Clinical trial of a herbal topical cream in treatment of Acne vulgaris

Abstract

The main anti-acne medicines including retinoids, systemic and topical antibiotics and hormones
have several severe side effects. According to the literature, it is expected that Calendula
officinalis, Rosa canina, Zataria multiflora and Trigonella foenum graecum which have
antibacterial, anti-inflammatory and anti -oxidant effects, and Glycin max that have
phytosterogenic property, heal the inflamed lesions of the disease. The purpose of the present
study was to formulate and clinically evaluate an anti-acne cream using the extract of the above
mentioned plants.A general formula of a cleansing cream containing bees wax, spermaceti,
borax, liquid paraffin and water was considered and then corrected. The best formula was chosen
according to its physicochemical properties. The stability of the formulation was evaluated. After
diagnosis of acne vulgaris, the number of inflamed lesions and severity of the disease were
determined. After two weeks of administration of the coded formulations, the process of
improvement and possible complications were reviewed. If necessary, the treatment was
continued for another two weeks. Finally, the patients were examined for the number of lesions
and the severity again. The significance of data was analyzed statistically using SP SS. The final
product was water in oil cream with suitable appearance and acceptable physico -chemical
stability. Clinical trial results were significantly illustrated the effectiveness of the formulation
mainly against papular and pustular lesions; disease severity and acne-induced inflammation,
when compared to placebo. Previous researches have shown the presence of steroid saponins in
Trigonella foenum graecum that make it a good anti-inflammatory agent. Also, anti-oxidant and
anti-microbial effect of the other above mentioned plants are reported. Considering the results of
the present study, when compared with placebo, a formulation containing the mixed extract of
the plants has a good efficacy in the treatment of acne vulgaris by reduction of the number of
lesions and inflamed area.
Key words: Acne vulgaris, Herbal extract, Cream

Full text

RESEARCH ARTICLE Am. J. PharmTech Res. 2012; 2(2) ISSN: 2249-3387
Please cite this article in press as: Moghimipour E et al., Clinical trial of a herbal topical cream in
treatment of Acne vulgaris. American Journal of PharmTech Research 2012.
Clinical trial of a herbal topical cream in treatment of Acne vulgaris
Eskandar Moghimipour 1,2*, Amir Siahpoosh1,2, Reza Yaghoobi3, Alireza Malayeri2,
Fatemeh Faramarzi2
1 Medicinal Plant Research Center, Jundishapur University of Medical Sciences, Ahvaz, Iran
2 Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
3 Departments of Dermatology, Faculty of Medicine, Ahvaz Jundishapur University of Medical
Sciences, Ahvaz, Iran
ABSTRACT
The main anti-acne medicines including retinoids, systemic and topical antibiotics and hormones
have several severe side effects. According to the literature, it is expected that Calendula
officinalis, Rosa canina, Zataria multiflora and Trigonella foenum graecum which have
antibacterial, anti-inflammatory and anti-oxidant effects, and Glycin max that have
phytosterogenic property, heal the inflamed lesions of the disease. The purpose of the present
study was to formulate and clinically evaluate an anti-acne cream using the extract of the above
mentioned plants.A general formula of a cleansing cream containing bees wax, spermaceti,
borax, liquid paraffin and water was considered and then corrected. The best formula was chosen
according to its physicochemical properties. The stability of the formulation was evaluated. After
diagnosis of acne vulgaris, the number of inflamed lesions and severity of the disease were
determined. After two weeks of administration of the coded formulations, the process of
improvement and possible complications were reviewed. If necessary, the treatment was
continued for another two weeks. Finally, the patients were examined for the number of lesions
and the severity again. The significance of data was analyzed statistically using SPSS. The final
product was water in oil cream with suitable appearance and acceptable physico-chemical
stability. Clinical trial results were significantly illustrated the effectiveness of the formulation
mainly against papular and pustular lesions; disease severity and acne-induced inflammation,
when compared to placebo. Previous researches have shown the presence of steroid saponins in
Trigonella foenum graecum that make it a good anti-inflammatory agent. Also, anti-oxidant and
anti-microbial effect of the other above mentioned plants are reported. Considering the results of
the present study, when compared with placebo, a formulation containing the mixed extract of
the plants has a good efficacy in the treatment of acne vulgaris by reduction of the number of
lesions and inflamed area.
Key words: Acne vulgaris, Herbal extract, Cream
*Corresponding Author Email: moghimipour@yahoo.com
Received 5 January 2012, Accepted 18 January 2012
Journal home page: http://www.ajptr.com/

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INTRODUCTION:
Acne is a common skin disease that affects nearly 80% of adolescents and young adults aged 11
to 30 years. The development of inflammatory lesions often drives acne patients to seek
treatment.
Topical medicaments used in acne treatment, e.g. tretinoin may cause primarily mild dermatitis.
Bleaching of skin and hair is the common side effect of benzoyl peroxide. Although there is no
proved documentation regarding systemic absorption of retionids through skin, they should be
avoided in pregnancy and lactation1. Cosmoceuticals and pharmaceuticals are growing areas of
interest and controversy in acne therapy and developing new active botanical extracts and
compounds to provide them as anti–acne agents is still a field with great potential2.
Calendula officinalis is a short-lived aromatic perennial plant in the family Asteraceae that is
used for the treatment of skin disorders and pain, and as a bactericide, antiseptic and anti-
inflammatory agent. Its main components are carotenoids, flavenoids, triterpene saponins and
essential oil with anti inflammatory, antibacterial and antiviral effects. It has been demonstrated
that a formulation containing the plant extract stimulates epithelization and renewing of wounded
skin 3, 4.
Trigonella foenum (Fenugreek) is a plant in the family Fabaceae containing saponins, mucilage
and alkaloids. The mucilage component of the plant is responsible for its protecting, emollient
and healing effects. The saponin component has antimicrobial and anti-inflammatory activities.3,4
Rosa canina is a rose species normally ranging in height from 1-5 m which is high in certain
antioxidants. It mainly contains fatty components, vit-E and, essential oil and protein substances.
It has been shown that galactolipid component of the plant, with anti- inflammatory effect may
prevent chemotatic reactions in peripheral circulation 5.
Glycin max is a species of legume native to East Asia and mainly consisting of phospholipids
and phytoesterols with anti-tumor and phytoestrogenic properties3, 4.
Zataria multiflora Boiss. is a plant belonging to the Laminaceae family that geographically
grows in Iran, Pakistan and Afghanistan 6. Its main compositions are flavenoids, essential oil,
tanin and polyphenols with anti-inflammatory, antiallergic and angioprotective effects. The main
constituents of the essential oil of this plant are phenolic compounds such as carvacrol and
thymol 7. Its antifungal, and antihelmintic effects are well demonstrated 8. The essential oil of
Z.multiflora has been traditionally used for its antiseptic, anti spasm and local anesthetic
effects6,9. The present article includes formulation of a topical cream using above mentioned herb
extracts and a controlled clinical trial using this multicomponent dermatological cream.

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MATERIALS AND METHODS
Plant materials
Calendula officinalis was collected from Kolah-Ghazi Mountain, Isfahan, and identified in the
department of botany Isfahan University, Iran. Soy beans were purchased from Behshahr, Iran.
The other plant materials were kindly donated by research and natural resources center, Isfahan,
Iran. Aerial parts of C. officinalis and Z. multiflora, hips of R. canina and soybeans were dried in
the shadow at room temperature for 72 hours, then weighed and stored in dark, cool and dry
place. Then, they were grounded to fine powder and separately suspended in methanol in a
beaker to avoid solvent evaporation, the container was closed using parafilm and stored in
shadow for 48 hours. The extracts were then filtered and concentrated in rotary evaporator to a
completely dried film. Z. multiflora essential oil was extracted from 250 g powder in a BP
extraction apparatus for 2 hours.
Formulation design.
Due to occlusion characteristics of fatty bases, the minimum amounts of fats were used. Also,
lactic acid was added to adjust the cream pH to 5.3. To enhance wash ability of the final product,
polyethylene glycol 2000 was added. Since creams and other water containing dosage forms
should be preserved from microbial contamination, methyl and propyl paraben (3:1 ratio) were
added to the formulation. Finally, herbal material was incorporated to cream base by levigation.
The amounts of herbal materials are listed in Table 1
Table 1. The amount of herbal material used in formulation design.
Plant Material
Amount
Z. multiflora Extract
1g
R. canina Extract
1g
C. officinalis Extract
2g
T. foenum Extract
2g
G. max Extract
1g
Z.multiflora Essential oil
0.3ml
Clinical trial
A clinical trial was initiated to evaluate the efficacy of final formulation in comparison to
placebo in patients with acne vulgaris. Sixty patients with mild to moderate acne vulgaris were
selected and inspected for the disease. After identification by dermatologist, informed consent
was obtained from patients. The purpose of the study, nature of the herbal preparation, the
procedures to be carried out and the potential risks and benefits were explained to the study
subjects in detail. The Ahvaz Jundishapur University of Medical Sciences Ethical committee
permission was obtained before performing the experiment. Patients who had systemic

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complications, menopausal disorders and patients with drug induced acne were excluded from
the study. Also, pregnant and lactating patients and some who had systemic topical treatment
during last 3 month were not included. The final population of each group was about 30 persons.
During the initial visit, the patients were assigned to two treatment groups. The first group was
contained patients with less than 10 papulo-pustular lesions which were considered as mild. The
second was contained patients with more than 10 papulo–pustular lesions that were considered as
moderate 10.
Then, patients were assigned to two herbal cream and placebo control groups and were
recommended to apply once daily. Also they were advised to come for follow up 2 and 4 weeks
after treatment. Inflammation index and the total number of lesions were recorded at every
follow up. The severity index was determined by dermatologist according to the lesion counts,
the diameter and the depth of lesions for each patient.
Statistical Method
After data collection, labels were decoded and results analyzed using SPSS software program.
All the results were analyzed statistically using Student-t test for paired data of different levels of
significance. The results were presented as mean± standard error.
RESULTS AND DISCUSSION
The mean age of patients for treatment and control group were 21.97 and 22.13 yeas
respectively. The pretreatment records are shown in Table 2.
Table 2. Comparison of acne vulgaris indices in treatment and control groups before
treatment (Mean± SE, n=30)
Index
The number
of papules
The number
of pustules
The number
of nodules
Inflammation
Severity
index
Control
19.03±2.26
1.13±0.36
0.40±0.28
17.93±2.95
1.73±0.11
Test
16.40±3.05
2.51±0.57
0.68±0.36
22.24±0.12
1.73±0.08
P value
0.49
0.098
0.589
0.27
1.00
The results indicated that the incidence of papular lesions before treatment in treatment and
control group were 16.40 ± 3.05 and 19.03 ± 2.26 respectively and There was no significant
difference between them (P= 0.49). Also, there were no significant differences regarding the
other disease indices between the two groups (p> 0.1)
The results indicated that the inflammatory acne index and total counts were significantly
reduced after 2-4 weeks of treatment with the herbal cream (Figure 2). It was showed that the
rate of healing of papular and pustular lesions in test and control groups were significantly
different (p < 0.01, and p < 0.05, respectively). Although, there was no significant difference
between the number of nodular lesions in treatment and control groups (p = 0.240), the

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inflammation index in the treatment group was significantly reduced (p < 0.0001). The results of
administration of herbal and placebo cream are tabulated in table 3 and plotted in Figure 1.
Table 3. Comparison of acne vulgaris indices in treatment and control groups after
treatment (Mean ± SE, n=30)
Index
The number
of papules
The number
of pustules
The number
of nodules
Inflammation
Severity
index
Control
13.46 ±2.80
0.60 ± 0.22
0.13 ± 0.08
14.20 ± 2.76
1.6 ± 0.12
Test
4.51 ± 0.90
0.62± 0.25
0.1 ± 0.07
5.03 ± 1.03
1.2 ± 0.07
P value
<0.0001
0.104
0.66
<0.0001
<0.0001
Figure 1. Comparison of the number of acne lesions, inflammation and severity indices in
test and control groups (Mean ± SE, n=30).
Figure2. A woman with acne vulgaris: before treatment with the herbal cream (left), and 2
weeks after treatment (right). Clinical improvement was observed after treatment with the
formulated mixed herbal cream.
Regarding prevalence of acne specifically in teenagers, and side effects of oral systemic
medications, it was decided to incorporate a mixture of some medicinal plants with proved
antimicrobial, anti- inflammatory, anti–oxidant and phytoestrogenic activities in a proper
formula for treatment of acne vulgaris. Extract of T. foenum seeds which contains steroid
saponins have shown wound healing property 11. Also, anti-oxidative, anti–inflammatory and
0
2
4
6
8
10
12
14
16
18
Control
Test

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hepatoprotective properties of the plant are previously reported 12, 13, 14. Recent studies suggested
NSAID – like mechanism for its anti-inflammatory, analgesic and anti-hyperthermic effects 15, 16.
It has been reported that the plant is a potent antimicrobial agent against s. aureus 17 which is the
main cause of skin infections. Hips of Rosa canina contain vast amount of ascorbic acid with
well known anti oxidant property 18. The plant is also used for its antimicrobial, wound healing
and anti-inflammatory properties 5, 19, 20. Calendula officinalis is a plant with known wound
healing effect and have been used in traditional medicine for treatment of burns and other dermal
disorders 21. It has been also used for its anti-inflammatory and anti-oxidant properties 22. Soy
bean is a phytoestrogenic anti-inflammatory plant with known anti microbial and anti–oxidant
effects 23, 24. There are many reports regarding antifungal and antibacterial effects of Z. multiflora
25. The results of the present clinical trial indicated that treatment with the formulated herbal
cream resulted in clinical efficacy in comparison with negative control (Figure. 1). A 3 fold
smaller reduction in papula number was found after 3 weeks of treatment with the herbal cream
and control groups. Also comparing the number of pustules and nodules after 3 weeks, the
reduction of inflammation and severity indices in test group were highly significant (p <0.0001).
Many herbs with antioxidant, anti–inflammatory and soothing properties have been used as anti-
acne therapeutic alone or in combination. Among them, oat (Avena sativa), chamomile, turmeric,
licorice, Aloe vera, curcumin, soybean, coffee, mushrooms, green tea and pine bark extract are
more emphasized. Although there is not any crucial evidence of their activity, but theoretically,
their healing and anti-inflammatory properties help the reduction of acne-induced erythma. Also
their soothing effect is a positive factor for reduction of acne induced dried skin 26. However, the
result of the study of pocrolimus cream in the treatment of moderate acne vulgaris indicated that
there was no significant improvement in the number of inflammatory lesions 10. Nevertheless,
the results of a controlled phase II clinical trial showed the effectiveness of an Ayurvedic
multicomponent dermatological formulation, containing the soft extract of some Indian endemic
herbs in the treatment of acne vulgaris 26, 27.
According to the results of an in vitro study, extracts of rose, duzhong (Eucummia ulmodes) and
yerba mate (ilex paraguarensis) mate exhibited notable antimicrobial activity against
propionibacterium acne . They were also able to suppress the secretion of tumor necrosis factor
(TNF) and some other inflammation mediators 2.
CONCLUSION
Although the empiric evidences for the efficacy of herbal medicines in acne is not strongly
enough, the results of the present study suggest that the combination of the herbs are useful in the

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treatment of acne vulgaris. However, long-term clinical trials are still needed to assess the
probable adverse effects of the formulation.
ACKNOWLEDGEMENT
The paper is issued from Pharm.D. Thesis of Fatemeh Faramarzi and financially supported by
Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
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