Construction of Polyethyleneimine-modified Dual-mesoporous Silica Gene Carrier

doi:10.15541/jim20190099

Abstract

Abstract:

To develop a new kind of non-viral inorganic gene vector, embedded dual-mesoporous silica spheres (EDMSNs) was modified with polyethyleneimine (PEI) or 3-aminopropyltriethoxysilane (APTMS). Then gene loading and transfection ability of the obtained EDMSNs-PEI and EDMSNs-NH₂ was tested by choosing the pCMV-EGFP-N1 plasmids (pDNA) as a gene model. Particle morphologies, particle sizes, zeta potentials, and pore structure parameters of EDMSNs-NH₂ and EDMSNs-PEI were characterized by transmission electron microscopy, dynamic light scattering and nitrogen adsorption-desorption. Both EDMSNs-NH₂ and EDMSNs-PEI display obvious dual-mesoporous structure and well-defined spherical morphology. Average dynamic sizes of EDMSNs-NH₂ and EDMSNs-PEI are 343.2 and 338.9 nm, while zeta potentials of EDMSNs-NH₂ and EDMSNs-PEI are +18 and +43 mV, respectively. The results of gel electrophoresis, CCK-8 assay, and fluorescence microscope show that the pDNA loading amount of EDMSNs-PEI is 8%, much higher than that (1%) of EDMSNs-NH₂. In addition, EDMSNs-PEI exhibits lower cytotoxicity than that of pure PEI polymers and lipofectamine2000. When the mass ratio of EDMSNs- PEI/pDNA is 33 : 1, the transfection rate of the 293T cells by EDMSNs-PEI/pDNA reaches a maximum at 72 h. In conclusion, EDMSNs-PEI, compared with EDMSNs-NH2, has a higher positive potential and pDNA loading capacity, which can be used as a promising non-viral inorganic gene vector for the treatment of diseases such as malignant glioma.

Key words: embedded dual-mesoporous silica spheres, polyethyleneimine, gene therapy, malignant glioma

CLC Number:

TQ174

FANG Mei-Rong,QIN Li-Mei,JIA Xiao-Bo,LI Yong-Sheng,NIU De-Chao,HU Ze-Lan. Construction of Polyethyleneimine-modified Dual-mesoporous Silica Gene Carrier[J]. Journal of Inorganic Materials, 2020, 35(2): 187-192.

Figures/Tables 6

Fig. 1 TEM images of EDMSNs-PEI (a) and EDMSNs- NH2 (b)

Fig. 2 Zeta potentials (a) and particle size distributions (b) of EDMSNs, EDMSNs-PEI and EDMSNs-NH2

Fig. 3 N2 adsorption-desorption isotherms and the corresponding DFT pore size distributions of EDMSNs-PEI (a-b) and EDMSNs-NH2 (c-d)

Fig. 4 Electrophoresis diagram of EDMSNs-PEI/pDNA and EDMSNs-NH2/pDNA nanoparticles at different weight ratios (a), DNaseI digestion electrophoresis diagram (b) and zeta potentials (c) of EDMSNs-PEI/pDNA with M indicating DNA marker and C indicating naked pDNA Weight ratio=weight ratios of EDMSNs-PEI and pDNA.1, 2, 3, 4, 5, 6, 7: weight ratios of 5 : 1, 12.5 : 1, 25 : 1, 33 : 1, 50 : 1, 100 : 1, 200 : 1; 8, 9, 10, 11, 12, 13: weight ratios of 200 : 1 ,100 : 1, 50 : 1, 33 : 1, 25 : 1, 12.5 : 1 for EDMSNs-PEI/pDNA nanocomplexes treated with DnaseI

Fig. 5 Cytotoxicity assay of PEI 25kD, LIP2K and EDMSNs-PEI in 293T (a) and u87(b). * p<0.01, EDMSNs-PEI

Fig. 6 Transfection efficacy of EDMSNs-PEI/pDNA nanoparticle in 293T cell lines.

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