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Bioactivity and toxicity evaluation of nutraceuticals using in vitro cell-based models: A review

Tran Hung Son
Published 09/20/2022

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How to Cite
Tran Hung Son. "Bioactivity and toxicity evaluation of nutraceuticals using in vitro cell-based models: A review". Vietnam Journal of Food Control. vol. 5, no. 4 (en), pp. 553-574, 2022

Main Article Content


Nutraceutical is one of the new concepts appearing in recent years, referring to food products derived from natural sources that benefit human health in preventing and treating diseases, besides providing nutritional value. During the development of these products, it is important to evaluate the toxicity and bioactivity of potential compounds and to study the mechanism of action of these nutraceutical compounds at the cellular and molecular levels. Among the many different experimental models, the in vitro cell-based model has emerged as a model with many advantages for preliminary screening of nutraceuticals activity and toxicity before conducting further studies on in vivo models. This review summarizes some basic techniques commonly used to screen nutraceuticals' toxicity. Recent studies of the bioactivity of nutraceuticals in various areas, such as antioxidant, anti-inflammation, antiobesity, neuroprotection, and gut health improvement, are also reviewed to introduce the application of the cell-based model in nutraceutical bioactivity research. New modern techniques using in vitro cell-based models have been applied in this field, such as highthroughput screening, 3D-cell culture, and organ-on-a-chip are also discussed in this paper.


Bioactivity, toxicity, nutraceuticals, in vitro, cell-based models.


[1]. C. M. Dominguez, N. Oturan, A. Romero, A. Santos, and M. A. Oturan, "Removal of lindane wastes by advanced electrochemical oxidation," Chemosphere, vol. 202, pp. 400- 409, 2018.
[2]. ThermoFisher Scientific, "Cell culture basic Handbook,” 2020.
[3]. A. Adan, Y. Kiraz, and Y. Baran, "Cell Proliferation and Cytotoxicity Assays," Current Pharmaceutical Biotechnology, vol. 17, no. 14, pp. 1213-1221, 2016.
[4]. W. Strober, "Trypan Blue Exclusion Test of Cell Viability," Current Protocols in Immunology, 111, pp. A3 B 1-A3 B 3, 2015.
[5]. F. Denizot and R. Lang, "Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability," Journal of Immunological Methods, vol. 89, no. 2, pp. 271-277, 1986.
[6]. P. Ngamwongsatit, P. P. Banada, W. Panbangred, and A. K. Bhunia, "WST-1-based cell cytotoxicity assay as a substitute for MTT-based assay for rapid detection of toxigenic Bacillus species using CHO cell line," Journal of Microbiological Methods, vol. 73, no. 3, pp. 211-215, 2008.
[7]. T. Decker and M. L. Lohmann-Matthes, "A quick and simple method for the quantitation of lactate dehydrogenase release in measurements of cellular cytotoxicity and tumor necrosis factor (TNF) activity," Journal of Immunological Methods, vol. 115, no. 1, pp. 61-69, 1988.
[8]. J. Uggeri, R. Gatti, S. Belletti, R. Scandroglio, R. Corradini, B. M. Rotoli, and G. Orlandini, "Calcein-AM is a detector of intracellular oxidative activity," Histochemistry and Cell Biology, vol. 122, no. 5, pp. 499-505, 2004.
[9]. T. L. Riss, R. A. Moravec, A. L. Niles, et al., "Cell Viability Assays,”, Editors, Assay Guidance Manual, Bethesda (MD), 2004.
[10]. D. H. Phillips and V. M. Arlt, "Genotoxicity: damage to DNA and its consequences,” EXS, vol. 99, pp. 87-110, 2009.
[11]. J. Alejandra Izquierdo-Vega, J. A. Morales-González, M. Sánchez-Gutiérrez, G. Betanzos-Cabrera, S. M. Sosa-Delgado, M. T. Sumaya-Martínez, Á. MoralesGonzález,4 R. Paniagua-Pérez, E. Madrigal-Bujaidar, and E. Madrigal-Santillán, "Evidence of Some Natural Products with Antigenotoxic Effects. Part 1: Fruits and Polysaccharides," Nutrients, vol. 9, no. 2, 2017.
[12]. A. R. Collins, "The comet assay for DNA damage and repair: principles, applications, and limitations," Molecular Biotechnology, vol. 26, no. 3, pp. 249-261, 2004.
[13]. E. B. Kurutas, "The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state," Nutrition Journal, vol. 15, no. 1, pp. 71, 2016.
[14]. L. A. Pham-Huy, H. He, and C. Pham-Huy, "Free radicals, antioxidants in disease and health," International Journal of Biomedical Science, vol. 4, no. 2, pp. 89-96, 2008.
[15]. C. Furger, "Live cell assays for the assessment of antioxidant activities of plant extracts," Antioxidants (Basel), vol. 10, no. 6, 2021.
[16]. P. Liu, W. Wang, J. Tang, R. P. Bowater, and Y. Bao, "Antioxidant effects of sulforaphane in human HepG2 cells and immortalised hepatocytes," Food and Chemical Toxicology, vol. 128, pp. 129-136, 2019.
[17]. X. She, F. Wang, J. Ma, and X. Chen, "In vitro antioxidant and protective effects of corn peptides on ethanol-induced damage in HepG2 cells," Food Agricultural Immumology, vol. 27, no. 1, pp. 1-12, 2015.
[18]. J. Y. Lee and C. H. Kang, "Probiotics Alleviate Oxidative Stress in H2O2-Exposed Hepatocytes and t-BHP-Induced C57BL/6 Mice," Microorganisms, vol. 10, no. 2, 2022.
[19]. R. Scrivo, M. Vasile, I. Bartosiewicz, and G. Valesini, "Inflammation as "common soil" of the multifactorial diseases," Autoimmunity Reviews, vol. 10, no. 7, pp. 369-374, 2011.
[20]. N. Q. C. Thanh ,T. D. Binh, P. L. A. Tuan, N. D. H. Yen, D. T. X. Trang, N. T. Tuan, K. Kanaori, and K. Kamei, "Anti-Inflammatory Effects of Lasia spinosa Leaf Extract in Lipopolysaccharide-Induced RAW 264.7 Macrophages," International Journal of Molecular Sciences, vol. 21, no. 10, 2020.
[21]. L. Dong, L. Yin, Y. Zhang, X. Fu, and J. Lu, "Anti-inflammatory effects of ononin on lipopolysaccharide-stimulated RAW 264.7 cells," Molecular Immunology, vol. 83, pp. 46- 51, 2017.
[22]. Y. Tian, S. Zhou, R. Takeda, K. Okazaki, M. Sekita, and K. Sakamoto, "Antiinflammatory activities of amber extract in lipopolysaccharide-induced RAW 264.7 macrophages," Biomedicine and Pharmacotherapy, vol. 141, p. 111854, 2021.
[23]. M. Cargnello and P. P. Roux, "Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases," Microbiology and Molecular Biology Reviews, vol. 75, no. 1, pp. 50-83, 2011.
[24]. M. R. Guimaraes, F. R. Leite, L. C. Spolidorio, K. L. Kirkwood, and C. Rossa, Jr., "Curcumin abrogates LPS-induced pro-inflammatory cytokines in RAW 264.7 macrophages. Evidence for novel mechanisms involving SOCS-1, -3 and p38 MAPK," Archives of Oral Biology, vol. 58, no. 10, pp. 1309-1317, 2013.
[25]. F. Xue, X. Nie, J. Shi, Q. Liu, Z. Wang, X. Li , J. Zhou, J. Su, M. Xue, W-D. Chen, YD. Wang , "Quercetin Inhibits LPS-Induced Inflammation and ox-LDL-Induced Lipid Deposition," Fronties in Pharmacology, vol. 8, pp. 40, 2017.
[26]. S. J. Hwang, Y. W. Kim, Y. Park, H. J. Lee, and K. W. Kim, "Anti-inflammatory effects of chlorogenic acid in lipopolysaccharide-stimulated RAW 264.7 cells," Inflammation Research, vol. 63, no. 1, pp. 81-90, 2014.
[27]. Y. H. Choi, G. Y. Kim, and H. H. Lee, "Anti-inflammatory effects of cordycepin in lipopolysaccharide-stimulated RAW 264.7 macrophages through Toll-like receptor 4-mediated suppression of mitogen-activated protein kinases and NF-kappaB signaling pathways," Drug Design, Development and Therapy, vol. 8, pp. 1941-1953, 2014.
[28]. S. S. Ranaweera, C. Y. Dissanayake, P. Natraj, Y. J. Lee, and C. H. Han, "Antiinflammatory effect of sulforaphane on LPS-stimulated RAW 264.7 cells and ob/ob mice," Journal of Veterinary Science, vol. 21, no. 6, pp. e91, 2020.
[29]. S. H. Choi, S. H. Lee, M. G. Kim, H. J. Lee, and G. B. Kim, "Lactobacillus plantarum CAU1055 ameliorates inflammation in lipopolysaccharide-induced RAW264.7 cells and a dextran sulfate sodium-induced colitis animal model," Journal of Dairy Science, vol. 102, no. 8, pp. 6718-6725, 2019.
[30]. M. Griet, H. Zelaya, M. V. Mateos, S. Salva, G. E. Juarez, G. F. de Valdez, J. Villena, G. A. Salvador, and A. V. Rodriguez , "Soluble factors from Lactobacillus reuteri CRL1098 have anti-inflammatory effects in acute lung injury induced by lipopolysaccharide in mice," PLoS One, vol. 9, no. 10, pp. e110027, 2014.
[31]. H. E. Park, K. H. Do, and W. K. Lee, "The immune-modulating effects of viable Weissella cibaria JW15 on RAW 264.7 macrophage cells," Journal of Biomedical Research, vol. 34, no. 1, pp. 36-43, 2019.
[32]. N. Lee, S. Lee, S. Lee, S. W. Jang, H. S. Shin, J-H. Park, M. S. Park, and B-H. Lee, "Lysed and disrupted Bifidobacterium bifidum BGN4 cells promote anti-inflammatory activities in lipopolysaccharide-stimulated RAW 264.7 cells," Saudi Journal of Biology Science, vol. 28, no. 9, pp. 5115-5118, 2021.
[33]. A. C. Archer, N. K. Kurrey, and P. M. Halami, "In vitro adhesion and anti-inflammatory properties of native Lactobacillus fermentum and Lactobacillus delbrueckii spp," Journal of Applied Microbiology, vol. 125, no. 1, pp. 243-256, 2018.
[34]. Y. M. Kim, I. H. Kim, J. W. Choi, M. K. Lee, and T. J. Nam, "The anti-obesity effects of a tuna peptide on 3T3-L1 adipocytes are mediated by the inhibition of the expression of lipogenic and adipogenic genes and by the activation of the Wnt/beta-catenin signaling pathway," International Journal of Molecule Medicine, vol. 36, no. 2, pp. 327-334, 2015.
[35]. F. J. Ruiz-Ojeda, A. I. Ruperez, C. Gomez-Llorente, A. Gil, and C. M. Aguilera, "Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review," International Journal of Molecule Science, vol. 17, no. 7, 2016.
[36]. Y. S. Jeong, H. K. Jung, K-H. Cho, K-S. Youn, and J-H. Hong "Anti-obesity effect of grape skin extract in 3T3-L1 adipocytes," Food Science and Biotechnology, vol. 20, no. 3, pp. 635-642, 2011.
[37]. K. J. Han, N. K. Lee, H. S. Yu, H. Park, and H. D. Paik, "Anti-adipogenic Effects of the Probiotic Lactiplantibacillus plantarum KU15117 on 3T3-L1 Adipocytes," Probiotics Antimicrob Proteins, vol. 14, no. 3, pp. 501-509, 2022.
[38]. M. Miyoshi, A. Ogawa, S. Higurashi, and Y. Kadooka, "Anti-obesity effect of Lactobacillus gasseri SBT2055 accompanied by inhibition of pro-inflammatory gene expression in the visceral adipose tissue in diet-induced obese mice," European Journal of Nutrition, vol. 53, no. 2, pp. 599-606, 2014.
[39]. K-H. Lee, J-L. Song, E-S. Park, J. Ju, H-Y. Kim, and K-Y. Park, "Anti-Obesity Effects of Starter Fermented Kimchi on 3T3-L1 Adipocytes," Preventive Nutrition and Food Science, vol. 20, no. 4, pp. 298-302, 2015.
[40]. X. Chen, C. Guo, and J. Kong, "Oxidative stress in neurodegenerative diseases," Neural Regeneration Research, vol. 7, no. 5, pp. 376-385, 2012.
[41]. A. Gonzalez-Sarrias, M. A. Nunez-Sanchez, F. A. Tomas-Barberan, and J. C. Espin, "Neuroprotective Effects of Bioavailable Polyphenol-Derived Metabolites against Oxidative Stress-Induced Cytotoxicity in Human Neuroblastoma SH-SY5Y Cells," Journal of Agricultural and Food Chemistry, vol. 65, no. 4, pp. 752-758, 2017.
[42]. S. Thummayot, C. Tocharus, D. Pinkaew, K. Viwatpinyo, K. Sringarm, and J. Tocharus, "Neuroprotective effect of purple rice extract and its constituent against amyloid beta-induced neuronal cell death in SK-N-SH cells," Neurotoxicology, vol. 45, pp. 149-158, 2014.
[43]. M. J. Cheon, S. M. Lim, N. K. Lee, and H. D. Paik, "Probiotic Properties and Neuroprotective Effects of Lactobacillus buchneri KU200793 Isolated from Korean Fermented Foods," International Journal of Molecule Science, vol. 21, no. 4, 2020.
[44]. J. F. Cryan, K. J. O'Riordan, C. S. M. Cowan, et al., "The microbiota-gut-brain axis," Physiology Review, vol. 99, no. 4, pp. 1877-2013, 2019.
[45]. I. Hubatsch, E. G. Ragnarsson, and P. Artursson, "Determination of drug permeability and prediction of drug absorption in Caco-2 monolayers," Nature Protocols, vol. 2, no. 9, pp. 2111-2119, 2007.
[46]. C. Stolfi, C. Maresca, G. Monteleone, and F. Laudisi, "Implication of Intestinal Barrier Dysfunction in Gut Dysbiosis and Diseases," Biomedicines, vol. 10, no. 2, 2022.
[47]. J. Y. Kim, T. A. N. Le, S. Y. Lee, et al., "3,3'-Diindolylmethane improves intestinal permeability dysfunction in cultured human intestinal cells and the model animal caenorhabditis elegans," Journal of Agricultural and Food Chemistry, vol. 67, no. 33, pp. 9277-9285, 2019.
[48]. K. K. Putt, R. Pei, H. M. White, and B. W. Bolling, "Yogurt inhibits intestinal barrier dysfunction in Caco-2 cells by increasing tight junctions,” Food and Function, vol. 8, no. 1, pp. 406-414, 2017.
[49]. L. Rhayat, M. Maresca, C. Nicoletti, et al., "Effect of Bacillus subtilis strains on intestinal barrier function and inflammatory response,” Frontier in Immunology, vol. 10, p. 564, 2019.
[50]. C. Gong, Z. Ni, C. Yao, et al., "A high-throughput assay for screening of natural products that enhanced tumoricidal Aactivity of NK cells,” Biological Procedures Online, vol. 17, p. 12, 2015.
[51]. J. L. Tan, F. Li, J. Z. Yeo, et al., "New high-throughput screening identifies compounds that reduce viability specifically in liver cancer cells that express high levels of SALL4 by inhibiting oxidative phosphorylation,” Gastroenterology, vol. 157, no. 6, pp. 1615-1629 e1617, 2019.
[52]. J. Perez Del Palacio, C. Diaz, M. de la Cruz, et al., "High-throughput screening platform for the discovery of new immunomodulator molecules from natural product extract libraries,” Journal of Biomolecular Screening, vol. 21, no. 6, pp. 567-578, 2016.
[53]. G. Alzeeb, J. P. Metges, L. Corcos, and C. Le Jossic-Corcos, "Three-Dimensional Culture Systems in Gastric Cancer Research,” Cancers (Basel), vol. 12, no. 10, 2020.
[54]. S Chenchula, S Kumar, and S Babu, "Comparitive efficacy of 3dimensional (3D) cell culture organoids vs 2dimensional (2D) cell cultures vs experimental animal models in disease modeling, drug development, and drug toxicity testing,” International Journal of Current Research and Review, 2019.
[55]. M. R. Kim, S. Y. Cho, H. J. Lee, et al., "Schisandrin C improves leaky gut conditions in intestinal cell monolayer, organoid, and nematode models by increasing tight junction protein expression,” Phytomedicine, vol. 103, p. 154209, 2022.
[56]. T. Cai, Y. Qi, A. Jergens, et al., "Effects of six common dietary nutrients on murine intestinal organoid growth,” PLoS One, vol. 13, no. 2, pp. e0191517, 2018.
[57]. Y. Wang, T. Wei, Q. Wang, et al., "Resveratrol's neural protective effects for the injured embryoid body and cerebral organoid,” BMC Pharmacology and Toxicology, vol. 23, no. 1, p. 47, 2022.
[58]. M. S. Jeon, Y. Y. Choi, S. J. Mo, et al., "Contributions of the microbiome to intestinal inflammation in a gut-on-a-chip,” Nano Convergence, vol. 9, no. 1, pp. 8, 2022.
[59]. I. Raimondi, L. Izzo, M. Tunesi, et al., "Organ-On-A-Chip in vitro Models of the Brain and the Blood-Brain Barrier and Their Value to Study the Microbiota-Gut-Brain Axis in Neurodegeneration,” Frontiers in Bioengineering and Biotechnology, vol. 7, pp. 435, 2019.
[60]. Min-Hyeok Kim, D. Kim, and J. Hwan, "A gut-brain axis-on-a-chip for studying transport across epithelial and endothelial barriers,” Journal of Industrial and Engineering Chemistry, vol. 101, pp. 126-134, 2021.