{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,7]],"date-time":"2026-06-07T08:29:45Z","timestamp":1780820985384,"version":"3.54.1"},"reference-count":42,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2021,10,4]],"date-time":"2021-10-04T00:00:00Z","timestamp":1633305600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["aap.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Periodontology"],"published-print":{"date-parts":[[2022,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Recently we have generated recombinant human osteopontin (rhOPN) using a plant platform (<jats:italic>Nicotiana benthamiana<\/jats:italic>) and demonstrated, when coated on culture plates, its osteogenic induction capacity of human periodontal ligament (PDL) cells. The aim of this study is to elucidate the molecular mechanism underlying the rhOPN\u2010induced osteogenic differentiation of human PDL cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Full length rhOPN (FL\u2010OPN) and three constructs of OPN containing integrin binding domain (N142), calcium binding domain (C122) and mutated calcium\u2010binding domain (C122\u03b4) were generated from <jats:italic>N. benthamiana<\/jats:italic>. Human PDL cells were isolated from extracted third molars and cultured on FL\u2010OPN, N142, C122, or C122\u03b4\u2010coated surfaces. Real\u2010time PCR and Western blot analyses were used to determine mRNA and protein expression. In vitro calcification was determined by Alizarin red staining. A chemical inhibitor and RNAi silencing were used to elucidate signaling pathways. In silico analyses were performed to predict the protein\u2010protein interaction. In vivo analysis was performed using a rat calvaria defect model.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Human PDL cells seeded on FL\u2010OPN and C122\u2010coated surfaces significantly increased both mRNA and protein expression of osterix (OSX) and enhanced in vitro calcification. Soluble FL\u2010OPN as well as a surface coated with N142 did not affect <jats:italic>OSX<\/jats:italic> expression. Inhibition of activin receptor\u2010like kinase (ALK\u20101) abolished the induction of osterix expression. In silico analysis suggested a possible interaction between the calcium binding domain (CaBD) of OPN and ALK\u20101 receptor. C122, but not C122\u03b4 coated surfaces, induced the expression of p\u2010Smad\u20101 and this induction was inhibited by an ALK\u20101 inhibitor and RNAi against ALK\u20101. In vivo data showed that 3D porous scaffold containing C\u2010122 enhanced new bone formation as compared to scaffold alone.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The results suggest that next to full length OPN, the CaBD of OPN, if coated to a surface, induces osteogenic differentiation via interaction with ALK\u20101 receptor.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jper.21-0184","type":"journal-article","created":{"date-parts":[[2021,8,28]],"date-time":"2021-08-28T10:47:16Z","timestamp":1630147636000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Osteopontin induces osteogenic differentiation by human periodontal ligament cells via calcium binding domain\u2013ALK\u20101 interaction"],"prefix":"10.1002","volume":"93","author":[{"given":"Daneeya","family":"Chaikiawkeaw","sequence":"first","affiliation":[{"name":"Center of Excellence in Regenerative Dentistry, Faculty of 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