J Korean Acad Prosthodont.  2015 Jul;53(3):198-206. 10.4047/jkap.2015.53.3.198.

Effect of titanium surface microgrooves and thermal oxidation on in vitro osteoblast responses

• Seo JH
• Leesungbok R
• Ahn SJ
• Park SJ
• Lee MH
• Lee SW

• KMID: 2195249
• DOI: http://doi.org/10.4047/jkap.2015.53.3.198

Abstract

PURPOSE
We aimed to investigate the effect of combined various microgrooves and thermal oxidation on the titanium (Ti) and to evaluate various in vitro responses of human periodontal ligament cells (PLCs).
MATERIALS AND METHODS
Grade II titanium disks were fabricated. Microgrooves were applied on titanium discs to have 0/0 microm, 15/3.5 microm, 30/10 microm, and 60/10 microm of respective width/depth by photolithography. Thermal oxidation was performed on the microgrooves of Ti substrata for 3 h at 700degrees C in air. The experiments were divided into 3 groups: control group (ST), thermal oxidation group (ST/TO), and combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO). Surface characterization was performed by field-emission scanning microscopy. Cell adhesion, osteoblastic differentiation, and mineralization were analyzed using the bromodeoxyurdine (BrdU), Alkaline phosphatase (ALP) activity, and extracellular calcium deposition assays, respectively. Statistical analysis was performed using the oneway analysis of variance and Pearson's bivariate correlation analysis (SPSS Version 17.0).
RESULTS
In general, the combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO) showed significantly higher levels compared with the control (ST) or thermal oxidation (ST-TO) groups in the BrdU expression, ALP activity, and extracellular calcium deposition. Gr60-TO group induced highest levels of cell adhesion and osteoblastic differentiation.
CONCLUSION
Within the limitation of this study, we conclude that the Ti surface treatment using combined microgrooves and thermal oxidation is highly effective in inducing the cell adhesion andosteoblastic differentiation. The propose surface is also expected to be effective in inducing rapid and strong osseointegration of Ti oral implants.

Keyword

Titanium; Microgrooves; Thermal oxidation; Osteoblastic differentiation

MeSH Terms

Alkaline Phosphatase
Analysis of Variance
Bromodeoxyuridine
Calcium
Cell Adhesion
Humans
Microscopy
Osseointegration
Osteoblasts*
Periodontal Ligament
Titanium*
Alkaline Phosphatase
Bromodeoxyuridine
Calcium
Titanium

Figure

• 

  Fig. 1. Field emission scanning electron microscopic images of (A) ST (×500),(B) Gr15 (×200), (C) Gr30 (×200) and (D) Gr60 (×200).

• 

  Fig. 2. Field emission scanning electron microscopic images of the smooth titanium (ST) and thermally oxidized ST (ST-TO) groups at various magnifications.(A) ST (×1,000), (B) ST (×50,000), (C) ST-TO (×1,000), (D) ST-TO (×50,000). Note that polished texture at nano- to submicroscale widths appear in (A) and (B).

• 

  Fig. 3. Comparison result of the cell adhesion of human periodontal ligament cells on ST, ST-TO, Gr15-TO, Gr30-TO and Gr60-TO titanium substrata after 16 h of culture using bromodeoxyuridine assay. One-way ANOVA (n = 4). ∗∗∗: significant difference (P<.001).

• 

  Fig. 4. Comparison result of the alkaline phosphatase activity of human periodontal ligament cells on ST, ST-TO, Gr15-TO, Gr30-TO and Gr60-TO titanium substrata after 7 and 14 days of osteogenic culture. One-way ANOVA (n = 4). ∗∗∗: significant difference (P<.001).

• 

  Fig. 5. Comparison result of the osteoblast differentiation of human periodontal ligament cells on ST, ST-TO, Gr15-TO, Gr30-TO and Gr60-TO titanium substrata after 24 days of osteogenic culture using extracellular calcium deposition assay. One-way ANOVA (n = 4). ∗∗∗: significant difference (P<.001).

• 

  Fig. 6. Scatter-plot results from the Pearson' s correlation analysis. Significant correlations were present for (A), (B), (C), (D), (E) and (F) (P<.01) (n = 20).

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