The effect of blasting and anodizing-combined treatment of implant surface on response of osteoblast-like cell

Bo-Yong Seo; Young-Min Kim; Jae-Won Choi; Mi-Jung Yun; Young-Chan Jeon; Chang-Mo Jeong; Gyu-Cheon Kim; Jung-Bo Huh

doi:10.4047/jkap.2015.53.1.9

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Seo, Kim, Choi, Yun, Jeon, Jeong, Kim, and Huh: The effect of blasting and anodizing-combined treatment of implant surface on response of osteoblast-like cell

Original Article

J Korean Acad Prosthodont 2015;53(1):9-18.

Published online: 9 February 2015

DOI: https://doi.org/10.4047/jkap.2015.53.1.9

The effect of blasting and anodizing-combined treatment of implant surface on response of osteoblast-like cell

Bo-Yong Seo^1,^a, Young-Min Kim^2,^a, Jae-Won Choi¹, Mi-Jung Yun¹, Young-Chan Jeon¹, Chang-Mo Jeong¹, Gyu-Cheon Kim², Jung-Bo Huh^1,^∗

¹Department of Prosthodontics, Pusan National University, Yangsan, Republic of Korea

²Department of Oral Anatomy, School of Dentistry, Pusan National University, Yangsan, Republic of Korea

^∗Corresponding Author: Jung-Bo Huh Department of Prosthodontics, School of Dentistry, Pusan National University 20 Geumo-ro, Mulgeum-eup, Yangsan 626-770, Republic of Korea +82 55 360 5130: e-mail,huhjb@pusan.ac.kr

^a These authors are equally contributed to this work.

^∗ This study was supported by Pusan National University research grant (2 years).

Received 7 November 201418 December 2014 Accepted 7 January 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) 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 examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell).

Materials and methods

Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05.

Results

In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained 0.08 ± 0.04 ㎛ in the control group, 0.52 ± 0.14 ㎛ in ASD group and 1.45 ± 0.25 ㎛ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase.

Conclusion

In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.

Keywords: Anodizing treatment, RBM treatment, Surface roughness, Osteoblast-like cell, Adhesion, Proliferation, Differentiation

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Fig. 1.

FE-SEM images of each group. (A) Control group (×100), (B) Control group (×500), (C) ASD group (×100), (D) ASD group (×500), (E) RBM/ ASD group (×100), (F) RBM/ ASD group (×500).

Fig. 2.

The results of the energy dispersive spectrometry. (A) Control group, (B) ASD group, (C) RBM/ASD group.

Fig. 3.

The degree of cell adhesion by crystal violet staining assay after 3 hours incubation time (n=7 in each group). ∗ P<.05, ∗∗ P<.01.

Fig. 4.

Adhesion pattern images of MG-63 cells by SEM (×40,000 magnification). (A) Control group, (B) ASD group, (C) RBM/ASD group.

Fig. 5.

Confocal laser scanning microscopy images of MG-63 cells (×400 magnification). (A) Control group (incubation for 2 hours), (B) Control group (incubation for 24 hours), (C) ASD group (incubation for 2 hours), (D) ASD group (incubation for 24 hours), (E) RBM/ASD group (incubation for 2 hours), (F) RBM/ASD group (incubation for 24 hours).

Fig. 6.

The degree of cell proliferation by XTT assay after 3 days incubation time (n=7 in each group). ∗P<.05.

Fig. 7.

The Results of electrophoresis after RT-PCR. Osteoclacin, osteopontin and collagen type I were to show a similar expression levels in all three groups, ALP showed the highest expression levels in RBM / ASD group.

Fig. 8.

The degree of cell calcification by Alizarin red S assay after 21 days incubation time (n=7 in each group). ∗∗P<.01.

Table 1.

Surface roughness by stereo-SEM in each group

Group	Sa (㎛) ± SD	Sq (㎛) ± SD	Rt (㎛) ± SD
Control	0.08 ± 0.04	0.09 ± 0.02	1.95 ± 0.11
ASD	0.52 ± 0.14	0.68 ± 0.19	6.44 ± 1.51
RBM/ASD	1.45 ± 0.25	1.85 ± 0.33	13.76 ± 2.69

Sa: Average roughness, Sq: Root Mean Square roughness, Rt: Distance between the highest and the lowest point.

Table 2.

The percentage of components in each group's surface

Element (%)	Carbon	Oxide	Phosphate	Titanium
Control	2.21	6.27	0.74	90.77
ASD	2.66	43.09	7.34	46.91
RBM/ASD	3.03	48.60	7.63	40.74

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