J Adv Prosthodont.  2014 Apr;6(2):96-102. 10.4047/jap.2014.6.2.96.

Cell attachment and proliferation of bone marrow-derived osteoblast on zirconia of various surface treatment

• Pae A
• Lee H
• Noh K
• Woo YH

• KMID: 2233336
• DOI: http://doi.org/10.4047/jap.2014.6.2.96

Abstract

PURPOSE
This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture.
MATERIALS AND METHODS
Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA).
RESULTS
From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating.
CONCLUSION
The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.

Keyword

Zirconia; Calcium phosphate; Bone marrow-derived osteoblasts; Ion beam assisted deposition; Aerosol deposition

MeSH Terms

Alkaline Phosphatase
Calcium
Cell Adhesion
Cell Proliferation
Chemistry
Durapatite
Microscopy, Electron, Scanning
Osteoblasts*
Photoelectron Spectroscopy
Pseudopodia
Alkaline Phosphatase
Calcium
Durapatite

Figure

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  Fig. 1 Evaluation of cell proliferation using MTT assay for the ZS group, CaP group, and HA group. The data are expressed as the mean values (MV) ± standard deviation (SD) of three independent experiments. No significant differences were observed between the groups (P>.05).

• 

  Fig. 2 SEM observations of osteoblast cultures on surface-modified zirconia after 24 h incubation (×5,000). (A) ZS group at 24 h, (B) CaP group at 24 h, (C) HA group at 24 h. The cells were sporadically triangular or spread out in shape with long filopodia on all three groups of specimens.

• 

  Fig. 3 SEM observations of osteoblast cultures on surface-modified zirconia after 48 h incubation (×5,000). (A) ZS group at 48 h, (B) CaP group at 48 h, (C) HA group at 48 h. The cells showed more contact with each other and in intimate contact with the surface on all three groups of specimens.

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  Fig. 4 Evaluation of the cellular differentiation by using ALP analysis after 14 days incubation. The ALP activity of osteoblasts on calcium phosphate coated zirconia was the highest. No significant differences were observed among the groups (P>.05).

• 

  Fig. 5 XPS survey spectrum of ZS, CaP, HA groups.

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  Fig. 6 The level of mRNA for type I collagen, osteocalcin, and osteonectin. GAPDH was used as an internal control.

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  Fig. 7 Concentrations of the released (A) Ca2+ and (B) P- from the coatings obtained.

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