Journal of Pharmacy & Bioallied Sciences. Apr/30/2019; 11(Suppl 2): S198-S202

PMID: 31198337

Preparation of Mouthwash and Gel from Rosa damascena Mill and Evaluating Its Effectiveness—An In Vivo Analysis

Abstract

Background:

Rosa damascena Mill, commonly known as Damascus rose and garland rose, belongs to Rosaceae family, which has around 200 different types of rose species. R. damascena Mill is famous for its fragrance and beauty, and hence it is highly cultivated all over the world. In addition, this plant is also used for its medicinal properties.

Objectives:

The objectives of this study are to prepare mouth wash and gel from the oil of R. damacena Mill, to evaluate the effectiveness of the medicinal properties of prepared mouth wash and gel, to evaluate its anti-inflammatory and analgesic properties, and to evaluate the adverse effects.

Materials and Methods:

At the JSS College of Pharmacy, Ooty, the oil of R. damacena Mill was converted into mouth wash and gel. To evaluate the effectiveness of the prepared rose gel and mouthwash, 24 Wister and 24 Albino rats were subjected to acetic acid induced writhing, hot plate method, and Carragennin induced paw edema test.

Results:

Acetic acid induced writhing in mice and hot plate method were carried out. Both methods proved presence of analgesic activity, and Carragennin induced paw edema test showed the anti-inflammatory property of R. damascena mill.

Conclusion:

These two in vivo studies proved that the presence of analgesic and anti-inflammatory properties in prepared R. damascena mill mouth wash and gel.

INTRODUCTION

Rosa damascena Mill is one among the 200 species of Rosaceae family of thorny shrub plants. It is commonly known as garland rose.[1 2] It is a shrub with 1–2 m in height from the Rosaceae family. Flowers of this plant are colorful, shinny, and large. Because of its fragrance and beautiful flowers it is widely cultivated all over the world.[3 4] In addition to these qualities it is also being used for its medical properties.[5] This study was undertaken to check the medicinal values of this plant specifically the anti-inflammatory and analgesic.

For the study purpose commercially available rose oil was used, and according to the manufacturer oil was derived from dried petals of the flower by steam distillation method.[6]

Aim

To evaluate the medicinal properties of prepared mouthwash and gel from of R. damascena Mill oil.

Objectives

The objectives of the present study were to check anti-inflammatory, analgesic, and allergic reaction properties of mouthwash and gel prepared from R. damascena Mill oil.

MATERIALS AND METHODS

For application convenience, the rose oil was converted into mouthwash and gel. This was done at JSS Pharmacology College, Ooty, Tamil Nadu, India.

The procedure followed to prepare gel and mouthwash are explained in the following sections.

Preparation of mouth gel

The ingredients for the formulation of the gel were properly weighed. Hydroxypropyl methylcellulose (HPMC K14) was dispersed in 50 mL distilled water maintained at 70°C for 20 minutes using a magnetic stirrer. Then polyethylene glycol-400, sucrose, citric acid, and preservatives (methylparaben/propylparaben) were added with stirring, and required amount of sodium citrate was dissolved in 10 mL of distilled water and added to the mixture. Finally, rose oil was added slowly while mixing and speed was maintained appropriately. The mixture was allowed to cool to room temperature to form gel.

The composition of gel is shown in Table 1.

Table 1

Formula of gel

S. No.	Ingredients	F1 (%)	F2 (%)
1	Rose oil	5	10
2	HPMC K14	3	3
3	PEG 400	10	10
4	Sucrose	2	2
5	Sodium citrate	0.05	0.05
6	Citric acid	0.2	0.2
7	Methylparaben	0.2	0.2
8	Water	Q.S.	Q.S.

HPMC K14 = hydroxypropyl methylcellulose, PEG 400 = polyethylene glycol-400, Q.S. = quantity sufficient

Preparation of mouthwash

To make mouthwash, rose oil and tergitol-N9 was taken and added to the water while mixing at appropriate speed.

Composition used for mouthwash is as follows: Triton X-100 (TX-100) (hydrophillic nonionic surfactant), Tween-40 (T-40) (water-soluble nonionic surfactant), Span-80 (s-80) (oil-soluble nonionic surfactant), and Tergitol-N9 (N-9) (water-soluble nonionic surfactant). All are based on U.S. Food & Drug Administration (USFDA) approved standards.

To evaluate the effects of the prepared rose gel and mouthwash, both were tested on 24 Wistar rats and 24 Albino mice with the approval of animal ethical committee of JSS Pharmacy College, Ooty, Tamil Nadu, India.

Analgesic activity

Acetic acid–induced writhing in mice

Procedure

Acetic acid–induced writhing method was used for evaluating the analgesic activity. Any type of writhing movement was considered as a positive response.

Drugs used

Acetic acid: 1% v/v (1 mL/100 g) inject 1 mL/100 g body weight of mice (i.p.)
Aspirin: Dose 25 mg/kg i.p. stock solution containing 2.5 mg/mL of the drug
Test drugs: A, B, C, D (1 mL/100 g) inject 1 mL/100 g body weight of mice (i.p.)

To evaluate analgesic activity, Swiss Albino mice weighing between 25 and 30 kg were used.

They were divided in to six groups (six animals each)
Group I: Acetic acid treated group
Group II: Acetic acid and Aspirin treated group
Group III–VI: A, B, C, and D and acetic acid treated groups
The approximate volume of acetic acid solution was administered to first group (control) placed individually for observation [Table 2].
The number of abdominal contractions, trunk twist response, and extension of hind limbs and the number of animals showing such response were recorded during a period of 10 minutes.
In II–VI groups, aspirin and test drugs A, B, C, and D were injected. Acetic acid solution was administered to these animals after 15 minutes. The onset and severity of writhing response was noted.
The mean of writhing scores and percentage of inhibition of control of aspirin and A, B, C, and D treated group were calculated [Table 3].

Table 2

Formula of mouthwash

S. No.	Ingredients	F1 (%)	F2 (%)
1	Rose oil	5	10
2	Tergitol-N9	2	2
3	Water	Q.S.	Q.S.

Q.S. = quantity sufficient

Table 3

Analgesic activity by acetic acid induced writhing in mice

Treatment	Dose (mg/kg)	No. of wriths in 10 minutes	Inhibition (%)
Control	—	25.17 ± 0.70	—
Aspirin	25	11.67 ± 0.42*	56
A	5	17.24 ± 0.36*	41
B	10	15.36 ± 0.25*	40
C	5	14.83 ± 0.31*	44
D	10	13.50 ± 0.42*	48

Values are expressed as mean ± SEM (n = 6). Data were analyzed by using one-way analysis of variance followed by Tukey’s test

*P < 0.05 compared with control

Hot plate method

Procedure

The responses of the mice to the heat, at temperatures that do not damage the skin, were jumping, withdrawal of the paws, and licking of the paws. The commercially available electric hot plate was used. The temperature was controlled at 55°C to 56°C. The mice were placed on the hot plate and the time until either licking or jumping occurs was recorded by a stopwatch.

Drugs used

Acetic acid: 1% v/v (1 mL/100 g) inject 1 mL/100 g body weight of mice (i.p.)
Aspirin: Dose 25 mg/kg i.p. stock solution containing 2.5 mg/ mL of the drug
Test drugs: A, B, C, D (1 mL/100 g) inject 1 mL/100 g body weight of mice (i.p.)

For evaluation of analgesic property, Swiss Albino mice weighing between 25 and 30 g were used.

The animals were divided in to six groups of six animals each: Group I: Control (normal saline), Group II: Aspirin, and Group III–VI: A, B, C, D.

The basal reaction time was noted by observing licking or jump response of hind paw (which one occur first) in animals, when placed on the hot plate maintained at a constant temperature (55°C). Normally animals show such response in 6–8 seconds. To avoid damage to the paws a cutoff period of 15 seconds was fixed.

Aspirin and test drugs ABCD were injected to the respective group of animals and the reaction times of animals on hot plate were noted at 30, 60, 90, and 120 seconds. When the reaction time increased, 15 seconds was taken as maximum analgesia and the animals were removed from the hot plate to avoid injury to paws. The results are provided in Table 4.

Table 4

Analgesic activity by hot plate method in mice

Treatment	Dose (mg/kg)	Reaction time in seconds

		0	30	60	90	120
Control	—	7.33 ± 0.33	7.00 ± 0.36	6.83 ± 0.30	7.12 ± 0.36	7.16 ± 0.37
Aspirin	25	7.16 ± 0.3	5.83 ± 0.37*	5.00 ± 0.25*	3.14 ±0.41*	3.33 ± 0.211*
A	5	7.16 ± 0.16	5.33 ± 0.21*	4.33 ± 0.21*	3.64 ± 0.31*	3.83 ± 0.30*
B	10	7.83 ± 0.16	5.56 ± 0.22*	4.16 ± 0.16*	3.66 ± 0.21*	3.83 ± 0.16*
C	5	7.43 ± 0.18	5.50 ± 0.26*	4.50 ± 0.34*	3.16 ± 0.30*	3.66 ± 0.21*
D	10	7.15 ± 0.12	5.54 ± 0.36*	3.66 ± 0.21*	3.00 ± 0.25*	3.50 ± 0.22*

Values are expressed as mean ± SEM (n = 6). Data were analyzed by using one-way analysis of variance followed by Tukey’s test

*P < 0.05 compared with control

Anti-inflammatory activity

Carrageenan-induced paw edema test

The Wistar Albino rats were divided into six groups (six rats for each group):

Group I: Solvent control 1 mL/kg 0.3% carboxy methyl cellulose (CMC) orally
Group II: Indomethacin 10 mg/kg in 0.3% CMC orally
Group III–VI: A, B, C, D (1 mL/100 g) inject 1 mL/100 g body weight of mice (p.o.)

By injecting carrageenan 1% w/w (0.1 mL) into the subplantar region of the left hind paw in the rats had produced the acute paw edema. One hour before testing, the test drugs A, B, C, and D and indomethacin 10 mg/kg were administered orally. The control group received vehicle 0.1 mL/100 g. By using digital plethysmometer (UGO Basile, Italy) the paw volume was measured at 0, 1, 2, 3, 4, and 6 hours after carrageenan test. The percent increase in the paw edema (paw volume) was calculated by comparing it with zero minute reading. The percentage of inhibition of edema was calculated after fourth hour assuming that 100% inflammation is attained in vehicle group.

RESULTS

Analgesic activity

Acetic acid–induced writhing in mice

All the standard and test drugs significantly (P < 0.05) reduced the number of abdominal constriction and stretching of hind limbs induced by the injection of acetic acid compared to the control. The test drugs A, B, C, and D showed 41%, 40%, 44%, and 48% of inhibition, respectively, against control; however standard drug showed 56% inhibition.

All the test and standard drugs showed significant change (P < 0.05) in reaction time compared to the control. Treatment groups showed decrease in reaction time from 30 to 90 minutes and thereafter showed increase in reaction time to 120 minutes. The same effect was also observed with standard drug aspirin.

Anti-inflammatory activity

Carrageenan (0.1 mL) induced paw edema test showed that test drugs have significant (P < 0.05) anti-inflammatory property. The anti-inflammatory activity of test drugs had significant activity from third hour onward and it was maintained up to sixth hour compared to the control group [Table 5].

Table 5

Inhibitory effects of test drugs on carrageenan-induced edema of the hind paw in rats

Group	Dose (mg/kg)	Swelling volume (mL)

		1 hour	2 hour	3 hour	4 hour	6 hour
Control	—	1.53 ± 0.07	2.33 ± 0.09	3.31 ± 0.05	3.65 ± 0.06	3.38 ± 0.07
Indomethacin	10	1.54 ± 0.05	1.86 ± 0.10	2.13 ± 0.08	2.11 ± 0.12	1.89 ± 0.06
A	5	1.56 ± 0.21	2.29 ± 0.21	2.98 ± 0.11	2.59 ± 0.43	2.46 ± 0.04
B	10	1.52 ± 0.19	2.14 ± 0.06	2.58 ± 0.44	2.49 ± 0.07	2.43 ± 0.25
C	5	1.57 ± 0.68	1.98 ± 0.23	2.16 ± 0.51	2.02 ± 0.14	1.93 ± 0.08
D	10	1.57 ± 0.58	1.96 ± 0.43	2.15 ± 0.51	2.01 ± 0.14	1.93 ± 0.08

Values are expressed in terms of mean ± SEM; P < 0.05 vs. control group

DISCUSSION

To reduce the adverse effects of the synthetically derived drugs or to replace with alternative the pharma companies started exploring herbs. One among the traditionally used herb for medicinal purpose is R. damascena Mill.[5]

The anti-inflammatory property

Previous studies have shown that R. damascena Mill has anti-inflammatory effect.[6 7] In this study the effect of anti-inflammatory property of the gel has been demonstrated by carrageenan-induced paw edema test. According to previous studies, essential oil had no anti-inflammatory effect while the extract could significantly reduce edema, which may be due to inhibition of the mediators of acute inflammation.[8 9] In addition, R. damascena contains vitamin C which has antioxidant and anti-inflammatory effects.[10 11]

The analgesic effect

In our study we used acetic acid–induced writhing test and hot plate method to evaluate the analgesic effect of R. damascena Mill in mice. Test results have shown promising effect (P < 0.05) in comparison with other drugs [Table 4]. There was increased response time in both the tests. In previous studies also effects of R. damascena Mill have been demonstrated. In one study, the effect of aqueous, ethanolic, and chloroformic extracts in mice on hot plate and tail flick was evaluated and only ethanolic extract showed analgesic effect.[12] However, studies have shown that hydroalcoholic extract has a potent analgesic effect in acetic and formalin tests[13] Based on this study’s results it can be derived that ingredients of the plant that are not soluble in water may be responsible for the analgesic effect. The nonwater-soluble components of rose oil quercetin and kaempferol may be responsible for the analgesic effect.

CONCLUSION

Since ancient times herbs are being used for their medicinal properties in various forms. They are available easily, cost-effective and have their proven medicinal validity. In dentistry derivatives of herbs are in the form of mouthwash, gel, and tooth paste. In our study, the results had shown that the mouthwash and gel prepared from rose oil have the properties of analgesic and anti-inflammatory. These can be used in dentistry after some more in vivo studies.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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