Formulation and Evaluation of Antimicrobial and Healing Activities of a Herbal Gel Against Mouth Ulcer Causing Bacteria

Aphthous ulcers, or mouth ulcer are common oral mucous membrane disorder which characterized by appearance of painful lesion on the mucous membrane of mouth. These ulcer may induced by various factor, including mechanical trauma, stress, hormonal variations, nutritional deficiency and microbial infections. While they generally benign, mouth ulcer can affect person’s quality of life due to pain, discomfort and difficulty in eating or speaking.[3] Ulcers frequently occur in the mouth area, prompting individuals to consult their doctor or dental specialist. The symptoms that are typically reported include inflammation, a burning feeling, or discomfort. They can present in any part of the oral cavity but may be painful. if it occurs in the movable area. Mouth ulcers are yellowish or whitish depressions with Red margination in the mucous lining of the mouth cavity, characterized by inflammation and pain.[4]

Herbal medicines are getting increasing patient compliance as they are avoiding typical side effects of allopathic medicines. It is no wonder that the world’s one-fourth population i.e. 1.42 billion people, are dependent on traditional medicines for the treatment of various diseases. Medicinal plants have been a major source of cure for human diseases since time immemorial. Herbal formulations have now a day’s undergone more thorough investigation for their potential in preventing and cure oral disease [6]. Herbs have long been used traditionally for routine cleaning of teeth and dental disease and to treat various oral diseases [7]. Oral diseases like oral cancer, dental caries and periodontal diseases among the most important oral health problems. There is a well-established link between the activities of microbial species that form part of the micro biota of the oral cavity and oral diseases. The big need for alternative treatment, products and prevention options for oral diseases that are safe, economical and effective comes from the rise in disease incidence particularly in developing countries, increased resistance by pathogenic bacteria to currently used chemotherapeutics and antibiotics opportunistic infections in immune compromised individuals and financial that is economical considerations in developing countries.[2]

1.1 Mouth Ulcer Gel

Mouth ulcers, also called as aphthous ulcer, is a common oral mucosa disorder affecting people worldwide. The conventional treatment option for mouth ulcer often involves synthetic medication which may causing several side effect. Therefore, the development of safe, effective and natural remedies is warranted. This research focuses on developing and assessing a gel for mouth ulcers made from herbs as a different treatment method which characterized by appearance of painful lesion on the mucous membrane of mouth. These ulcer may induced by various factor, including mechanical trauma, stress, hormonal variations, nutritional deficiency and microbial infections. While they generally benign, mouth ulcer can affect person’s quality of life due to pain, discomfort and difficulty in eating or speaking. These plant products may be used in their powdered form, crude form, purified extracts, or derivative form. It is extremely difficult to prepare a herbal gel using a single natural material that would be milder and safer than the synthetic ones.[5]

Fig.-1:  Mouth ulcer

1.2 Types of  Ulcer

Mouth ulcers are categorized as Major, Minor, or Herpetiform based on the size and number of ulcers [8]. The following are the most common forms of mouth ulcers:

1. Minor ulcers: Minor aphthous ulcers is the most common type, making almost 80% of all the cases. They are usually around 2–8 mm in size and mostly get cleared within 10 days to 2 weeks, but sometimes healing may feels slower. These ulcers are quite superficial and small (mostly less than 1 cm), and they appear either single or in small groups. They usually heal on their own and not leaving any scar afterward.[9]

Fig.-2:  Minor ulcer

2. Major ulcers: Major aphthous ulcers are the type that comes much bigger in size, usually more than 5–10 mm, and they are mostly more painful compared to the minor ones. These ulcers not heal quickly; instead, they can stay for many weeks or even some months before getting better. Because they goes deeper into the tissue, they often leaving behind a scar after healing.

Fig:-3:  Major ulcers

3. Herpetiform ulcers: Herpetiform ulcers, the third category, is a term that refers to the clustered appearance of lesions. This ulcer consists of a cluster of dozens of tiny lesions the size of pinheads. It has nothing to do with the herpes virus. These appear in enormous numbers, ranging from 10 to 100 at a time, and are made up of several tiny lesions that eventually join to form larger plaques.

Fig.- 4: Herpetiform ulcer

1. 3. Drug profile

1. Guava leaves

Fig.-5:  Guava leaves

Botanical Profile

• • Botanical name: Psidium guajava
  • Family: Myrtaceae
  • Part used: Leaves (young and mature)
  • Morphology: Opposite, simple, elliptic to oblong leaves with prominent veins and aromatic odor when crushed.

Phytochemical Constituents

• • Tannins – responsible for astringent and antidiarrheal activity.
  • Flavonoids (e.g., quercetin, kaempferol, rutin) – potent antioxidants, anti-inflammatory, and antimicrobial.
  • Pentacyclic triterpenoids (e.g., oleanolic acid, ursolic acid) – hepatoprotective, anti-inflammatory.
  • Saponins – immunomodulatory and antimicrobial.

Pharmacological Actions

• • Antimicrobial: Effective against Staphylococcus aureus, Escherichia coli, and fungi.
  • Antidiarrheal: Tannins reduce intestinal motility and fluid secretion.
  • Antioxidant: Flavonoids and phenolics scavenge free radicals.
  • Anti-inflammatory: Triterpenoids and flavonoids reduce inflammation.
  • Antidiabetic: Leaf extracts help lower blood glucose by inhibiting carbohydrate-digesting enzymes.
  • Wound healing: Promotes tissue regeneration due to tannins and flavonoids.

Traditional & Ethnomedicinal Uses

• • Decoction of leaves used for diarrhea, dysentery, and gastroenteritis.

2. Fitkari

Fig.-6:  Fitkari

• • Name: Alum (Potassium Aluminum Sulfate)
  • Chemical Name / Formula: Chemical Name: Potassium aluminum sulfate dodecahydrate Formula: Kal (SO?)?·12H?O
  • Synonyms: Potash alum Common alum Tartar emetic (in some contexts, though slightly different)
  • Category / Class: Inorganic salt Astringent / Styptic / Antiseptic

3. Neem

Fig.- 7: Neem

Botanical Profile

• • Botanical name: Azadirachta indica A. Juss.
  • Family: Meliaceae
  • Part used: Leaves (fresh or dried)
  • Morphology: Pinnate leaves, serrated margins, bitter taste, characteristic strong odor.

Phytochemical Constituents

• • Limonoids: Nimbin, nimbidin, nimbinin, azadirachtin – insecticidal, antimicrobial, anti-inflammatory.
  • Flavonoids: Quercetin – antioxidant, anticancer potential.
  • Tannins & Glycosides: Astringent, antimicrobial, wound healing.
  • Sterols & Coumarins: Support hepatoprotective and immunomodulatory activity.

Pharmacological Actions

• • Antimicrobial & Antiparasitic: Effective against bacteria, fungi, viruses, and parasites.
  • Antidiabetic: Helps regulate blood glucose by modulating carbohydrate metabolism.
  • Anti-inflammatory: Limonoids reduce inflammation and pain.
  • Hepatoprotective: Protects liver from toxins and oxidative stress.
  • Anticancer potential: Modulates cell signaling pathways, showing promise in cancer prevention.

Traditional & Ethnomedicinal Uses

• • Used in Ayurveda, Unani, Siddha for skin diseases, fever, diabetes, and infections.
  • Neem leaf decoction for malaria, intestinal worms, and oral health.
  • Applied topically for eczema, acne, and wound healing.

4. Tulsi

Fig.-8: Tulsi

Botanical Profile

• Botanical name: Ocimum sanctum Linn
  • Family: Lamiaceae
  • Part used: Leaves (fresh or dried)
  • Morphology: Small branched aromatic herb (~70 cm tall), opposite green leaves, serrated margins, purple/white flowers.

Phytochemical Constituents

• • Volatile oils (eugenol, methyl eugenol, carvacrol, linalool) – antimicrobial, aromatic.
  • Ursolic acid & oleanolic acid – anti-inflammatory, hepatoprotective.
  • Rosmarinic acid – antioxidant, neuroprotective.
  • Flavonoids (orientin, vicenin, luteolin, apigenin) – antioxidant, adaptogenic.
  • Tannins & saponins – astringent, immunomodulatory.

Pharmacological Actions

• • Adaptogen: Helps the body resist stress.
  • Antimicrobial: Effective against bacteria, fungi, and viruses.
  • Anti-inflammatory: Reduces swelling and pain.
  • Antioxidant: Neutralizes free radicals, protects organs.
  • Immunomodulatory: Enhances immune response.
  • Antidiabetic: Improves glucose metabolism.
  • Neuroprotective: Supports memory and cognition.

Traditional & Ethnomedicinal Uses

• • Sacred plant in Ayurveda and Hindu rituals.
  • Decoctions for cough, cold, fever, bronchitis, asthma.
  • Chewing leaves for oral health.
  • Used in skin diseases, wound healing, and digestive disorders

5. Aloevera

Fig.-9: Aloe Vera Pharmacognosy of Aloe Vera (Aloe barbadensis Miller)

Botanical Profile

• Botanical name: Aloe barbadensis Miller
  • Family: Asphodelaceae (formerly Liliaceae)
  • Part used: Fresh leaves (gel and latex)
  • Morphology: Succulent plant with thick, fleshy, lance-shaped leaves containing mucilaginous gel.

 Phytochemical Constituents

• • Anthraquinones: Aloin, emodin – laxative, antimicrobial.
  • Polysaccharides: Acemannan – immunomodulatory, wound healing.
  • Sterols: Campesterol, β-sitosterol, lupeol – anti-inflammatory.
  • Vitamins: A, C, E, B12, folic acid – antioxidant.

Pharmacological Actions

• • Wound healing: Stimulates fibroblast activity and collagen synthesis.
  • Anti-inflammatory: Bradykinase and sterols reduce swelling.
  • Antimicrobial: Effective against bacteria, fungi, and viruses.
  • Immunomodulatory: Acemannan enhances immune response.
  • Digestive aid: Latex (aloin) acts as a laxative; gel soothes gastric irritation.
  • Antioxidant: Vitamins and minerals protect against oxidative stress.

Traditional & Ethnomedicinal Uses

• • Used in Ayurveda, Unani, and folk medicine for burns, wounds, constipation, and skin diseases.
  • Gel applied topically for sunburn, acne, eczema, psoriasis.
  • Juice consumed for digestive health, immunity, and detoxification.

Excipients profile

1. Carbapol 934

Fig.-10: Carbapol934

Chemical & Structural Features

• • Type: Cross-linked polyacrylic acid polymer (carbomer).
  • Synonyms: Carbomer 934P, carboxypolymethylene.
  • Polymerization: Traditionally benzene-based, though non-benzene grades (e.g., Ultrez 10, Carbopol 5984) are recommended.

Functional Properties

• • Thickener & Rheology Modifier: Provides medium to high viscosity in gels, emulsions, and suspensions.
  • Suspending Agent: Stabilizes dispersed particles in formulations.
  • Bioadhesive: Enhances adhesion to mucosal surfaces, useful in topical and oral drug delivery.
  • Flow Characteristics: Exhibits short flow and creamy sensory profile.

2. Propylene Glycol

3. Propyl paraben

Propyl paraben have preservative properties and is valued for its ability to inhibits the growth of microorganisms, including bacteria, fungi, and molds. It is effective against a wide ranges of pathogens and help prevent spoilage in many consumer products.[5]

4. Distilled water

• • Chemical formula: H2O
  • Molar mass :18.02g/mol
  • Shelf life :5 year

Use: Distilled water is used in practical experiments because it's free of impurities and minerals that could interfere with the

Fig.-11:  Distilled water

Procedure

1. Extraction of Herbal Ingredients (maceration methods)-
   1. Fresh guava, neem, and Tulsi leaves were washed thoroughly with distilled water.
   2. The leaves were shade-dried and powdered final.
   3. Each powdered plant material was subjected to maceration with distilled water or 70% methanol for 24 hours.
   4. The mixtures were filtered using Whatman filter paper to obtain clear extracts
   5. The filtrates were concentrated on a water bath at low temperature to obtain thick herbal extracts.
2. Preparation of Gel Base-
   1. The required quantity of Carbopol 940 was dispersed slowly in distilled water and allowed to hydrate for 2–3 hours
   2. Propyl paraben was dissolved in warm water and added to the hydrated Carbopol solution.
   3. Propylene glycol was added as a humectant and co-solvent, and the mixture was stirred until a smooth, uniform gel base was formed.
3. Incorporation of Herbal Extracts-
   1. The concentrated extracts of Guava, Neem, Tulsi, and Aloe vera gel were gradually added to the prepared gel base
   2. Alum (Fitkari) was dissolved separately in a small amount of water and added to the formulation
   3. The entire mixture was stirred continuously using a mechanical stirrer until a uniform and homogeneous herbal gel was obtained.

4. Adjustment of pH-
   1. The pH of the gel was measured (desired pH: 6.0–6.8, suitable for oral mucosa.
   2. Triethanolamine (TEA) was added dropwise to adjust the pH to the required level

Evaluation of herbal gel

1. Visual Appearance

The prepared gels were tested for colour, clarity, texture, transparency and presence of any gritty particles.

2. Measurement of pH

The pH of herbal gel formulations were determined by using paper of pH meter. 0.5 gm of gel was taken and dispersed in 2 ml of distilled water and keep aside for 15-20 minutes. The measurement of pH of formulation was carried out in three times and the average values are 6.0- 6.5

3. Homogeneity

All developed gel formulations were tested for homogeneity by visual inspection after the gels have been set in to the container. They were tested for their presence and appearance of any aggregates.[10]

4. Spread ability

Spread ability is expressed in terms of time in seconds taken by two slides to slip off from gel that is placed in between the slides under the direction of certain load. If the time taken for separation of two slides is less then better the spread ability.[10]

Spread ability is calculated by using the formula.

S = M × L / T

Where M = weight tied to upper slide

L = length of glass slides

T = time taken to separate the slides

Spread ability of gel formulations were reported

Advantages

The gels are having many advantages such as.[10]

1. Gel can avoid gastrointestinal drug absorption difficulties caused by gastrointestinal pH.
2. Gels are having property to avoid enzymatic activity and drug interaction with food and drinks.
3. They can substitute for oral administration of medication when the route is unsuitable
4. They can avoid the first pass effect, that is, the initial pass of drug substance through the human body.
5. They avoid systemic and portal circulation following gastrointestinal absorption.
6. Gels are not deactivated by liver enzymes because the liver is bypassed.
7. They are non-invasive.
8. They have good patient compliance.
9. They are applied over skin for slow and prolonged absorption.
10. Gels have also been applied in pharmacy to some viscous suspension for oral use for
11. example Aluminium hydroxide gel.
12. They have localized effect with

Disadvantages

The gels are having many disadvantages such as.[40]

1. Variable Potency: The concentration of bioactive compounds in herbal extracts can vary depending on factors such as plant species, growth conditions, and extraction methods, leading to inconsistency in efficacy.
2. Limited Scientific Evidence: While traditional use of guava leaves and fitkari for oral health is well-documented, scientific evidence supporting their efficacy as mouth ulcer treatments is still limited, warranting further research.
3. Regulatory Challenges: Herbal products may face regulatory hurdles related to standardization, quality control, and labeling requirements, which can impact their market availability and acceptance.
4. Potential Allergic Reactions: Despite being natural, herbal ingredients can still cause allergic reactions or sensitivities in some individuals, necessitating caution in formulation and usage.
5. Cultural Acceptance: Herbal remedies may not be widely accepted or trusted in certain cultures or communities, leading to reluctance in adopting them as treatment options for mouth ulcers.

RESULT & DISCUSSION