Comparative Evaluation of the Anti-Plaque and Anti-Gingivitis Effects of Nanobiofusion Gingival Gel (NBF), Placebo Gel, and Commercial Toothpaste

Participants in this group serve as the test group and will use the Nano-Bio Fusion (NBF) gingival gel, which is a toothpaste-like gel with bioadhesive properties created using nanotechnology. The gel uses nano-sized particles to improve adhesion and cellular uptake in the moist oral environment, and it is formulated with naturally derived active ingredients, including antioxidants (vitamins C and E), bee propolis, aloe vera, green tea extract, olive oil, and mastic oil.

Participants in this group serve as the test group and will use the Nano-Bio Fusion (NBF) gingival gel, which is a toothpaste-like gel with bioadhesive properties created using nanotechnology. The gel uses nano-sized particles to improve adhesion and cellular uptake in the moist oral environment, and it is formulated with naturally derived active ingredients, including antioxidants (vitamins C and E), bee propolis, aloe vera, green tea extract, olive oil, and mastic oil.

They will apply a pea-sized amount of the gel to a standardized soft-bristled toothbrush and brush all tooth surfaces and the gumline using the Modified Bass technique. Participants are required to brush thoroughly twice daily (in the morning after breakfast and at night before bed) for 2 minutes per session over a 3-month follow-up period.

Participants in this group serve as the placebo control to isolate the specific effects of the active ingredients. They will use a custom-formulated placebo gel developed by NanoCureTech that exactly matches the color, consistency, and flavor of the NBF gel, but lacks the active nano-bio fusion ingredients.

Participants in this group serve as the placebo control to isolate the specific effects of the active ingredients. They will use a custom-formulated placebo gel developed by NanoCureTech that exactly matches the color, consistency, and flavor of the NBF gel, but lacks the active nano-bio fusion ingredients. Participants in this arm will apply a pea-sized amount of the placebo gel to a standard soft-bristled toothbrush and brush all tooth surfaces and the gumline using the Modified Bass technique. Participants are required to brush thoroughly twice daily (in the morning after breakfast and at night before bed) for 2 minutes per session over a 3-month follow-up period.

Participants in this group serve as the positive control, representing the current standard of oral hygiene. They will use a commercial dentifrice (Colgate maximum cavity protection) that contains 1450 ppm fuoride (1.1% sodium monofluorophosphate), calicium, and arginine, along with standard toothpaste excipients. To maintain blinding, it will be dispensed in identical opaque tubes.

Participants in this group serve as the positive control, representing the current standard of oral hygiene. They will use a commercial dentifrice (Colgate maximum cavity protection) that contains 1450 ppm fuoride (1.1% sodium monofluorophosphate), calicium, and arginine, along with standard toothpaste excipients. To maintain blinding, it will be dispensed in identical opaque tubes. Participants in this arm will apply a pea-sized amount of the toothpaste to a standard soft-bristled toothbrush and brush all tooth surfaces and the gumline using the Modified Bass technique. Participants are required to brush thoroughly twice daily (in the morning after breakfast and at night before bed) for 2 minutes per session over a 3-month follow-up period.

A disclosing agent (EMS biofilm doscloser) will be applied on all teeth and all surfaces to make the plaque visible. Clinical intra-oral photos will be taken for all teeth and all surafces (from 2nd molar on the right side to the 2nd molar on the other left side) for the buccal and palatal/lingual surfaces. The photos will be assessed by a blinded examiner to record plaque on the mesial, middle, and distal sites on both the buccal and lingual surfaces of the teeth. Each site will be given a score according to the following index:

Score Description 0 No plaque

1. Separate flecks of plaque at the cervical (gingival) margin
2. Thin continuous band of plaque (≤1 mm) at the cervical margin
3. Band of plaque >1 mm wide but covering <1/3 of the crown
4. Plaque covering ≥1/3 but <2/3 of the crown
5. Plaque covering ≥2/3 of the crown

Scores will be recorded at the mesial, middle, and distal sites on both the buccal and lingual surfaces of teeth (from 2nd molar on the right side to the 2nd molar on the left side). Each site will be scored according to the following index:

Score 0: Normal gingiva: Healthy appearance, No inflammation, No bleeding on probing Score 1: Mild inflammation: Slight change in color, Slight edema (swelling), No bleeding on probing Score 2: Moderate inflammation: Redness, Edema and glazing, Bleeding on probing present Score 3: Severe inflammation: Marked redness and edema, Ulceration, Spontaneous bleeding (may occur without probing)

This measure serves as an objective and sensitive indicator of gingival inflammation, used to complement the Gingival Index. It is assessed by gently probing the gingiva with periodontal probe and recording bleeding as present or absent. The outcome is reported as a full-mouth Bleeding on Probing percentage for each participant. The score ranges from 0% to 100%, where 0% is equivalent to completely healthy gums with no bleeding upon probing, and higher percentages indicate more widespread gingival inflammation and poorer gum health.

Extrinsic teeth staining will be evaluated. Before the application of disclosing agent to assess plaque, clinical intra-oral photos will be taken for all teeth and all surafces (from 2nd molar on the right side to the 2nd molar on the other left side) for the buccal and palatal/lingual surfaces. The photos will be assessed by a blinded examiner to record stains on both the buccal and lingual surfaces of the teeth. Each site will be given a score according to the following index:

Score 0: No discoloration, clean and polished tooth surface, natural appearance in color Score1: Slight yellowing discoloration, yellowish film over the entire extent of the clinical crown, slight brownish discoloration along the gingival margin.

Score 2: Moderate brownish discoloration on the interproximal surfaces and in the apical third of the clinical crown.

Score 3: Heavy brown and black discoloration over the entire extent of the tooth surface, black discoloration predomina.

Participants will rate the following criteria for the used product on numeric rating scale from 0 to 10:

• Taste (0 very bad taste to 10 very good taste).
• Texture (0 very unpleasant to 10 very pleasant).
• Foam (0 very unpleasant to 10 very pleasant).
• Ease of use (0 very difficult to use to 10 very easy to use).

Participants will also be asked to choose the most appropriate answer for the following questions:

• Are you going to use the product after study completion? (will not ever use it again, might use it again, or will definitely use it again).
• Have you noticed any teeth discoloration after using the product? (the teeth became cleaner and whiter, no change in tooth colour, teeth became more yellowish in colour).
• Have you felt any gum pain or discomfort while using the product? (yes, no, sometimes).
• Have you experienced any allergy after using the product? (yes, no).
• Would you recommend using the product to family and friends? (yes, no, maybe).