Evaluation of the cytotoxic effects of hyperthermia and 5-fluorouracil-loaded magnetic nanoparticles on human colon cancer cell line HT-29

Figures & data

Table 1. Measurement of hydrodynamic size and zeta potential of nanoparticles.

Samples	Mean diameter (nm)	Zeta potential ± S.D. (mV)
5-FU-loaded PLGA nanoparticles with iron  oxide core	40.6	−22.51 ± 1.62
PLGA nanoparticles with iron oxide core	61.4	−20.82 ± 1.16
5-FU-loaded PLGA nanoparticles without iron  oxide core	36.2	−27.12 ± 0.29
PLGA nanoparticles without iron oxide core	83.9	−15.44 ± 1.33

Figure 1. Transmission electron micrographs of (A): 5-FU-loaded PLGA nanoparticles with iron oxide cores with magnification ×167 000, (B): PLGA nanoparticles without iron oxide core with magnification ×60 000, (C) SAED image associated with the sample from figure (A).

Table 2. Iron content of the cells after 74 h incubation with free and different concentrations of 5-FU-loaded PLGA-coated iron oxide nanoparticles. N(0) mean free nanoparticles and N(1), N(5), N(10) mean the nanoparticles release 1, 5 and 10 μM 5-FU respectively during 74 h, Mean ± STD of three independent experiments. The particle number per cell has been quantitatively calculated.

Sample	Fe (ppm) ± SD	Particle number per cell
N (0)	0.034 ± 0.002	0
N (1)	0.242 ± 0.005	3.24 × 10^4
N (5)	1.259 ± 0.012	1.68 × 10^5
N (10)	2.64 ± 0.134	3.56 × 10^5

Figure 2. In vitro release profiles of 5-FU from PLGA nanoparticles with and without iron oxide core. The plot represents the mean ± standard deviation of the results, performed in triplicates.

Figure 3. Survival fraction of HT-29 cells in spheroid culture versus different concentrations of 5-FU in the presence and absence of hyperthermia at 43 °C for 1 h. Mean ± SEM of three independent experiments (n = 9).

Figure 4. Effects of different concentrations of 5-FU released from (A) PLGA nanoparticles with iron oxide core, (B) PLGA nanoparticles without iron oxide core, alone and in combination with hyperthermia at 43 °C for 1 h, on the survival fraction of HT-29 cells in spheroid culture. Mean ± SEM of three independent experiments (n = 9).

Figure 5. Survival fraction of HT-29 cells in spheroid culture versus different concentrations of free 5-FU or 5-FU-loaded PLGA nanoparticles with and without iron oxide core. Mean ± SEM of three independent experiments (n = 9).

Figure 6. survival fraction of HT-29 cells in spheroid culture versus different concentrations of 5-FU-loaded PLGA nanoparticles with and without iron oxide core in the presence of hyperthermia at 43 °C for 1 h. Mean ± SEM of three independent experiments (n = 9).

Table 3. The extent of cytotoxic effects as compared to the control observed in HT-29 cells on treatment with all forms of 5-FU, both types of nanoparticles alone or in combination with hyperthermia. Mean ± SD of 3 independent experiments (n = 9).

5-FU concentration Groups
Groups	1 μM	5 μM	10 μM
5-FU	31.6% ± 2.3	49% ± 2.6	75.5% ± 1.5
5-FU + Hyperthermia	53.1% ± 1.4	74.0% ± 1.2	86.0% ± 0.0
5-FU-loaded PLGA nanoparticle without iron oxide core	50.0% ± 1.7	68.0% ± 2.7	82.6% ± 0.6
5-FU-loaded PLGA nanoparticle without iron oxide core + Hyperthermia	68.6% ± 4.0	82.7% ± 4.2	88.3% ± 3.1
5-FU-loaded PLGA nanoparticle with iron oxide core	49.0% ± 4.4	70.3% ± 1.2	85.3% ± 2.5
5-FU-loaded PLGA nanoparticle with iron oxide core + Hyperthermia	67.0% ± 6.2	81.0% ± 6.1	97.3% ± 1.2