Antibacterial effects of elastomeric ligatures coated with Kombucha-synthesized bacterial nanocellulose against streptococcus mutans Randomized Clinical
Determination the Antibacterial effects of elastomeric ligatures coated with Kombucha-synthesized bacterial nanocellulose against streptococcus mutans
Design
A three-way blind clinical trial with a block method and a control group, the sample size in each group was calculated to be 36. A total of 72 patients between 15 and 30 years of age
Settings and conduct
A three-way blind clinical trial with the قشدیخئهظشفهخد block method.
Among the people who refer to the Department of Orthodontics, Faculty of Dentistry, Shiraz University of Medical Sciences, who need permanent orthodontic treatment.
Participants/Inclusion and exclusion criteria
Male and female؛Age ۱۵ to ۳۰ years / non pregnant؛No significant medical history or drug use؛No anti-inflammatory or antibiotic medications taken within 3 months prior to the study؛habit of brushing twice daily with fluoride toothpaste؛No history of periodontal therapy؛Probing depth less than 4 mm؛No gingivitis or active carious lesion؛Gingival and plaque index value <۱
Diseases and medication affect dental biofilm؛Pregnancy؛Poor oral hygiene؛Active periodontal disease؛Severe crowding of anterior teeth or malposition of the lateral incisor؛Smoking؛History of mouth breathing
Intervention groups
A type of brackets , composite and adhesive is used for bonding in all patients. After the initial sorting and place of placing the 18 steel wire, the basic saliva sample is collected from the intervention and control group patients. The dental plaque sample (T0) is taken from the upper lateral incisors bilaterally using a sterile periodontal probe. PEPs obtained plaque samples at T0, BNC release ligatures were used for experimental group patients and conventional ligatures were used for control patients. Saliva and plaque samples were collected again at 7, 14, 21, and 28 days after placement of ligatures is collected.
Main outcome variables
Bacterial colony counting
General information
Reason for update
Acronym
bacterial nanocellulose = BNC
IRCT registration information
IRCT registration number:IRCT20181121041713N5
Registration date:2023-10-22, 1402/07/30
Registration timing:prospective
Last update:2023-10-22, 1402/07/30
Update count:0
Registration date
2023-10-22, 1402/07/30
Registrant information
Name
neda babanouri
Name of organization / entity
Country
Iran (Islamic Republic of)
Phone
+98 71 3628 0801
Email address
babanouri@sums.ac.ir
Recruitment status
Recruitment complete
Funding source
Expected recruitment start date
2023-10-28, 1402/08/06
Expected recruitment end date
2023-11-25, 1402/09/04
Actual recruitment start date
empty
Actual recruitment end date
empty
Trial completion date
empty
Scientific title
Antibacterial effects of elastomeric ligatures coated with Kombucha-synthesized bacterial nanocellulose against streptococcus mutans Randomized Clinical
Public title
Antibacterial effects of elastomeric ligatures coated with bacterial nanocellulose
Purpose
Diagnostic
Inclusion/Exclusion criteria
Inclusion criteria:
Male and female patients
Age 15 to 30 years
non pregnant
No significant medical history or drug use
No anti-inflammatory or antibiotic medications taken within ۳ months prior to the study
No chewing gum or mouthwash use in the week before and during the study
habit of brushing twice daily with fluoride toothpaste
No history of periodontal therapy or current periodontal disease Probing depth less than ۴ mm across the entire dentition
No gingivitis or active carious lesion
Gingival index and plaque index (Silness-Löe) value <1
Exclusion criteria:
Diseases and medication use likely to affect dental biofilm
Pregnancy
Poor oral hygiene
Active periodontal disease
Severe crowding of anterior teeth or malposition of the lateral incisor likely to interfere with the cleaning of the tooth surface
Smoking
History of mouth breathing
Age
From 15 years old to 30 years old
Gender
Both
Phase
3
Groups that have been masked
Participant
Care provider
Data analyser
Sample size
Target sample size:
72
Randomization (investigator's opinion)
Randomized
Randomization description
The selected 72 patients will be randomly divided into test and control groups (n=36). Block randomization method will be used with block size of 6, used online software RANDOM.ORG to allocate patients to intervention and control groups with a ۱:۱ ratio. Subsequently, the random sequences to the study groups will be concealed in opaque envelopes and shuffled before the intervention to increase the unpredictability of the random allocation sequence. Each patient will be asked to pick a sealed envelope to assign to either test or control .Allocation concealment will be done to avoid selection bias.
Blinding (investigator's opinion)
Triple blinded
Blinding description
triple-blind clinical trial
Due to the color change in elastomeric ligatures from gray to gold following BNC coating, the clinician could not be blind to the study groups. However, the patients, the assessor who collect the biofilm samples and the statistician will be blind to the study groups
Placebo
Used
Assignment
Parallel
Other design features
Secondary Ids
empty
Ethics committees
1
Ethics committee
Name of ethics committee
Ethics committee of Shiraz University of Dentistry
Street address
Mehr St.- Qasr Al-Dasht St.- Faculty of Dentistry
City
Shiraz
Province
Fars
Postal code
7195615878
Approval date
2023-08-13, 1402/05/22
Ethics committee reference number
IR.SUMS.DENTAL.REC.1402.055
Health conditions studied
1
Description of health condition studied
Antibacterial effects of elastomeric ligatures coated with Kombucha-synthesized bacterial nanocellulose against streptococcus mutans
ICD-10 code
ICD-10 code description
Primary outcomes
1
Description
Bacterial colony counting
Timepoint
The beginning of the study and days 7, 14, 21 and 28
Method of measurement
Counting the number of bacterial colonies after sampling
Secondary outcomes
empty
Intervention groups
1
Description
Intervention group: A minimum of ۲۰ brackets will be used for patients. fixed pre adjusted edgewise appliances (Mini Master Bracket, ۰.۰۲۲-in MBT prescription; American Orthodontics, USA) with a slot size of ۰.۰۲۲x۰.۰۲۸ inch will be used in all patients. The same type of composite and adhesive will be used for bracket bonding in all patients. After primary leveling and alignment and at placing of steel wire ۱۸, baseline saliva samples will be collected.ntervention group receive coated elastomeric ligatures.Dental plaque sample (T۰) will be obtained from the upper lateral incisor site bilaterally using a sterile periodontal probe. manually delete the supra-gingival plaque , dries and then inter the sterile paper points with tweezers in selected sites. The tip left in situ for about one minute , then removed and inserted into a sterile container for sending in the laboratory (۱۵). The plaque sample will be transferred into a microtube containing ۱ mL saline. After obtaining the plaque samples in T۰, BNC releasing ligatures will be used for patients in the test group while conventional ligatures will be used for control patients. The samples will be placed next to dry ice and will be sent to a microbiology lab in less than ۲ h. Saliva and plaque samples will be collected again at ۷, ۱۴, ۲۱ and ۲۸ days after placement of ligatures (T۱, T۲, T۳, T۴)Preparation of coating solution and Preparation of elastomeric ligatures:The Kombucha SCOBY starter culture was obtained from an Iranian origin and activated every ۱۴ days. The raw cellulose material in the study was Kombucha membranes, an outgrowth of a symbiotic culture of bacteria and yeast fermenting tea broth. The process of making tea infusion involved steeping ۵ grams of green tea (Basilur green tea, Ceylon) in boiling sterile water for ۱۵ min followed by adding ۱۲% high fructose corn syrup as sweetener (Hungrasweet F۵۰, HungranaKft., Ipartelep, Hungary). The sweetened infusion, amounting of two liters, was aseptically poured into a ۳-liter sterile brown glass bottle. Then, ۱۰% of Kombucha tea previously fermented was added as inoculum. The symbiotic culture of bacteria and yeast (SCOBY), originating from a Romanian culture, contained acetobacteria from Komagataeibacter and Gluconobacter genera, as well as yeasts from several genera such as Zygosaccharomyces, Brettanomyces/Dekkera and Pichia, and lactobacteria. A cotton towel was wrapped around the brown glass bottle and secured at the neck of the bottle. The Kombucha culture was allowed to ferment at around ۲۳ °C for ۳۰ days in order to yield Kombucha vinegar. The initial brown Kombucha membranes, referred to as K_۰ sample, weighed ۴۶۱ g. NaOH was tested in ۱ and ۴ M concentrations (۱:۲ solid:liquid ratio) to extract KM from proteins, saccharides, and amino acids, resulting in samples encoded K_۱M and K_۴M. With respect to the thickness of initial membranes, they were washed between ۱۰ to ۳۰ times (۱۰ min each time) by using ۱ M NaOH solutions. The washing process was made more effective by using an Elmasonic P ultrasonic bath (Elma, Singen, Germany). Following the alkaline treatment and ultrasonication process, the purified KMs underwent multiple washes with distilled water until reaching a neutral pH. The beige white membranes (K_۱M), weighing ۳۴۷ g, underwent additional treatment with ۴ M NaOH before being neutralized with distilled water to produce K_۴M. Purified K_۴M never-dried membranes, weighing ۳۳۰ g (۱% w dried cellulose, all concentrations will refer to dry-cellulose content), were ground with a blender (۱۰۰۰ W, ۱۰ series of ۵ min) and diluted with ۵.۵ L water (۵.۷ × ۱۰-۲% w/v) to produce the K_B samples. The K_B samples were subjected to ۳ hours of wet deep grinding by a recirculating colloidal mill (۲۵۰ µm blade space and ۲۵ L/min flow rate), representing about ۸۰۰ passes. Then, the following approach weas experimented:The colloidal mill sample was diluted ۱۰× (۵.۷ × ۱۰-۳% w/v) and then atomized (or spray-dried) by using a Mini Spray dryer B-۲۹۰ (Buchi, Flawil, Switzerland) to attain BNC in dried form. The dried form of nanocellulose is crucial for various bionanocomposites. The optimized operating parameters for nano-atomization were a flow rate of ۴ mL/min for the cellulose suspension, an air flow of ۵۰۰ L/h, with inlet and outlet temperatures of respectively ۱۷۵ °C and ۹۰ °C
Category
Prevention
2
Description
Control group: A minimum of ۲۰ brackets will be used for patients. fixed pre adjusted edgewise appliances (Mini Master Bracket, ۰.۰۲۲-in MBT prescription; American Orthodontics, USA) with a slot size of ۰.۰۲۲x۰.۰۲۸ inch will be used in all patients. The same type of composite and adhesive will be used for bracket bonding in all patients. After primary leveling and alignment and at placing of steel wire ۱۸, baseline saliva samples will be collected from both the control group receive non coated elastomeric ligatures
Category
Placebo
Recruitment centers
1
Recruitment center
Name of recruitment center
Orthodontic Department of Shiraz Dental Faculty
Full name of responsible person
dr. Fatemeh Lavayi mashhadi
Street address
, Mehr St.,Qasr al-Dasht St. Faculty of Dentistry
City
Shiraz
Province
Fars
Postal code
7195615878
Phone
+98 71 3626 3193
Email
Lavaeef@sums.ac.ir
Sponsors / Funding sources
1
Sponsor
Name of organization / entity
Shiraz University of Medical Sciences
Full name of responsible person
Dr. Mohammadhashem Hashempur
Street address
central building of Shiraz University of Medical Sciences - 7th floor- Zand St. - next to Red Crescent
City
Shiraz
Province
Fars
Postal code
7195615878
Phone
+98 71 3235 2782
Email
hashempurm@sums.ac.ir
Grant name
Grant code / Reference number
Is the source of funding the same sponsor organization/entity?
Yes
Title of funding source
Shiraz University of Medical Sciences
Proportion provided by this source
100
Public or private sector
Public
Domestic or foreign origin
Domestic
Category of foreign source of funding
empty
Country of origin
Type of organization providing the funding
Academic
Person responsible for general inquiries
Contact
Name of organization / entity
Shiraz University of Medical Sciences
Full name of responsible person
Dr. Neda Babanouri
Position
Associate Professor
Latest degree
Specialist
Other areas of specialty/work
Dentistry
Street address
Mehr St.,Qasr al-Dasht St. Faculty of Dentistry
City
Shiraz
Province
Fars
Postal code
7195615878
Phone
+98 71 3626 3193
Email
babanouri@sums.ac.ir
Person responsible for scientific inquiries
Contact
Name of organization / entity
Shiraz University of Medical Sciences
Full name of responsible person
Dr. Neda Babanouri
Position
Associate professor
Latest degree
Specialist
Other areas of specialty/work
Dentistry
Street address
Mehr St.,Qasr al-Dasht St. Faculty of Dentistry
City
Shiraz
Province
Fars
Postal code
7195615878
Phone
+98 71 3626 3193
Email
hajmohammadi1996@yahoo.com
Person responsible for updating data
Contact
Name of organization / entity
Shiraz University of Medical Sciences
Full name of responsible person
Amir Hossein Hajmohammadi
Position
قثسهیثدف
Latest degree
Medical doctor
Other areas of specialty/work
Dentistry
Street address
Mehr St.,Qasr al-Dasht St. Faculty of Dentistry
City
Shiraz
Province
Fars
Postal code
7195615878
Phone
+98 71 3626 3193
Email
hajmohammadi1996@yahoo.com
Sharing plan
Deidentified Individual Participant Data Set (IPD)
Yes - There is a plan to make this available
Study Protocol
Yes - There is a plan to make this available
Statistical Analysis Plan
Yes - There is a plan to make this available
Informed Consent Form
Yes - There is a plan to make this available
Clinical Study Report
Yes - There is a plan to make this available
Analytic Code
Yes - There is a plan to make this available
Data Dictionary
Yes - There is a plan to make this available
Title and more details about the data/document
General data including age, sex, medical and dental status, along with outcomes and results can be shared.
When the data will become available and for how long
Access starts 6 months after results are published
To whom data/document is available
Researchers working in academic and scientific institutions
Under which criteria data/document could be used
Access for researchers and people who intend to continue scientific research on this matter.