A Novel Combination of Pre-, Pro-, and Postbiotics Demonstrates Protective Effects on Intestinal Permeability in an In Vitro Gut Barrier Model

OBJECTIVES: To test the effects of a novel product combining probiotics (Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12), a prebiotic (bacteriophage blend), and a postbiotic (tributyrin), and its individual components, on intestinal permeability in an in vitro gut barrier model. METHODS: Short term colonic incubation was performed for the individual components, the full product, and a blank control under conditions representative of the proximal large intestine, using the fecal microbiota of a single healthy human donor. Colonic batch suspensions were collected after 48 hours of incubation and filter sterilized. Caco-2 cell monolayers were placed on top of PMA-differentiated THP1-Blue(TM) cells, creating an in vitro model of intestinal barrier disruption. Colonic fermentation samples were diluted in complete medium and given apically to Caco-2/THP1-Blue(TM) co-cultures for 24 hours. Intestinal permeability was assessed with transepithelial electrical resistance (TEER). TEER was measured in Caco-2 monolayers at baseline and 24 hours after pre-treatment of Caco-2/THP-1 Blue(TM) co-cultures. Each final TEER value was normalized to its corresponding initial value. RESULTS: Application of colonic suspensions from the combination of pre-, pro-, and postbiotics significantly increased TEER values compared to control (p < 0.05). Probiotics and tributyrin were also found to increase TEER (p < 0.05) but no significant effect was observed for prebiotic phages (p > 0.05). CONCLUSIONS: Colonic batch fermentations of a novel combination of pre-, pro-, and postbiotics showed improved gut barrier function in vitro, indicated by increased TEER values. Results suggest that probiotic strains and tributyrin likely modulate these effects, consistent with previous research linking the probiotic LGG and short chain fatty acid butyrate to gut barrier protection. FUNDING SOURCES: The study was conducted by an independent lab, ProDigest, and funded and sponsored by Ritual (Natals, Inc.). Compound Solutions, Inc. also financially supported the study.