Formulation and Evaluation of Polyherbal Gel for Anti-Acne Treatment

1. 1. Acne

One frequent chronic inflammatory skin disorder is acne vulgaris. Approximately 80% of young adults and adolescents have it. It is a condition that affects the skin's pilosebaceous units and can cause lesions that are either inflammatory or not. Acne is characterized by open comedones (blackheads), closed comedones (whiteheads), and inflammatory lesions such nodules, pustules, and papules. According to Thiboutot et al, acne ought to be treated like a chronic condition that can have psychological effects on its sufferers.^[1] Acne vulgaris is caused by three microorganisms: Propionibacterium acnes, Staphylococcus aureus, and Staphylococcus epidermis. Acne develops from the fast proliferation of microorganisms.

0. 2. Types of acne– [2]
1. Whiteheads -

Remain beneath the skin's surface and are very small.

2. Blackheads –

Though they do not originate from dirt, they have a vivid black appearance and rise to the skin's surface. The colour of black heads is black; it isn't black because of the dirt. In most cases, keratin is an oxidized protein.

3. Papules -

They are tiny, sensitive pink pimples that are easily noticeable on the skin.

4. Nodules –

 Evidently apparent on the skin's surface. These are big, painful pimples that are visible on the skin's surface and are embedded deeply in the skin.

5.  Cysts –

Easily noticeable on the skin's surface. They are painful, pus-filled, highly ingrained, and readily scar.

6. Pustules –

Pustules, often known as zits or pimples, are visible on the skin's surface and are red at the base with pus at the top.

1. 1. Moringa Oleifera

Moring tree shows the great potential and has passed every test for nutritional value, medicinal qualities, environmental suitability, and safety for food is the evergreen, softwood, perennial "Moringa oleifera" of the Monogenetic Moringaceae family.^[3]

2.3.1 Uses of Moringa as a skin care –

Moringa has many healthy nutrients for skin such as – Vitamin A & Vitamin E. Moringa leaves contain antioxidant and antibacterial properties. Due to these properties, moringa is helpful in preventing acne, it also helps in removing dark spots, pimples and blackheads.

1. 1. Ashwagandha

Common names for ashwagandha (W. somnifera, fam. Solanaceae) include "Indian Winter cherry" and "Indian Ginseng." Due to its numerous health advantages, it has been used as a Rasayana for millennia and is considered one of the most significant herbs in Ayurveda, the traditional medical system of India. Ashwagandha belonging to the family Solanaceae.

2.4.1Uses of Ashwagandha as a skin care –

Ashwagandha also having antibacterial and anti-inflammatory properties which makes ashwagandha is a natural remedy for reducing acne and pimples. Ashwagandha helps to moisturizes and nourishes your skin, reducing the wrinkles.

1. Experimental Work:
   1. MATERIALS

Distilled water, HPMC K4M, Carbopol 934, Methyl Paraben, Propyl paraben, triethanolamine,70% ethanol, rose oil. Above all the ingredients used were obtained from Satara college of Pharmacy, Satara.

1. 2. METHODOLOGY

Collection of plant material

1. Moringa oleifera

Fresh moringa leaves were collected from farm. Leaves were dried under sunlight. Then dried leaves were powdered coarsely. The plant was authenticated by the botany department of LBS College of arts, science, and commerce, Satara.

Extraction of moringa leaves powder –

Powdered moringa oleifera leaves (100 gm each; 1:10 w/v) extracted by infusion (30 min) under room temperature (25-30 ?) and boiling water (100 ?), classified as cold extraction.^[5]

2. Ashwagandha

Ashwagandha powder is collected from local market of Satara. (Waghdole Ayurvedics, Satara.)

Extraction of Ashwagandha powder –

100-gram W. Somnifera dried root powder exhaustively extracted with ethanol, using drug-solvent ratio 1: 10 by maceration method (10 hours). Then the extract was filtered and concentrated.^[6]

Method For Preparation of Anti-Acne Gel:

Procedure ^[6]

Evaluation Of Herbal Gel:

Organoleptic evaluation

The resulting gel’s organoleptic properties, such as color, odor and state have been evaluated. The appearance of the gel was judged by its color and roughness and graded.^[7]

Homogeneity

After the generated gel were placed in the container, they were all visually inspected for homogeneity. They had examinations to check for aggregates and to see how they looked.^[8]

Measurement of pH

pH of the gel was measured by using pH meter.^[9]

Spreadability

It indicates the extent of area to which gel readily spreads on application to skin or affected part. The therapeutic potency of a formulation also depends upon its spreading value. Spread ability is expressed in terms of time in seconds taken by two slides to slip off from gel which is placed in between the slides under the direction of certain load. Lesser the time taken for the separation of two slides, better the spreadability. It is calculated by using the formula :

 S = M. L / T where,

M= weight tied to upper slide

L= length of glass slide

T = Time taken to separate the slide^[10]   

Washability

Formulation was applied on the skin and then ease and extent of washing with water were checked observations manually.^[11]

Skin Irritation test

Make a 1 square centimetre mark on the dorsal surface of the left hand. When    applying the gel, the designated area was noted along with the time. At regular intervals for up to 24 hours, irritability, erythema, and edema were assessed and reported.^[5]

Viscosity

Viscosity of the gel is measured by Brookfield viscometer.^[12]

Antimicrobial activity of prepared gel against Staphylococcus aureus.

1. Preparation of Bacterial Culture: Obtain a culture of Staphylococcus aureus from a reliable source. Grow the bacteria in a suitable medium until reaching the desired cell density.
2. Inoculation of the Gel: Apply the gel containing moringa and ashwagandha onto agar plates using sterile techniques. Ensure even distribution of the gel.
3. Incubation: Incubate the agar plates at an appropriate temperature (usually around 37°C) for a specific period (typically 24 hours) to allow bacterial growth.
4. Measurement of Bacterial Growth: After the incubation period, assess the growth of Staphylococcus aureus on the agar plates. This can be done by observing colony formation or by measuring turbidity using a spectrophotometer.
5. Comparison: Compare the growth of Staphylococcus aureus on agar plates containing the gel with those without the gel (control). This allows you to determine if the gel has a bacteriostatic effect by inhibiting bacterial growth.

RESULT AND DISCUSSION:

I. Organoleptic evaluation

Organoleptic evaluation revealed that formulation of herbal gel has semisolid in nature, greenish & smooth in appearance.

Table 4: Organoleptic Evaluation

Homogeneity

The prepared gel was visually inspected, and it was found that homogeneity is good.

III. Measurement of pH

The pH of gel was found to be in the range of 5.5 to  5.9 which is desirable because studies show that the pH of topical formulations must be close to that of skin.

IV. Spreadability

Spreadability = Mass * length / time

Where, m =20 gm

l = 5 cm

t =6 sec

S = 20 * 5 /6 = 16.66 gm.cm/sec

Spreadability of the gel was found to be 16.66 gm.cm/sec

V. Washability

Washability test was carried out by applying a small amount of gel on the hand then washing it with help of tap water. Formulation was easily washable.

Figure No. 1: Before Gel Wash              Figure No. 2: After Gel Wash

VI. Skin irritation test

The skin shows no redness, edema, inflammation and irritation after application of gel. It indicates the formulation is safe to use.

VII. Viscosity

Viscosity of the prepared gel was found to be 4470 cp.

Antimicrobial activity of prepared gel against Staphylococcus aureus.

According to microbiological study, the gel has positive effects on microbial development, and the zone reader was used to determine the zone of inhibition for Staphylococcus aureus the zone of inhibition measured 15 mm.

Figure no. 3: Formulated gel showing zone of inhibition against Staphylococcus aureus