Abasic study for cell based gene therapy for dental implant

牙种植体治疗已被证明是一种可预测的修复缺失牙齿的治疗方法，然而，在将种植体植入牙槽骨脊后，骨与钛之间的漫长愈合时间可能是遗留问题之一。基因治疗加速种植体-骨整合可能是解决这一问题的新策略之一。我们用脂质体转染法、超声转染法和这三种方法相结合的方法检测了含有β-半乳糖苷酶报告基因编码的质粒DNA-非病毒载体复合体的转染率和细胞毒性。实验采用贴壁于细胞培养板表面的原代骨髓细胞。对超声功率和超声持续时间的条件进行了测试，结果表明，脂质体法和声孔法的转染率均高于对照组。随着脂质体-DNA复合体试剂浓度的增加，细胞毒作用增强。超声微泡显示细胞毒性较低，与对照组相当。采用脂质体介导法和超声造影法相结合，可检测到试剂的沉积。结果表明，超声强度和超声时间可能存在最佳条件，这与最适的质粒-DNA浓度相对应。当条件较差时，细胞存活率和转染率变差。

Dental implant therapy has been proven as a predictable treatment method for replacement of missing teeth Nevertheless, the long healing period to make integration between bone and titanium after insertion of implants into the alveolar ridge may be one of a remaining problem. Gene therapy to accelerate on bone-implant integration might be a one of a new strategy to resolve the problem. We tested transfection efficiency and cytotoxicity of a plasmid DNA-nonviral vector complex including the code of beta-galactosidase reporter gene using lipofection, sonoporation and combination of these methods. Primaly bone marrow cells attached on cell culture plate surface were used for these experiments. The conditions of ultrasound power and duration were also tested for the sonoporation method.Lipofedion and sonoporation methods showed better transfection efficiencies than the control group. Cytotoxicity was higher with concentration of the reagents of liposome-DNA complex. Low cytotoxidty was shown on sonoporation with micro bubble equal to the control group. When the combination method of lipofection and sonoporation was used, deposition of the reagents was detected. It was suggested that optimal condition may exist on the ultrasound intensity and duration, which be correspondenced on the optimal plasmid-DNA concentration. Cell viability and transfection efficiency became worse when the condition was bad.