J Korean Acad Prosthodont.  2015 Jan;53(1):9-18. 10.4047/jkap.2015.53.1.9.

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

• Seo BY
• Kim YM
• Choi JW
• Yun MJ
• Jeon YC
• Jeong CM
• Kim GC
• Huh JB

• KMID: 2057202
• DOI: http://doi.org/10.4047/jkap.2015.53.1.9

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 microm in the control group, 0.52 +/- 0.14 microm in ASD group and 1.45 +/- 0.25 microm 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.

Keyword

Anodizing treatment; RBM treatment; Surface roughness; Osteoblast-like cell; Adhesion; Proliferation; Differentiation

MeSH Terms

Cell Adhesion
Cell Differentiation
Cell Proliferation
Control Groups
Enzyme-Linked Immunosorbent Assay
Evaluation Studies as Topic
Gentian Violet
Ions
Microscopy, Confocal
Oxygen
Spectrum Analysis
Titanium
Gentian Violet
Ions
Oxygen
Titanium

Figure

• 

  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.

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  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.

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  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).

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  Fig. 6. The degree of cell proliferation by XTT assay after 3 days incubation time (n=7 in each group). ∗P<.05.

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  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.

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  Fig. 8. The degree of cell calcification by Alizarin red S assay after 21 days incubation time (n=7 in each group). ∗∗P<.01.

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