Mineral trioxide aggregate promotes the odonto/osteogenic differentiation and dentinogenesis of stem cells from apical papilla via nuclear factor kappa B signaling pathway.
BACKGROUND
Mineral trioxide aggregate (MTA) has been widely used in clinical apexification and apexogenesis. However, the effects of MTA on the stem cells from apical papilla (SCAPs) and the precise mechanism of apexogenesis have not been elucidated in detail.


METHODS
Multiple colony-derived stem cells were isolated from the apical papillae, and the effects of MTA on the proliferation and differentiation of SCAPs were investigated both in vitro and in vivo. Activation of nuclear factor kappa B (NFκB) pathway in MTA-treated SCAPs was analyzed by immunofluorescence assay and Western blot.


RESULTS
MTA at the concentration of 2 mg/mL did not affect the proliferation activity of SCAPs. However, 2 mg/mL MTA-treated SCAPs presented the ultrastructural changes, up-regulated alkaline phosphatase, increased calcium deposition, up-regulated expression of odontoblast markers (dentin sialoprotein and dentin sialophosphoprotein) and odonto/osteoblast markers (runt-related transcription factor 2 and osteocalcin), suggesting that MTA enhanced the odonto/osteoblastic differentiation of SCAPs in vitro. In vivo results confirmed that MTA can promote the regular dentinogenesis of SCAPs. Moreover, MTA-treated SCAPs exhibited the up-regulated cytoplasmic phos-IκBα and phos-P65, enhanced nuclear P65, and increased nuclear translocation of P65. When co-treated with BMS345541 (the specific NFκB inhibitor), MTA-mediated odonto/osteoblastic differentiation was significantly attenuated.


CONCLUSIONS
MTA at the concentration of 2 mg/mL can improve the odonto/osteogenic capacity of SCAPs via the activation of NFκB pathway.
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China; Endodontic Department, School of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
Ming
M
Yan
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China; Endodontic Department, School of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
Jintao
J
Wu
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
Yan
Y
Yu
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China; Department of Endodontics, Suzhou Stomatological Hospital, Suzhou, Jiangsu, China.
Yanping
Y
Wang
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
Lizhe
L
Xie
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China; Endodontic Department, School of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China.
Guangdong
G
Zhang
Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, China; Department of Endodontics, Suzhou Stomatological Hospital, Suzhou, Jiangsu, China. Electronic address: yuziyi_yjh@hotmail.com.
Jinhua
J
Yu
Comprehensive Dental Care, Faculty of Dentistry, University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Chengfei
C
Zhang
D000293:Adolescent; D000469:Alkaline Phosphatase / Q000187:drug effects; D017607:Aluminum Compounds / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D017610:Calcium Compounds / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000187:drug effects; D003810:Dentinogenesis / Q000187:drug effects*; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins / Q000187:drug effects; D006801:Humans; D020672:I-kappa B Proteins / Q000187:drug effects; D007093:Imidazoles / Q000494:pharmacology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000187:drug effects*; D009805:Odontogenesis / Q000187:drug effects*; D015675:Osteocalcin / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects*; D010087:Oxides / Q000494:pharmacology*; D010750:Phosphoproteins / Q000187:drug effects; D011810:Quinoxalines / Q000494:pharmacology; D012387:Root Canal Filling Materials / Q000494:pharmacology*; D012795:Sialoglycoproteins / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects*; D017640:Silicates / Q000494:pharmacology*; D013234:Stem Cells / Q000187:drug effects*; D019227:Tooth Apex / Q000166:cytology*; D051996:Transcription Factor RelA / Q000187:drug effects; D015854:Up-Regulation; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
D000469:Alkaline Phosphatase; D017607:Aluminum Compounds; D002118:Calcium; D017610:Calcium Compounds; D050659:Core Binding Factor Alpha 1 Subunit; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins; D020672:I-kappa B Proteins; D007093:Imidazoles; D016328:NF-kappa B; D015675:Osteocalcin; D010087:Oxides; D010750:Phosphoproteins; D011810:Quinoxalines; D012387:Root Canal Filling Materials; D012795:Sialoglycoproteins; D017640:Silicates; D051996:Transcription Factor RelA; D000099311:Dentin Sialophosphoprotein; C471109:4(2'-aminoethyl)amino-1,8-dimethylimidazo(1,2-a)quinoxaline; C493709:RUNX2 protein, human; C086631:mineral trioxide aggregate
Apical papilla; dentinogenesis; nuclear factor kappa B; odontoblast; stem cell