{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:10:19Z","timestamp":1790748619763,"version":"4.1.0"},"reference-count":62,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2021,12,19]],"date-time":"2021-12-19T00:00:00Z","timestamp":1639872000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deutsche Gesellschaft f\u00fcr Kieferorthop\u00e4die e.V","award":["Hennig 2018-11"],"award-info":[{"award-number":["Hennig 2018-11"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Periodontitis is characterized by bacterially induced inflammatory destruction of periodontal tissue. This also affects fibroblasts of the human periodontal ligaments (HPdLF), which play a coordinating role in force-induced tissue and alveolar bone remodeling. Excessive inflammation in the oral tissues has been observed with simultaneous stimulation by pathogens and mechanical forces. Recently, elevated levels of growth differentiation factor 15 (GDF15), an immuno-modulatory member of the transforming growth factor (TGFB) superfamily, were detected under periodontitis-like conditions and in force-stressed PdL cells. In view of the pleiotropic effects of GDF15 in various tissues, this study aims to investigate the role of GDF15 in P. gingivalis-related inflammation of HPdLF and its effect on the excessive inflammatory response to concurrent compressive stress. To this end, the expression and secretion of cytokines (IL6, IL8, COX2\/PGE2, TNF\u03b1) and the activation of THP1 monocytic cells were analyzed in GDF15 siRNA-treated HPdLF stimulated with P. gingivalis lipopolysaccharides alone and in combination with compressive force. GDF15 knockdown significantly reduced cytokine levels and THP1 activation in LPS-stimulated HPdLF, which was less pronounced with additional compressive stress. Overall, our data suggest a pro-inflammatory role for GDF15 in periodontal disease and demonstrate that GDF15 partially modulates the force-induced excessive inflammatory response of PdLF under these conditions.<\/jats:p>","DOI":"10.3390\/ijms222413608","type":"journal-article","created":{"date-parts":[[2021,12,19]],"date-time":"2021-12-19T20:37:27Z","timestamp":1639946247000},"page":"13608","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["GDF15 Supports the Inflammatory Response of PdL Fibroblasts Stimulated by P. gingivalis LPS and Concurrent Compression"],"prefix":"10.3390","volume":"22","author":[{"given":"Albert","family":"Stemmler","sequence":"first","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9347-0172","authenticated-orcid":false,"given":"Judit","family":"Symmank","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Steinmetz","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katrin","family":"von Brandenstein","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph-Ludwig","family":"Hennig","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Collin","family":"Jacobs","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, University Hospital Jena, Leutragraben 3, 07743 Jena, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11514","DOI":"10.1073\/pnas.94.21.11514","article-title":"Mic-1, a novel macrophage inhibitory cytokine, is a divergent member of the tgf-beta superfamily","volume":"94","author":"Bootcov","year":"1997","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"103","DOI":"10.3389\/fcvm.2020.00103","article-title":"Serum gdf15, a promising biomarker in obese patients undergoing heart surgery","volume":"7","author":"Sarkar","year":"2020","journal-title":"Front. Cardiovasc. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3410","DOI":"10.1073\/pnas.0530278100","article-title":"Large-scale delineation of secreted protein biomarkers overexpressed in cancer tissue and serum","volume":"100","author":"Welsh","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"111228","DOI":"10.1016\/j.exger.2021.111228","article-title":"Gdf15 as a biomarker of ageing","volume":"146","author":"Liu","year":"2021","journal-title":"Exp. Gerontol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1007\/s004410051337","article-title":"Expression of a novel member of the tgf-beta superfamily, growth\/differentiation factor-15\/macrophage-inhibiting cytokine-1 (gdf-15\/mic-1) in adult rat tissues","volume":"297","author":"Bottner","year":"1999","journal-title":"Cell Tissue Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3742","DOI":"10.1128\/MCB.20.10.3742-3751.2000","article-title":"Characterization of growth-differentiation factor 15, a transforming growth factor beta superfamily member induced following liver injury","volume":"20","author":"Hsiao","year":"2000","journal-title":"Mol. Cell. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4983","DOI":"10.1158\/0008-5472.CAN-05-4067","article-title":"Role of macrophage inhibitory cytokine-1 in tumorigenesis and diagnosis of cancer","volume":"66","author":"Bauskin","year":"2006","journal-title":"Cancer Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"187","DOI":"10.3109\/08977194.2011.607137","article-title":"The tgf-beta superfamily cytokine, mic-1\/gdf15: A pleotrophic cytokine with roles in inflammation, cancer and metabolism","volume":"29","author":"Breit","year":"2011","journal-title":"Growth Factors"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"272","DOI":"10.7150\/ijbs.2929","article-title":"Tgf-beta and bmp signaling in osteoblast differentiation and bone formation","volume":"8","author":"Chen","year":"2012","journal-title":"Int. J. Biol. Sci."},{"key":"ref_10","first-page":"213","article-title":"Growth\/differentiation factor-15 inhibits differentiation into osteoclasts\u2014A novel factor involved in control of osteoclast differentiation","volume":"78","author":"Vanhara","year":"2009","journal-title":"Differ. Res. Biol. Divers."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1002\/jbmr.1538","article-title":"Positive regulation of osteoclastic differentiation by growth differentiation factor 15 upregulated in osteocytic cells under hypoxia","volume":"27","author":"Hinoi","year":"2012","journal-title":"J. Bone Miner. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e511","DOI":"10.3324\/haematol.2015.124511","article-title":"Growth differentiation factor 15 (gdf15) promotes osteoclast differentiation and inhibits osteoblast differentiation and high serum gdf15 levels are associated with multiple myeloma bone disease","volume":"100","author":"Westhrin","year":"2015","journal-title":"Haematologica"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11516","DOI":"10.1038\/s41598-019-47639-x","article-title":"Mechanically-induced gdf15 secretion by periodontal ligament fibroblasts regulates osteogenic transcription","volume":"9","author":"Symmank","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"111745","DOI":"10.1016\/j.yexcr.2019.111745","article-title":"Gdf15 induced by compressive force contributes to osteoclast differentiation in human periodontal ligament cells","volume":"387","author":"Li","year":"2020","journal-title":"Exp. Cell Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1111\/j.1600-0757.2005.00141.x","article-title":"Structure of periodontal tissues in health and disease","volume":"40","author":"Nanci","year":"2006","journal-title":"Periodontology 2000"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1089\/scd.2019.0025","article-title":"Periodontal ligament stem cells: Current knowledge and future perspectives","volume":"28","author":"Trubiani","year":"2019","journal-title":"Stem Cells Dev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2058738419828613","DOI":"10.1177\/2058738419828613","article-title":"Effect of short peptides on neuronal differentiation of stem cells","volume":"33","author":"Caputi","year":"2019","journal-title":"Int. J. Immunopathol. Pharmacol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1093\/ejo\/19.6.615","article-title":"Osteoblast-like properties of human periodontal ligament cells: An in vitro analysis","volume":"19","author":"Basdra","year":"1997","journal-title":"Eur. J. Orthod."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.kjms.2018.01.007","article-title":"Orthodontic tooth movement: The biology and clinical implications","volume":"34","author":"Li","year":"2018","journal-title":"Kaohsiung J. Med. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1111\/j.1600-0765.2010.01331.x","article-title":"The human periodontal ligament cell: A fibroblast-like cell acting as an immune cell","volume":"46","author":"Jonsson","year":"2011","journal-title":"J. Periodontal. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/s00056-014-0212-1","article-title":"Effects of mechanical and bacterial stressors on cytokine and growth-factor expression in periodontal ligament cells","volume":"75","author":"Proff","year":"2014","journal-title":"J. Orofac. Orthop. Fortschr. Kieferorthopadie"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Yamaguchi, M., and Fukasawa, S. (2021). Is inflammation a friend or foe for orthodontic treatment?: Inflammation in orthodontically induced inflammatory root resorption and accelerating tooth movement. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22052388"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1016\/j.ajodo.2009.06.021","article-title":"Root resorption associated with orthodontic tooth movement: A systematic review","volume":"137","author":"Weltman","year":"2010","journal-title":"Am. J. Orthod. Dentofac. Orthop."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1111\/adj.12486","article-title":"Potential risks of orthodontic therapy: A critical review and conceptual framework","volume":"62","author":"Wishney","year":"2017","journal-title":"Aust. Dent. J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.ajodo.2017.08.015","article-title":"Incisor malalignment and the risk of periodontal disease progression","volume":"153","author":"Alsulaiman","year":"2018","journal-title":"Am. J. Orthod. Dentofac. Orthop."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"17038","DOI":"10.1038\/nrdp.2017.38","article-title":"Periodontal diseases","volume":"3","author":"Kinane","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"101","DOI":"10.4103\/jisp.jisp_385_17","article-title":"Polymicrobial synergy and dysbiosis: An overview","volume":"22","author":"Shaikh","year":"2018","journal-title":"J. Indian Soc. Periodontol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jdsr.2017.06.001","article-title":"Molecular mechanisms of porphyromonas gingivalis-host cell interaction on periodontal diseases","volume":"53","author":"Nakayama","year":"2017","journal-title":"Jpn. Dent. Sci. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"284","DOI":"10.3390\/jcm9010284","article-title":"Porphyromonas gingivalis load is balanced by 0.20% chlorhexidine gel. A randomized, double-blind, controlled, microbiological and immunohistochemical human study","volume":"9","author":"Traini","year":"2020","journal-title":"J. Clin. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1038\/s41522-020-0120-7","article-title":"Polymicrobial periodontal disease triggers a wide radius of effect and unique virome","volume":"6","author":"Gao","year":"2020","journal-title":"npj Biofilms Microbiomes"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.bbrc.2019.11.074","article-title":"Porphyromonas gingivalis triggers inflammatory responses in periodontal ligament cells by succinate-succinate dehydrogenase-hif-1alpha axis","volume":"522","author":"Su","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Symmank, J., Appel, S., Bastian, J.A., Knaup, I., Marciniak, J., Hennig, C.L., Doding, A., Schulze-Spate, U., Jacobs, C., and Wolf, M. (2021). Hyperlipidemic conditions impact force-induced inflammatory response of human periodontal ligament fibroblasts concomitantly challenged with p. Gingivalis-lps. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22116069"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"15789","DOI":"10.1038\/s41598-017-16190-y","article-title":"Porphyromonas gingivalis lipopolysaccharides act exclusively through tlr4 with a resilience between mouse and human","volume":"7","author":"Nativel","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5041","DOI":"10.1128\/IAI.72.9.5041-5051.2004","article-title":"Porphyromonas gingivalis lipopolysaccharide contains multiple lipid a species that functionally interact with both toll-like receptors 2 and 4","volume":"72","author":"Darveau","year":"2004","journal-title":"Infect. Immun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"12281","DOI":"10.1073\/pnas.1918508117","article-title":"Cxcl5-mediated recruitment of neutrophils into the peritoneal cavity of gdf15-deficient mice protects against abdominal sepsis","volume":"117","author":"Santos","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"119356","DOI":"10.1016\/j.lfs.2021.119356","article-title":"Atf4-mediated gdf15 suppresses lps-induced inflammation and muc5ac in human nasal epithelial cells through the pi3k\/akt pathway","volume":"275","author":"Li","year":"2021","journal-title":"Life Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"L514","DOI":"10.1152\/ajplung.00324.2017","article-title":"Overproduction of growth differentiation factor 15 promotes human rhinovirus infection and virus-induced inflammation in the lung","volume":"314","author":"Wu","year":"2018","journal-title":"Am. J. Physiol. Lung Cell. Mol. Physiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1161\/01.RES.0000207919.83894.9d","article-title":"Gdf15, a cardioprotective tgf-beta superfamily protein","volume":"98","author":"Ago","year":"2006","journal-title":"Circ. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1712","DOI":"10.3389\/fphys.2018.01712","article-title":"Gdf15 and growth control","volume":"9","author":"Emmerson","year":"2018","journal-title":"Front. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/j.cell.2019.07.033","article-title":"Gdf15 is an inflammation-induced central mediator of tissue tolerance","volume":"178","author":"Luan","year":"2019","journal-title":"Cell"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1007\/s00424-020-02459-1","article-title":"Gdf15, an update of the physiological and pathological roles it plays: A review","volume":"472","author":"Assadi","year":"2020","journal-title":"Pflug. Arch."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1038\/s41598-017-00902-5","article-title":"Growth differentiation factor-15 deficiency augments inflammatory response and exacerbates septic heart and renal injury induced by lipopolysaccharide","volume":"7","author":"Abulizi","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"641851","DOI":"10.1155\/2013\/641851","article-title":"Nag-1\/gdf15 transgenic mouse has less white adipose tissue and a reduced inflammatory response","volume":"2013","author":"Kim","year":"2013","journal-title":"Mediat. Inflamm."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"978","DOI":"10.1038\/s41598-018-37425-6","article-title":"Solid stress-induced migration is mediated by gdf15 through akt pathway activation in pancreatic cancer cells","volume":"9","author":"Kalli","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"992","DOI":"10.3389\/fonc.2019.00992","article-title":"Mechanical compression regulates brain cancer cell migration through mek1\/erk1 pathway activation and gdf15 expression","volume":"9","author":"Kalli","year":"2019","journal-title":"Front. Oncol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3796","DOI":"10.1172\/JCI91561","article-title":"Nf-kappab regulates gdf-15 to suppress macrophage surveillance during early tumor development","volume":"127","author":"Ratnam","year":"2017","journal-title":"J. Clin. Investig."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1038\/icb.2013.15","article-title":"Regulation of tnf-alpha with a focus on rheumatoid arthritis","volume":"91","author":"Moelants","year":"2013","journal-title":"Immunol. Cell Biol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.advms.2018.03.001","article-title":"Interactions between the growth hormone and cytokines\u2014A review","volume":"63","author":"Szalecki","year":"2018","journal-title":"Adv. Med. Sci."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"984","DOI":"10.1016\/j.jaci.2016.06.033","article-title":"Interleukins (from il-1 to il-38), interferons, transforming growth factor beta, and tnf-alpha: Receptors, functions, and roles in diseases","volume":"138","author":"Akdis","year":"2016","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1002\/eji.1830240524","article-title":"Interleukin-10 controls interferon-gamma and tumor necrosis factor production during experimental endotoxemia","volume":"24","author":"Marchant","year":"1994","journal-title":"Eur. J. Immunol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2898","DOI":"10.4049\/jimmunol.158.6.2898","article-title":"Il-13 protects mice from lipopolysaccharide-induced lethal endotoxemia: Correlation with down-modulation of tnf-alpha, ifn-gamma, and il-12 production","volume":"158","author":"Muchamuel","year":"1997","journal-title":"J. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1007\/s00784-019-03111-8","article-title":"Orthodontic cell stress modifies proinflammatory cytokine expression in human pdl cells and induces immunomodulatory effects via tlr-4 signaling in vitro","volume":"24","author":"Marciniak","year":"2020","journal-title":"Clin. Oral Investig."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"8172","DOI":"10.1038\/s41598-020-65089-8","article-title":"Immunorthodontics: In vivo gene expression of orthodontic tooth movement","volume":"10","author":"Klein","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/j.archoralbio.2010.09.017","article-title":"Mechanical stress enhances production of cytokines in human periodontal ligament cells induced by porphyromonas gingivalis","volume":"56","author":"Yamamoto","year":"2011","journal-title":"Arch. Oral Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1038\/s41591-020-0945-x","article-title":"Antibody-mediated inhibition of gdf15-gfral activity reverses cancer cachexia in mice","volume":"26","author":"Suriben","year":"2020","journal-title":"Nat. Med."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1002\/jcsm.12077","article-title":"Map3k11\/gdf15 axis is a critical driver of cancer cachexia","volume":"7","author":"Lerner","year":"2016","journal-title":"J. Cachexia Sarcopenia Muscle"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1016\/j.tem.2020.10.004","article-title":"Insights into mechanisms of gdf15 and receptor gfral: Therapeutic targets","volume":"31","author":"Rochette","year":"2020","journal-title":"Trends Endocrinol. Metab. TEM"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1439","DOI":"10.1177\/0022034510378684","article-title":"Tgf-ss1 and fak regulate periostin expression in pdl fibroblasts","volume":"89","author":"Wen","year":"2010","journal-title":"J. Dent. Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1182\/blood-2016-01-696617","article-title":"Gdf-15 inhibits integrin activation and mouse neutrophil recruitment through the alk-5\/tgf-betarii heterodimer","volume":"128","author":"Artz","year":"2016","journal-title":"Blood"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"14751","DOI":"10.1038\/s41598-017-15281-0","article-title":"Valid gene expression normalization by rt-qpcr in studies on hpdl fibroblasts with focus on orthodontic tooth movement and periodontitis","volume":"7","author":"Kirschneck","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"15706","DOI":"10.1038\/s41598-020-72736-7","article-title":"Distinguish fatty acids impact survival, differentiation and cellular function of periodontal ligament fibroblasts","volume":"10","author":"Symmank","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_62","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(-delta delta c(t)) method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/24\/13608\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:51:54Z","timestamp":1760169114000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/24\/13608"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,19]]},"references-count":62,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["ijms222413608"],"URL":"https:\/\/doi.org\/10.3390\/ijms222413608","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,19]]}}}