Formulation and evaluation of topical herbal gel for the treatment of arthritis in animal model

Aiyalu, Rajasekaran; Govindarjan, Arulkumaran; Ramasamy, Arivukkarasu

doi:10.1590/S1984-82502016000300015

Rajasekaran Aiyalu ^**Correspondence: R. Aiyalu. Professor of Pharmaceutical Chemistry. KMCH College of Pharmacy. Kovai Estate, Kalapatti Road, Coimbatore - 641048. Tamilnadu, India. E-mail: rsekaran2001in@yahoo.co.in

KMCH College of Pharmacy, Coimbatore, Tamilnadu, India

Arulkumaran Govindarjan

KMCH College of Pharmacy, Coimbatore, Tamilnadu, India

Arivukkarasu Ramasamy

KMCH College of Pharmacy, Coimbatore, Tamilnadu, India

*Correspondence: R. Aiyalu. Professor of Pharmaceutical Chemistry. KMCH College of Pharmacy. Kovai Estate, Kalapatti Road, Coimbatore - 641048. Tamilnadu, India. E-mail: rsekaran2001in@yahoo.co.in

CONFLICT OF INTEREST

Authors declare no conflict of interest.

Figures (13)
Tables (13)

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FIGURE 1
In vitrodiffusion profile of topical herbal gels (F1-F6) and diclofenac sodium gel.

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FIGURE 2
Zero order plot for F4 topical herbal gel formulation.

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FIGURE 3
First order plot for F4 topical herbal gel formulation.

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FIGURE 4
Higuchi diffusion plot for F4 topical herbal gel formulation.

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FIGURE 5
Herbal gel formulation F4 in FCA induced arthritic rats.

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FIGURE 6A
Histopathological section of ankle joint of control group of rat under 10 X showing normal joint surface.

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FIGURE 6B
Histopathological section of ankle joint of control group of rat under . 40 X showing normal cartilage and cortex.

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FIGURE 6C
Histopathological section of ankle joint of arthritic control group of rat under 10 X showing inflammation.

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FIGURE 6D
Histopathological section of ankle joint of arthritic control group of rat under 40 X showing inflammation.

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FIGURE 6E
Histopathological section of ankle joint of diclofenac gel treated group of rat under 10 X showing normal cartilage and cortex and marrow.

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FIGURE 6F
Histopathological section of ankle joint of diclofenac gel treated group of rat under 40 X showing normal cartilage and cortex and marrow.

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FIGURE 6G
Histopathological section of ankle joint of herbal gel treated group Of rat under 10 X showing mild inflammation.

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FIGURE 6H
Histopathological section of ankle joint of herbal gel treated group of rat under 40 X showing mild .inflammation.

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TABLE I
Gel formulations with carbopol 934 and carbopol 940

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TABLE II
Evaluation parameters for topical herbal gel formulation made with 1.5% Carbopol 934

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TABLE III
In vitro release kinetic study of topical herbal gel formulated with Carbopol 934

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TABLE IV
Primary skin irritation test for herbal gel formulation 940 and 934

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TABLE V
Stability studies of topical herbal gel formulation

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TABLE VI
Effect of diclofenac sodium, F4 herbal gel formulation on body weight changes in FCA Induced arthritic rats

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TABLE VII
Evaluation of anti-arthritic activity of herbal gel formulation F4 in FCA induced arthritic rats

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TABLE VIII
Alterations in various pain test scores in FCA Induced arthritis in rats

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TABLE IX
Effect of formulation F4 on hematological parameters in FCA induced arthritic rats

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TABLE X
Effect of formulation F4 on biochemical parameters in FCA induced arthritic rats

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TABLE XI
In vitro determination of serum TNFα level in FCA induced arthritic rats

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TABLE XII
In vitro determination of serum Interleukin levels (IL-1ß and IL-6) in FCA induced arthritic rats

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TABLE XIII
Effect of F4 formulation on thymus and spleen weight changes in FCA induced arthritic rats

Gel code	Cardiospermum halicacabum extract (g)	Vitex negundo extract (g)	Carbopol 934 (g)	Carbopol 940 (g)	Triethanol amine (g)	Disodium EDTA (g)	Propylene Glycol (g)	D.M. water (100 g)
F1	0.5	0.5	1.5	---	1.5	0.005	5	q.s
F2	1	1	1.5	----	1.5	0.005	5	q.s
F3	1.5	1.5	1.5	----	1.5	0.005	5	q.s
F4	2	2	1.5	----	1.5	0.005	5	q.s
F5	2.5	2.5	1.5	---	1.5	0.005	5	q.s
F6	3	3	1.5	----	1.5	0.005	5	q.s
F7	0.5	0.5	---	1.5	1.5	0.005	5	q.s
F8	1	1	---	1.5	1.5	0.005	5	q.s
F9	1.5	1.5	----	1.5	1.5	0.005	5	q.s
F10	2	2	----	1.5	1.5	0.005	5	q.s
F11	2.5	2.5	----	1.5	1.5	0.005	5	q.s
F12	3	3	---	1.5	1.5	0.005	5	q.s

Code	Conc (%)	pH*	Viscosity* (poise)	Spreadability* g cm/sec	Net content* % w/w	Extrudability*	Physical appearance
F1	0.5	7.57	0.3850	32.19	99.7	Good	Greenish, smooth and translucent
F 2	1.0	7.57	0.3862	45.05	105	Excellent	Dark green, smooth, homogenous, translucent
F 3	1.5	7.79	0.3873	56.39	105	Good	Dark green, smooth, homogenous, translucent
F 4	2.0	7.60	0.3882	64.00	105	Excellent	Dark green, smooth, homogenous, translucent
F 5	2.5	7.88	0.3891	71.38	101	Excellent	Dark green, smooth, homogenous, translucent
F 6	3.0	7.42	0.3906	75.74	101	Excellent	Dark green, smooth, homogenous, translucent

Formulation code	Zero order R²	First Order R²	Higuchi diffusion model R²	Best fitted model
F1	0.969	0.911	>1	Zero order
F2	0.954	0.947	0.936	Zero order
F3	0.924	0.933	>1	First order
F4	0.989	0.913	>1	Zero order
F5	0.900	0.907	0.909	Higuchi
F6	0.912	0.892	0.917	Higuchi

	Rabbit Numbers			Rabbit		Combined index
	1	2	3	Control	Average
1 h
Erythema Score	0	0	0	0	0.00	0.00
Edema Score	0	0	0	0	0.00
24 h
Erythema Score	0	0	0	0	0.00	0.00
Edema Score	0	0	0	0	0.00
48 h
Erythema Score	0	0	0	0	0.00	0.00
Edema Score	0	0	0	0	0.00
72 h
Erythema Score	0	0	0	0	0.00	0.00
Edema Score	0	0	0	0	0.00
7 days
Erythema Score	0	0	0	0	0.00	0.00
Edema Score		0	0	0	0	0.00
10 days
Erythema Score	0	0	0	0	0.00	0.00
Edema Score	0	0	0	0.00

Groups	Initial body weight (g)	Body wt after 21 days of FCA induction	Body wt after treatment 25^th day	Body wt after treatment 29^th day	Body wt after treatment 35 ^th day	Body wt after treatment 42^nd day	Weight gain (g)
Normal Control	146.3 ± 0.84	172.2± 1.70	173.2 ± 1.89	177.50 ± 1.54	184.50 ± 1.12	192.70 ± 1.25	20.50±1.89
Arthritic control	145.3 ± 0.84	142.80 ± 0.79^a	141.20 ± 0.94 ^a	138.30 ± 1.05 ^a	136.00 ± 1.13 ^a	132.70 ± 1.26 ^a	-11.00±1.34
Diclofenac sodium topical gel (1% w/w)	145.00± 1.15	142.20 ± 1.25 ^a	143.00± 1.41 ^ns	144.20 ± 1.35 ^b	146.50 ± 1.83 ^a	149.80 ± 1.66 ^c	7.66±0.66
Topical herbal gel formulation (2% w/w)	145.5 ± 1.05	141.70 ± 1.38 ^a	142.20 ± 1.49 ^ns	143.00 ± 1.34 ^b	144.80 ± 1.56^b	147.50 ± 1.71^c	5.83±0.47

Groups	Rat paw volume (mm)
	Before treatment			After treatment
	Initial	After 21 days	25^th day	29^thday	35^th day	42^nd day
Normal control	4.85± 0.14	5.16± 0.23	5.09 ± 0.20	5.39±0.10	5.63 ± 0.12	5.75 ± 0.78^d
Arthritic control	4.81± 0.12	10.62± 0.15^a	10.65 ± 0.26 ^a	10.68 ± 0.17 ^a	10.74 ± 0.23 ^a	10.79 ± 0.11 ^a
Diclofenac sodium topical gel (1 % w/w)	5.10± 0.13 ^a	10.40± 0.15 ^a	10.47 ± 0.22^ns	9.81 ± 0.16^b	8.93 ± 0.23^c	8.19 ± 0.07^c
Topical herbal gel formulation F4 (2 % w/w)	4.95± 0.11^a	10.38± 0.14 ^a	10.33 ± 0.14^ns	9.74 ± 0.13^c	9.15 ± 0.15^c	8.73 ± 0.17^c

Groups	Pain test		Mobility score	Stance score
Groups	Extension	Flexion	Mobility score	Stance score
Arthritic control	9.5 ± 0.34	8.33 ± 0.33	1.33 ± 0.21	1.50 ± 0.22
diclofenac sodium topical gel (1 % w/w)	5.16 ± 0.16^d	4.66 ± 0.21 ^d	2.66 ± 0.21^d	2.33 ± 0.21^d
Topical herbal gel formulation (2% w/w)	4.66 ± 0.21^d	3.66 ± 0.33 ^d	3.16 ± 0.21^d	2.83 ± 0.16^d

Groups	Hb (mg%)		WBC (x10³/mm³)		RBC (x10⁶/mm³)		ESR (mm/h)
Groups	Before treatment (21^st day)	After treatment (42^nd day)	Before treatment (21^st day)	After treatment (42^nd day)	Before treatment (21^st day)	After treatment (42^nd day)	Before treatment (21^st day)	After treatment (42^nd day)
Normal control	15.92±0.18	16.38±0.15	6.53±0.15	6.85±0.12	9.64±0.06	10.22±0.20	5.50±0.42	5.16±0.40
Arthritic control	11.02 ± 0.30^a	9.98±0.17^a	11.97±0.22^a	10.88±0.15^a	7.99±0.20	6.31±0.14^a	34.67±1.22^a	42.83±1.35^a
Diclofenac sodium topical gel 1% w/w	11.23±0.21^a	14.97±0.26^b	11.28±0.19 ^a	8.10±0.09^b	7.36±0.25 ^a	8.86±0.21^b	38.17±0.83^a	27.17±2.02^b
Topical herbal gel formulation 2% w/w	12.22± 0.14^a	13.48±0.18^b	11.93±0.40^a	9.41 ±0.04^b	7.18 ±0.23^a	8.16 ±0.10^b	40.67± 0.55^a	31.33±1.83^b

Group	Urea (mg/dL)		Uric acid (mg /dL)
Group	Before treatment (21^st day)	After treatment (42^nd day)	Before treatment (21^st day)	After treatment (42^nd day)
Normal control	16.05 ± 0.60	16.42±0.59	2.85±0.05	2.91±0.07
Arthritic control	44.70±0.92^a	46.57±0.94^a	7.16±0.15^a	7.40±0.18^a
Diclofenac sodium topical gel (1% w/w)	45.33 ± 0.7 ^a	28.17 ±1.17^b	6.63 ±0.58 ^a	5.16±0.24^b
Topical herbal gel formulation (2% w/w )	44.17±0.70^a	34.97±0.76^b	7.15±0.14 ^a	5.45±0.31^b

Groups	TNFα (pg/mL)
Normal control	1.36 ± 0.23
Arthritic control	44.90 ± 2.36^a
Diclofenac sodium topical gel 1% w/w	6.28 ± 0.71^b
Topical herbal gel formulation 2% w/w	19.79 ± 0.81^b

Groups	Serum interleukin (pg/mL)
Groups	IL-1β	IL-6
Normal control	1.38 ± 0.24	19.58 ± 1.14
Arthritic control	67.70 ± 1.09^a	728.61 ± 31.99^a
Diclofenac sodium topical gel (1% w/w)	20.99 ± 0.60^b	313.95±14.66 ^b
Topical herbal gel formulation (2% w/w)	36.55 ± 1.25^b	428.12 ± 10.17^b

Group	Thymus weight (g)	Spleen weight (g)
Normal control	0.15 ± 0.010	0.58 ± 0.011
Arthritic control	0.65 ± 0.009^a	1.33 ± 0.064^a
Diclofenac sodium topical gel (1% w/w)	0.37 ± 0.009^b	0.74 ± 0.011^b
Topical herbal gel formulation (2 % w/w)	0.43 ± 0.007^b	0.89 ± 0.015^b

[1] *Correspondence: R. Aiyalu. Professor of Pharmaceutical Chemistry. KMCH College of Pharmacy. Kovai Estate, Kalapatti Road, Coimbatore - 641048. Tamilnadu, India. E-mail: rsekaran2001in@yahoo.co.in

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Formulation and evaluation of topical herbal gel for the treatment of arthritis in animal model

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