J Adv Prosthodont.  2011 Sep;3(3):152-160. 10.4047/jap.2011.3.3.152.

Effects of the immobilization of heparin and rhPDGF-BB to titanium surfaces for the enhancement of osteoblastic functions and anti-inflammation

• Huh JB
• Lee JY
• Lee KL
• Kim SE
• Yun MJ
• Shim JS
• Shim JS
• Shin SW

• KMID: 2118125
• DOI: http://doi.org/10.4047/jap.2011.3.3.152

Abstract

PURPOSE
This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity.
MATERIALS AND METHODS
The anodized titanium discs, not coated with any material, were used as a control group. In heparinized- Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at *P<.05 and **P<.001.
RESULTS
The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells (*P<.05 and **P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti (**P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-alpha and IL-6 of proinflammatory cytokine (*P<.05 and **P<.001).
CONCLUSION
The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

Keyword

Titanium; Heparin; rhPDGF-BB; Anti-inflammation; Osteoblastic function

MeSH Terms

Alkaline Phosphatase
Calcium
Cell Proliferation
Dopamine
Electrons
Heparin
Humans
Immobilization
Inflammation
Interleukin-6
Kinetics
Osteoblasts
Platelet-Derived Growth Factor
Proto-Oncogene Proteins c-sis
RNA, Messenger
Titanium
Tumor Necrosis Factor-alpha
Alkaline Phosphatase
Calcium
Dopamine
Heparin
Interleukin-6
Platelet-Derived Growth Factor
Proto-Oncogene Proteins c-sis
RNA, Messenger
Titanium
Tumor Necrosis Factor-alpha

Figure

• 

  Fig. 1 SEM of various Ti surfaces: (A) anodized Ti; (B) heparinized Ti; and (C) rhPDGF-BB-immobilized Ti.

• 

  Fig. 2 Release kinetic of rhPDGF-BB from the heparinized-Ti surface.

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  Fig. 3 Cytotoxicity tests of the anodized Ti, heparinized Ti, and rhPDGF-BB-immobilized Ti during culture periods of up to 48 h.

• 

  Fig. 4 Live/dead assay. (A) anodized Ti; (B) heparinized Ti; and (C) rhPDGF-BB-immobilized Ti after 48 h incubation.

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  Fig. 5 Proliferation of the MG-63 cells cultured on anodized Ti, heparinized Ti, and rhPDGF-BB-immobilized Ti after one-, three-, and seven-day culture.

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  Fig. 6 ALP activity of the MG-63 cells cultured on anodized Ti, heparinized Ti, and rhPDGF-BB-immobilized Ti after 7-, 14-, and 21-day incubation (*P<.05 and **P<.001).

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  Fig. 7 Calcium deposition of the MG-63 cells grown on the anodized Ti, heparinized Ti, and rhPDGF-BB-immobilized Ti after 21-day incubation (**P<.001).

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  Fig. 8 Gene expressions of the (A) osteopontin and (B) osteocalcine of the MG-63 cells grown on the anodized Ti, heparinized Ti, and rhPDGF-BB-immobilized Ti using real-time PCR after 21-day incubation (*P<.05 and **P<.001).

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  Fig. 9 Real-time PCR analysis of the anti-inflammatory effect using the transcript levels of the TNF-α and IL-6 of the MG-63 cells grown on the anodized Ti and heparinized Ti after 24 h incubation of LPS (**P<.001).

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