Department of Chemical and Materials Engineering, University of Kentucky, 177 F. Paul Anderson Tower, Lexington, KY, 40506, USA.
Naprogenix Inc., Lexington, KY, 40506, USA.
Adv Biol (Weinh). 2021 Mar;5(3):e2000173. doi: 10.1002/adbi.202000173. Epub 2021 Feb 9.
Cellular uptake and expulsion mechanisms of engineered mesoporous silica nanoparticles (MSNPs) are important in their design for novel biomolecule isolation and delivery applications such as nanoharvesting, defined as using nanocarriers to transport and isolate valuable therapeutics (secondary metabolites) out of living plant organ cultures (e.g., hairy roots). Here, temperature-dependent MSNP uptake and recovery processes in hairy roots are examined as a function of surface chemistry. MSNP uptake into hairy roots and time-dependent expulsion are quantified using Ti content (present for biomolecule binding) and fluorescence spectroscopy of fluorescently tagged MSNPs, respectively. The results suggest that functionalization and surface charge (regulated by amine group attachment) play the biggest role in the effectiveness of uptake and recovery. Comparison of MSNP interactions with hairy roots at 4 and 23 °C shows that weakly charged MSNPs functionalized only with Ti are taken up and expelled by thermally activated mechanisms, while amine-modified positively charged particles are taken up and expelled mainly by direct penetration of cell walls. Amine-functionalized MSNPs move spontaneously in and out of plant cells by dynamic exchange with a residence time of 20 ± 5 min, suggesting promise as a biomolecule nanoharvesting platform for plant organ cultures.
工程化介孔硅纳米粒子(MSNPs)的细胞摄取和排出机制对于其新型生物分子分离和递药应用的设计非常重要,例如纳米收获,这是指利用纳米载体将有价值的治疗剂(次生代谢物)从活体植物器官培养物(如发根)中运输和分离出来。在这里,研究了表面化学性质对发根中 MSNP 摄取和回收过程的温度依赖性。通过 Ti 含量(用于生物分子结合)和荧光标记 MSNPs 的荧光光谱,分别定量测定了 MSNP 进入发根和随时间排出的情况。结果表明,功能化和表面电荷(通过胺基附着调节)在摄取和回收的有效性中起着最大的作用。在 4°C 和 23°C 下比较 MSNP 与发根的相互作用表明,仅用 Ti 功能化的带弱电荷的 MSNPs 通过热激活机制被摄取和排出,而带正电荷的经胺修饰的颗粒则主要通过细胞壁的直接渗透被摄取和排出。胺功能化的 MSNPs 通过与细胞的动态交换自发地进入和离开植物细胞,停留时间为 20±5min,这表明它有希望成为植物器官培养物的生物分子纳米收获平台。