Pharmacological evaluation of nasal delivery of selegiline hydrochloride-loaded thiolated chitosan nanoparticles for the treatment of depression

Figures & data

Figure 1. Schematic representation of TCS synthesis. Covalent attachment was achieved by the formation of amide bonds between the primary amino groups of CS and the carboxylic acid groups of TGA-mediated EDC chemistry.

Table I. Experimental protocol design for forced swim- and tail suspension-induced depression.

Force swim (depressive condition) treatment period
1st day 3rd day 5th day 7th day [FX]	14th day
Alternate days of force swim	Behavioural assessment
Tail suspension	Biochemical estimation
	Brain tissue drug concentration

Figure 2. FT-IR spectrum of TCS.

Table II. Optimization of polymer/TPP ratio, homogenization speed, homogenization time, and drug optimization.

Formulation code	Polymer conjugate TPP ratio	Homogenization speed	Homogenization time	Drug	EE	Avg. particle size	PDI
F1	2.5:1	5000	10	10	–	105 ± 21.47	0.328 ± 0.048
F2	5:1	5000	10	10	–	186 ± 21.45	0.214 ± 0.041
F3	7.5:1	5000	10	10	–	454 ± 19.54	0.319 ± 0.041
F4	10:1	5000	10	10	–	690 ± 45.21	0.327 ± 0.015
F5	5:1	5000	10	10	–	129 ± 15.24	0.341 ± 0.014
F6	5:1	6000	10	10	–	179 ± 12.37	0.217 ± 0.016
F7	5:1	7000	10	10	–	245 ± 19.14	0.364 ± 0.014
F8	5:1	8000	10	10	–	451 ± 15.17	0.045 ± 0.042
F9	5:1	6000	8	10	–	287 ± 16.91	0.251 ± 0.043
F10	5:1	6000	9	10	–	260 ± 23.54	0.264 ± 0.041
F11	5:1	6000	10	10	–	245 ± 20.78	0.287 ± 0.041
F12	5:1	6000	11	10	–	216 ± 15.61	0.284 ± 0.051
F13	5:1	6000	12	10	–	397 ± 31.84	0.213 ± 0.0519
F14	5:1	6000	11	4	54 ± 2.41	140 ± 24.34	0.341 ± 0.0457
F15	5:1	6000	11	8	61 ± 3.31	173 ± 24.34	0.214 ± 0.049
F16	5:1	6000	11	12	70 ± 2.71	215 ± 34.71	0.214 ± 0.0421
F17	5:1	6000	11	16	69 ± 3.14	359 ± 29.85	0.341 ± 0.0271

The bold values under “Average particle size” and “PDI” were found to be optimum at optimized polymer conjugate TPP ratio (5:1), optimized homogenized speed (6000 rpm), optimized homogenization time (910 min) and optimum drug concentration (12 mg).

Figure 3. In vitro drug release profile of the TCs and the non-modified CNPs.

Figure 4. Effect of TCNs on the immobility time in depression-induced rats. Values were expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs; ^ep ≤ 0.05 as compared to CNPs.

Figure 5. Effect of TCNs on the sucrose uptake in depression-induced rats. Values are expressed as mean ± SEM. **p ≤ 0.05 as compared to 7th day SPT; ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low); ^dp ≤ 0.05 as compared to TCNs (high).

Figure 6. Effect of TCNs on the locomotor activity in depression-induced rats. Values are expressed as mean ± SEM. *p ≤ 0.05 as compared to 7th day locomotor activity; ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline.

Figure 7. Effect of TCNs on nitrite level in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low).

Figure 8. Effect of TCNs on GSH in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low); ^dp ≤ 0.05 as compared to TCNs (high); ^ep ≤ 0.05 as compared to CNPs.

Figure 9. The effect of TCNs on LPO in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low); ^dp ≤ 0.05 as compared to TCNs (high).

Figure 10. The effect of TCNs on catalase in forced swim-induced depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low).

Figure 11. The effect of TCNs on SOD in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to selegiline; ^cp ≤ 0.05 as compared to TCNs (low); ^dp ≤ 0.05 as compared to TCNs (high).

Figure 12. Effect of TCNs on the level of complex I in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to TCNs (low); ^cp ≤ 0.05 as compared to TCNs (high).

Figure 13. Effect of TCNs on the level of complex II in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to TCNs (low); ^cp ≤ 0.05 as compared to TCNs (high).

Figure 14. The effect of TCNs on the level of complex III in depression-induced rats. Values are expressed as mean ± SEM. ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to TCNs (low); ^cp ≤ 0.05 as compared to TCNs (high).

Figure 15. The effect of TCNs on the level of complex IV in depression-induced rats. Values are expressed as mean ± SEM ^ap ≤ 0.05 as compared to disease control; ^bp ≤ 0.05 as compared to TCNs (low); ^cp ≤ 0.05 as compared to TCNs (high); ^dp ≤ 0.05 as compared to CNPs.

Figure 16. The amount of various formulations found in the brain, following intranasal administration. The values of drug concentration in the brain are expressed as mean ± SEM.