Abstract
Akkermansia muciniphila is a mucin-degrading intestinal bacterium widely recognized for its beneficial effects on metabolic health, particularly in type 2 diabetes mellitus (T2DM).In the present study, the safety and antidiabetic-related functional properties of live and pasteurized A. muciniphila were first evaluated in vitro.Cytotoxicity, intestinal epithelial barrier integrity, and inhibition of carbohydrate-hydrolyzing enzymes (α-amylase and α-glucosidase) were assessed using Caco-2 cell models and enzyme-based assays.Pasteurized A. muciniphila demonstrated no cytotoxic effects, preserved epithelial barrier integrity, and showed enhanced inhibition of α-amylase and αglucosidase compared with the live form.Based on these findings, pasteurized A. muciniphila was incorporated into a chocolate-based functional food matrix.The resulting formulation was further characterized for its polyphenol and flavonoid content and antioxidant activity using DPPH, ABTS, and Ferric Reducing Antioxidant Power (FRAP) assays.The antidiabetic potential of the functional chocolate was evaluated in vivo using a high-fat diet/streptozotocin-induced T2DM rat model.Supplementation with chocolate enriched with pasteurized A. muciniphila significantly improved fasting blood glucose, insulin sensitivity, lipid profile, liver enzyme activity, and oxidative stress markers compared with diabetic controls.Overall, this study demonstrates that pasteurized A. muciniphila combined with a polyphenol-rich chocolate matrix represents a safe and effective functional food strategy for improving metabolic disturbances associated with T2DM.