Prebiotic and Anti-inflammatory Effects of a Pectin Polysaccharide

Study Overview
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Brief Summary
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The aim of this study is to determine, quantify and understand the potential prebiotic and anti-inflammatory effects of the pecticpolysaccharide rhamnogalacturonan I (RG-I). The effects of these dietary fibre fractions on barrier function will also be investigated.

Detailed Description
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The dietary fibre fractions from RG-I will be tested first for their prebiotic potential. Batch culture fermentation systems containing basal growth medium will be inoculated with faecal homogenates obtained from different time points (Task I) and will be used for analyzing microbiota composition and microbiota-associated metabolites. Subsequently, the collection of colon biopsies through sigmoidoscopy procedure of the same subjects will take place and the collected biopsies will be mounted in Ussing Chambers. Supernatants collected from Task I will be added to the mucosal side of the biopsy together with a stressor and two permeability markers, in order to investigate the effects of the fibre fractions on both paracellular and transcellular permeability.Throughout the study subjects will complete questionnaires related to their gastrointestinal health and dietary habits

Official Title
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Towards a Food Ingredient Clinically Proven to Benefit Gut Health: Novel RG-I Variants

Conditions
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Healthy

Intervention / Treatment
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* Other: Fermented product of carrot derived rhamnogalacturonan I


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Signed consent prior to any study related procedures Age 18-65 years Willing to abstain from regular consumption of prebiotics/probiotics products or medication known to alter gastrointestinal functions at least 4 weeks prior to the study visits Exclusion Criteria: Previous complicated gastrointestinal surgery Presence of gastrointestinal disorder or any disorder which the principal investigator considers to affect the results of the study Current diagnosis of psychiatric disease Current and past diagnosis inflammatory gastrointestinal disease (e.g. Irritable Bowel Disease) Systemic use of antibiotics or steroids medications in the last 3 months Frequent use of NSAID (Non Steroidal Anti Inflammatory Drugs) the last 2 months prior to study visits Regular consumption of prebiotic/probiotic products for the past 4 weeks Abuse of alcohol or drugs Frequent use of laxatives, anti-diarrheal, anti-cholinergic within last 12 weeks prior to study visits Pregnancy and breast-feeding

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Gut barrier function treatments<br>Stressor, fibre, combination of treatments. | Other: Fermented product of carrot derived rhamnogalacturonan I<br>* Stimulation of human colonic biopsies with the fermented product of carrot derived rhamnogalacturonan I<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change from baseline of barrier function after 90 minutes of ex vivo stimulation of the colonic biopsies. | Barrier function will be evaluated with the use of markers related to paracellular and transcellular permeability, through immunofluoresence and ELISA tecnhique. | Barrier function will be measured at baseline and after 90 minutes of ex vivo stimulation of the colonic biopsies. | 
| Change from baseline to the effect on the intestinal microbial populations and their metabolic products [i.e. Short-Chain Fatty Acids (SCFA)] (prebiotic effect) by the end of the in vitro fermentation process. | Microbial populations will be quantified with 16SRNA sequencing and quantitative Polymerase Chain Reaction (PCR). SCFAs will be quantified with Gas chromatography. | The levels of microbial populations and their metabolic products will be measured at baseline and after 6 hours of in vitro fermentation procedure. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Effect of RG-I fractions on immune system reinforcement in the end of the ex vivo stimulation of the colonic biopsies. | Selected pro- and anti- inflammatory cytokines levels will be measured on the colonic biopsies tissues by ELISA techniques. | Immune system reinforcement will be evaluated after 90 minutes of ex vivo stimulation of the colonic biopsies. | 
| Gastrointestinal health status prior to the initiation of the study. | Gastrointestinal health will be evaluated once with the use of a 1-day questionnaire consisting of 13 items concerning satiety, abdominal pain, diarrhoea, constipation and bloating.The intensity of each parameter will be assessed on a scale of 0-7, where '0' represents absence of symptoms and '7' severe symptoms. | Gastrointestinal health will be measured at baseline prior to the initiation of the study as background information. | 
| Dietary intake prior to the initiation of the study. | Dietary intake will be evaluated with the use of 3-days diet intake records. | Dietary intake will be measured at baseline prior to the initiation of the study as background information. | 
| Dietary habits prior to the initiation of the study. | Dietary habits will be evaluated with the use of food frequency questionnaire (FFQ). | Dietary habits will be measured at baseline prior to the initiation of the study as background information. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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paracellular permeability, transcellular permability, rhamnogalacturonan I, prebiotic, gut microbiota