Assessment of Spirulina Gel Prepared By Different Extraction Methods as an Adjunctive Therapy to Mechanical Debridement in the Treatment of Stage II Grade B Periodontitis

Assessment of Spirulina Gel Prepared By Different Extraction Methods as an Adjunctive Therapy to Mechanical Debridement in the Treatment of Stage II Grade B Periodontitis (Clinical and Laboratory Study)

I) Patients:

the present study will be carried on periodontitis patients. The patients will be informed about the treatment steps. This includes the possible effects or risks, and other treatment options according to the rules of the ethical committee of Faculty of dentistry, Mansoura University.

Groups:

Group I (negative control): 15 periodontally healthy subjects . Group II (positive control): 15 patients will be treated with mechanical debridement only.

Group III (study group A): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel extracted by phosphate buffered saline (PBS).

Group IV (study group B): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.

Group V (study group C): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by ethanol.

In group III, IV and V extract of spirulina algae powder (Emtenan Health Shop,2019 © Egypt,TEL 16246)

Periodontal evaluation will be performed for all patients at baseline and after three months including the following parameters:

• Plaque index (PI).
• Gingival index.
• Probing pocket depth (PPD).
• Clinical attachment level (CAL). Gingival crevicular fluid sampling. Local drug delivery Adequate amount of prepared ozonated sunflower and sunflower oil gel will be delivered into the selected periodontal pockets.

Immunological analysis of TNF-α in GCF

Study Overview

• Status: Completed
• Conditions: Periodontal Disease; Periodontal Therapy; Periodontitis Stage II
• Intervention / Treatment: Other: Mechanical debridement will be performed for 15 stage II periodontitis patient; Drug: spirulina gel extracted by phosphate buffered saline (PBS).; Drug: spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.; Drug: spirulina gel exracted by ethanol.

Detailed Description

I) Patients:

The patient will be selected from Out Patient Clinic, Department of Oral Medicine, Periodontology, Diagnosis and Oral Radiology, Faculty of Dentistry, Mansoura University. The patients will be informed about the treatment steps. This includes the possible effects or risks, and other treatment options according to the rules of the ethical committee of Faculty of dentistry, Mansoura University. The patients must be legally competent to provide written consent before performing any required steps.

A)Inclusion Criteria

1. Patients with Stage II Periodontitis grade B periodontitis (horizontal bone loss with maximum probing depth ≤ 5 mm and with 3-4 mm clinical attachment loss).
2. Minimum of 20 remaining teeth.
3. Age range (25 - 50) years.
4. Good compliance with the plaque control instructions following initial therapy.
5. Availability for follow up and maintenance program. B) Exclusion Criteria

1. Systemic diseases / drugs which could influence the outcome of the therapy (According to Cornell Medical Index-Health Questionnaire).

2. Receiving periodontal therapy in past 6 month. 3. Pregnant and lactating females. 4. Vulnerable groups of patients (e.g handicapped patients and decisionally impaired individuals).

5. Smoking.

D) Groups:

Group I (negative control): 15 periodontally healthy subjects . Group II (positive control): 15 patients will be treated with mechanical debridement only.

Group III (study group A): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel extracted by phosphate buffered saline (PBS).

Group IV (study group B): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.

Group V (study group C): 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by ethanol.

In group III, IV and V extract of spirulina algae powder (Emtenan Health Shop,2019 © Egypt,TEL 16246)

II) Clinical Procedures:

• Medical and dental histories will be taken from patients.
• Mechanical debridement will be performed using suitable ultrasonic and hand instruments for positive control and study groups.

A) Periodontal assessment:

Periodontal evaluation will be performed for all patients at baseline and after three months including the following parameters:

• Plaque index (PI).(16)
• Gingival index.(17)
• Probing pocket depth (PPD).(18)
• Clinical attachment level (CAL).(19) B) Gingival crevicular fluid sampling. The GCF samples will be collected from all subjects at baseline and three months after treatment.(20) C) Local drug delivery Adequate amount of prepared ozonated sunflower and sunflower oil gel will be delivered into the selected periodontal pockets.

III) Laboratory procedures

A) Pharmaceutics Section: Formulation development:

1. Extraction of Spirulina The study will involve extracting active components from spirulina using various solvents under controlled conditions to identify the most efficient extracting solvent and the effect of the solvent on the active components extraction. The oils will be used in preparation of gel formulation for applications in the mouth cavity using different formulation techniques.
2. Development of Formulations:

Based on the oil formulation, various gels will be developed with a focus on creating formulations suitable for clinical application.

• Different gel bases will be evaluated to optimize their physicochemical characteristics.
• Physicochemical Parameters of the gel formulations will be assessed to ensure the desired properties.
• The developed gels will undergo in-vitro testing under different conditions.
• Once optimized, the selected gel formulation(s) will be prepared for testing in clinical trials.

B) Immunological analysis of TNF-α in GCF Assessment of crevicular fluid TNF-α will be performed using enzyme-linked immunosorbent assay technique.

C)Sterilization and Disinfection Protocol for Laboratory Procedures To maintain aseptic conditions and prevent contamination during the laboratory phase, all work surfaces will be disinfected before and after procedures using 70% ethanol or a commercial disinfectant. Personnel will adhere to strict hygiene measures, including wearing disposable gloves, lab coats, and face masks. Glassware and metal instruments will be autoclaved before and after use, while plastic consumables will be pre-sterilized and discarded after single use. The active constituent formulation will be conducted under sterile conditions in a biosafety cabinet, with filtration through 0.22 µm or 0.45 µm Millipore filters to ensure sterility before gel incorporation. The final gel formulations will be packaged in sterile, sealed containers and stored under controlled conditions. Microbiological safety checks, including sterility testing on nutrient agar plates incubated at 37°C for 24-48 hours, will be performed alongside physicochemical assessments (pH and viscosity) to confirm formulation stability. Biohazardous waste will be autoclaved before disposal, and all sharp or glass waste will be discarded in designated biohazard containers following institutional guidelines. These sterilization measures will ensure product integrity, compliance with laboratory safety protocols, and the reliability of experimental results.

• Study Type: Interventional
• Enrollment (Actual): 75
• Phase: Early Phase 1

Contacts and Locations

Study Locations

• Egypt
  • Dakahlia Governorate
    • Al Mansurah, Dakahlia Governorate, Egypt, 00000
      • Mansoura University Faculty of Dentistry

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• 1. Patients with Stage II Periodontitis grade B periodontitis (horizontal bone loss with maximum probing depth ≤ 5 mm and with 3-4 mm clinical attachment loss).

  2. Minimum of 20 remaining teeth. 3. Age range (25 - 50) years. 4. Good compliance with the plaque control instructions following initial therapy.

  5. Availability for follow up and maintenance program.

Exclusion Criteria:

1. Systemic diseases / drugs which could influence the outcome of the therapy (According to Cornell Medical Index-Health Questionnaire).
2. Receiving periodontal therapy in past 6 month.
3. Pregnant and lactating females.
4. Vulnerable groups of patients (e.g handicapped patients and decisionally impaired individuals).

5. Smoking.

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Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Double

Number of Arms

5

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
No Intervention: negative control periodontally healthy subjects; periodontally healthy subjects no intervention
Experimental: positive control; patients will be treated with mechanical debridement only.	Other: Mechanical debridement will be performed for 15 stage II periodontitis patient; Mechanical debridement will be performed for 15 stage II periodontitis patient
Experimental: study group A; patients will be treated with mechanical debridement followed by delivery of spirulina gel extracted by phosphate buffered saline (PBS).	Drug: spirulina gel extracted by phosphate buffered saline (PBS).; 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel extracted by phosphate buffered saline (PBS).
Experimental: study group B; patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.	Drug: spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.; 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by 1.5% calcium chloride (CaCl2) aqueous solution.
Experimental: study group C; patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by ethanol.	Drug: spirulina gel exracted by ethanol.; 15 patients will be treated with mechanical debridement followed by delivery of spirulina gel exracted by ethanol

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
assess the clinical effectiveness of the locally delivered spirulina gel prepared by different pharmaceutical extraction methods as an adjunctive therapy to mechanical debridement in stage II grade B periodontitis.	1-Plaque index: Evaluates the presence of plaque in a dichotomous manner. Each buccal, lingual, mesial, and distal surface of all present teeth is recorded as positive if it presents soft accumulations at the dentogingival junction. 2-Bleeding on Probing : Recorded from all present teeth (excluding third molars) at four sites per tooth: mesial, distal, buccal, and lingual. BOP is assessed in a dichotomous manner, indicating the presence or absence of bleeding following gentle probing.3-Gingival index:score of 0 represents normal gingiva score of 1 mild inflammation score of 2 moderate inflammation score of 3 severe inflammation • 4-Probing pocket depth:It is defined as the distance, measured in millimeters, from the free gingival margin to the base of the gingival sulcus.5-• Clinical attachment level:the distance in millimeters from the cementoenamel junction to the base of the periodontal pocket.	3months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
assess the impact of the locally delivered spirulina gel prepared by different pharmaceutical extraction methods as an adjunctive therapy to mechanical debridement on the GCF level of TNF-α in stage II grade B periodontitis.	GCF sample will be taken to measure the TNF-alpha TNF-alpha is inflammatory marker will be assessed by using the enzyme-linked immunosorbent assay technique.	3 months

Collaborators and Investigators

• Sponsor: Mansoura University
• Investigators: Principal Investigator: Faisal Mohammed Alsharif, Bachelor's degree, Mansoura University

Study record dates

Study Major Dates

• Study Start (Actual): March 10, 2025
• Primary Completion (Actual): April 25, 2026
• Study Completion (Actual): April 25, 2026

Study Registration Dates

• First Submitted: June 3, 2026
• First Submitted That Met QC Criteria: June 9, 2026
• First Posted (Actual): June 15, 2026

Study Record Updates

• Last Update Posted (Actual): June 15, 2026
• Last Update Submitted That Met QC Criteria: June 9, 2026
• Last Verified: March 1, 2025

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Mouth Diseases; Stomatognathic Diseases; Periodontitis; Periodontal Diseases; Inorganic Chemicals; Chlorine Compounds; Calcium Compounds; Chlorides; Hydrochloric Acid; Calcium Chloride
• Other Study ID Numbers: M02020250M

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No