Li J Z, Li H, Sasaki T, Holman D, Beres B, Dumont R J, Pittman D D, Hankins G R, Helm G A
Molecular Neurosurgery Lab, Department of Neurosurgery, University of Virginia Health System, Charlottesville, VA 22908, USA.
Gene Ther. 2003 Sep;10(20):1735-43. doi: 10.1038/sj.gt.3302075.
Bone morphogenetic protein (BMP) adenoviral vectors for the induction of osteogenesis are being developed for the treatment of bone pathology. However, it is still unknown which BMP adenoviral vector has the highest potential to stimulate bone formation in vivo. In this study, the osteogenic activities of recombinant human BMP-2, BMP-4, BMP-6, BMP-7, and BMP-9 adenoviruses were compared in vitro, in athymic nude rats, and in Sprague-Dawley rats. In vitro osteogenic activity was assessed by measuring the alkaline phosphatase activity in C2C12 cells transduced by the various BMP vectors. The alkaline phosphatase activity induced by 2 x 10(5) PFU/well of BMP viral vector was 4890 x 10(-12) U/well for ADCMVBMP-9, 302 x 10(-12) U/well for ADCMVBMP-4, 220 x 10(-12) U/well for ADCMVBMP-6, 45 x 10(-12) U/well for ADCMVBMP-2, and 0.43 x 10(-12) U/well for ADCMVBMP-7. The average volume of new bone induced by 10(7) PFU of BMP vector in athymic nude rats was 0.37+/-0.03 cm(3) for ADCMVBMP-2, 0.89+/-0.07 cm(3) for ADCMVBMP-4, 1.02+/-0.07 cm(3) for ADCMVBMP-6, 0.24+/-0.05 cm(3) for ADCMVBMP-7, and 0.63+/-0.07 cm(3) for ADCMVBMP-9. In immunocompetent Sprague-Dawley rats, no bone formation was demonstrated in the ADCMVBMP-2, ADCMVBMP-4, and ADCMVBMP-7 groups. ADCMVBMP-6 at a viral dose of 10(8) PFU induced 0.10+/-0.03 cm(3) of new bone, whereas ADCMVBMP-9 at a lower viral dose of 10(7) PFU induced more bone, with an average volume of 0.29+/-0.01 cm(3).
用于诱导骨生成的骨形态发生蛋白（BMP）腺病毒载体正在研发中，用于治疗骨疾病。然而，目前仍不清楚哪种BMP腺病毒载体在体内刺激骨形成的潜力最大。在本研究中，比较了重组人BMP-2、BMP-4、BMP-6、BMP-7和BMP-9腺病毒在体外、无胸腺裸鼠和Sprague-Dawley大鼠中的成骨活性。通过测量各种BMP载体转导的C2C12细胞中的碱性磷酸酶活性来评估体外成骨活性。2×10⁵ PFU/孔的BMP病毒载体诱导的碱性磷酸酶活性，ADCMVBMP-9为4890×10⁻¹² U/孔，ADCMVBMP-4为302×10⁻¹² U/孔，ADCMVBMP-6为220×10⁻¹² U/孔，ADCMVBMP-2为45×10⁻¹² U/孔，ADCMVBMP-7为0.43×10⁻¹² U/孔。在无胸腺裸鼠中，10⁷ PFU的BMP载体诱导的新骨平均体积，ADCMVBMP-2为0.37±0.03 cm³，ADCMVBMP-4为0.89±0.07 cm³，ADCMVBMP-6为1.02±0.07 cm³，ADCMVBMP-7为0.24±0.05 cm³，ADCMVBMP-9为0.63±0.07 cm³。在具有免疫活性的Sprague-Dawley大鼠中，ADCMVBMP-2、ADCMVBMP-4和ADCMVBMP-7组未显示骨形成。病毒剂量为10⁸ PFU的ADCMVBMP-6诱导了0.10±0.03 cm³的新骨，而病毒剂量较低为10⁷ PFU的ADCMVBMP-9诱导出更多的骨，平均体积为0.29±0.01 cm³。
