Oral and Gut Microbiota Changes During Long-term Fasting in Humans (OralFast)

The aim of this longitudinal trial is to investigate the effects of a 10±3-day fasting period in 30 subjects. Changes in the oral and gut microbiota, halitosis, as well as inflammatory and antioxidative parameters and biomarkers of sulfur metabolism will be analysed.

Study Overview

• Status: Completed
• Conditions: Healthy; Periodontitis
• Intervention / Treatment: Procedure: Long-term fasting

Detailed Description

Fasting has been linked with changes in gut microbiota as part of the physiological adaptation. Furthermore, associations have been described between oral disease and nutrition (diet rich in carbohydrates). The aim of the present longitudinal clinical study is to explore the effect of a 10±3-day fasting period on the oral and gut microbiota. 30 systemically healthy subjects attending the Buchinger Wilhelmi Clinics for fasting, will participate. Changes in oral microbiota will be assessed in plaque and saliva samples and a number of inflammatory markers will be analyzed in Gingival Crevicular fluid (GCF) and blood obtained before, during and after the fasting period. The changes in the oral microbiota will be associated to the composition of the gut microbiota. Changes in periodontal parameters and halitosis related to the fasting period will be further evaluated. Given the known association between sulfur metabolism and the redox status, parameters from the antioxidant status and oxidative damages will also be assessed.

• Study Type: Interventional
• Enrollment (Actual): 36
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Germany
  • Überlingen, Germany, 88662
    • Buchinger Wilhelmi Clinic

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 80 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Both subjects with periodontitis and healthy periodontium will be included.
• Signed informed consent

Exclusion Criteria:

• Not able to sign the informed consent
• diagnosticated with cachexia, anorexia, nervosa, advanced kidney, liver or cerebrovascular insufficiency
• Smoking
• the intake of antibiotics within the last 8 weeks, as well as the intake of probiotics within the last 4 weeks
• periodontal treatment in the last 6 months
• chronic manifest psychical and psychiatric diseases
• participation in another study
• pregnancy or breastfeeding

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Prevention
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Long-term fasting; Long-term fasting according to the Buchinger Wilhelmi fasting program.	Procedure: Long-term fasting; The participants will undergo a fasting program that includes the daily intake of 250 kcal under medical supervision.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in halitosis	Concentration of volatile sulfide compounds (hydrogen sulfide, methylmercaptan and dimethyl sulphide) using the OralChrome devise	daily from baseline to the 3rd day of food reintroduction
Changes in microbiota compositions of subgingival microbial and plaque sampling	Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Parvimonas micra (Pm) and Prevotella intermedia (Pi) analysed by RT-q-PCR	Baseline, fasting day 10, food reintroduction day 3
Changes in microbiota composition in saliva	16 rRNA gene amplicon sequencing	Baseline, fasting day 10, food reintroduction day 3, 1 and 3 months afterwards
Changes in microbiota composition in faeces	16 rRNA gene amplicon sequencing	Baseline, first spontaneous stool after food reintroduction (up to day 3), 1 and 3 months afterwards
Changes in total antioxidant capacity (TAC)	(mmol DPPH/L)	Baseline, fasting day 3, 5, 10
Changes in hydrogen sulfide production capacity	measured in serum and urine using the lead acetate assay	at the beginning and the end of fasting, on day 3 and 5, as well as at the food reintroduction

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in glutathione (GSH)	(µmol/g Hb)	Baseline, fasting day 3, 5, 10
Changes in thiobarbituric acid reactive substances (TBARS)	(µmol/L)	Baseline, fasting day 3, 5, 10
Changes in malondialdehyde (MDA)	nM/L	Baseline, fasting day 3, 5, 10
Changes in protein carbonyls	(nmol/mg protein)	Baseline, fasting day 3, 5, 10
Changes in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase	(µL)	Baseline, fasting day 3, 5, 10
Changes in glutathione reductase (GR) activity	(U/g Hb)	Baseline, fasting day 3, 5, 10
Changes in glutathione peroxidase (GPx) activity	(U/g Hb)	Baseline, fasting day 3, 5, 10
Changes in peroxiredoxin activity	(U/g Hb)	Baseline, fasting day 3, 5, 10
Changes in catalase (CAT) activity	(U/mg Hb)	Baseline, fasting day 3, 5, 10
Changes in superoxide dismutase (SOD) activity	(U/g Hb)	Baseline, fasting day 3, 5, 10
Changes in hydroxyl radical scavenging	(mmol deoxyribose/ml)	Baseline, fasting day 3, 5, 10
Changes in superoxydradical scavenging	(mmol NBT/L plasma)	Baseline, fasting day 3, 5, 10
Changes in ABTS scavenging	(mmol/L)	Baseline, fasting day 3, 5, 10
Changes in reducing power	(mmol/L plasma)	Baseline, fasting day 3, 5, 10
Changes in thiosulfate	measured using the monobromobimane method using HPLC and LC-MS/MS	Baseline, fasting day 10, food reintroduction day 3
Changes in persulfidation	persulfidation in serum and urine	Baseline, fasting day 10, food reintroduction day 3
Changes in cystein	using the S-sulfocysteine (SSC) method which is a HPLC-based method using automated precolumn derivatization with OPA and UV detection at 338 nm in serum and urine	Baseline, fasting day 10, food reintroduction day 3
Changes in amino acid metabolites	using a targeted approach in serum and urine	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-10 in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in G-CSF in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IFN-γ in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in TNF-α in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-1β in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-6 in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-8 in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-17 in blood	analysed in blood with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-10 in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in G-CSF in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IFN-γ in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in TNF-α in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-1β in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-6 in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-8 in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3
Changes in IL-17 in gingival crevicular fluid	analysed in gingival crevicular fluid with Bio-Plex 200 Suspension array System	Baseline, fasting day 10, food reintroduction day 3

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in Rosenberg scale	organoleptic intensity scale for measuring oral malodor: 0 = no odor to 5 = intense odor	daily from baseline to the 3rd day of food reintroduction
Changes in Seemann scale	organoleptic assessment: 1 = odor noticable from 10 cm to 3 = odor noticeable from 1m	daily from baseline to the 3rd day of food reintroduction
Changes in Ayurvedic diagnosis of the tongue	examination of the tongue	daily from baseline to the 3rd day of food reintroduction
Changes in urinary ketone bodies	using ketostix	daily from baseline to the 3rd day of food reintroduction
Changes in capillary blood ketones	using GlucoMen areo 2k	daily from baseline to the 3rd day of food reintroduction
Changes in capillary blood glucose	using GlucoMen areo 2k	daily from baseline to the 3rd day of food reintroduction
Changes in ph value of the urin	using test strips	daily from baseline to the 3rd day of food reintroduction
Changes in weight	kg	daily from baseline to the 3rd day of food reintroduction as well as 1 and 3 months after
Changes in blood pressure	systolic and diastolic in mmHg	daily from baseline to the 3rd day of food reintroduction as well as 1 and 3 months after
Changes in pulse	beats per minute	daily from baseline to the 3rd day of food reintroduction as well as 1 and 3 months after
Plaque Index score	Sillness & Loe, 1964: from 0 = no plaque to 3 = abundant plaque along the gingival margin, interdental spaces filled with plaque	baseline
Gingival index score	Löe & Sillness, 1963: from 0 = normal gingiva to 3 = severe inflammation	baseline
Probing pocket depth	Bleeding on probing, recorded as present within 30 sec (1) or absent (0)	baseline
clinical attachment level	distance between the cemento-enamel junction and the tip of the graduated steel	baseline
recession	distance between CEJ and marginal edge of the free gingiva	baseline
Changes in leucocytes	(thousands/µl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in erythrocytes	(millions/µl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in haemoglobin	(g/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in haematocrit	(in %)	Baseline, fasting day 10, and food reintroduction day 3
Changes in MCV	(fl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in MCH	(pg)	Baseline, fasting day 10, and food reintroduction day 3
Changes in MCHC	(g/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in thrombocytes	(thousands/µl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in international normalized ratio	ratio	Baseline, fasting day 10, and food reintroduction day 3
Changes in Quick	(in %)	Baseline, fasting day 10, and food reintroduction day 3
Changes in partial thromboplastin time	(sec)	Baseline, fasting day 10, and food reintroduction day 3
Changes in aspartate aminotransferase (AST/GOT)	(U/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in alanine aminotransferase (ALT/GPT)	(U/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in gamma-glutamyl transferase (GGT)	(U/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in alkaline phosphatase	(U/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in uric acid	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in urea	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in glomerular Filtration Rate (GFR)	(ml/min/1.73m2KOF)	Baseline, fasting day 10, and food reintroduction day 3
Changes in creatinine	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in thyroid-stimulating hormone (TSH)	(mE/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in total cholesterol	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in triglyceride	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in high-density lipoprotein (HDL)	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in low-density lipoprotein (LDL)	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in LDL / HDL - ratio	ratio	Baseline, fasting day 10, and food reintroduction day 3
Changes in glucose	(mg/dl)	Baseline, fasting day 10, and food reintroduction day 3
Changes in HbA1c	(mmol/mol)	Baseline, fasting day 10, and food reintroduction day 3
Changes in sodium	(mmol/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in potassium	(mmol/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in calcium	(mmol/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in magnesium	(mmol/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in erythrocyte sedimentation rate	(mm/h)	Baseline, fasting day 10, and food reintroduction day 3
Changes in C - reactive protein	(mg/l)	Baseline, fasting day 10, and food reintroduction day 3
Changes in pH value of saliva	(mol/l)	Baseline, fasting day 10, food reintroduction day 3, 1 and 3 months afterwards

Collaborators and Investigators

• Sponsor: Buchinger Wilhelmi Development & Holding GmbH
• Collaborators: King's College London; University of Geneva, Switzerland; ETH Zurich (Switzerland); Buchinger Wilhelmi Clinic
• Investigators: Principal Investigator: Françoise Wilhelmi de Toledo, MD, Buchinger Wilhelmi

Study record dates

Study Major Dates

• Study Start (Actual): September 1, 2022
• Primary Completion (Actual): October 8, 2022
• Study Completion (Actual): January 6, 2023

Study Registration Dates

• First Submitted: June 29, 2022
• First Submitted That Met QC Criteria: July 4, 2022
• First Posted (Actual): July 8, 2022

Study Record Updates

• Last Update Posted (Estimate): January 13, 2023
• Last Update Submitted That Met QC Criteria: January 12, 2023
• Last Verified: January 1, 2023

More Information

Terms related to this study

• Keywords: intestinal microbiota; halitosis; oral microbiota; inflammatory status; redox status; sulfur metabolism
• Additional Relevant MeSH Terms: Stomatognathic Diseases; Periodontal Diseases; Mouth Diseases; Periodontitis
• Other Study ID Numbers: BuWi_OralFast_2022

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

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