Biological effects of rAAV-caAlk2 coating on structural allograft healing
Biological effects of rAAV-caAlk2 coating on structural allograft healing
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DOI：
10.1016/j.ymthe.2005.02.026
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发表时间：
2005-08-01
影响因子：
12.4
通讯作者：
Schwarz, EM
中科院分区：
文献类型：
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作者：
Koefoed, M;Ito, H;Schwarz, EM
结构性同种异体骨移植物由于缺乏成骨和重建能力而经常发生断裂。为了克服这些局限性，我们利用了重组腺相关病毒（rAAV）介导的体内基因转移包被的同种异体移植物。使用β-半乳糖苷酶作为报告基因，我们表明，4毫米小鼠股骨同种异体移植物与rAAV-LacZ包被能够转导相邻的炎性细胞和成骨细胞在骨折骨痂移植后。虽然这种LacZ载体对同种异体移植物愈合没有影响，但是通过rAAV-caAlk 2涂层递送的骨形态发生蛋白信号在第14天直接在同种异体移植物的皮质表面上诱导软骨内骨形成。到第28天，仅存在rAAV-caAlk 2涂覆的同种异体移植物的皮质表面的新编织骨的重塑和大量骨吸收的证据。愈合42天后，rAAV-LacZ-与rAAV-caAlk 2涂层同种异体移植物的显微CT分析显示新骨形成显著增加（0.67 ± 0.21 vs 2.49 ± 0.40 mm（3）; P < 0.005）。此外，用rAAV-Alk 2包被的同种异体移植物移植的股骨的3D显微CT图像提供了第一个证据，即皮质同种异体移植物周围的骨完全桥接是可能的。这些结果表明，无细胞的rAAV包被的同种异体移植物具有移植后体内再生的潜力。
Structural bone allografts often fracture due to their lack of osteogenic and remodeling potential. To overcome these limitations, we utilized allografts coated with recombinant adeno-associated virus (rAAV) that mediate in vivo gene transfer. Using beta-galactosidase as a reporter gene, we show that 4-mm murine femoral allografts coated with rAAV-LacZ are capable of transducing adjacent inflammatory cells and osteoblasts in the fracture callus following transplantation. While this LacZ vector had no effect on allograft healing, bone morphogenetic protein signals delivered via rAAV-caAlk2 coating induced endochondral bone formation directly on the cortical surface of the allograft by day 14. By day 28 there was evidence of remodeling of the new woven bone and massive osteoclastic resorption of the cortical surface of the rAAV-caAlk2-coated allografts only. Micro-CT analysis of rAAV-LacZ- vs rAAV-caAlk2-coated allografts after 42 days of healing demonstrated a significant increase in new bone formation (0.67 +/- 0.21 vs 2.49 +/- 0.40 mm(3); P < 0.005). Furthermore, the 3D micro-CT images of femurs grafted with rAAV-Alk2-coated allografts provided the first evidence that complete bridging of bone around a cortical allograft is possible. These results indicate that cell-free, rAAV-coated allografts have the potential to revitalize in vivo following transplantation.