Cationization of Inulin via Atom Transfer Radical Polymerization for Gene Delivery†

doi:10.7503/cjcu20140087

Abstract

Abstract:

In order to develop new biocompatible gene delivery system, in the present study, a novel non-virus gene delivery vector PDIN was synthesized by grafting poly(DMAEMA) onto the backbone of inulin oligosaccharide via atom transfer radical polymerization(ATRP). ¹H NMR, particle size analysis and zeta-potential measurements, TEM and gel electrophoresis were performed to characterize PDIN and PDIN/pDNA complexes. At different N/P ratios(2, 1, 0.5 and 0.25), PDIN could condense pDNA into complex nanoparticles of 75.55 nm to 104 nm in sizes. Gel electrophoresis retardation assay demonstrated that PDIN could interact with pDNA via electrostatic interaction excellently even in the presence of high concentration heparin. The results of MTT assay displayed that the PDIN/pDNA complexes showed comparable cytotoxicity against MCF-7, HeLa, COS7 and HepG2 cells with that of Lipofectamine2000/pDNA complex. PDIN showed good biocompatibility due to its low hemolytic rate. The activity of β-galactosidase expressed in COS7 cells revealed that the highest transfection efficiency[(3.36±0.74) U/mg protein] of PDIN as a new gene delivery carrier was obtained at N/P ratio of 1, which was comparable with that of Lipofectamine2000[(4.33±0.77) U/mg]. These results suggested that inulin oligosaccharide-grafted poly(DMAEMA)(PDIN) might be an excellent biocompatible candidate for gene delivery systems.

Key words: Inulin, Atom transfer radical polymerization, pGFP, pβ, gal, Non-virus gene vector

CLC Number:

TrendMD:

WANG Jingyun, Fu Xue, BAO Yongming. Cationization of Inulin via Atom Transfer Radical Polymerization for Gene Delivery^†[J]. Chem. J. Chinese Universities, 2014, 35(10): 2124.

Figures/Tables 7

Scheme 1 Synthetic routes of PDIN

Table 1 Particle size and ζ potential of PDIN/pDNA complexes

N/P	Mean diameter/nm(PDI)	ζ potential/mV	N/P	Mean diameter/nm(PDI)	ζ potential/mV
0.25	104±4.08(0.183)	2.36±0.12	1	85.4±5.42(0.236)	5.32±0.14
0.5	96.59±8.51(0.192)	4.24±0.28	2	75.58±0.43(0.232)	6.13±0.37

Fig.1 TEM image of PDIN polymer

Fig.2 Hemolysis test results for PDIN and PDIN/pDNA (A) PDIN at different concentrations. a. 1 mg/mL PDIN; b. 2 mg/mL PDIN; c. 1 mg/mLPEI 25K; d. 2 mg/mL PEI 25K; (B) PDIN/pDNA complex(N/P=2) at 1 mg/mL of PDIN. a. PDIN/pDNA; b. PEI 25K/pDNA.

Fig.3 Cell viability of PDIN/pDNA complexes on different cells Cells were treated with PDIN/pDNA or Lipo2000/pDNA complexes for 48 h at 37 ℃. Cell viability was measured by the MTT method. * P<0.05; ** P<0.01. a. Control; b. N/P=0.25; c. N/P=0.5; d. N/P=1; e. N/P=2; f. Lipo2000.

Fig.4 Transfection efficiency of PDIN/pGFP at different N/P ratios in COS7 cells(200×)

Fig.5 Transfection efficiency of PDIN/pβgal at different N/P ratios a. N/P=0.25; b. N/P=0.5; c. N/P=1; d. N/P=2; e. pDNA; f. Lipo2000/pDNA.

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