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

Hyun-min Chang; Seong-Joo Heo; Seong-Kyun Kim; Jai-Young Koak

doi:10.4047/jkap.2019.57.2.142

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Chang, Heo, Kim, and Koak: Inhibition of Osteoclast differentiation based on precipitation time of titanium surfaces immersed in modified simulated body fluid

Original Article

The Journal of Korean Academy of Prosthodontics 2019; 57(2): 142-149.

Published online: 25 April 2019

DOI: https://doi.org/10.4047/jkap.2019.57.2.142

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

Hyun-min Chang

, Seong-Joo Heo

, Seong-Kyun Kim

, Jai-Young Koak

Department of Prosthodontics, School of Dentistry, Seoul National University, Seoul, Republic of Korea.

Corresponding Author: Jai-Young Koak. Department of Prosthodontics, School of Dentistry, Seoul National University 101, Daehak-ro, Jongno-gu, Seoul 03080, Republic of Korea. +82 (0)2 2072 2661: young21c@snu.ac.kr

Received 25 March 2019 Revised 15 April 2019 Accepted 18 April 2019

The Korean Academy of Prosthodontics

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Modified simulated body fluid (mSBF), Titanium alloy disk, Osteoclast differentiation, TRAP, NFATc1

Figures and Tables

Fig. 1

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

Fig. 2

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

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.

Table 1

Dual solution preparation of modified SBF (amounts to a final mixture of 2L)

SBF: simulated body fluid.

Table 2

4 Groups according to treatment method

mSBF: Modified simulated body fluid, DW: distilled water.

Notes

This study was supported by Seoul National University Dental Hospital Research Center (04-2014-0079).

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