Raval Yash S, Samstag Anna, Taylor Cedric, Huang Guohui, Mefford Olin Thompson, Tzeng Tzuen-Rong Jeremy
Department of Biological Sciences, Clemson University, Clemson, SC 29634, USA.
Department of Materials Science and Engineering, Clemson University, Clemson, SC 29634, USA.
Nanomaterials (Basel). 2021 Sep 22;11(10):2465. doi: 10.3390/nano11102465.
We have previously demonstrated that iron oxide nanoparticles with dopamine-anchored heterobifunctional polyethylene oxide (PEO) polymer, namely PEO-IONPs, and bio-functionalized with sialic-acid specific glycoconjugate moiety (Neu5Ac(α2-3)Gal(β1-4)-Glcβ-sp), namely GM3-IONPs, can be effectively used as antibacterial agents against target . In this study, we evaluated the biocompatibility of PEO-IONPs and GM3-IONPs in a normal human colon cell line CCD-18Co via measuring cell proliferation, membrane integrity, and intracellular adenosine triphosphate (ATP), glutathione GSH, dihydrorhodamine (DHR) 123, and caspase 3/7 levels. PEO-IONPs caused a significant decrease in cell viability at concentrations above 100 μg/mL whereas GM3-IONPs did not cause a significant decrease in cell viability even at the highest dose of 500 μg/mL. The ATP synthase activity of CCD-18Co was significantly diminished in the presence of PEO-IONPs but not GM3-IONPs. PEO-IONPs also compromised the membrane integrity of CCD-18Co. In contrast, cells exposed to GM3-IONPs showed significantly different cell morphology, but with no apparent membrane damage. The interaction of PEO-IONPs or GM3-IONPs with CCD-18Co resulted in a substantial decrease in the intracellular GSH levels in a time- and concentration-dependent manner. Conversely, levels of DHR-123 increased with IONP concentrations. Levels of caspase 3/7 proteins were found to be significantly elevated in cells exposed to PEO-IONPs. Based on the results, we assume GM3-IONPs to be biocompatible with CCD-18Co and could be further evaluated for selective killing of pathogens in vivo.
我们之前已经证明,具有多巴胺锚定的异双功能聚环氧乙烷(PEO)聚合物的氧化铁纳米颗粒,即PEO-IONPs,并用唾液酸特异性糖缀合物部分(Neu5Ac(α2-3)Gal(β1-4)-Glcβ-sp)进行生物功能化,即GM3-IONPs,可以有效地用作针对靶标的抗菌剂。在本研究中,我们通过测量细胞增殖、膜完整性以及细胞内三磷酸腺苷(ATP)、谷胱甘肽(GSH)、二氢罗丹明(DHR)123和半胱天冬酶3/7水平,评估了PEO-IONPs和GM3-IONPs在正常人结肠细胞系CCD-18Co中的生物相容性。当浓度高于100μg/mL时,PEO-IONPs导致细胞活力显著下降,而即使在最高剂量500μg/mL时,GM3-IONPs也未导致细胞活力显著下降。在存在PEO-IONPs的情况下,CCD-18Co的ATP合酶活性显著降低,但在存在GM3-IONPs时未降低。PEO-IONPs还损害了CCD-18Co的膜完整性。相比之下,暴露于GM3-IONPs的细胞显示出明显不同的细胞形态,但没有明显的膜损伤。PEO-IONPs或GM3-IONPs与CCD-18Co的相互作用导致细胞内GSH水平以时间和浓度依赖性方式大幅下降。相反,DHR-123的水平随着IONP浓度的增加而升高。在暴露于PEO-IONPs的细胞中,发现半胱天冬酶3/7蛋白水平显著升高。基于这些结果,我们认为GM3-IONPs与CCD-18Co具有生物相容性,并可进一步评估其在体内选择性杀死病原体的能力。