Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 126, Issue 33, Pages 6338-6344Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpcb.2c04154
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Funding
- National Science Foundation [ECCS-1542152]
- Stanford EDGE
- Bio-XBowes fellowship
- ChEM-HCBI fellowship
- National Institutes of Health(NIH) [T32GM120007]
- NIH [K08EY028176, P30-EY026877]
- Research to Prevent Blindness(RPB)
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This work develops a tunable in vitro model of the mucin layer of the human tear film. The model involves supported lipid bilayers and biotinylated bovine submaxillary mucins tethered onto the bilayers. The model can be used with interferometry to monitor film thickness and investigate tear film breakup. Higher mucin coverage in the model leads to delayed breakup times, potentially due to a thicker hydrated layer that protects film stability.
In this work, we describe the development of a tunable, acellular in vitro model of the mucin layer of the human tear film. First, supported lipid bilayers (SLBs) comprised of the phospholipid DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) and biotinyl cap PE (1,2-dioleoyl-sn-glycero-3-phosphoethanol-amine-N-(cap biotinyl)) are created on the surface of a glass dome with radius of curvature comparable to the human eye. Next, biotinylated bovine submaxillary mucins (BSM) are tethered onto the SLB using streptavidin protein. The mucin presentation can be tuned by altering the concentration of biotinylated BSM, which we confirm using fluorescence microscopy. Due to the optically smooth surface that results, this model is compatible with interferometry for monitoring film thickness. Below a certain level of mucin coverage, we observe short model tear film breakup times, mimicking a deficiency in membrane-associated mucins. In contrast, the breakup time is significantly delayed for SLBs with high mucin coverage. Because no differences in mobility or wettability were observed, we hypothesize that higher mucin coverage provides a thicker hydrated layer that can protect against external disturbances to thin film stability. This advance paves the way for a more physiological, interferometry-based in vitro model for investigating tear film breakup.
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