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[Application of polytetrafluoroethylene as VEGF gene carrier]
1The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China. taosifeng@yahoo.com.cn
Objective:
To investigate the feasibility of vascular graft material polytetrafluoroethylene (PTFE) as gene carrier of vascular endothelial growth factor (VEGF).
Methods:
The PTFE strips were soaked in pCDI-hVEGF(121) solution,marked with EB solution, and then observed under ultraviolet light. The PTFE strips carrying pCDI-hVEGF(121) were soaked in normal saline and the value of optical density at 260 nm on different time was measured, then the controlled release curve was made. The PTFE strips carrying pCDI-hVEGF(121) were implanted into the left thigh muscles of rabbits and the PTFE strips without VEGF gene were implanted into the right. The expression of VEGF(121) mRNA versus beta-actin mRNA in muscles around vascular graft materials was evaluated by reverse transcriptase polymerase chain reaction(RT-PCR). The expression of VEGF(121) protein in muscle was determined by Western blot method.
Result:
The fluorescence on PTFE strips carrying pCDI-hVEGF(121) was observed under ultraviolet light. The controlled release curve demonstrated that the gene release was fast during 0.5 -4 h, then slowed down afterwards,and lasted for 72 h. RT-PCR and Western blot showed VEGF(121)/beta-actin mRNA ratios were 1.053+/-0.356,1. 718+/-0.404, 2.021+/-0.303, 1.872+/-0.231, 0.986+/-0.254, 0.340+/-0.116 and VEGF(121)protein expression levels were 0.328+/-0.088, 1.019+/-0.105, 2.249 +/-0.203, 2.036+/-0.079, 1.670+/-0.132, 0.636+/-0.107 at 1, 7, 14, 28, 42, and 56 days after implantation, respectively.
Conclusion:
The PTFE graft can be used as carrier of VEGF gene and VEGF gene can be transferred to skeletal muscle by this method.
