In vitro cytotoxicity and transfection efficiency of pDNA encoded p53 gene-loaded chitosan-sodium deoxycholate nanoparticles

Figures & data

Table 1 Mean particle size, polydispersity index (PDI), zeta potential, and entrapment efficiency % (EE%) of pDNA-loaded chitosan-sodium deoxycholate (CS-DS) nanoparticles

Formula	CS:DS (w:w)	pDNA/CS (w:w%)	Particle size ± SD (nm)	PDI ± SD	Zeta potential ± SD(mV)	EE% ± SD
F1	2:1	2.5	391.5±51.24	0.53±0.06	58.40±5.13	75.56±7.25
F2	2:1	5	405.1±46.39	0.56±0.09	53.31±4.52	82.88±6.89
F3	2:1	10	383.3±41.72	0.52±0.10	55.34±6.12	90.63±8.78
F4	2:1	20	377.3±45.84	0.60±0.09	57.22±4.83	91.67±7.43
F5	2:1	30	371.4±27.26	0.52±0.08	53.32±4.24	94.89±5.67
F6	1:1	2.5	264.5±21.45	0.41±0.09	47.15±3.87	72.23±6.88
F7	1:1	5	275.1±16.34	0.26±0.07	44.16±4.81	84.94±7.18
F8	1:1	10	223.9±17.82	0.32±0.06	44.34±3.64	86.84±5.54
F9	1:1	20	204.4±12.63	0.34±0.04	39.20±2.78	91.54±8.77
F10	1:1	30	198.6±19.22	0.24±0.07	40.50±3.84	92.88±6.39
F11	1:2	2.5	192.8±14.51	0.44±0.06	38.60±2.92	48.46±3.12
F12	1:2	5	173.3±15.36	0.32±0.06	37.42±3.33	54.54±5.64
F13	1:2	10	177.2±19.87	0.46±0.05	38.34±3.18	70.88±4.78
F14	1:2	20	154.9±11.48	0.35±0.04	41.56±5.11	76.74±6.12
F15	1:2	30	158.8±16.53	0.37±0.04	44.45±3.82	74.78±6.54
F16	1:3	2.5	131.3±18.34	0.42±0.08	38.60±2.74	38.25±3.25
F17	1:3	5	127.2±9.49	0.37±0.05	37.42±2.69	49.47±3.13
F18	1:3	10	105.3±16.73	0.52±0.04	34.34±2.84	53.73±3.27
F19	1:3	20	96.5±11.31	0.56±0.07	34.91±4.15	52.54±5.44
F20	1:3	30	116.1±9.84	0.41±0.06	35.20±3.67	55.76±5.33

Figure 1 Transmission electron microscopy image of pDNA-loaded chitosan-sodium deoxycholate nanoparticles (A) F10 and (B) F15.

Figure 2 Electrophoretic analysis of the free pDNA and pDNA loaded chitosan-sodium deoxycholate (CS-DS) nanoparticles, lane1: free pDNA, lane 2–6: pDNA-loaded CS-DS nanoparticles (F6–F10, respectively).

Figure 3 In vitro release profiles of pDNA from different formulations prepared from chitosan-sodium deoxycholate nanoparticles with 30% of pDNA (F5, F10, F15, and F20).

Table 2 Mean particle size, zeta potential, and entrapment efficiency % (EE%) of F10 and F15 before and after storage at 4°C

Storage period (month)	Formula	Mean particle size (nm) ± SD, n=3	Mean zeta potential (mV) ± SD, n=3	Mean EE% ± SD, n=3
0	F10	198.6±19.22	+40.5±3.84	92.88±6.39
	F15	158.9±16.53	+44.2±3.82	74.78±6.54
1	F10	191.2±16.50	+41.1±3.25	91.91±7.83
	F15	162.9±14.48	+46.1±4.94	72.42±8.24
3	F10	215.4±19.80	+41.6±4.76	88.32±5.54
	F15	171.6±15.62	+47.2±5.21	71.31±9.07

Figure 4 In vitro cytotoxicity on Caco-2 cells of (A) plain chitosan-sodium deoxycholate (CS-DS) nanoparticles (represented as CS concentration) and (B) pDNA-loaded CS-DS nanoparticles (F10) compared to naked pDNA.

Figure 5 Fold expression of p53 gene in Caco-2 cells treated with naked pDNA and pDNA-loaded chitosan-sodium deoxycholate nanoparticles (F10).

Figure 6 Mean p53 protein concentrations (±SD, n=3) in Caco-2 cells treated with naked pDNA and pDNA-loaded chitosan-sodium deoxycholate nanoparticles (F10).