Abasic study for cell based gene therapy for dental implant

牙科种植体治疗已被证明是一种可预测的治疗方法，用于替换缺失的牙齿。然而，在将种植体插入牙槽嵴后，骨和钛之间的愈合期较长，这可能是遗留的问题之一。通过基因治疗促进骨-种植体结合可能是解决这一问题的新策略之一。我们使用脂质体转染、声致穿孔和这些方法的组合来测试质粒DNA-非病毒载体复合物的转染效率和细胞毒性，所述质粒DNA-非病毒载体复合物包括β-半乳糖苷酶报告基因的编码。将贴附在细胞培养板表面上的原代骨髓细胞用于这些实验。对声孔法的超声功率和时间条件进行了实验研究，结果表明，Lipofedion法和声孔法的转染效率明显高于对照组。脂质体-DNA复合物的细胞毒性随试剂浓度的增加而增加。声致孔微泡组细胞毒性与对照组相当。当使用脂质体转染和声穿孔的组合方法时，检测到试剂的沉积。结果表明，超声波强度和作用时间可能存在最佳条件，并与最佳质粒DNA浓度相对应。细胞存活率和转染效率随条件的恶化而降低。

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.