Objective This study aimed to explore biotinylated liposomes (BLPs) as novel carriers to enhance the oral delivery of insulin.Methods The hypothesis of specific receptor-ligand interaction was investigated using the Caco-2 cell model.Biotin labeled with 5-aminofluorescein was used to specifically bind the biotin receptors,and this event was photographed using a confocal laser scanning microscope (CLSM).Caco-2 cell monolayers and Caco-2/Raji co-cultured monolayers with M cell characteristics were used to assess the transport of the INS preparations.The cellular uptake of fluorescent isothiocyanate-labeled insulin (FITC-INS)-loaded liposomes under different conditions was visualized and quantified by CLSM and flow cytometry,respectively.Results The significance of the biotinylation was confirmed by the facilitated absorption of the BLPs through receptor-mediated endocytosis.Increased cellular uptake and quick gastrointestinal transport further verified the ability of the BLPs to enhance absorption.These results provide a proof of concept that BLPs can be used as potential carriers for the oral delivery of insulin.Conclusions The oral delivery of insulin was enhanced with BLPs.It is the biotin receptor mediated transcellular route rather than the paracellular route that INS was transported through the intestinal epithelial cells.