Nerve growth factor promotes the expression of vascular endothelial growth factor in the fracture healing process

doi:10.3969/j.issn.2095-4344.2014.24.016

Abstract

Abstract:

BACKGROUND: The fracture healing process is very complex, involving a variety of cytokines. At present, the existing studies focus on bone morphogenetic protein, fibroblast growth factor, transforming growth factor β, vascular endothelial cell growth factor and insulin-like growth factor. The role of nerve growth factor on vascular endothelial cell growth factor during the fracture healing process remains unclear.
OBJECTIVE: To investigate the effect of nerve growth factor on vascular endothelial growth factor expression during the process of fracture healing in rabbits.
METHODS: Radial fracture model established in white rabbits were divided into three groups, they were treated with nerve growth factor, nerve growth factor monoclonal antibody, and saline respectively, serving as nerve growth factor group, antagonizing nerve growth factor group, and control group.
RESULTS AND CONCLUSION: The vascular endothelial growth factor expression was detected with western blot analysis at 24 hours, 48 hours, 1 week, 3 weeks, 6 weeks, 8 weeks post-injury. The results showed that the expression level was the highest in nerve growth factor group, then in control group, and the lowest in antagonizing nerve growth factor group (P < 0.05). Experimental findings suggest that, nerve growth factor can promote the expression of vascular endothelial growth factor in the fracture healing process.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: nerve growth factor, vascular endothelial growth factor, fracture healing, rabbits, proteins

CLC Number:

R318

Liu Yu-peng, Zhao De-wei, Wang Wei-ming, Zhang Yao, Li Fang, Liu Zhen-gang. Nerve growth factor promotes the expression of vascular endothelial growth factor in the fracture healing process[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(24): 3863-3869.

Figures/Tables 2

Quantitative analysis of experimental animals All experimental rabbits were involved in the results analysis, without any loss. General observation All rabbits in the three groups survived with no deaths and ate properly. The surgical incisions exhibited no signs of infection or purulence and healed at approximately 1 postoperative week. At 24 hours postoperatively, the samples in the three groups all exhibited subcutaneous swelling at the incision sites, with no hematomas formed at the fracture sites. At 48 hours postoperatively, initial hematomas at the fracture sites were formed in all three groups. At 1 week postoperatively, hematoma organization was formed at the fracture sites (NGF group > control group > NGF antagonist group), with discontinuous fractures. At 3 weeks postoperatively, continuous fractures were observed, with primary callus formation, and strong bony connections were formed at the fracture sites (NGF group > control group > NGF antagonist group). At 6 weeks postoperatively, the callus sizes had shrunk compared with the primary calluses (NGF group < control group < NGF antagonist group). At 8 weeks postoperatively, the NGF group and the control group showed no obvious fracture lines, while the NGF antagonist group still showed a small amount of callus. Western blot analysis and statistical analysis results Images of immunoblots showing VEGF expression with an internal reference in the bone tissues at different time points in the three groups are shown in Figure 2. The absorbance ratios and the means with standard deviations of VEGF target protein expression versus the internal reference in the bone tissues at different time points in the three groups are shown in Table 1, and their changing trend over time is displayed in Figure 2. Finally, a statistical analysis of variance was conducted for the absorbance ratios of VEGF expression versus the internal reference in the bone tissues at different time points in the three groups (Table 1). In the fracture-healing process, VEGF expression in the callus continuously increased, reaching a peak at postoperative 3 weeks, and then gradually decreased (Figure 2). VEGF expression in the callus was higher in the NGF group than in the other two groups, while VEGF expression in the callus was lower in the NGF antagonist group than in the other two groups (Table 1). The one-way analysis of variance for the absorbance ratios of the target protein VEGF versus the internal reference protein in each group revealed that, compared with the NGF antagonist group and the control group, the results of the NGF group were higher at all time points, with statistically significant differences (P < 0.05). These results indicate that NGF had a positive role in promoting VEGF expression at the fracture site. The differences between the NGF antagonist group and the control group at the 24- and 48-hour and the 8-week time points were not statistically significant (P > 0.05), which may indicate that the NGF antibody concentrations had not yet reached effective levels at the early stages of fracture healing and that the anti-NGF antibody concentration was completely attenuated at the late stage of fracture healing (Table 1)."

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