Intercalation of shRNA-plasmid in Mg-Al layered double hydroxide nanoparticles and its cellular internalization for possible treatment of neurodegenerative diseases

被引:9
作者
Acharya, Rituparna [1 ,2 ]
Alsharabasy, Amir Mohammed [3 ]
Saha, Suman [1 ]
Rahaman, Sk Hasanur [1 ]
Bhattacharjee, Arnab [1 ]
Halder, Suranjana [4 ]
Chakraborty, Monisha [2 ]
Chakraborty, Jui [1 ]
机构
[1] Cent Glass & Ceram Res Inst, CSIR, Bioceram & Coating Div, 196 Raja SC Mullick Rd, Kolkata 700032, India
[2] Jadavpur Univ, Sch Biosci & Engn, 188 Raja SC Mullick Rd, Kolkata 700032, India
[3] Egyptian Atom Energy Author, Natl Ctr Radiat Res & Technol, Radiat Biol Dept, POB 29,3 Ahmed El Zomor St, Cairo, Egypt
[4] Jadavpur Univ, Dept Lifesci & Biotechnol, 188 Raja SC Mullick Rd, Kolkata 700032, India
关键词
Mg-Al layered double hydroxide (LDH); shRNA-plasmid-LDH nanoconjugate; TNF alpha assay; Storage and enzyme stability; Release kinetics; NONCODING RNAS; GENE; DNA; SYSTEM; EXPRESSION; STABILITY; INJECTION; CROSS; MICE;
D O I
10.1016/j.jddst.2019.05.008
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
In the present work, nanoconjugates of shRNA-plasmid and a non-viral nanoceramic vector, e.g., Mg-Al layered double hydroxide (Mg-Al LDH), were synthesized and intercalated. Subsequently, these particles with an average size of 40-60 nm, were transfected into mammalian neuroblastoma cells (SH-SY5Y). The as prepared Mg-Al LDH was able to protect the incorporated shRNA-plasmid against a range of pH values, DNaseI, endonucleases, and serum components. To test the applicability of the nanoconjugate for future in-vivo studies, serum from three different model experimental animals viz, mouse, rat and guinea pig was used for the serum protection study. Additionally, we showed that prolonged storage at different temperatures does not affect the quality of the nanoconjugate. Using this nanoconjugate to transform cells, a maximum internalization of similar to 26% at 24h was achieved. Lastly, we demonstrated effective and safe delivery of the plasmid by measuring GFP production and shRNA-induced knockdown of TNF alpha.
引用
收藏
页码:500 / 508
页数:9
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