Riboflavin immobilized Fe₃O₄ magnetic nanoparticles carried with n-butylidenephthalide as targeting-based anticancer agents

Figures & data

Figure 1. TEM images of (A) Fe₃O₄ MNPs and (B) Fe₃O₄@RFMP MNPs. Scale bar: 100 nm.

Figure 2. (A) Plot of the binding amount of BP (2.66 × 10^–4 M, 0.2 mL) onto Fe₃O₄@RFMP MNPs (0.1 mg) versus the incubation time. (B) The corresponding UV–Vis absorption spectra of the sample containing BP obtained before and after binding with Fe₃O₄@RFMP MNPs followed by rinse with the solution containing 6 × 10⁻³% TFA (pH ∼3) two times (W1 and W2). The incubation time was 10 min.

Figure 3. Examination of cell cytotoxicity of BP-Fe₃O₄@RFMP MNPs. Cell viability obtained by incubating BP-Fe₃O₄@RFMP MNPs (4 mg mL⁻¹) with (A) LNCaP cells (∼20000 cells mL^–1, 0.1 mL), (B) Hep G2 cells (∼20000 cells mL^–1, 0.1 mL), (C) T-47D cells (∼20000 cells mL^–1, 0.1 mL) and (D) NIH/3T3 cells (∼20000 cells mL^–1, 0.1 mL) for different times.

Figure 4. Bright field microscopic images obtained by treating LNCaP (A–H), Hep G2 (I–P), T-47D (Q–X) with media only (A, I and Q), Fe₃O₄ MNPs (∼4 mg mL⁻¹) (B, J and R), Fe₃O₄@RFMP MNPs (∼4 mg mL⁻¹) (C, K and S) and BP-Fe₃O₄@RFMP MNPs (∼4 mg mL⁻¹) (D, L and T) for 4 h followed by culturing another 68 h after renewing the media, then rinsed with aqueous NaCl (0.9%,1 mL ×3) and stained with Trypan blue and Hoechst 33342 dye before observation under microscopy. Panels (E)–(H), (M)–(P) and (U)–(X) were the corresponding images obtained under fluorescence microscopy. The scale bar is 10 μm. The exposure time was 6 ms.

Figure 5. Investigation of caspase 3 activity. Bar graphs of (A) LNCap, (B) Hep G2, (C) T-47D and (D) NIH/3T3 cells obtained by plotting the ratio of the fluorescence intensity at 505 nm of the resultant fluorescence spectra obtained after conducting different treatments followed by the caspase 3 activity assay to that obtained from the control samples (blank) according to the results shown in Supplementary Figure S6.