In Vitro Anticancer Effects of Doxorubicin Loaded BSA-dextran Nanoparticles with Different Molecular Weights of Dextran and Different Dextran Conjugation Degrees†

doi:10.7503/cjcu20130708

Abstract

Abstract:

Bovine Sercurn albumin-dextran(BSA-dextran) conjugates were prepared with different molecular weights of dextran and different molar ratios of BSA to dextran via Maillard reaction. Doxorubicin loaded BSA-dextran nanoparticles were fabricated by changing the pH and then temperature of the mixture. The nanoparticles with 5×10³ and 10×10³ dextran have a size about 60 nm, and the nanoparticles with 62×10³ dextran have a size about 200 nm. The doxorubicin loading efficiency is in the range of 81%—98%, and the loading amount is 7.4%—16.9%. In vitro cell viability investigation confirms the excellent biocompatibility of BSA-dextran conjugates. Compared with free doxorubicin, the nanoparticles can enhance the cellular internalization of the loaded doxorubicin and they show lower anticancer activity at lower doxorubicin concentrations because of the slower release of the loaded doxorubicin. In comparison with the nanoparticles with 62×10³ and 10×10³ dextran, the nanoparticles with 5×10³ dextran and higher dextran conjugation degree show better cellular internalization and better anticancer activity in vitro due to their smaller size, denser dextran brush surface, certain free doxorubicin concentration, and faster release rate of the loaded doxorubicin.

Key words: Dextran, Bovine Sercurn albumin(BSA), Drug delivery, Nanoparticles, Anticancer activity, Doxorubicin

CLC Number:

O636
O629

TrendMD:

HAO Hequn, YAO Ping. In Vitro Anticancer Effects of Doxorubicin Loaded BSA-dextran Nanoparticles with Different Molecular Weights of Dextran and Different Dextran Conjugation Degrees^†[J]. Chem. J. Chinese Universities, 2014, 35(3): 652.

Figures/Tables 8

Table 1 BSA-DEX conjugates produced from different molecular weights of dextran and different molar ratios of BSA to dextran

Sample	M_w(DEX)	n(BSA):n(DEX) in feed	m(BSA):m(DEX) in feed	Average DEX number conjugated to each BSA
B2-D1-62k	62×10³	2:1	1:0.47	0.484^[25]
B1-D1-62k		1:1	1:0.94
B1-D2-62k		1:2	1:1.88
B1-D3-10k	10×10³	1:3	1:0.45	2.4
B1-D4-10k		1:4	1:0.61	2.7
B1-D5-10k		1:5	1:0.76	3.1
B1-D6-5k	5×10³	1:6	1:0.45
B1-D7-5k		1:7	1:0.53
B1-D8-5k		1:8	1:0.61	5.9
B1-D9-5k		1:9	1:0.68	7.5

Table 2 Size distribution, loading efficiency(LE), and loading amount(LA) results of DOX loaded nanoparticles before and after 30 d storagea

Sample	Fresh prepared				After 30 d
Sample	D_h/nm	PDI	LE(%)	LA(%)	D_h/nm	PDI
DOX/B2-D1-62k	198±14	0.13±0.04	92.4±1.9	15.7±0.5	194±15	0.11±0.02
DOX/B1-D1-62k^b	140±6	0.15±0.01	91.0±1.8	11.7±0.2	—	—
DOX/B1-D2-62k^b	108±8	0.14±0.02	84.8±0.8	7.4±0.1	—	—
DOX/B1-D3-10k	106±8	0.24±0.04	98.0±0.2	16.9±0.1	104±8	0.24±0.02
DOX/B1-D4-10k^c	68±8	0.24±0.02	95.9±0.4	14.9±0.1	72±10	0.22±0.04
DOX/B1-D5-10k	58±6	0.28±0.08	95.6±0.6	13.7±0.1	58±4	0.29±0.04
DOX/B1-D6-5k	198±2	0.31±0.01	84.3±0.6	14.5±0.3	208±8	0.34±0.02
DOX/B1-D7-5k	114±2	0.25±0.02	83.9±0.4	14.0±0.2	118±6	0.27±0.01
DOX/B1-D8-5k	76±2	0.24±0.01	82.8±0.3	13.3±0.2	74±6	0.25±0.02
DOX/B1-D9-5k	66±2	0.37±0.01	81.1±0.7	12.5±0.1	70±4	0.36±0.03

Fig.1 TEM images of DOX/B2-D1-62k(A), DOX/B1-D5-10k(B) and DOX/B1-D9-5k(C) nanoparticles Samples (B) and (C) were negative stained with phosphotungstic acid.

Fig.2 Accumulative release of DOX from DOX/B2-D1-62k(a), DOX/B1-D5-10k(b) and DOX/B1-D9-5k(c) nanoparticles in PBS(A) and 0.1 mol/L pH=5.0 acetate buffer(B) Free DOX solution was assayed as a control(curve d).

Fig.3 Flow cytometry analysis of the cellular uptake of control(a), free DOX(b), DOX/B2-D1-62k(c), DOX/B1-D5-10k(d) and DOX/B1-D9-5k(e) nanoparticles with a fixed DOX concentration of 2 mg/mL after 2 h incubation at 37 ℃

Fig.4 Confocal fluorescence microscopy images of cellular uptake of free DOX(A, B) and DOX/B1-D9-5k nanoparticles(C, D) with a fixed DOX concentration of 2 μg/mL after 2 h incubation at 37 ℃(A, C) and 4 ℃(B, D)

Fig.5 Cell viability against B2-D1-62k, B1-D5-10k and B1-D9-5k conjugates after 48 h incubation with different BSA concentrations

Fig.6 Cell viability against free DOX solution as well as DOX/B2-D1-62k, DOX/B1-D5-10k and DOX/B1-D9-5k nanoparticles after 48 h(A) and 72 h(B) incubation with different DOX concentrations* P>0.05 compared with DOX/B2-D1-62k group; ** P<0.05 compared with DOX/B2-D1-62k group; *** P<0.01 compared with DOX/B2-D1-62k group.

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