on heptamethine cyanine (Cy820 and Cy820–IMC) were synthesized and their abilities to target and abolish tumor cells via photodynamic therapy (PDT) were explored. Some hepthamethine cyanine dyes can be transported into cancer cells via the organic anion transporting polypeptides (OATPs). In this study, we aimed to enhance the target ability of the sensitizer by conjugation Cy820 with indomethacin, a non-steroidal anti-inflammatory drug (NSAID), to obtain Cy820–IMC that aimed to target cyclooxygenase–2 (COX–2) which overexpresses in cancer cells. The results showed that Cy820–IMC internalized the cancer cells faster than Cy820 which was verified to be related to COX–2 level and OATPs. Based on PDT experiments, Cy820–IMC has higher photocytotoxicity index than Cy820, >7.13 and 4.90, respectively, implying that Cy820–IMC showed better PDT property than Cy820. However, Cy820 exhibits slightly higher normal-to-cancer cell toxicity ratio than Cy820–IMC, 6.58 and 3.63, respectively. Overall, Cy820–IMC has superior cancer targetability and enhanced photocytoxicity. These characteristics can be further improved towards clinically approved sensitizers for PDT. © 2022 Elsevier Inc. Keyword: Cancer targeting; COX-2; Cy820; Heptamethine cyanine; NIR photosensitizer; Photodynamic therapy Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85126985454&doi=10.1016%2fj.bioorg.2022.105758&partnerID=40&md5=0daf700c4dbc536705922600f9912cbf DOI: 10.1016/j.bioorg.2022.105758
2022	Enhancement of cordycepin production from Cordyceps militaris culture by epigenetic modification Kunhorm P., Chueaphromsri P., Chaicharoenaudomrung N., Noisa P.	Biotechnology Letters
Abstract: Cordycepin (3′-deoxyadenosine) is a nucleoside analogue and biosynthesised by Cordyceps militaris, an entomopathogenic fungus. In this study, an epigenetic modifier was applied to static liquid cultures to enhance cordycepin production. C. militaris was cultured in a static liquid culture, and valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, was supplemented in order to modifying the epigenetic status. Gene regulatory network was explored to understand the molecular mechanisms underlying cordycepin production. 50 micromolar of VPA enhanced cordycepin production by 41.187% via the upregulation of 5′-nucleotidase, adenylate kinase, phosphorybosyltransferase, Cns1, Cns2, Cnsa3, and Cns4 of C. militaris for at least 2 days after VPA treatment. The maximum production of cordycepin was 2,835.32 ± 34.35 mg/L in 400 mL-working volume. A scaled-up culture was established with a working volume of 10 L, which led to the slight decrease of cordycepin production. This might due to multifactorial effects, for instance limited aeration and an uneven dispersion of nutrients in the culture system. This scaled-up culture was still needed further optimization. The modification of epigenetic status by VPA significantly enhanced cordycepin production by altering key gene regulatory network of C. militaris. The strategy established in this study might be applicable to other microorganism culture in order to improving the production of bioactive compounds. Graphical abstract: This work aimed to enhance the production of cordycepin by modifying the epigenetic status of C. militaris, in which subsequently altered gene regulatory network of cordycepin biosynthesis pathway. The weekly supplementation of valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, significantly improve cordycepin production over 40%, compared to the untreated control, and the gene regulatory network of C. militaris was also adapted. [Figure not available: see fulltext.]. © 2022, The Author(s), under exclusive licence to Springer Nature B.V. Keyword: Cordycepin; Cordyceps militaris; Epigenetic modification; Static liquid culture Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85125834286&doi=10.1007%2fs10529-022-03241-2&partnerID=40&md5=2a71861d13f2cda05aa490aa50e7292e DOI: 10.1007/s10529-022-03241-2
2022	Transgenic Immortalization of Human Dermal Fibroblasts Mediated through the MicroRNA/SIRT1 Pathway Promjantuek W., Chaicharoenaudomrung N., Phonchai R., Kunhorm P., Noisa P.	In Vivo
Abstract: Background/Aim: Human dermal fibroblasts (HDFs) are widely used as a skin model in cosmetic and pharmaceutical industry due their advantages for the cosmetic industry and medical aspects. Telomeres are key players in controlling cellular aging, in which telomeres and the telomerase enzyme (hTERT) can maintain proliferative capacity and prolong cellular senescence. The primary aim of the study was to elucidate the underlying mechanisms of hTERT/SV40 immortalization of human dermal fibroblasts. Materials and Methods: Transgenic expression of hTERT and SV40 large antigen, as well as co-transfection of both factors was performed and their significance evaluated in terms of HDF immortalization efficiency. Results: The results showed that the immortalized fibroblasts of all conditions can be cultured in over 60 passages and maintain their telomere length. Further, key markers of skin cells, such as COL1A1, KRT18 and ELASTIN, were up-regulated in immortalized cells. In addition, p53 expression was enhanced in all immortalized cells, in accordance with activation of the SIRT1 gene upon transgenic immortalization. The significant role of SIRT1 in fibroblast proliferation was assessed by shRNA-knockdown, and it was found that SIRT1 silencing led to loss of Ki67, a proliferation marker. Moreover, miR-93, a SIRT1-targeted miRNA, also had a significantly reduced expression in the co-transfected immortalized cells, highlighting the linkage of the miRNA and SIRT1 pathway in the immortalization of human dermal fibroblasts. Conclusion: This evidence from this study could © 2022 International Institute of Anticancer Research. All rights reserved. Keyword: Human dermal fibroblasts; Immortalization; Micro-RNA; SIRT1 Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85123036670&doi=10.21873%2fINVIVO.12685&partnerID=40&md5=e3d25738bae91c43ed8d623842aba5f0 DOI: 10.21873/INVIVO.12685
2022	Nanoencapsulation of Cordycepin Induces Switching from Necroptosis to Apoptosis in Human Oral Cancer Cells (HSC-4) Through Inhibition of Receptor-Interacting Serine/Threonine-Protein Kinase 3 (RIPK3) and Autophagy Modulation Kaokaen P., Chaicharoenaudomrung N., Kunhorm P., Mesil K., Binlateh T., Noisa P., Jitprasertwong P.	Natural Product Communications
Abstract: Human oral squamous carcinoma is considered the most common oral cancer; it imposes multiple oral and dental consequences as a result of oral cancer treatment. We previously found that the nanoencapsulation of cordycepin (CS) could inhibit oral cancer cells. However, the mechanism of action was not understood. The aim of this study was to investigate the signaling pathway by which CS and encapsulated nanoparticles (NPs) activate the inhibition of cancer cell growth. We demonstrated that human oral cancer (HSC-4) cells underwent necroptosis when incubated with high concentrations of CS, but not when incubated with either low concentrations of CS or encapsulated CS NPs. High concentrations of CS-induced necroptosis of HSC-4 cells, demonstrated by a reduction in apoptotic (BAX, Caspase-3, Caspase-8, and Caspase-9) and autophagic genes (LC3, Atg5, and Atg12). However, low concentrations of CS significantly induced the expression of autophagic gene LC3. Interestingly, encapsulated CS NPs induced a significant increase in apoptotic genes (P53, BAX, Caspase-3, Caspase-8, and Caspase-9), but a significant decrease in autophagic (P62, Atg5, and Atg12) and necroptotic genes (receptor-interacting serine/threonine-protein kinase 3 [RIPK3]). We also found that encapsulated CS NPs enhanced the accumulation of cellular protein and decreased secreted supernatant protein levels. Moreover, encapsulated CS NPs had higher efficacy in terms of reactive oxygen species (ROS) generation-mediated inhibition of autophagy compared to nonencapsulated CS, suggesting that nanoencapsulation of CS can switch the program of HSC-4 cell death from necroptosis to apoptosis. In conclusion, HSC-4 cells have a defense strategy against CS-induced cell apoptosis, but this problem can be solved through the use of encapsulation combined with nanotechnology. © The Author(s) 2022. Keyword: apoptosis; bioactivity; cordycepin; human oral squamous carcinoma cells; nanoparticles; necroptosis Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85123675944&doi=10.1177%2f1934578X221074838&partnerID=40&md5=1291f5d6e493d984eba108ee45f2fe1a DOI: 10.1177/1934578X221074838
2021	Autophagy promoted neural differentiation of human placenta-derived mesenchymal stem cells SOTTHIBUNDHU A., MUANGCHAN P., PHONCHAI R., PROMJANTUEK W., CHAICHAROENAUDOMRUNG N., KUNHORM P., NOISA P.	In Vivo
Abstract: Background/Aim: Human placenta-derived mesenchymal stem cells (hPMSCs) are multipotent and possess neurogenicity. Numerous studies have shown that Notch inhibition and DNA demethylation promote neural differentiation. Here, we investigated the modulation of autophagy during neural differentiation of hPMSCs, induced by DAPT and 5-Azacytidine. Materials and Methods: HPMSCs were treated with DAPT to induce neural differentiation, and the autophagy regulating molecules were used to assess the impact of autophagy on neural differentiation. Results: The hPMSCs presented with typical mesenchymal stem cell phenotypes, in which the majority of cells expressed CD73, CD90 and CD105. hPMSCs were multipotent, capable of differentiating into mesodermal cells. After treatment with DAPT, hPMSCs upregulated the expression of neuronal genes including SOX2, Nestin, and βIII-tubulin, and the autophagy genes LC3I/II and Beclin. These genes were further increased when 5-Azacytidine was co-supplemented in the culture medium. The inhibition of autophagy by chloroquine impeded the neural differentiation of hPMSCs, marked by the downregulation of βIII-tubulin, while the activation of autophagy by valproic acid (VPA) instigated the emergence of βIII-tubulin-positive cells. Conclusion: During the differentiation process, autophagy was modulated, implying that autophagy could play a significant role during the differentiation of these cells. The blockage and stimulation of autophagy could either hinder or induce the formation of neural-like cells, respectively. Therefore, the refinement of autophagic activity at an appropriate level might improve the efficiency of stem cell differentiation. © 2021 International Institute of Anticancer Research. All rights reserved. Keyword: Autophagy; Human placenta; Mesenchymal stem cells; Neural differentiation; Notch signaling Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85113852610&doi=10.21873%2fINVIVO.12543&partnerID=40&md5=cca609ab494debeda17adad40d75569c DOI: 10.21873/INVIVO.12543
2021	Flavylium-based hypoxia-responsive probe for cancer cell imaging Pewklang T., Wet-Osot S., Wangngae S., Ngivprom U., Chansaenpak K., Duangkamol C., Lai R.-Y., Noisa P., Sukwattanasinitt M., Kamkaew A.	Molecules
Abstract: A hypoxia-responsive probe based on a flavylium dye containing an azo group (AZOFlav) was synthesized to detect hypoxic conditions via a reductase-catalyzed reaction in cancer cells. In in vitro enzymatic investigation, the azo group of AZO-Flav was reduced by a reductase in the presence of reduced nicotinamide adenine dinucleotide phosphate (NADPH) followed by fragmentation to generate a fluorescent molecule, Flav-NH2. The response of AZO-Flav to the reductase was as fast as 2 min with a limit of detection (LOD) of 0.4 μM. Moreover, AZO-Flav displayed high enzyme specificity even in the presence of high concentrations of biological interferences, such as reducing agents and biothiols. Therefore, AZO-Flav was tested to detect hypoxic and normoxic environments in cancer cells (HepG2). Compared to the normal condition, the fluorescence intensity in hypoxic conditions increased about 10-fold after 15 min. Prolonged incubation showed a 26-fold higher fluorescent intensity after 60 min. In addition, the fluorescence signal under hypoxia can be suppressed by an electron transport process inhibitor, diphenyliodonium chloride (DPIC), suggesting that reductases take part in the azo group reduction of AZO-Flav in a hypoxic environment. Therefore, this probe showed great potential application toward in vivo hypoxia detection. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. Keyword: Activity-based sensing; Azo dye; Flavylium; Hypoxia detection; Turn-on fluorescent sensor Scopus Link: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85113752428&doi=10.3390%2fmolecules26164938&partnerID=40&md5=d9da039db70f38ab4efa490bb16d0481 DOI: 10.3390/molecules26164938