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Effects of grafting cell penetrate peptide and RGD on endocytosis and biological effects of Mg-CaPNPs-CKIP-1 siRNA carrier system in vitro

接枝 TAT 和 RGD 对 Mg-CaPNPs-CKIP-1 siRNA 载体系统的体外胞吞和生物学效应的影响

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Abstract

Calcium phosphate nanoparticles (CaPNPs) have good biocompatibility as gene carriers; however, CaPNPs typically exhibit a low transfection efficiency. Cell penetrate peptide (TAT) can increase the uptake of nanoparticles but is limited by its non-specificity. Grafting adhesion peptide adhesion peptide on carriers can enhance their targeting. The Plekho1 gene encodes casein kinase-2 interacting protein-1 (CKIP-1) , which can negatively regulate osteogenic differentiation. Based on the above, we produced a Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system via hydrothermal synthesis, silanization and adsorption. The effects of this carrier system on cell endocytosis and biological effects were evaluated by cell culture in vitro. The results demonstrate that CaPNPs with 7% Mg (60 nm particle size, short rod shape and good dispersion) were suitable for use as gene carriers. The carrier system boosted the endocytosis of MG63 cells and was helpful for promoting the differentiation of osteoblasts, and the dual-ligand system possessed a synergistic effect. The findings of this study show the tremendous potential of the Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system for efficient delivery into cells and osteogenesis inducement.

摘要

磷酸钙纳米粒 (CaPNPs) 作为基因载体具有良好的生物相容性;然而, CaPNPs 通常表现出较低的转染效率。细胞穿透肽 (TAT) 可以增加纳米颗粒的摄取, 但由于其非特异性而受到限制。纳米载体表面接枝细胞黏附肽 (精氨酸-甘氨酸-天冬氨酸, RGD) 可增强其靶向性。Plekho1 基因编码酪蛋白激酶-2 相互作用蛋白-1 (CKIP-1) 可负调控成骨分化。在此基础上, 本研究通过水热合成法、硅烷化法和吸附法制备了 Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA 载体系统。通过体外细胞培养, 评价了该载体系统对细胞胞吞及生物学效应的影响。结果表明, 掺镁7%的CaPNPs (粒径 60 nm, 短棒形, 分散性好) 适合作为基因载体。载体系统促进了 MG63 细胞的内吞作用, 有利于促进成骨细胞的分化, 且双配体系统具有协同作用。结果表明, Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA 载体系统在高效转运进入细胞和诱导成骨方面具有巨大的潜力。

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Authors and Affiliations

  1. Department of Stomatology, the Third Xiangya Hospital, Central South University, Changsha, 410013, China

    Man-fei Yi  (易曼菲), Liang-jian Chen  (陈良建), Hui-li He  (贺惠莉), Lei Shi  (石蕾), Chun-sheng Shao  (邵春生) & Bo Zhang  (张博)

  2. State Key Laboratory of Powder Metallurgy, Central South University, Changsha, 410083, China

    Liang-jian Chen  (陈良建)

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  1. Man-fei Yi  (易曼菲)
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Correspondence to Liang-jian Chen  (陈良建).

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Foundation item

Project (81571021) supported by the National Natural Science Foundation of China; Project (2018zzts944) supported by the Graduate Student Independent Exploration Innovation Fund of the Central South University, China; Projects (2015WK3012, 2018SK2017) supported by the Hunan Provincial Science and Technology Department, China; Project (20160301) supported by New Talent Project of the Third Xiangya Hospital of Central South University, China

Contributors

YI Man-fei contributed to the paper writing, designed the project, carried out data processing, and performed data analysis. CHEN Liang-jian, HE Huili, and SHI Lei contributed to the conception of the study. HE Hui-li, SHI Lei, SHAO Chun-sheng, and ZHANG Bo performed data analysis and offered some valuable suggestions for the contents of the manuscript. YI Man-fei, CHEN Liang-jian replied to reviewers’ comments and revised the final version.

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The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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Yi, Mf., Chen, Lj., He, Hl. et al. Effects of grafting cell penetrate peptide and RGD on endocytosis and biological effects of Mg-CaPNPs-CKIP-1 siRNA carrier system in vitro. J. Cent. South Univ. 28, 1291–1304 (2021). https://doi.org/10.1007/s11771-021-4697-7

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