Neuroprotective study of Nigella sativa-loaded oral provesicular lipid formulation: in vitro and ex vivo study

Figures & data

Table 1. Composition of Nigella sativa-loaded proniosome formulation.

Code	S20:CH:PC	S40:CH:PC	S60:CH:PC	T20:CH:PC	T40:CH:PC	T80:CH:PC
NS1	9:1:1	–	–	–	–	–
NS2	–	9:1:1	–	–	–	–
NS3	–	–	9:1:1	–	–	–
NS4	–	–	–	9:1:1	–	–
NS5	–	–	–	–	9:1:1	–
NS6	–	–	–	–	–	9:1:1

S20 – Span20; S40 – Span40; S60 – Span60; T20 – Tween 20; T40 – Tween 40; T80 – Tween 80; CH – cholesterol; PC – phosphatidylcholine. Each formulation contains 4 mg/ml Nigella sativa.

Table 2. Micromeritic and physicochemical evaluation.

Formulation	Angle of repose (h)^a	Carr's index^a	Hausner’s ratio^a	PDI	No. of vesicles Per mm^3× 10^3
NS1	29.1	13.9	1.13	0.215	2.92
NS2	27.5	11.1	1.12	0.243	2.71
NS3	26.9	10.0	1.11	0.225	3.21
NS4	24.8	10.9	1.12	0.196	2.98
NS5	25.4	8.20	1.12	0.226	3.01
NS6	27.1	7.51	1.13	0.274	2.88

Figure 1. Size distribution of optimized NS-loaded niosome.

Table 3. Quality evaluation parameters.

Code	Size (nm)	Entrapmentefficiency (%)	Flux (µg/cm^2/h)	Enhancementratio
NS1	378.54 ± 8.65	77.71 ± 3.56	6.11 ± 5.43	1.82
NS2	391.22 ± 8.38	79.23 ± 5.24	6.87 ± 6.36	2.05
NS3	417.98 ± 9.12	89.15 ± 4.98	7.11 ± 4.64	2.12
NS4	388.95 ± 7.65	81.54 ± 6.65	5.38 ± 7.54	1.61
NS5	435.43 ± 5.72	85.59 ± 5.71	6.21 ± 6.12	1.72
NS6	406.76 ± 6.48	86.89 ± 6.22	7.23 ± 7.08	2.16
Control			3.34 ± 4.17

Figure 2. TEM image of niosome dispersion from reconstituted proniosomal formulation.

Figure 3. In vitro release of Nigella sativa from proniosome.

Table 4. Effect of Nigella sativa proniosome on locomotor count and grip strength in middle cerebral artery occludes rats.

Groups (n = 6)	Treatment	Locomotor activity (count/10 min)	Grip strength (kg/unit)
I	Normal control	31.5 ± 0.62	0.62 ± 0.01
II	Sham operated	38.83 ± 0.71	0.52 ± 0.04
III	MCAO	09.50 ± 0.65^a,b	0.10 ± 0.01^a,b
IV	Asp + MCAO	24.16 ± 0.49^c	0.49 ± 0.03^c
V	NS6 + MCAO	15.83 ± 0.61^d	0.38 ± 0.02^d

MCAO – middle cerebral artery occlusion; Asp – Aspirin.

Data represented as mean ± SEM.

Significance by one-way ANOVA followed by Dunnett’s t test.

^ap < 0.001 versus normal control.

^bp < 0.001 versus Sham-operated.

^cP < 0.01 versus the MCAO group.

^dP < 0.05 versus the MCAO group.

Table 5. Effect of Nigella sativa proniosome on TBARS, GSH, SOD and catalase in middle cerebral artery occludes rats.

Groups (n = 6)	Treatment	TBARS (nmoles MDA formed/mg protein)	GSH (µmol/mg protein)	SOD (U/mg protein)	Catalase (nmoles of H2O2/ min/mg protein)
I	Normal Control	4.32 ± 0.051	2.15 ± 0.023	10.21 ± 0.51	12.21 ± 0.10
II	Sham Operated	3.98 ± 0.35	3.08 ± 0.059	12.01 ± 0.32	15.50 ± 0.31
III	MCAO	7.11 ± 0.11^a,b	0.89 ± 0.01^c,d	3.93 ± 0.41^c,d	5.12 ± 0.10^c,d
IV	Asp + MCAO	3.21 ± 0.14^e	1.29 ± 0.03^e	8.18 ± 0.43^e	8.40 ± 0.08^e
V	NS6 + MCAO	3.01 ± 0.11^e	1.10 ± 0.01^e	7.18 ± 0.62^e	8.01 ± 0.1^e

Significance by one-way ANOVA followed by Dunnett’s t test.

^ap < 0.05 versus control.

^bp < 0.05 versus Sham.

^cp < 0.01 versus control.

^dp < 0.01 versus Sham.

^ep < 0.05 versus MCAO.

Table 6. Effects of Nigella sativa proniosome on infarct volume in middle cerebral artery occlusion occluded rats.

Groups (n = 6)	Treatment	Infarction volume (mm^3)
I	MCAO	65.21 ± 0.01
II	Aspirin + MCAO	39.70 ± 0.03^a
III	NS6 + MCAO	48.01 ± 0.05^b

Data represented as mean ± SEM.

Significance by one-way ANOVA followed by Dunnett’s t test.

^ap < 0.01 versus MCAO.

^bp < 0.05 versus MCAO.