Department of Ophthalmology, The Second Affiliated Hospital of Xi'an Medical University, Xi'an 710021, Shaanxi, PR China.
Xi'an Medical University, Xi'an 710021, Shaanxi, PR China.
J Biomed Nanotechnol. 2021 Aug 1;17(8):1679-1689. doi: 10.1166/jbn.2021.3141.
We established a self-decomposable SiO₂ encapsulated mitochondrial targeting short peptide SS31 drug loading system (SiO₂@SS31) to determine its nano-sustained release characteristics in eukaryotic cells. We explored the protection of SiO₂@SS31 on the 661W cells after oxidative injury by H₂O₂. After the drug loading, we detected the morphology of SiO₂@SS31 by transmission electron microscopy (TEM). Moreover, high-pressure liquid chromatography (HPLC) was used to determine the drug capacity and encapsulation efficiency of the nanoparticles. Then, the release curve in vitro was drawn. The 661W cells were cultured in vitro to allow the detection of cytotoxicity by the MTT assay. The SS31loaded nanoscale microspheres labeled with fluorescein isothiocyanate (SiO₂@FITC-SS31) were prepared, and their sustained release effect was detected with intracellular endocytosis, using confocal microscopy and flow cytometry. Within 15 days, the SiO2@SS31 nanoparticles were completely decomposed and simultaneously released the SS31 peptide in deionized water and normal saline. Nonetheless, the process was faster in simulated body fluid and serum. The MTT assay suggested that SiO₂@SS31 has sustained protection compared with SS31 in the 661W cells at 48 h. Flow cytometry proved SiO₂@FITC-SS31 could maintain a high level and last longer after 24 h. The SS31 peptide, which has excellent medical application prospects, can be slowly and continuously released from self-decomposable SiO₂ and targeted to concentrate on mitochondria.
我们构建了一种自分解的 SiO₂ 包裹的靶向线粒体的短肽 SS31 药物载药系统（SiO₂@SS31），以确定其在真核细胞中的纳米持续释放特性。我们探讨了 SiO₂@SS31 对 H₂O₂ 氧化损伤后 661W 细胞的保护作用。载药后，我们通过透射电子显微镜（TEM）观察 SiO₂@SS31 的形态。此外，还使用高压液相色谱（HPLC）测定纳米颗粒的载药量和包封率。然后绘制体外释放曲线。体外培养 661W 细胞，通过 MTT 法检测细胞毒性。制备了标记异硫氰酸荧光素的 SS31 载纳米微球（SiO₂@FITC-SS31），并用共聚焦显微镜和流式细胞术检测其细胞内内吞作用的持续释放效果。在 15 天内，SiO2@SS31 纳米颗粒在去离子水和生理盐水完全分解，同时释放 SS31 肽。然而，在模拟体液和血清中，这个过程更快。MTT 法检测结果表明，SiO₂@SS31 在 48 h 时对 661W 细胞的保护作用优于 SS31。流式细胞术证明，SiO₂@FITC-SS31 在 24 h 后仍能保持高水平并持续更长时间。具有良好医学应用前景的 SS31 肽可从自分解的 SiO₂ 中缓慢、持续地释放，并靶向聚集在线粒体。
