Preparation and characterization of magnetic nanoparticles containing Fe₃O₄-dextran-anti-β-human chorionic gonadotropin, a new generation choriocarcinoma-specific gene vector

Figures & data

Figure 1 Transmission electron micrograph of magnetic iron oxide (Fe₃O₄)-dextran-anti-β-HCG nanoparticles.

Abbreviations: HCG, human chorionic gonadotropin.

Table 1 Inhibitory rate of Fe₃O₄-dextran-anti-β-HCG nanoparticle to JEG-3 cells

	Control group	18.75 (μg/mL)	30 (μg/mL)	37.5 (μg/mL)	56.25 (μg/mL)	75 (μg/mL)
A570	0.374 ± 0.025	0.372 ± 0.031	0.368 ± 0.027	0.361 ± 0.023	0.310 ± 0.037	0.293 ± 0.034
Inhibitory rate (%)	0	0.5	1.6	3.5	17.1	21.7

Abbreviations: HCG, human chorionic gonadotropin; Fe₃O₄, magnetic iron oxide.

Table 2 Transfection efficiency of anti-β-HCG monoclonal antibody bound to nanoparticle

Antibody concentration (μg/mL)	Anti-β-HCG antibody (OD data)	Aldehyde nanoparticle (OD data)	Preservation rate (%)	Nonaldehyde nanoparticle (OD data)	Preservation rate (%)
150	0.441	0.385	87.3	0.058	13.2
100	0.372	0.341	91.7	0.050	13.4
80	0.316	0.282	89.2	0.041	13.0
50	0.281	0.253	90.0	0.039	13.9
10	0.227	0.208	91.6	0.031	13.7

Abbreviations: HCG, human chorionic gonadotropin; OD, optical density.

Figure 2 Immunofluorescence staining of A) aldehyde Fe₃O₄-dextran-anti-β-HCG nanoparticles, B) anti-β-HCG antibody, and C) nonaldehyde Fe₃O₄-dextran-anti-β-HCG nanoparticle.

Abbreviations: Fe₃O₄, magnetic iron oxide; HCG, human chorionic gonadotropin.

Figure 3 Electrophoresis of simple GAPDH AS-ODN (second track), nonaldehyde Fe₃O₄-dextran-anti-β-HCG-GAPDH AS-ODN (third track), aldehyde Fe₃O₄-dextran-anti-β-HCG-GAPDH AS-ODN (fourth track), and Fe₃O₄-dextran-anti-β-HCG nanoparticles (fifth track).

Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; AS-ODN, antisense oligodeoxynucleotides; Fe₃O₄, magnetic iron oxide; HCG, human chorionic gonadotropin.

Figure 4 Electrophoresis of simple GAPDH AS-ODN (second track), nonaldehyde Fe₃O₄-dextran-anti-β-HCG-GAPDH AS-ODN (third track), and aldehyde Fe₃O₄-dextran-anti-β-HCG-GAPDH AS-ODN (fourth track) that has been digested by DNase I.

Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; AS-ODN, antisense oligodeoxynucleotides; Fe₃O₄, magnetic iron oxide; HCG, human chorionic gonadotropin; DNase I, deoxyribonuclease I.

Table 3 Fluorescence intensity of nanoparticle mixtures in JEG-3 cells

Groups	Fluorescence intensity
I. Control group	3.31 ± 0.37
II. Fe3O4-dextran-anti-β-HCG nanoparticle complexes	9.69 ± 0.81
III. GAPDH AS-ODN	8.41 ± 0.72
IV. Fe3O4-dextran-anti-β-HCG-GAPDH AS-ODN complexes	26.57 ± 2.38
V. Liposome-GAPDH AS-ODN complexes	15.36 ± 1.25

Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; AS-ODN, antisense oligodeoxynucleotides; HCG, human chorionic gonadotropin; Fe₃O₄, magnetic iron oxide.

Table 4 Fluorescence intensity in different cells

Groups	Fluorescence intensity
JEG-3 cells	26.57 ± 2.38
JAR cells	26.35 ± 2.27
RL95-2 cells	17.24 ± 1.06
Hela cells	16.91 ± 1.51
HepG2 cells	13.26 ± 1.20
Lovo cells	13.77 ± 1.18

Table 5 Distribution of Fe₃O₄-dextran-anti-β-HCG nanoparticles in nude mice (mg/g)

Group	Heart	Liver	Spleen	Lung	Kidney	Choriocarcinoma tumor
I	0.614 ± 0.05	1.351 ± 0.16	0.753 ± 0.07	0.663 ± 0.04	0.356 ± 0.02	5.959 ± 0.38
II	0.598 ± 0.05	2.994 ± 0.19	1.805 ± 0.15	0.586 ± 0.05	0.394 ± 0.03	4.112 ± 0.39
III	0.718 ± 0.09	6.398 ± 0.65	2.991 ± 0.24	0.697 ± 0.07	0.297 ± 0.02	1.262 ± 0.11

Abbreviations: HCG, human chorionic gonadotropin; Fe₃O₄, magnetic iron oxide.

Figure 5 Theory of how Fe₃O₄-dextran-anti-β-HCG nanoparticles are prepared.

Abbreviations: Fe₃O₄, magnetic iron oxide; HCG, human chorionic gonadotropin; BSA, bovine serum albumin.