J Korean Orthop Assoc.  2011 Oct;46(5):357-363.

Enhancement of Vasculogenesis and Osteogenesis Using Granulocyte-Colony Stimulating Factor in the Rat Model of Tibial Distraction Osteogenesis

• Shin SJ
• Lee DY
• Lee HR
• Kim JI
• Yoo WJ
• Cho TJ
• Choi IH

Abstract

PURPOSE
Proper speed of distraction is critical for successful new bone formation in distraction osteogenesis. The purpose of this study was to evaluate the effect of granulocyte-colony stimulating factor (G-CSF) on formation of new blood vessels and new bones in the rat model of tibial distraction osteogenesis (DO) to develop enhancement method of bone formation while increasing the distraction speed.
MATERIALS AND METHODS
Forty two rat-tibial DO models were included in this study, and were divided into 3 groups; group I (rapid distraction), group II (rapid distraction with G-CSF), and group III (slow distraction). The amount of bone formation and relative blood flow were analyzed by sequential radiographs and laser Doppler perfusion imaging (LDPI). Blood sampling was done before G-CSF injection, at 2 weeks and 5 weeks after G-CSF injection and surface expression such as Scal-1+ and C-kit+ of endothelial progenitor cells (EPCs) was analyzed by fluorescence-activated cell sorter (FACS) for the effects of G-CSF in inducing mobilization of EPCs.
RESULTS
The amount of new bone formation in the distraction gap on serial radiographs was higher during the consolidation period in groups II and III than in group I but, the difference was not significant (p>0.05). The relative blood flow in the distraction gap in groups II and III increased more significantly than in group I (p<0.05). FACS analysis showed an increased EPCs fraction after G-CSF injection.
CONCLUSION
We demonstrated that G-CSF administration ameliorated bone formation and blood flow during rapid distraction in the rat model of tibial distraction osteogenesis. We think that G-CSF has an effect on mobilization of EPCs resulting in an increase in the blood flow.

Keyword

endothelial progenitor cell; granulocyte-colony stimulating factor; distraction osteogenesis

MeSH Terms

Animals
Blood Vessels
Granulocyte Colony-Stimulating Factor
Osteogenesis
Osteogenesis, Distraction
Perfusion Imaging
Rats
Stem Cells
Granulocyte Colony-Stimulating Factor

Figure

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  Figure 1 This photo shows bilateral mono-fixators at the right tibia in rat DO (distraction ostegenesis) model.

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  Figure 2 Serial radiographs during consolidation period in groups 1 (A), 2 (B), and 3 (C).

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  Figure 3 This graph shows average of relative bone density value during consolidation period after cessation of distraction in 3 groups. Relative bone densities of groups 2, and 3 were higher than that of group 1 during 6 weeks and that was not statistically significant (p>0.05, by student t-test).

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  Figure 4 This graph shows average of relative blood flow after operation (corticotomy) in 3 groups. Relative blood flows of groups 2, and 3 were higher than that of group 1 during 6 weeks and that was statistically significant (*p=0.04, †p=0.03, by student t-test).

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  Figure 5 G-CSF induced the mobilization of Sca-1 and C-kit positive cell fractions (right shift in FACS analysis, solid line) before the start of distraction (dash line). Sca-1 and C-kit positive cell fractions maintained during distraction and it decreased after 3 weeks of consolidation.

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