Synthesis, characterization and anticancer activity of vincristine loaded folic acid-chitosan conjugated nanoparticles on NCI-H460 non-small cell lung cancer cell line

Figures & data

Fig. 1 Vincristine loaded folic acid-chitosan conjugated nanoparticles of 4:25 formulation (a) average size and polydispersity index, and, (b) and zeta potential.

Table 1 Average size, polydispersity index and zeta potential of vincristine loaded folic acid-chitosan conjugated nanoparticles at pH 5.

Formulations	Average size (nm)	Polydispersity index	Zeta potential (mV)
Blank FA-CS NPs	95.59 ± 0.52	0.229 ± 0.31	+8.91 ± 0.36
1:25	121.9 ± 0.95	0.271 ± 0.76	+9.11 ± 0.29
2:25	129.8 ± 0.30	0.250 ± 0.02	+9.38 ± 0.61
3:25	124.9 ± 0.96	0.249 ± 0.02	+8.96 ± 0.50
4:25	130.0 ± 0.89	0.221 ± 0.01	+9.83 ± 1.41

Table 2 Encapsulation efficiency (%) and loading capacity (%) of vincristine loaded folic acid-chitosan conjugated nanoparticles.

S. No.	Formulations	Encapsulation efficiency (%)	Loading capacity (%)
1.	1:25	50 ± 0.58	10.90 ± 0.45
2.	2:25	70 ± 0.50	12.24 ± 0.51
3.	3:25	80 ± 0.45	25.40 ± 0.38
4.	4:25	95 ± 0.35	48.65 ± 0.47

± Standard deviation (n = 3).

Table 3 Effect of storage on average size, polydispersity index and zeta potential of blank and vincristine loaded folic acid-chitosan conjugated nanoparticles in sodium phosphate buffer (pH 7.4) at 4 °C after 10 days.

Formulations	After 10 days
	Average size (nm)	Polydispersity index	Zeta potential (mV)
Blank FA-CS NPs	203.0 ± 0.50	0.329 ± 0.10	+3.94 ± 0.25
1:25	220.0 ± 0.96	0.259 ± 0.45	+2.76 ± 0.35
2:25	341.1 ± 0.58	0.611 ± 0.09	+3.74 ± 0.40
3:25	225.3 ± 0.34	0.547 ± 0.04	+4.25 ± 0.80
4:25	208.2 ± 0.43	0.336 ± 0.03	+3.83 ± 0.28

± Standard deviation (n = 3).

Fig. 2 FTIR spectra showing shifting in peaks due to loading of vincristine (a) blank FA-CS NPs and vincristine loaded FA-CS NPs at different formulations, (b) 1:25, (c) 2:25, (d) 3:25, (e) 4:25

Fig. 3 SEM images of nanoparticles showing spherical shaped (a) blank FA-CS NPs and vincristine loaded FA-CS NPs at different formulations, (b) 1:25, (c) 2:25, (d) 3:25, (e) 4:25.

Fig. 4 Transmission electron microphotographs of blank and vincristine loaded FA-CS nanoparticles showing encapsulation of vincristine (arrows) (a) blank and different formulations, (b) 1:25, (c) 2:25, (d) 3:25, (e) 4:25.

Fig. 5 Cumulative release (%) behavior of vincristine from folic acid-chitosan conjugated nanoparticles for different formulations (1:25, 2:25, 3:25, 4:25) showing slow and sustained release in phosphate buffered saline (pH 6.7).

Fig. 6 Cellular uptake of nanoparticles in NCI-H460 cells 40× (a) control, (b) enhanced fluorescence with vincristine loaded folic acid-chitosan conjugated nanoparticles.

Table 4 Cell viability (%) of blank and vincristine loaded FA-CS nanoparticles at different concentrations.

S. No.	Concentration (mg/ml)	Cell viability (%)
		Blank FA-CS NPs	Vincristine loaded FA-CS NPs
1.	5	87.14	46.47
2.	10	71.49	32.78
3.	15	52.89	43.15
4.	20	40.25	38.17
5.	25	47.25	36.51

Fig. 7 Fluorescent micrographs showing effect of nanoparticles in NCI-H460 cells (i) intracellular ROS level, (ii) mitochondrial transmembrane potential, (iii) apoptotic morphological changes.

Fig. 8 Light microphotographs of erythrocytes aggregation (a) control, (b) vincristine loaded folic acid-chitosan conjugated nanoparticles.