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Original Research

Preparation of biocompatible heat-labile enterotoxin subunit B-bovine serum albumin nanoparticles for improving tumor-targeted drug delivery via heat-labile enterotoxin subunit B mediation

, , , , &
Pages 2149-2156 | Published online: 06 May 2014

Figures & data

Figure 1 Schematic formation of 5-FU-loaded BSA-LTB nanoparticles.

Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 1 Schematic formation of 5-FU-loaded BSA-LTB nanoparticles.Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 2 Fluorescence microscope images of identification of LTB with immunofluorescence.

Notes: (A) BSA-LTB nanoparticles; (B) BSA nanoparticles.

Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 2 Fluorescence microscope images of identification of LTB with immunofluorescence.Notes: (A) BSA-LTB nanoparticles; (B) BSA nanoparticles.Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 3 Morphological shapes (A) and particle sizes (B) of 5-FU-loaded BSA-LTB nanoparticles.

Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 3 Morphological shapes (A) and particle sizes (B) of 5-FU-loaded BSA-LTB nanoparticles.Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 4 In vitro release profile of BSA-LTB nanoparticles in phosphate-buffered saline (pH 5.4 and pH 7.4 at 37°C) for 48 hours.

Note: Data are expressed as the mean ± standard deviation (n=3).

Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 4 In vitro release profile of BSA-LTB nanoparticles in phosphate-buffered saline (pH 5.4 and pH 7.4 at 37°C) for 48 hours.Note: Data are expressed as the mean ± standard deviation (n=3).Abbreviations: BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B.

Figure 5 In vitro uptake ability of NPs.

Notes: Fluorescent microscopy analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells. The scale bar is 50 μm (A). Fluorescence spectrum analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells (n=3, *P<0.05, versus the corresponding BSA NPs) (B). Fluorescence spectrum analysis of the uptake of NPs labeled by FITC incubated with SMMC-7721 cells for 6 hours in media containing different amounts of free LTB (n=3, *P<0.05, versus the corresponding NP groups in media containing 5 μg/mL of LTB) (C).

Abbreviations: BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NP, nanoparticle.

Figure 5 In vitro uptake ability of NPs.Notes: Fluorescent microscopy analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells. The scale bar is 50 μm (A). Fluorescence spectrum analysis of the uptake of FITC-labeled BSA NPs and FITC-labeled BSA-LTB NPs in SMMC-7721 cells (n=3, *P<0.05, versus the corresponding BSA NPs) (B). Fluorescence spectrum analysis of the uptake of NPs labeled by FITC incubated with SMMC-7721 cells for 6 hours in media containing different amounts of free LTB (n=3, *P<0.05, versus the corresponding NP groups in media containing 5 μg/mL of LTB) (C).Abbreviations: BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NP, nanoparticle.

Figure 6 Viability of cells after incubation with either free 5-FU, 5-FU-loaded BSANPs, or 5-FU-loaded BSA-LTB NPs, at various 5-FU concentrations (3.75–30 μg/mL).

Notes: (A) After 24 hours; (B) after 48 hours; (C) after 72 hours (n=3)

Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles.

Figure 6 Viability of cells after incubation with either free 5-FU, 5-FU-loaded BSANPs, or 5-FU-loaded BSA-LTB NPs, at various 5-FU concentrations (3.75–30 μg/mL).Notes: (A) After 24 hours; (B) after 48 hours; (C) after 72 hours (n=3)Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles.

Figure 7 Cell apoptosis determined by Annexin V-FITC staining.

Notes: Flow cytometer analysis of the apoptotic and necrotic cells (Q1: necrotic; Q2: late apoptotic; Q3: live; Q4: early apoptotic) after 48 hours of incubation with the free 5-FU, 5-FU-loaded BSA NPs, and 5-FU-loaded BSA-LTB NPs, respectively. Results are expressed as means ± standard deviation (n=3).

Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles; PI, propidium iodide.

Figure 7 Cell apoptosis determined by Annexin V-FITC staining.Notes: Flow cytometer analysis of the apoptotic and necrotic cells (Q1: necrotic; Q2: late apoptotic; Q3: live; Q4: early apoptotic) after 48 hours of incubation with the free 5-FU, 5-FU-loaded BSA NPs, and 5-FU-loaded BSA-LTB NPs, respectively. Results are expressed as means ± standard deviation (n=3).Abbreviations: 5-FU, 5-fluorouracil; BSA, bovine serum albumin; FITC, fluorescein isothiocyanate; LTB, heat-labile enterotoxin subunit B; NPs, nanoparticles; PI, propidium iodide.