J Korean Acad Prosthodont.  2019 Apr;57(2):142-149. 10.4047/jkap.2019.57.2.142.

Inhibition of Osteoclast differentiation based on precipitation time of titanium surfaces immersed in modified simulated body fluid

• Chang HM
• Heo SJ
• Kim SK
• Koak JY

• KMID: 2444183
• DOI: http://doi.org/10.4047/jkap.2019.57.2.142

Abstract

PURPOSE
The purpose of this study is to investigate the changes of osteoclast differentiation inhibition according to the period of precipitation when titanium disks were immersed in Modified simulated body fluid (mSBF).
MATERIALS AND METHODS
Titanium alloy (Ti grade III) disks with machined surfaces and anodized surfaces were immersed in distilled water and mSBF, respectively. The immersion periods were 7 days, 14 days, 21 days and 28 days, and the control group was immersed in distilled water for each period. RAW 264.7 cells capable of differentiating into osteoclasts were used to measure the number of adherent cells, the measurement of TRAP activity, and the expression pattern of NFATc1 by western blotting.
RESULTS
The degree of inhibition of osteoclast differentiation was found to be statistically significant when the disks were immersed in mSBF for more than 14 days on both machined surfaces and anodized surfaces. There was no correlation between immersion time and cell attachment. When the disks were immersed for more than 14 days, TRAP activity was decreased and NFATc1 expression was inhibited. Futhermore, the decrease in TRAP activity and the inhibition of NFATc1 expression remained unchanged.
CONCLUSION
Immersion of titanium disks in mSBF for more than 14 days can prevent RAW 264.7 cells from differentiating into osteoclasts. Inhibition activity does not change even if the immersion period is for more than 14 days.

Keyword

Modified simulated body fluid (mSBF); Titanium alloy disk; Osteoclast differentiation; TRAP; NFATc1

MeSH Terms

Alloys
Blotting, Western
Body Fluids*
Immersion
Osteoclasts*
RAW 264.7 Cells
Titanium*
Water
Alloys
Titanium
Water

Figure

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  Fig. 1 Cell counting test. There is no significant difference among the groups. Deposition period dose not affect cell adhesion.

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  Fig. 2 TRAP activity on Ti disc. More than 14 days deposition period, TRAP activity was reduced regardless of surface roughness. *P < .05

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  Fig. 3 Western blot of NFATc1, β-actin. Group 2, 4 show decreased expression of NFATc1 after 14 days deposition. NFATc1 expression of 0, 7 days deposition were not different among group 1 – 4. Group 1, 3 show constant expression of NFATc1. NFATc1 is a final signal transducer of osteoclast differentiation. β-actin is a control of protein expression.

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