Department of Urology, Peking University Third Hospital, Peking University, Beijing, China.
Reproductive Medicine Center, Peking University Third Hospital, Peking University, Beijing, China.
Investig Clin Urol. 2022 May;63(3):359-367. doi: 10.4111/icu.20220016.
Our previous studies showed that nanotechnology improves derived adipose-derived stem cells (ADSCs) therapy for erectile dysfunction (ED). In this study, the Neuregulin-1(NRG1) gene was transfected into ADSCs with superparamagnetic iron oxide nanoparticles (SPION) further to improve the therapeutic effect of ADSCs on ED.
ADSCs were isolated from epididymal adipose tissue of Sprague-Dawley rats. The optimal concentration of PEI-SPION (SPION modified with polyethyleneimine) was selected to construct the gene complex. After electrostatic binding of PEI-SPION and DNA, a PEI layer was wrapped to make the PEI-SPION-NRG1-PEI gene transfection complex. Different groups were set up for transfection tests. Lipo2000 transfection reagent was used as the control. PEI-SPION-NRG1-PEI in the experimental group was transfected under an external magnetic field.
When the concentration of PEI-SPION was 10 µg/mL, it had little cytotoxicity, and cell activity was not significantly affected. PEI-SPION-NRG1-PEI forms positively charged nanocomposites with a particle size of 72.6±14.9 nm when N/P ≥8. The PEI-SPION-NRG1-PEI gene complex can significantly improve the transfection efficiency of ADSCs, reaching 26.74%±4.62%, under the action of the external magnetic field. PCR and Western blot showed that the expression level of the NRG1 gene increased significantly, which proved that the transfection was effective.
PEI-SPION can be used as a vector for NRG1 gene transfection into ADSCs. PEI-SPION-NRG1-PEI packaging has the highest transfection efficiency under the external magnetic field than the other groups. These findings may provide a new strategy for ADSCs therapy for ED.
我们之前的研究表明，纳米技术可改善衍生脂肪源性干细胞（ADSCs）治疗勃起功能障碍（ED）的效果。在这项研究中，我们进一步将神经调节蛋白 1（NRG1）基因转染到 ADSCs 中，使用超顺磁性氧化铁纳米粒子（SPION）提高 ADSCs 治疗 ED 的疗效。
从 Sprague-Dawley 大鼠附睾脂肪组织中分离 ADSCs。选择最佳的聚乙烯亚胺-SPION（SPION 经聚乙烯亚胺修饰）浓度构建基因复合物。PEI-SPION 与 DNA 静电结合后，包裹一层 PEI 以形成 PEI-SPION-NRG1-PEI 基因转染复合物。设置不同的转染测试组，以脂质体 2000 转染试剂作为对照组，在外部磁场下转染实验分组中的 PEI-SPION-NRG1-PEI。
当 PEI-SPION 的浓度为 10μg/ml 时，其细胞毒性较小，细胞活性无明显影响。当 N/P≥8 时，PEI-SPION-NRG1-PEI 形成粒径为 72.6±14.9nm 的带正电荷的纳米复合物。在外磁场的作用下，PEI-SPION-NRG1-PEI 基因复合物可显著提高 ADSCs 的转染效率，达到 26.74%±4.62%。PCR 和 Western blot 显示 NRG1 基因的表达水平显著提高，证明转染有效。
PEI-SPION 可作为 NRG1 基因转染 ADSCs 的载体。在外部磁场下，PEI-SPION-NRG1-PEI 包装的转染效率高于其他组。这些发现可能为 ADSCs 治疗 ED 提供新的策略。
