Angiopoiesis and bone regeneration via co expression of the hVEGF and hBMP genes from an adeno-associated viral vector in vitro and in vivo  

作　　者：Chen ZHANG Kun-zheng WANG Hui QIANG Yi-lun TANG Qian LI Miao LI Xiao-qian DANG 

机构地区：[1]Department of Orthopedic Surgery, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an710004, China [2]Department of Ultrasound, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an710004, China

出　　处：《Acta Pharmacologica Sinica》2010年第7期821-830,共10页中国药理学报（英文版）

基　　金：Acknowledgements This work was supported by the National Natural Science Foundation of China （No 30600624） and （No 30772189）. We acknowledge the support of Xi＇an Jiao Tong University. We are grateful to Dr Xiang-hui HUANG （Department of Orthopedics, Shaanxi Provincial People＇s Hospital, Xi＇an 710068, Shaanxi Province, China） for his help in the construction of the rAAV-hVEGF165-IRES-hBMP-7 viral vector.

摘　　要：Aim： To investigate the therapeutic potential of adeno-associated virus （AAV）-mediated expression of vascular endothelial growth factor （VEGF） and bone morphogenetic protein （BMP）. Methods： Four experimental groups were administered the following AAV vector constructs： rAAV-hVEGF165-internal ribosome entry site （IRES）-hBMP-7 （AAV-VEGF/BMP）, rAAV-hVEGF165-GFP （AAV-VEGF）, rAAV-hBMP-7-GFP （AAV-BMP）, and rAAV-IRES-GFP （AAV-GFP）. VEGF165 and BMP-7 gene expression was detected using RT-PCR. The VEGF165 and BMP-7 protein expression was determined by Western blotting and ELISA. The rabbit ischemic hind limb model was adopted and rAAV was administered intramuscularly into the ischemic limb. Results： Rabbit bone marrow-derived mesenchymal stem cells （BMSCs） were cultured and infected with the four viral vectors. The expression of GFP increased from the 7th day of infection and could be detected on the 28th day post-infection. In the AAV-VEGF/BMP group, the levels of VEGF165 and BMP-7 increased with prolonged infection time. The VEGF185 and BMP-7 secreted from BMSCs in the AAV-VEGF/BMP group enhanced HUVEC tube formation and resulted in a stronger osteogenic ability, respectively. In rabbit ischemic hind limb model, GFP expression increased from the 4th week and could be detected at 8 weeks post-injection. The rAAV vector had superior gene expressing activity. Eight weeks after gene transfer, the mean blood flow was significantly higher in the AAV-VEGF/BMP group. Orthotopic ossification was radiographically evident, and capillary growth and calcium deposits were obvious in this group. Conclusion： AAV-mediated VEGF and BMP gene transfer stimulates angiogenesis and bone regeneration and may be a new therapeutic technique for the treatment of avascular necrosis of the femoral head （ANFH）.Aim： To investigate the therapeutic potential of adeno-associated virus （AAV）-mediated expression of vascular endothelial growth fac- tor （VEGF） and bone morphogenetic protein （BMP�Aim： To investigate the therapeutic potential of adeno-associated virus （AAV）-mediated expression of vascular endothelial growth factor （VEGF） and bone morphogenetic protein （BMP）. Methods： Four experimental groups were administered the following AAV vector constructs： rAAV-hVEGF165-internal ribosome entry site （IRES）-hBMP-7 （AAV-VEGF/BMP）, rAAV-hVEGF165-GFP （AAV-VEGF）, rAAV-hBMP-7-GFP （AAV-BMP）, and rAAV-IRES-GFP （AAV-GFP）. VEGF165 and BMP-7 gene expression was detected using RT-PCR. The VEGF165 and BMP-7 protein expression was determined by Western blotting and ELISA. The rabbit ischemic hind limb model was adopted and rAAV was administered intramuscularly into the ischemic limb. Results： Rabbit bone marrow-derived mesenchymal stem cells （BMSCs） were cultured and infected with the four viral vectors. The expression of GFP increased from the 7th day of infection and could be detected on the 28th day post-infection. In the AAV-VEGF/BMP group, the levels of VEGF165 and BMP-7 increased with prolonged infection time. The VEGF185 and BMP-7 secreted from BMSCs in the AAV-VEGF/BMP group enhanced HUVEC tube formation and resulted in a stronger osteogenic ability, respectively. In rabbit ischemic hind limb model, GFP expression increased from the 4th week and could be detected at 8 weeks post-injection. The rAAV vector had superior gene expressing activity. Eight weeks after gene transfer, the mean blood flow was significantly higher in the AAV-VEGF/BMP group. Orthotopic ossification was radiographically evident, and capillary growth and calcium deposits were obvious in this group. Conclusion： AAV-mediated VEGF and BMP gene transfer stimulates angiogenesis and bone regeneration and may be a new therapeutic technique for the treatment of avascular necrosis of the femoral head （ANFH）.Aim： To investigate the therapeutic potential of adeno-associated virus （AAV）-mediated expression of vascular endothelial growth fac- tor （VEGF） and bone morphogenetic protein （BMP�

关 键 词：adeno-associated virus vascular endothelial growth factor bone morphogenetic protein （BMP） avascular necrosis of thefemoral head （ANFH） gene therapy 

分 类 号：Q786[生物学—分子生物学] Q51