{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T02:55:53Z","timestamp":1785466553629,"version":"3.56.0"},"reference-count":34,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2022,8,18]],"date-time":"2022-08-18T00:00:00Z","timestamp":1660780800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In animal models, the administration of ciliary neurotrophic factor (CNTF) was demonstrated to reduce bone mass and to participate in bone remodeling. Cementoblasts, a cell type embedded in the cementum, are the main cells to produce and mineralize the extracellular matrix. The effect of CNTF on cementoblasts has not yet been addressed. Thus, the goal of this in vitro study was to investigate possible influences of exogenous CNTF on cementogenesis, as well as autophagy regulation and subsequent mechanisms in cementoblasts. Cementoblasts (OCCM-30) were stimulated with exogenous CNTF. Alizarin Red staining was performed to analyze the functional differentiation (mineralization) of OCCM-30 cells. The release of OPG was quantified by ELISA. The expression of cementogenesis markers (RUNX-2, OCN, BMP-7, BSP, and SPON-2) was evaluated by RT-qPCR. Western blotting (WB) was performed for the protein expression of STAT3, COX-2, SHP-2, cPLA\u03b1, cPLA\u03b2; ERK1\/2, P38, and JNK. The autophagic flux was assessed using WB and RT-qPCR analysis of LC3A\/B, Beclin-1, and Atg-5, and the autophagosome was investigated by immunofluorescence staining (IF). The ERK1\/2 (FR180204) or STAT3 (sc-202818) antagonist was added, and the cellular response was analyzed using flow cytometry. Exogenous CNTF significantly attenuated mineralized nodule formation, impaired OPG release, and downregulated the mRNA levels of RUNX-2, OCN, BMP-7, and BSP. Moreover, CNTF induced the phosphorylation of STAT3 and activated a transient activation of SHP-2, cPLA\u03b2, ERK1\/2, P38, and JNK protein. CNTF also induced autophagosome formation and promoted autophagy-associated gene and protein expressions. Additionally, the inhibition of ERK1\/2 or STAT3 reversed a CNTF-induced mineralization impairment and had regulatory effects on CNTF-induced autophagosome formation. Our data revealed that CNTF acts as a potent inhibitor of cementogenesis, and it can trigger autophagy, in part by ERK1\/2 and STAT3 commitment in the cementoblasts. Thus, it may play an important role in inducing or facilitating inflammatory root resorption during orthodontic tooth movement.<\/jats:p>","DOI":"10.3390\/ijms23169311","type":"journal-article","created":{"date-parts":[[2022,8,18]],"date-time":"2022-08-18T21:39:21Z","timestamp":1660858761000},"page":"9311","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Ciliary Neurotrophic Factor (CNTF) Inhibits In Vitro Cementoblast Mineralization and Induces Autophagy, in Part by STAT3\/ERK Commitment"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3021-075X","authenticated-orcid":false,"given":"Jiawen","family":"Yong","sequence":"first","affiliation":[{"name":"Department of Orthodontics, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"},{"name":"Department of Periodontology, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"},{"name":"Stomatology Hospital, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Key Laboratory of Oral Biomedical Research of Zhejiang Province, Cancer Center of Zhejiang University, Hangzhou 310003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Gr\u00f6ger","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"},{"name":"Department of Periodontology, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"von Bremen","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Ruf","sequence":"additional","affiliation":[{"name":"Department of Orthodontics, Faculty of Medicine, Justus Liebig University Giessen, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"15529","DOI":"10.1038\/srep15529","article-title":"Ciliary neurotrophic factor has intrinsic and extrinsic roles in regulating B cell differentiation and bone structure","volume":"5","author":"Askmyr","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1042\/bj3090215","article-title":"Human ciliary neurotrophic factor: A structure-function analysis","volume":"309","author":"Kruttgen","year":"1995","journal-title":"Biochem. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1126\/science.2587985","article-title":"Purification, cloning, and expression of ciliary neurotrophic factor (CNTF)","volume":"246","author":"Lin","year":"1989","journal-title":"Science"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.cytogfr.2015.07.007","article-title":"Ciliary neurotrophic factor (CNTF): New facets of an old molecule for treating neurodegenerative and metabolic syndrome pathologies","volume":"26","author":"Pasquin","year":"2015","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/s00223-010-9337-4","article-title":"Ciliary neurotrophic factor inhibits bone formation and plays a sex-specific role in bone growth and remodeling","volume":"86","author":"McGregor","year":"2010","journal-title":"Calcif. Tissue Int."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"645005","DOI":"10.3389\/fcell.2021.645005","article-title":"Adiponectin as Well as Compressive Forces Regulate in vitro beta-Catenin Expression on Cementoblasts via Mitogen-Activated Protein Kinase Signaling Activation","volume":"9","author":"Yong","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"585346","DOI":"10.3389\/fphar.2020.585346","article-title":"Adiponectin Interacts In-Vitro with Cementoblasts Influencing Cell Migration, Proliferation and Cementogenesis Partly Through the MAPK Signaling Pathway","volume":"11","author":"Yong","year":"2020","journal-title":"Front. Pharmacol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1902\/jop.2010.100074","article-title":"Bone morphogenetic protein-7 enhances cementoblast function in vitro","volume":"81","author":"Hakki","year":"2010","journal-title":"J. Periodontol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Yong, J., Groeger, S., von Bremen, J., and Ruf, S. (2022). Ciliary Neurotrophic Factor (CNTF) and Its Receptors Signal Regulate Cementoblasts Apoptosis through a Mechanism of ERK1\/2 and Caspases Signaling. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23158335"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"716441","DOI":"10.3389\/fphys.2021.716441","article-title":"Autophagy Induces Expression of IL-6 in Human Periodontal Ligament Fibroblasts Under Mechanical Load and Overload and Effects Osteoclastogenesis in vitro","volume":"12","author":"Mayr","year":"2021","journal-title":"Front. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1016\/j.febslet.2010.02.053","article-title":"Autophagy","volume":"584","author":"Mizushima","year":"2010","journal-title":"FEBS Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1146\/annurev-pathol-020712-163918","article-title":"Autophagy as a stress-response and quality-control mechanism: Implications for cell injury and human disease","volume":"8","author":"Murrow","year":"2013","journal-title":"Annu. Rev. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5720","DOI":"10.1093\/emboj\/19.21.5720","article-title":"LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing","volume":"19","author":"Kabeya","year":"2000","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1111\/jcpe.12549","article-title":"Increased autophagy is required to protect periodontal ligament stem cells from apoptosis in inflammatory microenvironment","volume":"43","author":"An","year":"2016","journal-title":"J. Clin. Periodontol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1177\/0022034514553815","article-title":"Dual role of autophagy in lipopolysaccharide-induced preodontoblastic cells","volume":"94","author":"Pei","year":"2015","journal-title":"J. Dent. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"54","DOI":"10.31083\/j.fbl2702054","article-title":"MAPK and \u03b2-Catenin signaling: Implication and interplay in orthodontic tooth movement","volume":"27","author":"Yong","year":"2022","journal-title":"Front. Biosci. Landmark"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2631","DOI":"10.2741\/2260","article-title":"What triggers cell-mediated mineralization?","volume":"12","author":"Huitema","year":"2007","journal-title":"Front. Biosci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.bone.2014.03.053","article-title":"Myokines (muscle-derived cytokines and chemokines) including ciliary neurotrophic factor (CNTF) inhibit osteoblast differentiation","volume":"64","author":"Johnson","year":"2014","journal-title":"Bone"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1111\/j.1471-4159.2008.05759.x","article-title":"CNTF-evoked activation of JAK and ERK mediates the functional expression of T-type Ca2+ channels in chicken nodose neurons","volume":"108","author":"Trimarchi","year":"2009","journal-title":"J. Neurochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1007\/s00441-018-2890-2","article-title":"Inhibition of Stat3 signaling pathway decreases TNF-alpha-induced autophagy in cementoblasts","volume":"374","author":"Wang","year":"2018","journal-title":"Cell Tissue Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"118923","DOI":"10.1016\/j.bbamcr.2020.118923","article-title":"CXXC5 orchestrates Stat3\/Erk\/Akt signaling networks to modulate P. gingivalis-elicited autophagy in cementoblasts","volume":"1868","author":"Ma","year":"2021","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1080\/15548627.2015.1017192","article-title":"The role of STAT3 in autophagy","volume":"11","author":"You","year":"2015","journal-title":"Autophagy"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3436","DOI":"10.1038\/s41467-019-11373-9","article-title":"Increased autophagy in EphrinB2-deficient osteocytes is associated with elevated secondary mineralization and brittle bone","volume":"10","author":"Vrahnas","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.4161\/auto.36182","article-title":"Autophagy in osteoblasts is involved in mineralization and bone homeostasis","volume":"10","author":"Nollet","year":"2014","journal-title":"Autophagy"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"104960","DOI":"10.1016\/j.archoralbio.2020.104960","article-title":"Autophagy of periodontal ligament inhibits inflammation and reduces the decline of bone density during orthodontic tooth movement of mice","volume":"121","author":"Chen","year":"2021","journal-title":"Arch. Oral Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.redox.2016.11.004","article-title":"Inhibition of autophagy with bafilomycin and chloroquine decreases mitochondrial quality and bioenergetic function in primary neurons","volume":"11","author":"Redmann","year":"2017","journal-title":"Redox Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"5898","DOI":"10.1021\/nn5009879","article-title":"Bioactive silica nanoparticles promote osteoblast differentiation through stimulation of autophagy and direct association with LC3 and p62","volume":"8","author":"Ha","year":"2014","journal-title":"ACS Nano"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9710","DOI":"10.1111\/jcmm.16920","article-title":"Hypoxia-inducible factor 1-alpha acts as a bridge factor for crosstalk between ERK1\/2 and caspases in hypoxia-induced apoptosis of cementoblasts","volume":"25","author":"Yong","year":"2021","journal-title":"J. Cell. Mol. Med."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Yong, J., Gr\u00f6ger, S., Meyle, J., and Ruf, S. (2022). Immunorthodontics: Role of HIF-1\u03b1 in the Regulation of (Peptidoglycan-Induced) PD-L1 Expression in Cementoblasts under Compressive Force. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23136977"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Yong, J., Gr\u00f6ger, S., von Bremen, J., Meyle, J., and Ruf, S. (2022). Immunorthodontics: PD-L1, a Novel Immunomodulator in Cementoblasts, Is Regulated by HIF-1\u03b1 under Hypoxia. Cells, 11.","DOI":"10.3390\/cells11152350"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"62","DOI":"10.31083\/j.fbl2702062","article-title":"Effect of Low-Level Er: YAG (2940 nm) laser irradiation on the photobiomodulation of mitogen-activated protein kinase cellular signaling pathway of rodent cementoblasts","volume":"27","author":"Yong","year":"2022","journal-title":"Front. Biosci. (Landmark Ed.)"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Yong, J., Groeger, S., Ruf, S., and Ruiz-Heiland, G. (2021). Influence of leptin and compression in GAS-6 mediated homeostasis of periodontal ligament cell. Oral Dis., 1\u201312.","DOI":"10.1111\/odi.14092"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Yong, J., Groeger, S., Ruiz-Heiland, G., and Ruf, S. (2022). Selection and validation of reference gene for RT-qPCR studies in co-culture system of mouse cementoblasts and periodontal ligament cells. BMC Res. Notes, 15.","DOI":"10.1186\/s13104-022-05948-x"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1007\/s00441-019-03063-1","article-title":"Autophagy in periodontal ligament fibroblasts under biomechanical loading","volume":"378","author":"Memmert","year":"2019","journal-title":"Cell Tissue Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/16\/9311\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:11:45Z","timestamp":1760141505000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/16\/9311"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,18]]},"references-count":34,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["ijms23169311"],"URL":"https:\/\/doi.org\/10.3390\/ijms23169311","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,18]]}}}