Xu Leyuan, Zolotarskaya Olga Yu, Yeudall W Andrew, Yang Hu
Department of Biomedical Engineering, Virginia Commonwealth University, Richmond, VA, 23284, USA.
Adv Healthc Mater. 2014 Sep;3(9):1430-8. doi: 10.1002/adhm.201300515. Epub 2014 Feb 13.
Immobilizing highly branched polyamidoamine (PAMAM) dendrimers to the cell surface represents an innovative method of enhancing cell surface loading capacity to deliver therapeutic and imaging agents. In this work, hybridized immune cells, that is, macrophage RAW264.7 (RAW), with PAMAM dendrimer G4.0 (DEN) on the basis of bioorthogonal chemistry are clicked. Efficient and selective cell surface immobilization of dendrimers is confirmed by confocal microscopy. Viability and motility of RAW-DEN hybrids remain the same as untreated RAW cells according to WST-1 assay and wound closure assay. Furthermore, Western blot analysis reveals that there are no significant alterations in the expression levels of signaling molecules AKT, p38, and NFκB (p65) and their corresponding activated (phosphorylated) forms in RAW cells treated with azido sugar and dendrimer, indicating that the hybridization process neither induced cell stress response nor altered normal signaling pathways. Taken together, this work shows the feasibility of applying bioorthogonal chemistry to create cell-nanoparticle hybrids and demonstrates the noninvasiveness of this cell surface engineering approach.
将高度支化的聚酰胺-胺(PAMAM)树枝状大分子固定在细胞表面是一种提高细胞表面负载能力以递送治疗剂和成像剂的创新方法。在这项工作中,基于生物正交化学,将杂交免疫细胞,即巨噬细胞RAW264.7(RAW)与PAMAM树枝状大分子G4.0(DEN)进行点击反应。共聚焦显微镜证实了树枝状大分子在细胞表面的高效和选择性固定。根据WST-1测定法和伤口闭合测定法,RAW-DEN杂交细胞的活力和运动性与未处理的RAW细胞相同。此外,蛋白质印迹分析表明,在用叠氮糖和树枝状大分子处理的RAW细胞中,信号分子AKT、p38和NFκB(p65)及其相应的活化(磷酸化)形式的表达水平没有显著变化,这表明杂交过程既没有诱导细胞应激反应,也没有改变正常的信号通路。综上所述,这项工作展示了应用生物正交化学创建细胞-纳米颗粒杂交体的可行性,并证明了这种细胞表面工程方法的非侵入性。