Department of Otorhinolaryngology-Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kroener-Fresenius-Stiftung-Professorship, Universitätsklinikum Erlangen, 91054 Erlangen, Germany.
Department of Chemistry and Pharmacy, Organic Chemistry II, Friedrich-Alexander-University Erlangen-Nürnberg, 91058 Erlangen, Germany.
Molecules. 2022 Nov 1;27(21):7416. doi: 10.3390/molecules27217416.
Nanoformulations for delivering nucleotides into cells as vaccinations as well as treatment of various diseases have recently gained great attention. Applying such formulations for a local treatment strategy, e.g., for cancer therapy, is still a challenge, for which improved delivery concepts are needed. Hence, this work focuses on the synthesis of superparamagnetic iron oxide nanoparticles (SPIONs) for a prospective "magnetofection" application. By functionalizing SPIONs with an active catechol ester (CafPFP), polyethyleneimine (PEI) was covalently bound to their surface while preserving the desired nanosized particle properties with a hydrodynamic size of 86 nm. When complexed with plasmid-DNA (pDNA) up to a weight ratio of 2.5% pDNA/Fe, no significant changes in particle properties were observed, while 95% of the added pDNA was strongly bound to the SPION surface. The transfection in A375-M cells for 48 h with low amounts (10 ng) of pDNA, which carried a green fluorescent protein (GFP) sequence, resulted in a transfection efficiency of 3.5%. This value was found to be almost 3× higher compared to Lipofectamine (1.2%) for such low pDNA amounts. The pDNA-SPION system did not show cytotoxic effects on cells for the tested particle concentrations and incubation times. Through the possibility of additional covalent functionalization of the SPION surface as well as the PEI layer, Caf-PEI-SPIONs might be a promising candidate as a magnetofection agent in future.
纳米制剂作为疫苗将核苷酸递送到细胞中以及治疗各种疾病最近引起了极大的关注。将这种制剂应用于局部治疗策略,例如癌症治疗,仍然是一个挑战,需要改进的递送概念。因此,这项工作侧重于合成超顺磁氧化铁纳米粒子(SPIONs),用于有前途的“磁转染”应用。通过用活性儿茶酚酯(CafPFP)官能化 SPIONs,将聚乙烯亚胺(PEI)共价结合到其表面,同时保持所需的纳米级颗粒特性,水动力尺寸为 86nm。当与质粒 DNA(pDNA)复合时,重量比高达 2.5% pDNA/Fe 时,颗粒特性没有明显变化,而 95%的添加 pDNA 强烈结合到 SPION 表面。用低剂量(10ng)携带绿色荧光蛋白(GFP)序列的 pDNA 在 A375-M 细胞中转染 48 小时,转染效率为 3.5%。与低 pDNA 量(1.2%)的 Lipofectamine 相比,该值几乎高 3 倍。对于测试的颗粒浓度和孵育时间,pDNA-SPION 系统对细胞没有细胞毒性作用。通过 SPION 表面以及 PEI 层的进一步共价功能化的可能性,Caf-PEI-SPIONs 可能成为未来磁转染剂的有前途的候选物。