Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, B-9000 Ghent, Belgium.
Univ. Lille, CNRS, Centrale Lille, ISEN, Univ. Valenciennes, UMR 8520-IEMN, F-59000 Lille, France.
Int J Mol Sci. 2020 Feb 24;21(4):1540. doi: 10.3390/ijms21041540.
Nanoparticle mediated laser-induced photoporation is a physical cell membrane disruption approach to directly deliver extrinsic molecules into living cells, which is particularly promising in applications for both adherent and suspension cells. In this work, we explored surface modifications of graphene quantum dots (GQD) and reduced graphene oxide (rGO) with polyethylene glycol (PEG) and polyethyleneimine (PEI) to enhance colloidal stability while retaining photoporation functionality. After photoporation with FITC-dextran 10 kDa (FD10), the percentage of positive HeLa cells (81% for GQD-PEG, 74% for rGO-PEG and 90% for rGO-PEI) increased approximately two-fold compared to the bare nanomaterials. While for Jurkat suspension cells, the photoporation efficiency with polymer-modified graphene-based nanomaterial reached as high as 80%. Cell viability was >80% in all these cases. In addition, polymer functionalization proved to be beneficial for the delivery of larger macromolecules (FD70 and FD500) as well. Finally, we show that rGO is suitable for photoporation using a near-infrared laser to reach 80% FD10 positive HeLa cells at 80% cell viability. We conclude that modification of graphene-based nanoparticles with PEG and especially PEI provide better colloidal stability in cell medium, resulting in more uniform transfection and overall increased efficiency.
纳米颗粒介导的激光诱导光孔法是一种物理细胞膜破坏方法，可将外源分子直接递送至活细胞中，在黏附细胞和悬浮细胞的应用中特别有前景。在这项工作中，我们探索了将聚乙二醇（PEG）和聚乙烯亚胺（PEI）修饰石墨烯量子点（GQD）和还原氧化石墨烯（rGO），以增强胶体稳定性，同时保留光孔功能。用光孔处理 FITC-葡聚糖 10 kDa（FD10）后，与裸纳米材料相比，HeLa 细胞（GQD-PEG 为 81%，rGO-PEG 为 74%，rGO-PEI 为 90%）阳性细胞的百分比增加了约两倍。对于 Jurkat 悬浮细胞，聚合物修饰的基于石墨烯的纳米材料的光孔效率高达 80%。在所有这些情况下，细胞活力均>80%。此外，聚合物官能化被证明有利于较大的大分子（FD70 和 FD500）的传递。最后，我们表明 rGO 适合使用近红外激光进行光孔，在 80%细胞活力下可达到 80%的 FD10 阳性 HeLa 细胞。我们得出结论，用 PEG 特别是 PEI 修饰基于石墨烯的纳米颗粒可在细胞培养基中提供更好的胶体稳定性，从而导致更均匀的转染和整体效率提高。
