The in vitro fermentability of two resistant starch preparations type 2 (RS2) and type 3 (RS3) was investigated using human colonic microbiota. Prior to the fermentation experiments, samples were digested using two in vitro models, a batch (ba) and a dynamic (dy), as well as an in vivo method (il) for RS3. Digestion residues were fermented in vitro using a simple batch model lasting 24 h and a more sophisticated dynamic model enduring 72 h. During batch fermentation, metabolite productions and starch degradation rates were similar for RS2 and RS3 but higher for dy- compared to ba-digested samples. RS3il led to the lowest fermentability. Furthermore, increased butyrate ratios were observed for all preparations. The varying RS preparations behaved similarly in the dynamic fermentation but showed high SDs. Moreover, the fermentability was slow during the first 24 h, indicating that the microbiota needed an adaptation period to ferment RS. Propionate ratios increased at the expense of butyrate with exception for RS2dy showing an increase in acetate only. Differences in fermentability observed between the dynamic model, allowing a closer simulation of the in vivo behavior, and the batch model, recommended for screening purposes, could be due to the varying microbiota used.
Resistant starch type 2 (RS2) and type 3 (RS3) containing preparations were digested using a batch (a) and a dynamic in vitro model (b). Furthermore, in vivo obtained indigestible fractions from ileostomy patients were used (c). Subsequently these samples were fermented with human feces with a batch and a dynamic in vitro method. The fermentation supernatants were used to treat CACO2 cells. Cytotoxicity, anti-genotoxicity against hydrogen peroxide (comet assay) and the effect on barrier function measured by trans-epithelial electrical resistance were determined. Dynamically fermented samples led to high cytotoxic activity, probably due to additional compounds added during in vitro fermentation. As a consequence only batch fermented samples were investigated further. Batch fermentation of RS resulted in an anti-genotoxic activity ranging from 9-30% decrease in DNA damage for all the samples, except for RS2-b. It is assumed that the changes in RS2 structures due to dynamic digestion resulted in a different fermentation profile not leading to any anti-genotoxic effect. Additionally, in vitro batch fermentation of RS caused an improvement in integrity across the intestinal barrier by approximately 22% for all the samples. We have demonstrated that batch in vitro fermentation of RS2 and RS3 preparations differently pre-digested are capable of inhibiting the initiation and promotion stage in colon carcinogenesis in vitro.
The digestion resistant fraction obtained in vivo from ileostomy patients (59.4%) is similar to that obtained by the AOAC method for measuring retrograded resistant starch (59.7%). The relative glycemic response after consumption of 50 g of RRM was 58.5% compared to glucose set as 100%. When exposed to colonic microbiota, in vitro obtained indigestible fractions behave similarly to those obtained in vivo in ileostomy patients. Fermentation of RRM and production of butyric acid is negligible during the first months of life but develops subsequently during weaning. In adults, RRM fermentation results in a high yield of SCFA, with butyrate representing 21-31 mol % of total SCFA. The high yield of SCFA during colonic fermentation, observed from weaning age on, as well as the potential to help reduce glycemic load may be of benefit to a number of health-related functions in the host. Further study on clear clinical end points is warranted.
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