National Engineering Research Center for Healthcare Devices, Guangdong Institute of Medical Instruments, Biomedical Engineering Institute, Jinan University, Guangzhou, 510632, People's Republic of China.
Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of South China Structural Heart Disease, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, School of Medicine, South China University of Technology, Guangzhou, 510080, People's Republic of China.
Int J Nanomedicine. 2021 Jun 21;16:4197-4208. doi: 10.2147/IJN.S315444. eCollection 2021.
Intracellular protein delivery is emerging as a potential strategy to revolutionize therapeutics in the field of biomedicine, aiming at treating a wide range of diseases including cancer, inflammatory diseases and other oxidative stress-related disorders with high specificity. However, the current challenges and limitations are addressed to either synthetically or biologically through multipotency of engineering, such as protein modification, insufficient delivery of large-size proteins, deficiency or mutation of proteins, and high cytotoxicity.
We prepared the nanocomposites by mixing protein with PEI1200 at a certain molar ratio and demonstrated that it can deliver proteins into living cells in high efficiency and safety through the following experiments, such as dynamic light scattering, fluorescent detection, agarose gel electrophoresis, ß-Galactosidase activity detection, immunofluorescence staining, digital fluorescent detection, cell viability assay and flow cytometry.
The self-assembly of PEI1200/protein nanocomposites with appropriate molar ratio (4:1 and 8:1) could provide efficiently delivery of active proteins to a variety of cell types in the presence of serum. The nanocomposites could continuously release protein up to 96 h in their desired intracellular locations. In addition, these nanocomposites were able to preserve protein activity while maintain low cytotoxicity (when final concentration <1 μg/mL).
Collectively, PEI1200-based delivery system provided an alternative strategy to direct protein delivery in high efficiency and safety, offering increased potential applications in clinical biomedicine.
细胞内蛋白质递呈技术作为一种潜在的策略，正在引发生物医学治疗领域的革命，旨在针对癌症、炎症性疾病和其他与氧化应激相关的疾病进行高特异性治疗。然而，目前的挑战和限制主要通过多功能工程来解决，例如蛋白质修饰、大尺寸蛋白质的递呈效率不足、蛋白质的缺乏或突变以及高细胞毒性。
我们通过将蛋白质与 PEI1200 以一定的摩尔比混合来制备纳米复合材料，并通过以下实验证明它可以高效、安全地将蛋白质递呈到活细胞中，例如动态光散射、荧光检测、琼脂糖凝胶电泳、β-半乳糖苷酶活性检测、免疫荧光染色、数字荧光检测、细胞活力测定和流式细胞术。
具有适当摩尔比（4:1 和 8:1）的 PEI1200/蛋白质纳米复合材料的自组装可以在血清存在的情况下，有效地将活性蛋白质递呈到各种细胞类型中。纳米复合材料可以在其所需的细胞内位置持续释放蛋白质长达 96 小时。此外，这些纳米复合材料能够在保持低细胞毒性的同时（当最终浓度<1μg/mL 时）保持蛋白质的活性。
总的来说，基于 PEI1200 的递呈系统提供了一种高效、安全的蛋白质递呈替代策略，为临床生物医学提供了更多的潜在应用。
