{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T01:13:12Z","timestamp":1791594792237,"version":"4.3.4"},"reference-count":44,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2020,9,19]],"date-time":"2020-09-19T00:00:00Z","timestamp":1600473600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Bioactive compounds, such as organic acids (OA) and nature-identical compounds (NIC), can exert a role in the protection of intestinal mucosa functionality due to their biological properties. The aim of this study was to understand the role of 2 OA (citric and sorbic acid) and 2 NIC (thymol and vanillin), alone or combined in a blend (OA + NIC), on intestinal barrier functionality, either during homeostatic condition or during an inflammatory challenge performed with pro-inflammatory cytokines and lipopolysaccharides (LPS). The study was performed on the human epithelial cell line Caco-2, a well-known model of the intestinal epithelial barrier. The results showed how OA and NIC alone can improve transepithelial electrical resistance (TEER) and mRNA levels of tight junction (TJ) components, but OA + NIC showed stronger efficacy compared to the single molecules. When an inflammatory challenge occurred, OA + NIC blend was able both to ameliorate, and prevent, damage caused by the pro-inflammatory stimulus, reducing or preventing the drop in TEER and improving the TJ mRNA expression. The data support the role of OA + NIC in modulating gut barrier functionality and reducing the negative effects of inflammation in intestinal epithelial cells, thereby supporting the gut barrier functionality. <\/jats:p>","DOI":"10.3390\/molecules25184296","type":"journal-article","created":{"date-parts":[[2020,9,19]],"date-time":"2020-09-19T07:06:09Z","timestamp":1600499169000},"page":"4296","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Nature-Identical Compounds and Organic Acids Ameliorate and Prevent the Damages Induced by an Inflammatory Challenge in Caco-2 Cell Culture"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1584-7127","authenticated-orcid":false,"given":"Andrea","family":"Toschi","sequence":"first","affiliation":[{"name":"Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra n. 50, 40064 Ozzano dell\u2019Emilia (BO), Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Rossi","sequence":"additional","affiliation":[{"name":"Vetagro S.p.A., via Porro 2, 42124 Reggio Emilia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benedetta","family":"Tugnoli","sequence":"additional","affiliation":[{"name":"Vetagro S.p.A., via Porro 2, 42124 Reggio Emilia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Piva","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra n. 50, 40064 Ozzano dell\u2019Emilia (BO), Italy"},{"name":"Vetagro S.p.A., via Porro 2, 42124 Reggio Emilia, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ester","family":"Grilli","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra n. 50, 40064 Ozzano dell\u2019Emilia (BO), Italy"},{"name":"Vetagro Inc., 116W. Jackson Blvd., Suite #320, Chicago, IL 60604, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1007\/s005350070017","article-title":"First line of defense: The role of the intestinal epithelium as an active component of the mucosal immune system","volume":"35","author":"Pitman","year":"2000","journal-title":"J. Gastroenterol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"G413","DOI":"10.1152\/ajpgi.00304.2006","article-title":"Gastrointestinal dysfunction induced by early weaning is attenuated by delayed weaning and mast cell blockade in pigs","volume":"293","author":"Moeser","year":"2007","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"a002584","DOI":"10.1101\/cshperspect.a002584","article-title":"Physiology and function of the tight junction","volume":"1","author":"Anderson","year":"2009","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/35067088","article-title":"Multifunctional strands in tight junctions","volume":"2","author":"Tsukita","year":"2001","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Alaimo, A., and Rubert, J. (2019). The pivotal role of TRP channels in homeostasis and diseases throughout the gastrointestinal tract. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20215277"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1111\/j.1748-1716.2011.02338.x","article-title":"Cannabinoid actions at TRPV channels: Effects on TRPV3 and TRPV4 and their potential relevance to gastrointestinal inflammation","volume":"204","author":"Orlando","year":"2012","journal-title":"Acta Physiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Vitale, S., Strisciuglio, C., Pisapia, L., Miele, E., Barba, P., Vitale, A., Cenni, S., Bassi, V., Maglio, M., and Del Pozzo, G. (2017). Cytokine production profile in intestinal mucosa of paediatric inflammatory bowel disease. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0182313"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1186\/2049-1891-4-19","article-title":"The biological stress of early weaned piglets","volume":"4","author":"Campbell","year":"2013","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"864","DOI":"10.1016\/j.bbamem.2008.08.027","article-title":"Cytokine regulation of tight junctions","volume":"1788","author":"Capaldo","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Tugnoli, B., Giovagnoni, G., Piva, A., and Grilli, E. (2020). From acidifiers to intestinal health enhancers: How organic acids can improve growth efficiency of pigs. Animals, 10.","DOI":"10.3390\/ani10010134"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Rossi, B., Toschi, A., Piva, A., and Grilli, E. (2020). Single components of botanicals and nature-identical compounds as a non-antibiotic strategy to ameliorate health status and improve performance in poultry and pigs. Nutr. Res. Rev., 1\u201317.","DOI":"10.1017\/S0954422420000013"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1080\/09712119.2015.1079527","article-title":"Recent advances in the role of organic acids in poultry nutrition","volume":"44","author":"Khan","year":"2016","journal-title":"J. Appl. Anim. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Grilli, E., Tugnoli, B., Passey, J.L., Stahl, C.H., Piva, A., and Moeser, A.J. (2015). Impact of dietary organic acids and botanicals on intestinal integrity and inflammation in weaned pigs. BMC Vet. Res., 11.","DOI":"10.1186\/s12917-015-0410-0"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Swaggerty, C.L., Callaway, T.R., Kogut, M.H., Piva, A., and Grilli, E. (2019). Modulation of the immune response to improve health and reduce foodborne pathogens in poultry. Microorganisms, 7.","DOI":"10.3390\/microorganisms7030065"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Verhoeckx, K., Cotter, P., L\u00f3pez-Exp\u00f3sito, I., Kleiveland, C., Lea, T., Mackie, A., Requena, T., Swiatecka, D., and Wichers, H. (2015). Caco-2 cell line. The Impact of Food Bioactives on Health: In Vitro and Ex Vivo Models, Springer International Publishing.","DOI":"10.1007\/978-3-319-16104-4"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2013\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"361","DOI":"10.4081\/ijas.2007.361","article-title":"In vitro effects of some organic acids on swine cecal microflora","volume":"6","author":"Biagi","year":"2010","journal-title":"Ital. J. Anim. Sci."},{"key":"ref_18","unstructured":"Callaway, T.R., and Edrington, T.S. (2012). Organic acids and their role in reduce foodborne pathogens in food animals. On-Farm Strategies to Control Foodborne Pathogens, Advances in Food Safety and Food Microbiology."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2662","DOI":"10.3389\/fmicb.2019.02662","article-title":"Dietary supplementation with citrus extract altered the intestinal microbiota and microbial metabolite profiles and enhanced the mucosal immune homeostasis in yellow-feathered broilers","volume":"10","author":"Yu","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1046\/j.1365-2672.2001.01428.x","article-title":"A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol","volume":"91","author":"Lambert","year":"2001","journal-title":"J. Appl. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1111\/j.1365-2672.2004.02275.x","article-title":"Mode of antimicrobial action of vanillin against Escherichia coli, Lactobacillus plantarum and Listeria innocua","volume":"97","author":"Fitzgerald","year":"2004","journal-title":"J. Appl. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1016\/j.tiv.2015.02.006","article-title":"In vitro pro-oxidant\/antioxidant role of carvacrol, thymol and their mixture in the intestinal Caco-2 cell line","volume":"29","author":"Puerto","year":"2015","journal-title":"Toxicol. In Vitro"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1002\/ddr.10436","article-title":"Thymol analogues with antioxidant and L-type calcium current inhibitory activity","volume":"64","author":"Shen","year":"2005","journal-title":"Drug Dev. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"46395","DOI":"10.1039\/C7RA09302K","article-title":"Intracellular antioxidant effect of vanillin, 4-methylguaiacol and 4-ethylguaiacol: Three components in Chinese Baijiu","volume":"7","author":"Zhao","year":"2017","journal-title":"RSC Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1111\/nyas.13508","article-title":"Host\u2013microbiota interplay in mediating immune disorders","volume":"1417","author":"Felix","year":"2018","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1111\/jam.12053","article-title":"Development of a feed additive to reduce caecal Campylobacter jejuni in broilers at slaughter age: From in vitro to in vivo, a proof of concept","volume":"114","author":"Grilli","year":"2013","journal-title":"J. Appl. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"K2","DOI":"10.1016\/0014-5793(80)80564-3","article-title":"The role of citric acid in intermediate metabolism in animal tissues","volume":"117","author":"Krebs","year":"1980","journal-title":"FEBS Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/0378-5173(93)90195-L","article-title":"Cell-polarity dependent effect of chelation on the paracellular permeability of confluent caco-2 cell monolayers","volume":"90","author":"Noach","year":"1993","journal-title":"Int. J. Pharm."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"C978","DOI":"10.1152\/ajpcell.1990.259.6.C978","article-title":"Role of calcium in tight junction formation between epithelial cells","volume":"259","author":"Contreras","year":"1990","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2356","DOI":"10.2527\/jas.2010-3677","article-title":"Sorbic acid improves growth performance and regulates insulin-like growth factor system gene expression in swine","volume":"89","author":"Luo","year":"2011","journal-title":"J. Anim. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"E140","DOI":"10.2527\/jas.2007-0459","article-title":"Use of phytogenic products as feed additives for swine and poultry","volume":"86","author":"Windisch","year":"2008","journal-title":"J. Anim. Sci."},{"key":"ref_32","first-page":"543","article-title":"Thyme and cinnamon extracts induce anion secretion in piglet small intestine via cholinergic pathways","volume":"59","author":"Boudry","year":"2008","journal-title":"J. Physiol. Pharm."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.livsci.2010.06.087","article-title":"Thymol and trans-cinnamaldehyde reduce active nutrient absorption and chloride secretion in the pig jejunal Ussing chamber model","volume":"134","author":"Michiels","year":"2010","journal-title":"Livest. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1021\/cn500094a","article-title":"Transient receptor potential channels as targets for phytochemicals","volume":"5","author":"Premkumar","year":"2014","journal-title":"ACS Chem. Neurosci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.3389\/fonc.2019.01087","article-title":"Inflammation, cancer and immunity\u2014Implication of TRPV1 channel","volume":"9","author":"Bujak","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_37","first-page":"2765","article-title":"Mechanism of cytokine modulation of epithelial tight junction barrier","volume":"14","author":"Boivin","year":"2009","journal-title":"Front. Biosci."},{"key":"ref_38","first-page":"85","article-title":"Phytochemical evaluation, antioxidant assay, antibacterial activity and determination of cell viability (J774 and THP1 alpha cell lines) of P. sylvestris leaf crude and methanol purified fractions","volume":"15","author":"Sharma","year":"2016","journal-title":"EXCLI J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1080\/09168451.2014.951027","article-title":"Theaflavins enhance intestinal barrier of Caco-2 Cell monolayers through the expression of AMP-activated protein kinase-mediated Occludin, Claudin-1, and ZO-1","volume":"79","author":"Park","year":"2015","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1242\/bio.20149571","article-title":"TRPV1 mediates cellular uptake of anandamide and thus promotes endothelial cell proliferation and network-formation","volume":"3","author":"Hofmann","year":"2014","journal-title":"Biol. Open"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/nature00882","article-title":"TRPV3 is a calcium-permeable temperature-sensitive cation channel","volume":"418","author":"Xu","year":"2002","journal-title":"Nature"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"G1067","DOI":"10.1152\/ajpgi.00234.2005","article-title":"Normalizing genes for quantitative RT-PCR in differentiating human intestinal epithelial cells and adenocarcinomas of the colon","volume":"290","author":"Dydensborg","year":"2006","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"research0034.1","DOI":"10.1186\/gb-2002-3-7-research0034","article-title":"Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes","volume":"3","author":"Vandesompele","year":"2002","journal-title":"Genome Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1441","DOI":"10.1016\/j.tiv.2010.04.002","article-title":"Inflammatory parameters in Caco-2 cells: Effect of stimuli nature, concentration, combination and cell differentiation","volume":"24","author":"Hendrickx","year":"2010","journal-title":"Toxicol. In Vitro"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/18\/4296\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:11:33Z","timestamp":1760177493000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/18\/4296"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,19]]},"references-count":44,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["molecules25184296"],"URL":"https:\/\/doi.org\/10.3390\/molecules25184296","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,19]]}}}