优化靶向载基因超声纳米泡效能:实验研究北大核心CSCD

目的优选超声靶向微泡破坏(UTMD)技术介导载基因靶向纳米泡(T-NB)爆破的条件。方法采用薄膜水化法制备纳米泡,以生物素-亲和素-生物素连接方法制备pAd-AAV-9/miRNA-1 T-NB,观察其基本理化性质。基于机械指数(0.6、0.9、1.2、1.5)、T-NB溶液浓度(10^(5)/ml、10^(6)/ml、10^(7)/ml、10^(8)/ml)及辐照时间(30、45、60、120 s)设计三因素四水平正交实验,评价不同超声辐照参数对T-NB溶液信号强度的影响。结果pAd-AAV-9/miRNA-1 T-NB溶液为乳白色混悬液,浓度(0.74±0.10)×10^(8)/ml;T-NB为大小相近的球形,平均粒径(443.97±12.84)nm,平均电位(-7.33±0.09)mV。在不同辐照条件下,T-NB溶液的信号强度不同。机械指数、T-NB浓度及辐照时间均可显著影响信号强度(P均<0.05)。影响T-NB爆破的因素依次为机械指数、T-NB溶液浓度及辐照时间,机械指数0.6、T-NB浓度10^(6)/ml、辐照时间120 s为其最优组合。结论机械指数0.6、T-NB溶液浓度10^(6)/ml、辐照时间120 s为UTMD技术载基因T-NB的最优爆破条件。

Optimizing efficacy of targeted gene-carrying ultrasonic nanobubbles:An experimental study

Objective To optimize the blasting conditions for carrying gene-targeted nanobubble (T-NB) mediated with ultrasound-targeted microbubble destruction (UTMD) technology. Methods Nanobubbles were prepared with thin film hydration and pAd-AAV-9/miRNA-1 T-NB by using the biotin-avidin-biotin ligation methods, and the basic physicochemical properties were observed. Based on the mechanical index (0.6, 0.9, 1.2, 1.5), concentration of T-NB solution (10⁵/ml, 10⁶/ml, 10⁷/ml, 10⁸/ml) and irradiation time (30, 45, 60, 120 s), three-factor four-level orthogonal experiments were designed to evaluate the influence of different ultrasonic irradiation parameters on the signal intensity of T-NB solution. Results pAd-AAV-9/miRNA-1 T-NB solution was milky suspension with concentration (0.74±0.10)×10⁸/ml. T-NB present as similar balls with average particle size of (443.97±12.84)nm and average potential of (-7.33±0.09)mV. T-NB solution showed different signal intensities under different irradiation conditions. The mechanical index, T-NB concentration and irradiation time had significant influences on the signal intensity (all P<0.05). The impact factors of T-NB blasting included mechanical index, T-NB solution concentration and irradiation time in order, and the optimal combination was mechanical index of 0.6, T-NB concentration of 10⁶/ml and irradiation time of 120 s. Conclusion Mechanical index of 0.6, the concentration of T-NB solution of 10⁶/ml, and the irradiation time of 120 s were the optimal blasting conditions for carrying gene T-NB mediated with UTMD technology.