Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (24): 3863-3869.doi: 10.3969/j.issn.2095-4344.2014.24.016
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Liu Yu-peng1, 2, Zhao De-wei2, Wang Wei-ming2, Zhang Yao2, Li Fang2, Liu Zhen-gang2
Revised:
2014-05-19
Online:
2014-06-11
Published:
2014-06-11
Contact:
Liu Yu-peng, Master, Chief physician, Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, Liaoning Province, China; Department of Orthopaedic Surgery, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, Liaoning Province, China
About author:
Liu Yu-peng, Master, Chief physician, Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, Liaoning Province, China; Department of Orthopaedic Surgery, Affiliated Zhongshan Hospital of Dalian University, Dalian 116001, Liaoning Province, China
CLC Number:
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.
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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