Effect of Probiotics on Oral Wound Healing - Pilot Study (PROWOUND)
Primary Purpose
Wound
Status
Completed
Phase
Not Applicable
Locations
Study Type
Interventional
Intervention
L. reuteri DSM 17938/ATCC PTA
Placebo
Sponsored by
About this trial
This is an interventional supportive care trial for Wound focused on measuring oral wound healing, probiotics
Eligibility Criteria
Inclusion Criteria:
- Healthy individuals (no chronic compromising illnesses)
- Healthy oral conditions
- Good level of oral hygiene
Exclusion Criteria:
- Adults over 50 years old
- Previous allergic reaction to local anesthetics
- Compromised oral mucosa
- Systemic diseases
- Obesity
- Alcoholism
- Smoking habit
- Pregnancy
- Antibiotic therapy within the last two months
- Active infection that needs treatment with antibiotics
- Systemic medications (glucocorticoid steroids, non-steroidal anti-inflammatory drugs -NSAIDS, chemotherapy, anticoagulant therapy) Contraceptives allowed.
- Thrombocytopenia or any of various inherited coagulopathies.
- Patients undergoing radiotherapy
- Inability to provide informed consent by means of physical or mental handicap
Sites / Locations
Arms of the Study
Arm 1
Arm 2
Arm Type
Active Comparator
Placebo Comparator
Arm Label
L.reuteri
Placebo
Arm Description
L. reuteri DSM 17938/ATCC PTA L. reuteri DSM 17938/ATCC PTA lozenges three times daily for 16 days + L. reuteri DSM 17938/ATCC PTA L. reuteri DSM 17938/ATCC PTA probiotic oil, topically, once daily for 8 days.
Placebo lozenges three times daily for 16 days and placebo oil once daily for 8 days
Outcomes
Primary Outcome Measures
Percentage (%) of initial wound size area, measured in digital photographs
The reference value (100%) will be the area before intervention. The percentage is calculated with the second measure (photograph after the intervention)
Secondary Outcome Measures
Levels of salivary oxytocin and salivary IL-10, in pg/ml
As measured with standardized immuno-assays (ELISA kits)
Levels of TNF-α and Interleukin 1β in the wound exudate
As measured with standardized immuno-assays (ELISA kits)
Numerical estimation of S. aureus and β-haemolitic streptococci (CFU/ml)
As measured from bacterial cultures obtained from wound smears
Full Information
NCT ID
NCT03210779
First Posted
July 3, 2017
Last Updated
May 18, 2018
Sponsor
University of Copenhagen
Collaborators
Karolinska Institutet
1. Study Identification
Unique Protocol Identification Number
NCT03210779
Brief Title
Effect of Probiotics on Oral Wound Healing - Pilot Study
Acronym
PROWOUND
Official Title
Effect of Probiotic Tablets and Topical Probiotic Oil (Lactobacillus. Reuteri) on Oral Wound Healing (Pilot Study)
Study Type
Interventional
2. Study Status
Record Verification Date
May 2018
Overall Recruitment Status
Completed
Study Start Date
December 5, 2016 (Actual)
Primary Completion Date
June 30, 2017 (Actual)
Study Completion Date
June 30, 2017 (Actual)
3. Sponsor/Collaborators
Responsible Party, by Official Title
Principal Investigator
Name of the Sponsor
University of Copenhagen
Collaborators
Karolinska Institutet
4. Oversight
Studies a U.S. FDA-regulated Drug Product
No
Studies a U.S. FDA-regulated Device Product
No
Product Manufactured in and Exported from the U.S.
No
Data Monitoring Committee
No
5. Study Description
Brief Summary
This research will study the healing of standardized wounds created in the oral mucosa of volunteer participants during daily intake of tablets and topical application of oil containing the probiotic bacterium Lactobacillus reuteri. Our null hypothesis is that the consumption and topical application of probiotic supplements containing L. reuteri does not improve clinical healing of oral wounds in healthy participants when compared to controls.
Detailed Description
Intervention: after informed consent and baseline collection of clinical data (age,sex), the participants will be randomly assigned to one of the order of the treatments. As a run-in period, they will be given a supply of probiotic or placebo tablets enough for seven days. The participants are instructed to take two tablets daily (one in the morning and one in the evening, days 0-7) and are asked to refrain from any other foods or supplements containing probiotics. On day eight, the participant will attend to the clinics and a punch biopsy will be performed. Depending on the allocation group, probiotic or placebo oil will be supplied. The participant will be instructed to apply the oil locally on the wound with a plastic microbrush for 9 days every evening, and strongly reinforced to continue with the assigned tablets twice daily. The participant will attend the clinics on days two, five and eight post-wounding for follow-up photographies of wound healing. The first intervention period terminates with a wash-out period of four weeks, and then the second run-in an intervention period start following the same procedures.
The probiotic tablets and the probiotic oil (at the recommended dose) contain 2x10^8 CFU of Lactobacillus reuteri. Both the placebo tablets and oil are identical in size and composition but without the addition of the probiotic strains. The tablets and oil are packed in identical pots/bottles with color coding. The code will be kept from the investigators until the analysis are finished. Both the tablets and oil are provided by BIOGAIA AB.
Clinical procedures and sampling
All clinical procedures and sample collections will be conducted by a qualified and authorized dentist from the Institute of Odontology, Section for Cariology, Endodontics, Pediatric Dentistry and Clinical Genetics, Faculty of Health and Medical Sciences, University of Copenhagen. Collected samples (saliva, wound exudate and bacterial smear) will be stored in a freezer at -80 °C until all samples from all patients are gathered -for a maximum of six months. Running of all collected samples at the same time allows laboratory analyses to be carried out efficiently and avoids errors.
Punch biopsy: on day eight, a standardized circular wound of 3.5 mm diameter and approximately 1.5 mm deep will be created in the mucosa of the hard palate with aid of a biopsy punch. The biopsy is performed under aseptic conditions and local anesthesia using lidocaine 2% combined with epinephrine for local hemostasis. A scalpel is used to remove the surface epithelium and underlying superficial connective tissue, which will be discarded according to biosafety regulations from the Institute of Dentistry, University of Copenhagen. After wounding, the wound site will be photographed (blind-coded) with an intraoral camera, placing a standard-sized 6 mm diameter template around the wound as reference. The participant will attend to the dental clinics on days two, five and eight post-wounding to have a photograph of the wound taken by Ph.D student Gina Castiblanco. The wound-size on photographs will be measured by two raters and will be expressed as the ratio of the wound surface area relative to the standard label size with aid of software (Canvas 7, ACD Systems of America). The endpoint will be the percentage of initial wound size, measured at days two, five and eight post-wounding.
Saliva sampling: six samples of unstimulated whole saliva (UWS) fro m each subject will be collected (one at baseline, one right after the run-in period and one at the end of the intervention period). For USW collection, the participant will be sitting in a comfortable position with the head slightly bent. The participant is asked to keep the eyes open and make as few moves as possible, including swallowing. Then, he/she is asked to rinse the mouth in water and spit it out again. Thereafter, he/she is instructed to let saliva flow out of the mouth down a designated plastic cup for ten minutes. Aliquots of saliva (1.5 ml) will immediately be placed on ice and stored in a freezer at -80 °C until further analysis.
Collection of wound exudate: eight samples of wound exudate will be collected in total. The wound site will be dried with cotton pellets and periopaper strips (ProFlow, Amityville, NY, USA) will be gently pressed on the wound for 20 s. In case of blood contamination, the strip is discharged and the sampling is repeated after five min. The volume of exudate is recorded using a Peritron 8000 (Proflow) and expressed in µL. One strip holds approximately 0.1-0.4 µL of exudate. The strips are then transferred to plastic tubes and stored at -80 until further analysis.
Bacterial smear collection: four bacterial smears will be sampled. A sterile swab will be passed through the wound site, placed in a plastic tube and stored freezed at -80 °C until further analysis. The smears will be subjected to conventional microbial cultivation to detects the presence/absence and counts of bacterial species commonly associated with delayed wound healing (β-hemolytic streptococci and S. aureus).
Perception of pain and discomfort: a one-page logbook will be handed to the participants at the beginning of each run-in period so they can register the intake of tablets and their own perception of pain and discomfort according to a VAS-scale. The participant is also encouraged to register factors that could possibly influence the outcome (physical activity and number of alcoholic drinks). The participant will be asked to bring the filled logbook in the end of the intervention period.
Perceived stress and general health: in the end of the intervention period, the participant will be asked to fill the validated General Heath Questionnaire GHQ-12 (10) and the validated Perceived Stress Scale (11). These factors (self-perception of stress and general health) are known to be possible cofounders of wound healing outcomes.
Compliance: at the beginning of each intervention period, the participants will be provided with tablets enough for 16 days and oil for topical use enough for nine days application. Compliance will be checked through the participant´s completion of a logbook in which the daily intake of tablets and oil application is registered. For the last appointment, the participants will be asked to bring the remaining tablets and oil for counting/estimating.
Data analysis: all data will be processed with the open source statistics software "R" and controlled for normal distribution. Multivariate and univariate analysis of variance (MANOVAs and ANOVAs) will be used to assess within-subject change over time. A p-value less than 0.05 will be considered statistically significant. The obtained results will be published in an internationally recognized journal. If the results cannot be published in a journal, they will be available at the website of the University of the Odontology Institute. We make emphasis in that positive, negative and inconclusive results will be published.
Data management: all collected data will be treated in compliance with the Act on Processing of Personal Data and "The Health Law". At inclusion the participants will be assigned a unique number and a separate protocol without personal identification number. This study will not be reported to "Datatilsynet" as it is a private scientific health project
6. Conditions and Keywords
Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Wound
Keywords
oral wound healing, probiotics
7. Study Design
Primary Purpose
Supportive Care
Study Phase
Not Applicable
Interventional Study Model
Crossover Assignment
Model Description
Cross-over pilot trial
Masking
ParticipantCare Provider
Masking Description
Double Blind (Participant, Care Provider, Investigator)
Allocation
Randomized
Enrollment
10 (Actual)
8. Arms, Groups, and Interventions
Arm Title
L.reuteri
Arm Type
Active Comparator
Arm Description
L. reuteri DSM 17938/ATCC PTA L. reuteri DSM 17938/ATCC PTA lozenges three times daily for 16 days + L. reuteri DSM 17938/ATCC PTA L. reuteri DSM 17938/ATCC PTA probiotic oil, topically, once daily for 8 days.
Arm Title
Placebo
Arm Type
Placebo Comparator
Arm Description
Placebo lozenges three times daily for 16 days and placebo oil once daily for 8 days
Intervention Type
Dietary Supplement
Intervention Name(s)
L. reuteri DSM 17938/ATCC PTA
Intervention Description
Ingestion of active lozenge twice daily for 16 days and topical application of active oil once daily for 8 days
Intervention Type
Dietary Supplement
Intervention Name(s)
Placebo
Intervention Description
Ingestion of inactive lozenge twice daily for 16 days application of inactive oil once daily for 8 days
Primary Outcome Measure Information:
Title
Percentage (%) of initial wound size area, measured in digital photographs
Description
The reference value (100%) will be the area before intervention. The percentage is calculated with the second measure (photograph after the intervention)
Time Frame
8 days
Secondary Outcome Measure Information:
Title
Levels of salivary oxytocin and salivary IL-10, in pg/ml
Description
As measured with standardized immuno-assays (ELISA kits)
Time Frame
8 days
Title
Levels of TNF-α and Interleukin 1β in the wound exudate
Description
As measured with standardized immuno-assays (ELISA kits)
Time Frame
8 days
Title
Numerical estimation of S. aureus and β-haemolitic streptococci (CFU/ml)
Description
As measured from bacterial cultures obtained from wound smears
Time Frame
8 days
10. Eligibility
Sex
All
Minimum Age & Unit of Time
18 Years
Maximum Age & Unit of Time
50 Years
Accepts Healthy Volunteers
Accepts Healthy Volunteers
Eligibility Criteria
Inclusion Criteria:
Healthy individuals (no chronic compromising illnesses)
Healthy oral conditions
Good level of oral hygiene
Exclusion Criteria:
Adults over 50 years old
Previous allergic reaction to local anesthetics
Compromised oral mucosa
Systemic diseases
Obesity
Alcoholism
Smoking habit
Pregnancy
Antibiotic therapy within the last two months
Active infection that needs treatment with antibiotics
Systemic medications (glucocorticoid steroids, non-steroidal anti-inflammatory drugs -NSAIDS, chemotherapy, anticoagulant therapy) Contraceptives allowed.
Thrombocytopenia or any of various inherited coagulopathies.
Patients undergoing radiotherapy
Inability to provide informed consent by means of physical or mental handicap
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Svante Twetman, PhD
Organizational Affiliation
University of Copenhagen
Official's Role
Principal Investigator
12. IPD Sharing Statement
Plan to Share IPD
No
Citations:
PubMed Identifier
24675231
Citation
Erdman SE, Poutahidis T. Probiotic 'glow of health': it's more than skin deep. Benef Microbes. 2014 Jun 1;5(2):109-19. doi: 10.3920/BM2013.0042.
Results Reference
background
PubMed Identifier
30359300
Citation
Twetman S, Pedersen AML, Yucel-Lindberg T. Probiotic supplements containing Lactobacillus reuteri does not affect the levels of matrix metalloproteinases and interferons in oral wound healing. BMC Res Notes. 2018 Oct 25;11(1):759. doi: 10.1186/s13104-018-3873-9.
Results Reference
derived
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Effect of Probiotics on Oral Wound Healing - Pilot Study
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