Gene Transfection of Bioactive Glass Fibers

doi:10.15541/jim20140074

Abstract

Abstract:

Bioactive glasses (BGs) exhibit potential applications for gene transfection because of their composition including Ca and P. Here, bioactive glass fibers (BGFs) with mesopores or hierarchical nanopores, were prepared by electrospinning process using BGs Sol-Gel precursor and its effect on mediating gene transfection was investigated. The results indicate that BGF acts as a gene vector by releasing Ca²⁺ and PO₄^3-, and then reunioning them along with plasmid DNA in Dulbecco's Modified Eagle’s Medium (DMEM). BGFs have a dose-dependent manner in transfection efficiency. When using 1 μg/mL plasmid, the transfection efficiency of BGF with concentration at 1000 μg/mL is higher than 50% of lipofectamine LTX_PLUS. The transfection mechanism of BGF is similar to that of calcium phosphate (CaP) system. Furthermore, BGF’s sufficient ions releasing ensures stability and effectiveness to be applied in gene transfection, which makes BGF a promising candidate for gene delivery in replace of traditional gene carrier, the nano-calcium phosphate system.

Key words: bioactive glass, fiber, gene transfection, calcium phosphate

CLC Number:

TQ174

LIU Hui, CHEN Xiao-Feng, LI Xian, LI Yu-Li, LIN Ze-Feng. Gene Transfection of Bioactive Glass Fibers[J]. Journal of Inorganic Materials, 2014, 29(10): 1023-1028.

Figures/Tables 8

Fig. 1 Schematic of the BGF preparation process

Fig. 2 SEM image (a) and TEM images of different inner structure (b,c) and structure after mineralization for 6 h (d) of BGFs

Fig. 3 Agar gel electrophoresis of pAcGFP-OA after complexing with BGF A-D: BGF concentrations at 1000, 700, 400 and 100 μg/mL, respectively

Fig. 4 Fluorescent images of 293T cell after transfection with GFP plasmid-contained BGF at different concentrations a-i: BGF concentration at 100, 200, 400, 600, 800, 1000, 1300, 1650 and 2000 μg/mL, respectively;. j: Merged image from fluorescence and bright field images of 293T cell after transfection by GFP plasmid-contained BGF at 400 μg/mL; k: Image of positive control transfected by Lipofectamine. Bars in all images are 100 μm

Fig. 5 SEM images of fibers with different compositions and fluorescence microscope (FM) images of 293T cells transfected by equivalent fibers

Fig. 6 SEM images of different type of bioactive glasses and fluorescence microscope (FM) images of 293T cells transfected by equivalent fibers

Table 1 Transfection efficiency of 293T cells measured by flow cytometry

Concentration (BGF) /(μg· mL^-1)	0	200	400	600	800	1000	600	600	0(Lipofectamine)
Amount of GFP Plasmid /(ng·mL^-1)	1	1	1	1	1	1	2	4	1
Transfection efficiency /%	0	0.12	0.68	5.74	14.40	21.30	10.40	15.70	38.30

Fig. 7 Ion release profiles of different types of BGs after soaking in DMEM for 6 h F: Fiber; S: microsphere; I: Irregular

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