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Effects of Mineral Trioxide Aggregate on Cell Survival, Gene Expression Associated with Mineralized Tissues, and Biomineralization of Cementoblasts
Scopus
Toplam 115 atıf DOI
The purpose of this study was to investigate the effects of mineral trioxide aggregate (MTA) on survival, mineralization, and expression of mineralization-related genes of cementoblasts. Immortalized cementoblasts (OCCM) were maintained with Dulbecco modified Eagle medium containing 10% fetal bovine serum. Methyl-thiazol-diphenyl-tetrazolium experiments were performed at 24 and 72 hours to evaluate bioactive components released by MTA (0.002-20 mg/mL) on the cell survival of OCCM. Von Kossa staining was used to evaluate biomineralization of OCCM cells. Images of cementoblasts were taken on day 3 by using inverted microscopy. Gene transcripts for bone sialoprotein (BSP), OCN, collagen type I (COL I), and osteopontin (OPN) were evaluated on days 3 and 5 by using semiquantitative reverse transcriptase polymerase chain reaction. The 20 mg/mL concentration of MTA was toxic for OCCM cells, whereas other concentrations of MTA tested exhibited similar cell numbers when compared with control group, and the 0.02 mg/mL concentration of MTA increased OCCM cell survival at 72 hours. Although an apparent decrease in mineralization was observed in the highest 3 concentrations of MTA used, 0.02 and 0.002 mg/mL concentrations of MTA induced greater biomineralization of OCCM cells than seen in the control. Moreover, increased BSP and COL I mRNA expression was observed at 0.02 and 0.002 mg/mL concentrations of MTA. MTA did not have a negative effect on the viability and morphology of cementoblasts and induced biomineralization of cementoblasts at the concentrations of 0.02 and 0.002 mg/mL. Based on these results MTA can be considered as a favorable material regarding cell-material interaction. © 2009 American Association of Endodontists.
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Warm Gutta-Percha Techniques Regulate Cell Viability, Heat Shock, and Mineralized Tissue–associated Proteins of Cementoblasts
Scopus
Havuzumuzda 8 atıf almış
Introduction: The aim of this study was to determine the effect of the continuous wave of condensation technique (CWCT) and the thermoplastic gutta-percha injection (TGI) technique on the messenger RNA (mRNA) expressions of heat shock proteins (HSPs) and mineralized tissue–associated proteins of the immortalized mouse cementoblasts (OCCM.30). Methods: Crowns of human premolar teeth with single and straight canals were removed. The root canals were prepared up to the ProTaper Next X5 file (Dentsply Maillefer, Ballaigues, Switzerland) in combination with 2 mL 2.5% sodium hypochlorite solution. Roots (12 ± 2 mm height) were sterilized (121°C for 20 minutes) and placed vertically to the cell culture dishes using a tissue culture insert by opening holes according to the root diameter after the removal of 1 mm from the apex for appropriate adaptation to the petri dish surfaces. Six groups were created: control 1 (without teeth), control 2 (with teeth), AH Plus (Dentsply DeTrey, Konstanz, Germany), single-cone obturation (SC), CWCT, and thermoplastic gutta-percha injection technique (TGI). The viability of the OCCM.30 cells was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability experiments at 24 and 96 hours. The mRNA expression of HSP27, HSP70, and HSP90 and mineralized tissue markers (bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase) was evaluated by real-time polymerase chain reaction. Results: Reduced OCCM.30 cell viability was observed in all groups except the control groups. When the SC technique and CWCT and TGI groups were compared, it was observed that heat had a significant negative effect on cell viability (P <.05). A reduction in the mRNA expressions of HSP27, HSP70, and HSP90 was recognized in all test groups when compared with the control 1 group (P <.01). When the warm gutta-percha techniques were compared with the SC technique, a decrease in mRNA expression of HSP27 and HSP90 was noted (P <.01). The HSP70 transcript was similar in the CWCT group and the SC group. Higher HSP70 mRNA expression was observed in the TGI group compared with the SC group. In all groups except the control 1 group, bone sialoprotein, osteocalcin, runt-related transcription factor 2, type I collagen, and alkaline phosphatase mineralized tissue markers were affected, but this negative effect was higher in the heat-treated groups (P <.05). Conclusions: Within the limitations of this study, it was concluded that warm gutta-percha techniques reduced the mRNA expressions of the genes for HSPs and mineralized tissue–associated proteins of cementoblasts. Further animal studies are needed to clarify the effect of heat on the behavior of cementoblasts histologically in short- and long-term periods.
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Kurumlar (3)
Hacettepe Üniversitesi
Ankara, Turkey
Konya Research and Training Hospital
Konya, Turkey
Selçuk Üniversitesi
Selçuklu, Turkey