Brain targeted oral delivery of doxycycline hydrochloride encapsulated Tween 80 coated chitosan nanoparticles against ketamine induced psychosis: behavioral, biochemical, neurochemical and histological alterations in mice

Figures & data

Table 1. Composition, particle size and DEE of DCNP as per experimental design.

		Coded factor levels
Formulation code	Trial number	X1	X2	Particle size (nm)	DEE (%)
7	1	−1	1	248	75.15
12	2	0	0	209	78.09
5	3	0	0	213	79.12
6	4	1	0	237	82.34
9	5	1	1	228	85.67
10	6	0	0	198	77.45
2	7	0	−1	243	74.56
8	8	0	1	218	76.16
11	9	0	0	202	76.19
4	10	−1	0	236	72.38
13	11	0	0	215	79.02
3	12	1	−1	257	86.62
1	13	−1	−1	229	72.78
Coded level	−1	0	+1
X1: Chitosan (%)	0.05	0.15	0.25
X2: Tween 80 (%)	1.0	1.5	2.0

Figure 1. (a) 3D response surface graph of DCNP showing the effect of chitosan and Tween 80 on particle size. (b) 3 D response surface graph of DCNP showing the effect of chitosan and Tween 80 on DEE. (c) Overlay plot showing the location of optimized batch of DCNP.

Figure 2. TEM image of DCNP_opt.

Figure 3. FTIR spectra of (A) Chitosan (B) Doxycycline hydrochloride (C) DCNP_opt.

Table 2. Effect of doxycycline hydrochloride and DCNP_opt on various behavioral and biochemical parameters.

	Vehicle	Keta (50 mg/kg)	Keta + Olz (5 mg/kg)	Keta + Placebo (50 mg/kg)	Keta + Doxy (25 mg/kg)	Keta + Doxy (50 mg/kg)	Keta + DCNPopt (25 mg/kg)
Locomotor counts	283.33 ± 9.61	502.66 ± 7.46*	359.33 ± 10.44^a	490.50 ± 11.55	444.83 ± 19.61	394.16 ± 8.42^a	304.33 ± 23.4^a^,^x^,^q
Stereotyped behaviors	2.50 ± 0.34	27.66 ± 1.14*	7.83 ± 0.6^a	26.50 ± 0.84	21.16 ± 1.74^b	18.66 ± 1.47^a	11.16 ± 1.49^a^,^x^,^q
Immobility duration (s)	72.5 ± 1.61	149.83 ± 1.88*	81.83 ± 1.49^a	146.33 ± 1.99	125.16 ± 2.93^a	114.50 ± 3.08^a	81.16 ± 5.16^a^,^x^,^p
SDL (s)	237.66 ± 14.55	76.83 ± 3.98*	186 ± 2.22^a	74.16 ± 4.01	80.16 ± 4.52	80.50 ± 4.52	90.83 ± 3.17^#
TNF-α (ng/ml)	125.16 ± 3.65	499.66 ± 9.62*	128.00 ± 2.58^a	495.33 ± 12.02	378.66 ± 5.56^a	330.33 ± 15.51^a	147.33 ± 4.77^a^,^x^,^p
GSH (nmol/mg protein)	0.0051 ± 0.00020	0.0015 ± 0.00011*	0.0051 ± 0.00016^a	0.0016 ± 9.38E-50	0.0029 ± 2.72E-04^a	0.0038 ± 0.00011^a	0.0053 ± 1.69E-03^a^,^x^,^p
MDA (nmol/mg protein)	0.252 ± 0.014	0.563 ± 0.019*	0.228 ± 0.011^a	0.558 ± 0.020	0.424 ± 0.010^a	0.396 ± 0.007^a	0.208 ± 0.08^a^,^x^,^p
AchE (nmol/min/g protein)	38.82 ± 1.24	65.24 ± 2.05*	36.68 ± 1.92^a	64.79 ± 3.90	64.74 ± 1.68	64.25 ± 3.11	62.67 ± 1.41^#
Dopamine (pg/mg brain)	427.38 ± 27.11	981.73 ± 30.68*	429.62 ± 29.03^a	978.40 ± 25.40	688.33 ± 21.12^a	642.93 ± 27.21^a	435.16 ± 21.55^a^,^x^,^p
GABA (μg/g brain)	192.58 ± 6.75	107.80 ± 6.06*	208.88 ± 8.80^a	106.42 ± 6.11	147.50 ± 3.22^a	155.99 ± 3.56^a	211.30 ± 5.62^a^,^x^,^p

Values are expressed as mean ± SEM (n = 6).

* p < .001 vs vehicle group.

^a p < .001 vs ketamine group.

^x p < .001 vs Keta + Doxy (25 mg/kg).

^p p < .001 vs Keta + Doxy (50 mg/kg).

^q p < .01 vs Keta + Doxy (50 mg/kg).

^# p < .001 vs olanzapine.

Keta: Ketamine; Olz: Olanzapine; Doxy: Doxycycline hydrochloride; DCNP_opt: Optimized Doxycycline hydrochloride Chitosan Nanoparticles.

Figure 4. Effect of DCNP_opt on histopathological changes of brain. A = Vehicle treated brain, B = Ketamine treated brain, C = Doxycycline hydrochloride treated brain, D = DCNP_opt treated brain. ‘a’ – hyperchromatic nuclei, ‘b’ – perinuclear vacuolization, ‘c’ – dilated vascular channels.