INTRODUCTION:

Since the past timing for treating illness and any kind of skin related symptoms and to support healthy lifestyle human are depend over herbal or plant occurring medicines or natural herbs. Herbal remedies are employed in ancient system of medicines they have ability to heal any type of diseases. They are useful in various kinds of acute and chronic type of dermatological diseases.

It is universally acceptable medicines or formulations because of its therapeutic benefits for the treatment of infections, inflammatory conditions, eczema, contact dermatitis, psoriasis, leprosy and other major skin diseases.¹ Researchers have been focused on the development of Polyherbal formulations because of their benefits such as patient compliance, cost effective and rare produce any side effect or toxic effects on long duration of treatments. Trust of patients over herbal products or medicines increases day by day due to its safer use and more effectiveness than allopathic ones.^2,3

Acne is a skin disease, whose medical language is said Acne vulgaris, it is the common skin disorder which involves the oil glands at the base of hair follicles, this problem occurs when hair follicles becomes blocked with dead skin cells, sebum and bacteria. It affects teenagers, about 80% of the population between the age of 11 and 30 years. It is mostly distributed in lower facial area around the chin, mouth, and jaw line.^4,5 Microorganisms includes as Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes etc. those involve in the pathogenesis of this disease by producing metabolites that can react with sebum resulting the enhancement of the inflammatory process in body of acne suffered person.⁶

There are four main factors those are identified which can cause to the formation of acne lesions or being involved in the formation of acne lesions as given below.^7,8

a. Increased sebum production by sebaceous glands

b. Hyperkeratinization of the follicle

c. Bacterial growth or Colonization of the follicle by the anaerobic bacteria such as Propionibacterium acnes

d. Inflammatory reaction

Information of bacteria:⁹

Staphylococcus aureus, is a genus of gram-negative bacteria under microscope, form in grape like clusters and appears round in shape. They include approximately 40 species of this, 9 subspecies, one has three subspecies and one has four subspecies. They found mostly on the skin and mucous membrane of human and other organisms found all over the world.

The gel formulation is the best formulation when it applies over the skin it can easily spread without imparting greesiness or oilness and it is washable can be easily removed from the skin. This form is best possible form for safer delivery of medication through percutaneous and cutaneous. It has reaches to target tissue. The drug release from gel formulation depends on the physicochemical properties of the vehicle and the drug incorporated. Methods such as the selection of a suitable vehicle, co-administration of a chemical enhancer, etc. have been employed to increase the rate of drug release and skin permeation.¹⁰ This topical route of drug delivery has been used when oral route is not suitable to deliver medication then because it can avoid or bypass first pass metabolism, metabolic degradation and gastrointestinal irritation associated with oral delivery.¹¹

MATERIALS AND METHODS:

Selection and Authentication of plant materials:

Collection of the specimen such as leaves of Azadirachta indica (Meliaceae),¹² leaves of Aloe barbadensis (Asphodelaceae) were obtained from Charak Vanaushadhi udhyan, Mansarovar Ayurvedic medical college hospital and research centre, Bhopal (MP.). Seeds of Nigella sativa (Ranunculaceae) were purchased from local market of Bhopal city in April 2023, and Melaleuca alternifolia oil were purchased from local market. The specimens were collected and authenticated by Proff. Archana Sonare, Govt Jaywanti Haksar P G College, Betul (M.P.) for the proposed study.¹³

Chemicals:

All chemicals such as carbopol 934, carbopol 940, propylene glycol and methyl paraben used for research work were purchased from Indian Scientific Pvt. Ltd., Bhopal (M.P.) and remaining chemicals obtained from faculty of pharmacy laboratory, MGU., Sehore (M.P.).¹⁴

Extraction of plant materials:

Leaves of Azadirachta indica and seeds of Nigella sativa were Collected, Washed with purified water and air dried at room temperature. Both were examined for the presence of foreign matter, then crushed into small parts separately, The Solvents were prepared by added 50:50, Methanol: water into a beaker and filled into round bottom flask. The powdered drug of Azadirachta indica and hydroalcoholic solvent, 1:7 ratios was subjected to Soxhlet extraction at below 60 °C. Same extraction process carried out for Nigella sativa powder. The extracts were filtered with use of different filter papers, concentrated and finally dried to obtain dry powder. Both extracts were packed in a suitable container and then stored in refrigerator for further experimental process.^15,16,17 Aloe barbrdensis leaves were collected, Washed with purified water and air dried at room temperature. Leaves were cut transversely into small pieces and gel was obtained from center part of aloe vera leaves. The epidermis was removed and the inner gel like pulp present in the center of the aloe vera leaf was separated with the help of spatula and homogenized in a mixer. The extract was used immediately. Fresh aloe vera extract used for preparation of gel formulation.¹⁸ Tea tree oil (essential oil) was purchased from the local market.^19,20

Preparation of gel formulation:^18,19

The gel formulations for acne gel were formulated by cold mechanical method as per the composition given in Table 2. All the material required for preparation of gel was dispensed properly. Carbopol 940 or carbopol 934, gelling agent dispersed in an adequate quantity of water and add Aloe barbadensis decanted part in required quantity into it. Stirred it until all solid part got mixed uniformly with help of glass rod and kept a side refer as (PART A). Extract of Azadirachta indica, Nigella sativa mixed thoroughly into motor pestle, the required quantity of Melaleuca alternifolia oil was added. Preservative such as methyl paraben dissolved into remaining quantity of water filled in small beaker kept on water bath until solute get dissolved uniformly and propylene glycol or glycerin, humectants added subsequently into the solution containing methylparaben refer as (PART B). PART B was then mixed into PART A and mixed thoroughly with constant stirring and homogenous dispersion was obtained. The above mixture was stirred for 45 min at constant RPM by using mechanical stirrer. Finally, The PH was modified to neutral by dropwise addition of 10% sodium hydroxide solution, PH modifying agent was added into it upto uniform gel get formed. The formulation kept at room temperature for 24 hrs to observe its stability and consistency.

Table 1: List of Additives

Sr. No.	Ingredients	Supplier’s name
1.	Carbopol 940	Sun Labs
2.	Carbopol 934	Sun Labs
3.	Glycerin	Oxford Lab
4.	Propylene glycol	Sun Labs
5.	NaOH Solution (10%)	Oxford Lab
6.	Methyl paraben	Sun Labs
7.	Purified water	--

Table 2: Manufacturing Formula for formulation of 10% Polyherbal gel with different concentration of herbal plant extract

S. No.	Ingredients	Category	Quantity taken per 100 gm
S. No.	Ingredients	Category	F1	F2	F3	F4	F5	F6	F7	F8	F9	F10
1	Neem extract (Hydroalcholic), 1:7, 50:50	API	1.0%	1.5%	2.0%	2.5%	3.0%	1.0%	1.5%	2.0%	2.5%	3.0%
2	Kalonji extract (Hydroalcholic), 1:7, 50:50	API	4.0%	4.5%	5.0%	4.5%	4.0%	4.0%	4.5%	5.0%	4.5%	4.0%
3	Aloe vera decant	API	4.5%	3.5%	2.5%	2.5%	2.5%	4.5%	3.5%	2.5%	2.5%	2.5%
4	Teatree oil	API	0.5%	0.5%	0.5%	0.5%	0.5%	0.5%	0.5%	0.5%	0.5%	0.5%
5	Carbopol 934	Gelling agent	2%	2%	2%	2%	2%	--	--	--	--	--
6	Carbopol 940	Gelling agent	--	--	--	--	--	2%	2%	2%	2%	2%
7	Propylene glycol (ml)	Humectant	15ml	15ml	15ml	15ml	15ml	--	--	--	--	--
8	Glycerin (ml)	Humectant	--	--	--	--	--	15ml	15ml	15ml	15ml	15ml
9	Methyl paraben	Preservatives	0.1%	0.15	0.1%	0.1%	0.1%	0.1%	0.1%	0.1%	0.1%	0.1%
10	NaoH (10% w/v)	pH modifier	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.
11	Purified water (ml)	Vehicle	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.	Q.S.

Evaluation of Polyherbal gel formulation:^22,23

Figure 1: Blank gel formulation

Figure 2: Polyherbal gel formulation

Physical parameters: Colour, odour, appearance, consistency were evaluated by check it physically and recorded in Table 4 and Figure 1, 2 which shows the absence of aggregates.

Homogeneity Test: Prepared Polyherbal gel formulation applied over transparent type of glass, then checked it the proposed gel formulation was showed homogeneous, there were no any aggregates inside it.

pH determination: 1g of formulated gel was dissolved in 100ml of deionized water. The calibrated digital pH meter Advance lab equipment (Model No. 851 N) was used for the measurement of pH of each formulation. The values were recorded in Table 5.

Rheology (cP): The determination of rheological parameter of each gel formulation was done by using Brookfield DV-E Viscometer (Model No. RVDVE230) with spindle No. 7. The definite quantity of gel was added to a container. The gels were rotated at 100 rotation per minute (RPM) with spindle. At that speed the dial reading was recorded.

Spreadability (g.cm/sec): It showed that the extent of area to which gel readily spreads on application to skin or affected part. Firstly take two sets of glass plates of round shape, then prepared Polyherbal anti-acne gel formulation 500mg or 0.5g weighed was placed in the middle of glass plate that was given a millimeter block, before starting the experiment, weighed the other glass plate then placed the glass plate on the top of the gel mass and kept it for 5 minutes, After that add 50gm of weight then count for 5 minutes. Then add a load of upto 150gm then left for 5 minutes. The dispersion gel was recorded in Table 7. The following equation was used to calculate spreadability.

Spreadability = (Weight × Length) / Time

Extrudability: Polyherbal gel formulations were filled in collapsible tubes. Then extrudability of the each formulation was determined in term of weight in gm required to extrude a 0.5 cm. ribbon of gel in 10 sec.

Antimicrobial studies:^24,25,26

The Zone of inhibition test was performed for detection of antimicrobial property of prepared Polyherbal gel. Before initiating the antimicrobial testing, the test bacteria Propionibacterium acne (ATCC-11827) culture was revived in nutrient broth (HiMedia) while another test bacteria Staphylococcus epidermidis (ATCC-12228) and Staphylococcus aureus (ATCC-25923) were revived in nutrient broth (HiMedia) and all are maintained upto 0.5 McFarland. 20ml of sterile Muller-Hinton agar was poured and allowed to set the antibacterial property of different formulation was determined by using sterile Mueller-Hinton agar plates were prepared. for S. aureus and S. epidermidis. While Reinforced Clostridial agar plates were prepared for working with P. acnes. These agar plates were kept for some time to solidify. By using sterile swab Bacterial inoculums were taken from broth of revived cultures and rubbed over the solidified Mueller-Hinton agar media. Then leave it for 30 minutes on the surface of agar media that has been smeared with bacterial suspension. A sterile 6mm size of five wells was bored in each plate using a cork borer to cut wells of uniform distance in each plate. Antimicrobial susceptibility studies were performed by filled with approx 120mg of Polyherbal gel formulation were introduced into each well of culture plates. The culture plates with bacteria species were incubated in incubator for 24 hours at 37^oC and zones of inhibition were recorded. Then same process was applied with Reinforced Clostridial agar plates for P. acnes. The antibacterial activities were observed by measuring the zone of inhibition (in mm) after incubation.²¹ The results are shown in Table 8.

Table 3: List of Microorganism species employed for testing

S. No.	Test microbial species
1.	Propionibacterium acnes	ATCC-11827
2.	Staphylococcus aureus	ATCC-25923
3.	Staphylococcus epidermidis	ATCC-12228

RESULT:

All results of evaluation during investigation are showed and recorded.

Physical parameters: The physical parameters of prepared herbal gel formulation observed as per Table 4.

pH determination: The measurement of pH of each formulation was carried out by using calibrated digital pH meter, Advance lab equipment (Model No. 851 N) at constant temperature. And they obtained pH values of all samples are shown in Table 5.

Rheological studies (cP): Brookfield DV-E Viscometer (Model No. RVDVE230) with spindle No.7 was employed for determination of viscosity of prepared gel formulation. The obtained viscosity values of all samples are recorded in below Table 6.

Table 4: Organoleptic studies of Polyherbal gel

Formulation	Colour	Homogeneity	Phase seperation	Consistency
AGF1	brownish	Uniform	None	Excellent
AGF2	brownish	Uniform	None	Excellent
AGF3	brownish	Uniform	None	Excellent
AGF4	brownish	Uniform	None	Excellent
AGF5	brownish	Uniform	None	Excellent
AGF6	brownish	Uniform	None	Excellent
AGF7	brownish	Uniform	None	Excellent
AGF8	brownish	Uniform	None	Excellent
AGF9	brownish	Uniform	None	Excellent
AGF10	brownish	Uniform	None	Excellent

Table 5: pH studies of Polyherbal gel

Formulation	pH
AGF1	6.50
AGF2	6.70
AGF3	6.15
AGF4	6.40
AGF5	6.50
AGF6	6.35
AGF7	6.09
AGF8	6.40
AGF9	6.20
AGF10	6.35

Table 6: Viscocity studies of Polyherbal gel

Formulation	Viscocity (cP)
AGF1	1450
AGF2	1420
AGF3	1400
AGF4	1880
AGF5	1600
AGF6	1640
AGF7	1800
AGF8	1770
AGF9	1760
AGF10	1820

Table 7: Spreading Coefficient studies of Polyherbal gel

Formulation	Spreading Coefficient (g.cm/sec)
AGF1	10.12
AGF2	10.20
AGF3	10.50
AGF4	10.10
AGF5	10.00
AGF6	10.15
AGF7	10.50
AGF8	10.80
AGF9	10.12
AGF10	10.22

Table 8: Zone of inhibition studies of Polyherbal gel

S. No.	Sample code	Zone of inhibition against test microbial species
S. No.	Sample code	*Staphylococcus aureus* (ATCC-25923)	*Staphylococcus epidermidis* (ATCC-12228)	*Propionibacterium acnes* (ATCC-11827)
1.	AgF-1	14 mm	16 mm	17 mm
2.	AgF-2	14 mm	15 mm	17 mm
3.	AgF-3	15 mm	16 mm	16 mm
4.	AgF-4	16 mm	15 mm	17 mm
5.	AgF-5	15 mm	17 mm	17 mm
6.	AgF-6	15 mm	15 mm	18 mm
7.	AgF-7	15 mm	15 mm	17 mm
8.	AgF-8	14 mm	16 mm	17 mm
9.	AgF-9	16 mm	15 mm	16 mm
10.	AgF-10	16 mm	15 mm	16 mm

Spreadability (g.cm/sec): The 20g weight sample was placed and determines the values and recorded in below Table 7.

Zone of inhibition studies of Polyherbal gel: Zone of inhibition against three types of different bacteria’s were studied and are effective against microbial strains. All values are recorded in Table 8.

The antimicrobial activity test of prepared Polyherbal gel formulation containing hydroalcholic extracts was done by using bacterial species which are acne causing bacterial species such as Staphylococcus aureus (ATCC-25923), Staphylococcus epidermidis (ATCC-12228) and Propionibacterium acne (ATCC-11827) used in the present study for investigation and all are procured from HiMedia Laboratories Pvt. Ltd, india. The test samples coded as AgF1 to AgF10 were used for antimicrobial testing using dose approx 120mg of Polyherbal gel formulation were inserted in each well. All samples were able to impart inhibitory effect against test bacterial species used in current study. The polyphenol constituent contains in the extracts those are incorporated in the gel formulation that imparts antiacne activity due to the opening of mitochondrial permeability transition pore.

The gel sample AgF1 to AgF10 imparted a zone of inhibition of 15mm to 17mm diameter against Staphylococcus epidermidis and zone of inhibition of 16mm to 18mm diameter against Propionibacterium acnes whereas a zone of inhibition of 14mm to 16mm diameter was observed against bacterial species Staphylococcus aureus. From the above results, it was find that the prepared anti acne gel formulation showing a significant effect on zone of inhibition. The results are recorded and shown in Table 8. and Figure 3,4 and 5.

Figure 3: Gel AgF1 to AgF10 against Propionibacterium acnes

Figure 4: Gel AgF1 to AgF10 against Staphylococcus aureus

Figure 5: Gel AgF1 to AgF10 against Staphylococcus epidermidis

DISCUSSION:

Dermatological related issues are more common in now days. There are various kinds of products that are available in market among which synthetic products are majorly one. Synthetic products such as antibiotics for oral use and various topical products are effective and they are able to overcome these problems but they are harmful and having lot of side effects during long time treatment. Therefore it needs to focus on the herbal formulation. The natural product formulations that are effective on long duration therapy but not as similar to synthetic products but their side effects are negligible. Many naturally occurring herbal formulations that are prove to be effective against inching, acne, Psoriasis, skin care problem in both male and females. It is concluded on the basis of the results obtained in current studies, herbal formulation of Polyherbal gel shows satisfactory physicochemical parameters. Herbal product formulations are assumed to be safe for long period of application. The gel formulation apply topically and have greater benefits that is release of drug directly into the site of application at faster rate as compare to cream and ointment. The finding of Research shows that all medicinal plant part extracts used to formulate gel preparations (AgF1 to AgF10) in studies having antimicrobial activity.

CONCLUSION:

This study aimed to develop a Polyherbal gel for treatment of anti acne disease. The Ten Polyherbal gel formulations were prepared by varying proportion of herbal plant extracts and evaluated for their physicochemical properties. The methanolic extract of neem and kalonji exerts good antimicrobial effect, along with use aloe vera and tea tree oil showing synergistic effect during topical application. Carbopol of different grades (934 and 940) having gelling potency. Glycerin or propylene glycol as humectant, they help to keep skin moistened, and give emollient effect. methylparaben use as preservative to prevent deterioration cause by bacteria. Sodium hydroxide work as pH modifier added dropwise until gel consistency gets achieved. All invitro evaluation test of gel formulation like color, appearance, consistency, pH, Rheological parameters and antimicrobial test have showing satisfactory results.

Herbal formulations have most demand in the world market. The prepared herbal gel formulation are safe, They have property to cure dermatological problems like acne related issues without showing side effects on long duration of treatment. When apply topically it can easily spread over skin and not leave any scars or black spots. Gelling formulations are not sticky nature so it is easily remove after washing with water. It shows good antimicrobial properties. Hopefully it was concluded that the present studies brought advancement in the curing of acne related treatment using herbal plants extracts as well as developed formulations for safe and effective management of diseases.

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Received on 15.07.2024 Revised on 07.09.2024

Accepted on 10.10.2024 Published on 28.03.2025

Available online from April 01, 2025

Research J. Topical and Cosmetic Sci. 2025; 16(1):6-12.

DOI: 10.52711/2321-5844.2025.00002

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.