title: Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro.
abstract: Tricalcium silicate cements have been successfully employed in the biomedical field as bioactive bone and dentin substitutes, with widely acclaimed osteoactive properties. This research analyzed the effects of different tricalcium silicate cement formulations on the temporal osteoactivity profile of human bone marrow-derived mesenchymal stem cells (hMW-MSCs). These cells were exposed to four commercially available tricalcium silicate cement formulations in osteogenic differentiation medium. After 1, 3, 7 and 10 days, quantitative real-time polymerase chain reaction and Western blotting were performed to detect expression of the target osteogenic markers ALP, RUNX2, OSX, OPN, MSX2 and OCN. After 3, 7, 14 and 21 days, alkaline phosphatase assay was performed to detect changes in intracellular enzyme level. An Alizarin Red S assay was performed after 28 days to detect extracellular matrix mineralization. In the presence of tricalcium silicate cements, target osteogenic markers were downregulated at the mRNA and protein levels at all time points. Intracellular alkaline phosphatase enzyme levels and extracellular mineralization of the experimental groups were not significantly different from the untreated control. Quantitative polymerase chain reaction results showed increases in downregulation of RUNX2, OSX, MSX2 and OCN with increasing time of exposure to the tricalcium silicate cements, while ALP showed peak downregulation at day 7. For Western blotting, OSX, OPN, MSX2 and OCN showed increased downregulation with increased exposure time to the tested cements. Alkaline phosphatase enzyme levels generally declined after day 7. Based on these results, it is concluded that tricalcium silicate cements do not induce osteogenic differentiation of hBM-MSCs in vitro.
authors.affiliation: Department of Dental and Biomedical Material Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Ashraf A
authors.initials: AA
authors.lastname: Eid
authors.affiliation: Department of Oral Biology, Georgia Regents University, Augusta, GA, USA; Oral and Dental Research Division, Department of Surgery and Medicine, National Research Center, Egypt.
authors.forename: Khaled A
authors.initials: KA
authors.lastname: Hussein
authors.affiliation: Department of Prosthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, China.
authors.forename: Li-na
authors.initials: LN
authors.lastname: Niu
authors.affiliation: Department of Stomatology, Fuzhou Dongfang Hospital, Fuzhou, China.
authors.forename: Guo-hua
authors.initials: GH
authors.lastname: Li
authors.affiliation: Department of Dental and Biomedical Material Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Ikuya
authors.initials: I
authors.lastname: Watanabe
authors.affiliation: Department of Oral Biology, Georgia Regents University, Augusta, GA, USA; Department of Graduate Studies, Georgia Regents University, Augusta, GA, USA.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Al-Shabrawey
authors.affiliation: Department of Oral Biology, Georgia Regents University, Augusta, GA, USA; Department of Graduate Studies, Georgia Regents University, Augusta, GA, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: Pashley
authors.affiliation: Department of Oral Biology, Georgia Regents University, Augusta, GA, USA; Department of Graduate Studies, Georgia Regents University, Augusta, GA, USA. Electronic address: tayfranklin7@gmail.com.
authors.forename: Franklin R
authors.initials: FR
authors.lastname: Tay
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001843:Bone Cements; D001854:Bone Marrow Cells / Q000166:cytology*; D017610:Calcium Compounds; D002454:Cell Differentiation; D006801:Humans; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis; D060888:Real-Time Polymerase Chain Reaction; D017640:Silicates
chemical_list: D001843:Bone Cements; D017610:Calcium Compounds; D017640:Silicates; C506393:tricalcium silicate; D000469:Alkaline Phosphatase
keywords: Bone marrow; Human origin; Mesenchymal stem cells; Osteogenic differentiation; Tricalcium silicate cements
references.citation: J Biomed Mater Res. 1997 Dec 5;37(3):432-9
references.pmid: 9368148
references.citation: J Cell Biochem. 2006 Aug 15;98(6):1457-70
references.pmid: 16518860
references.citation: J Endod. 2014 Feb;40(2):255-60
references.pmid: 24461414
references.citation: Stem Cells. 2002;20(6):530-41
references.pmid: 12456961
references.citation: Blood Cells Mol Dis. 2001 May-Jun;27(3):677-90
references.pmid: 11482883
references.citation: J Endod. 2013 Jul;39(7):883-8
references.pmid: 23791256
references.citation: J Exp Med. 1951 May;93(5):415-26
references.pmid: 14832392
references.citation: Cell Biol Toxicol. 2006 Jan;22(1):39-46
references.pmid: 16463018
references.citation: Anal Biochem. 2004 Jun 1;329(1):77-84
references.pmid: 15136169
references.citation: J Dent. 2007 Aug;35(8):636-42
references.pmid: 17566626
references.citation: Acta Histochem. 2010 Sep;112(5):452-8
references.pmid: 19560800
references.citation: Sci STKE. 2005 Jul 26;2005(294):pe37
references.pmid: 16046665
references.citation: Dent Mater. 2006 May;22(5):434-40
references.pmid: 16182358
references.citation: J Endod. 2007 Apr;33(4):447-50
references.pmid: 17368336
references.citation: Biomaterials. 2013 Dec;34(38):10028-42
references.pmid: 24095251
references.citation: Int Endod J. 2013 Dec;46(12):1173-82
references.pmid: 23617276
references.citation: Science. 1997 Apr 4;276(5309):71-4
references.pmid: 9082988
references.citation: Nat Rev Mol Cell Biol. 2007 Dec;8(12):970-82
references.pmid: 18000526
references.citation: Stem Cells. 2004;22(5):823-31
references.pmid: 15342946
references.citation: Bone. 2004 May;34(5):818-26
references.pmid: 15121013
references.citation: J Biol Chem. 2003 Oct 17;278(42):41227-36
references.pmid: 12893825
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3213-8
references.pmid: 10725391
references.citation: Biomater Sci. 2013 Apr 5;1(4):379-392
references.pmid: 32481903
references.citation: PLoS One. 2013;8(3):e57564
references.pmid: 23483914
references.citation: Biochim Biophys Acta. 2012 Jun;1822(6):1004-18
references.pmid: 22387884
references.citation: Proc Natl Acad Sci U S A. 2002 Apr 2;99(7):4397-402
references.pmid: 11917104
references.citation: Transplant Proc. 2008 Oct;40(8):2649-54
references.pmid: 18929828
references.citation: Blood. 2004 Mar 1;103(5):1662-8
references.pmid: 14592819
references.citation: Blood. 2008 Jul 15;112(2):295-307
references.pmid: 18332228
references.citation: Stem Cells. 2010 Apr;28(4):788-98
references.pmid: 20127798
references.citation: J Dent Res. 2004 May;83(5):408-13
references.pmid: 15111634
references.citation: J Endod. 2013 Sep;39(9):1161-6
references.pmid: 23953291
references.citation: Dent Mater. 2002 Jun;18(4):318-23
references.pmid: 11992909
references.citation: J Endod. 2009 May;35(5):682-5
references.pmid: 19410082
references.citation: Cell. 2002 Jan 11;108(1):17-29
references.pmid: 11792318
references.citation: Methods Mol Biol. 2008;449:133-51
references.pmid: 18370089
references.citation: Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009 Apr;107(4):590-6
references.pmid: 19201220
references.citation: Nat Genet. 2000 Apr;24(4):391-5
references.pmid: 10742104
references.citation: Cell Tissue Kinet. 1970 Oct;3(4):393-403
references.pmid: 5523063
references.citation: J Biol Chem. 1946 Jul;164:321-9
references.pmid: 20989492
references.citation: Tissue Eng Part A. 2010 Oct;16(10):3185-97
references.pmid: 20590472
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: J Endod. 2010 Jul;36(7):1163-9
references.pmid: 20630291
references.citation: ACS Appl Mater Interfaces. 2011 Oct;3(10):4142-53
references.pmid: 21942767
references.citation: J Immunol. 2009 Jul 15;183(2):993-1004
references.pmid: 19561093
references.citation: Dev Cell. 2005 May;8(5):727-38
references.pmid: 15866163
references.citation: Hum Gene Ther. 2010 Oct;21(10):1226-38
references.pmid: 20804388
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7841-5
references.pmid: 11427725
references.citation: J Endod. 2008 Aug;34(8):970-4
references.pmid: 18634929