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Investigation of Bioactive Milk Phospholipid Liposomes and Soy Phospholipid Liposomes on Adipocyte Physiology

Kosmerl, Erica L
http://orcid.org/0000-0002-0751-4990
http://rave.ohiolink.edu/etdc/view?acc_num=osu1565776533223238

Abstract Details

2019, Master of Science, Ohio State University, Food Science and Technology.
Milk phospholipids (MPLs) are valuable dairy components and appear to impart human health benefits, including improved cognitive function in infants and adults. Furthermore, phospholipids (PLs) have been shown to lower LDL cholesterol and improve intestinal barrier function. The commercial food industry uses other dietary sources of PLs, such as soy lecithin. However, it remains unclear whether dissimilar composition of PLs from different dietary sources convey similar benefits. The hypothesis of this work was that milk PL (MPL) treatment will result in greater cellular SM content in 3T3-L1 adipocytes and enhance the ability to prevent a lipopolysaccharide (LPS)-induced inflammatory response compared to soy PLs (SPLs). To improve stability of PLs in cell culture media, liposome structures (LUVs) made from MPLs and SPLs (MPL-LUVs and SPL-LUVs, respectively) were prepared, optimized and characterized. Our objectives were then to quantify total cellular PL composition by high-performance liquid chromatography (HPLC), visualize the physiological effects of LUV treatment using brightfield and confocal laser scanning microscopy, and analyze the production of pro-inflammatory cytokines (IL-6 and MCP-1) after LUV treatment and subsequent LPS stimulation. Large and stable unilamellar vesicles were attained with diameters of 229.3 ± 7.0 and 235.3 ± 28.4 nm and zeta potentials of -15.63 ± 3.19 and -27.71 ± 2.67 mV for MPL- and SPL-LUVs, respectively. Cell viability and cytotoxicity assays showed that up to 0.5 mg PL/mL MPL- or SPL-LUVs was within the physiological range for fully-differentiated 3T3-L1 adipocytes. Cellular PLs after 0.5-mg PL/mL LUV treatments were then quantified using HPLC and resulted in a greater proportion of SM in MPL-LUV treated cells (17.6 μg/100 μg PL) than SPL-LUV treated cells (11.5 μg/100 μg PL). The morphological changes after LUV treatments were observed by brightfield microscopy, which demonstrate a disruption in normal adipocyte morphology by SPL-LUVs, but not MPL-LUVs. To visualize differences in localization and processing of LUVs, fluorescent LUVs (fLUVs) for both milk and soy were produced and treated to adipocytes for 24-h. Confocal micrographs of the adipocytes showed accumulation of fLUVs at the plasma membrane for both milk and soy, suggesting the plasma membrane is modified by LUV treatment. To assess whether the change in cellular SM, possibly at the plasma membrane, could result in physiological benefits, 24-h MPL- and SPL-LUV treatment was tested to prevent an NF-κB-mediated inflammatory response stimulated by 1 μg/mL lipopolysaccharide (LPS). IL-6 secretion did not differ between treatments; however, MCP-1 secretion was significantly reduced by 35% and 41% for MPL-LUV and SPL-LUV treatments, respectively (p<0.05). It is speculated that LUV treatments activate the PPAR⍺ transcription factor, preventing transcription of NF-κB pro-inflammatory genes. In summary, the most striking difference was the effects on cell morphology after LUV treatment, suggesting that milk and soy LUVs can alter plasma membrane composition and distribution of PLs in adipocytes, leading to various changes on membrane dynamics and, likely, cell signaling.
Rafael Jimenez-Flores (Advisor)
Ouliana Ziouzenkova (Committee Member)
Luis Rodriguez-Saona (Committee Member)
118 p.
Food Science
liposomes, nuclear factor kappa b, phospholipids, sphingomyelin

Recommended Citations

Citations

  • Kosmerl, E. L. (2019). Investigation of Bioactive Milk Phospholipid Liposomes and Soy Phospholipid Liposomes on Adipocyte Physiology [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1565776533223238

    APA Style (7th edition)

  • Kosmerl, Erica. Investigation of Bioactive Milk Phospholipid Liposomes and Soy Phospholipid Liposomes on Adipocyte Physiology. 2019. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1565776533223238.

    MLA Style (8th edition)

  • Kosmerl, Erica. "Investigation of Bioactive Milk Phospholipid Liposomes and Soy Phospholipid Liposomes on Adipocyte Physiology." Master's thesis, Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1565776533223238

    Chicago Manual of Style (17th edition)

osu1565776533223238
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This open access ETD is published by The Ohio State University and OhioLINK.