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.
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
Study Type
INTERVENTIONAL
Allocation
NA
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
17
Stimulation of human colonic biopsies with the fermented product of carrot derived rhamnogalacturonan I
Campus USÖ
Örebro, Sweden
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.
Time frame: 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.
Time frame: The levels of microbial populations and their metabolic products will be measured at baseline and after 6 hours of in vitro fermentation procedure.
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.
Time frame: 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.
Time frame: 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.
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Dietary intake will be evaluated with the use of 3-days diet intake records.
Time frame: 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).
Time frame: Dietary habits will be measured at baseline prior to the initiation of the study as background information.