Neuroprotective effects of granulocyte colony-stimulating factor and relationship to promotion of angiogenesis after spinal cord injury in rats

Laboratory investigation

Junko Kawabe M.D., Ph.D. 1 , Masao Koda M.D., Ph.D. 2 , Masayuki Hashimoto M.D., Ph.D. 1 , Takayuki Fujiyoshi M.D., Ph.D. 1 , Takeo Furuya M.D., Ph.D. 1 , Tomonori Endo M.D., Ph.D. 1 , Akihiko Okawa M.D., Ph.D. 1 , and Masashi Yamazaki M.D., Ph.D. 1
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  • 1 Department of Orthopaedic Surgery, Chiba University Graduate School of Medicine; and
  • 2 Department of Orthopaedic Surgery, Chiba Aoba Municipal Hospital, Chiba, Japan
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Object

Granulocyte colony-stimulating factor (G-CSF) has neuroprotective effects on the CNS. The authors have previously demonstrated that G-CSF also exerts neuroprotective effects in experimental spinal cord injury (SCI) by enhancing migration of bone marrow–derived cells into the damaged spinal cord, increasing glial differentiation of bone marrow–derived cells, enhancing antiapoptotic effects on both neurons and oligodendrocytes, and by reducing demyelination and expression of inflammatory cytokines. Because the degree of angiogenesis in the subacute phase after SCI correlates with regenerative responses, it is possible that G-CSF's neuroprotective effects after SCI are due to enhancement of angiogenesis. The aim of this study was to assess the effects of G-CSF on the vascular system after SCI.

Methods

A contusive SCI rat model was used and the animals were randomly allocated to either a G-CSF–treated group or a control group. Integrity of the blood–spinal cord barrier was evaluated by measuring the degree of edema in the cord and the volume of extravasation. For histological evaluation, cryosections were immunostained with anti–von Willebrand factor and the number of vessels was counted to assess revascularization. Real-time reverse transcriptase polymerase chain reaction was performed to assess expression of angiogenic cytokines, and recovery of motor function was assessed with function tests.

Results

In the G-CSF–treated rats, the total number of vessels with a diameter > 20 μm was significantly larger and expression of angiogenic cytokines was significantly higher than those in the control group. The G-CSF–treated group showed significantly greater recovery of hindlimb function than the control group.

Conclusions

These results suggest that G-CSF exerts neuroprotective effects via promotion of angiogenesis after SCI.

Abbreviations used in this paper: ANGPT = angiopoietin; BBB = Basso-Beattie-Bresnahan; Ct = threshold cycle; FGF2 = fibroblast growth factor 2; G-CSF = granulocyte colony-stimulating factor; HGF = hepatocyte growth factor; PBS = phosphate-buffered saline; PCR = polymerase chain reaction; RT = reverse-transcriptase; SCI = spinal cord injury; VEGF = vascular endothelial growth factor; VWF = von Willebrand factor.

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Contributor Notes

Address correspondence to: Masao Koda, M.D., Ph.D., Department of Orthopaedic Surgery, Chiba Aoba Municipal Hospital, 1273-2, Aoba-Cho, Chuo-Ku, Chiba 260-0852, Japan. email: masaomst@yahoo.co.jp.

Please include this information when citing this paper: published online July 1, 2011; DOI: 10.3171/2011.5.SPINE10421.

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