The Role of Food Matrices Supplemented with Milk Fat Globule Membrane in the Bioaccessibility of Lipid Components and Adaptation of Cellular Lipid Metabolism of Caco-2 Cells

Abstract

This study aimed to evaluate the digestive efficiency of food matrices supplemented with milk fat globule membrane isolated from buttermilk (BM-MFGM), using the INFOGEST in vitro digestion protocol hyphenated with the assessment of the digested material on the lipid profile of the Caco-2 cell culture model. First, we examined lipid profiles in food matrices supplemented with BM-MFGM and their subsequent digestion. The results showed distinct lipid profiles in different food matrices and micellar fractions. The presence of BM-MFGM lipids changed the cellular lipid profiles in Caco-2 cell cultures, with diverging contents in cholesteryl esters, triacylglycerides, and neutral lipids depending on the micellar food matrix factor. Hierarchical clustering analysis revealed patterns in cellular lipid responses to micellar stimuli, while volcano plots highlighted significant changes in cellular lipid profiles post-treatment. Thus, this study underscores the importance of in vitro digestion protocols in guiding food matrix selection for bioactive ingredient supplementation, elucidating intestinal epithelium responses to digested food stimuli.

Keywords: bioaccessibility; food matrix; in vitro digestion; intestinal cell model; milk fat globule membrane.

Figure 1

PLS−DA scores plot (A) and correlation loadings (B) for lipid profiles in LCM (blue circles), JM (red circles), and CM (yellow circles) supplemented with BM−MFGM considering the ‘Food matrix’ as the main factor. The complete list of lipids determined in the study is detailed in Tables S2–S4. Significant lipid features are identified with numbers: 3, triacylglyceride species carbon atom number (CN) CN28; 7, CN36; 8, CN38; 9, CN40; 10, CN42; 13, CN48; 14, CN50; 16, CN54; 17, Ʃneutral species; 18, Ʃpolar species; 19, cholesterol; 20, C10:0; 21, C10:1; 22, C12:0; 23, C14:0; 33, C18:0; 34, C18:1 c9; 37, C18:2; 38, C18:3; 43, C20:4 ω6 (AA); 48, Ʃsaturated fatty acids; 49, Ʃmonounsaturated fatty acids; 50, Ʃpolyunsaturated fatty acids; 51, ω3 fatty acids; 52, ω6 fatty acids; 53, Ʃshort chain fatty acids; 54, Ʃmedium chain fatty acids; 55, Ʃlong chain fatty acids; 56, cholesteryl esters; 57, triacylglycerides; 58, diacylglycerides; 59, free fatty acids plus cholesterol; 60, monoacylglycerides; 61, glucosylceramides; 62, gangliosides; 63, lactosylceramides; 65, phosphatidylethanolamine; 66, phosphatidylinositol; 68, phosphatidylcholine; 69, sphingomyelin; 70, lyso-phospholipids; 71, Ʃpolar lipids; 72, total polar lipids; 73, Ʃceramides; 74, Ʃneutral lipids. Performance of the model is denoted in Table S5.

Figure 2

PLS−DA scores plot (A) and correlation loadings (B) for lipid profiles in micellar fractions isolated from digested food matrices (LCM (blue circles), JM (red circles), and CM (yellow circles), respectively, supplemented with BM−MFGM considering the ‘Micelles class’ factor. The complete list of lipids determined in the study is detailed in Tables S2–S4. Significant lipid features are identified with numbers: 27, C16:0; 34, C18:1 c9; 48, Ʃsaturated fatty acids; 49, Ʃmonounsaturated fatty acids; 56, cholesteryl esters; 58, diacylglycerides; 63, lactosylceramides. Performance of the model is denoted in Table S5.

Figure 3

PLS−DA scores plot (A) and correlation loadings (B) for lipid profiles in cell cultures supplemented with micellar fractions isolated from digested food matrices (LCM (blue circles), JM (red circles), and CM (yellow circles), respectively) supplemented with BM−MFGM considering the ‘Micelles class’ factor. The complete list of lipids determined in the study is detailed in Tables S2–S4. Significant lipid features are identified with numbers: 1, triacylglyceride species carbon atom number (CN); 4, CN30; 5, CN32; 7, CN36; 9, CN40; 10, CN42; 11, CN44; 14, CN50, 15, CN52, 16, CN54; 17, Ʃneutral species; 18, Ʃpolar species; 27, C16:0; 32, C17:1 c10; 33, C18:0; 35, C18:1 c11; 37, C18:2; 43, C20:4 ω6 (AA); 48, Ʃsaturated fatty acids; 49, Ʃmonounsaturated fatty acids; 50, Ʃpolyunsaturated fatty acids; 52, ω6 fatty acids; 56, cholesteryl esters; 57, triacylglycerides; 59, free fatty acids plus cholesterol; 60, monoacylglycerides; 61, glucosylceramides; 68, phosphatidylcholine. Performance of the model is denoted in Table S5.

Figure 4

Heatmaps generated through hierarchical cluster analysis for paired comparisons of lipid profiles from micelles, isolated from digested food matrices (LCM, (A); JM, (B); CM, (C)) supplemented with BM-MFGM, and from the cell cultures treated with each micelle class. Significant lipid features are identified with numbers: 17, Ʃneutral species; 18, Ʃpolar species; 19, cholesterol; 23, C14:0; 27, C16:0; 29, C16:1 c9; 33, C18:0; 34, C18:1 c9; 35, C18:1 c11; 37, C18:2; 43, C20:4 ω6 (AA); 48, Ʃsaturated fatty acids; 49, Ʃmonounsaturated fatty acids; 50, Ʃpolyunsaturated fatty acids; 52, ω6 fatty acids; 54, Ʃmedium chain fatty acids; 55, Ʃlong chain fatty acids; 56, cholesteryl esters; 57, triacylglycerides; 58, diacylglycerides; 59, free fatty acids plus cholesterol; 60, monoacylglycerides; 61, glucosylceramides; 63, lactosylceramides; 65, phosphatidylethanolamine; 68, phosphatidylcholine; 69, sphingomyelin; 71, Ʃpolar lipids; 72, total polar lipids; 73, Ʃceramides; 74, Ʃneutral lipids.

Figure 5

Volcano plots showing changes in lipid features analyzed in cell cultures treated with micelles isolated from digested food matrices (LCM, (A); JM, (B); CM, (C)) supplemented with BM−MFGM. Lipid features with significance of fold change in experimental vs. control cells are indicated for downregulated (yellow circles) and upregulated (blue circles) lipid features. No significant fold change lipid classes are included (grey circles). Lipid features are identified with numbers: 17, ƩNeutral species; 18, Ʃpolar species; 19, cholesterol; 29, C16:1 c9; 33, C18:0; 34, C18:1 c9; 35, C18:1 c11; 37, C18:2; 48, Ʃsaturated fatty acids; 50, Ʃpolyunsaturated fatty acids; 53, Ʃshort chain fatty acids; 56, cholesteryl esters; 57, triacylglycerides; 58, diacylglycerides; 59, free fatty acids plus cholesterol; 60, monoacylglycerides; 61, glucosylceramides; 68, phosphatidylcholine; 71, Ʃpolar lipids; 74, Ʃneutral lipids. Lipid features with significance of fold change in experimental vs. cells.