Borchman, Foulks, Yappert
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1 Spectroscopic Characterization of Human Meibum Borchman, Foulks, Yappert
2 Prof. Marta Yappert Dr. Gary Foulks
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4 Composition Structure Function
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11 Composition Structure Function
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13 Tear Film Lipid Layer Function Decrease the rate of evaporation Reduce the free energy at the tear film surface and increase aqueous layer thickness Maintain Viscosity and surface tension
14 From Borchman et al. Eye & Contact Lens. In Press, Evaporation Rate (mg/min/cm 2 ) Phosphate Buffered Saline Doug's Tears
15 Evaporation Rate mg/min/cm plus bound wax plus wax on the surface plus wax emulsion Lactoglobulin (mg/ml): From Borchman et al. Eye & Contact Lens. In Press, 2009
16 Composition Structure Function
17 Composition There are over 30,000 molecular species of lipids in human meibum (Nicolaides and Santos, 1985)
18 TABLE 1 Comparison of the major lipid composition of normal human meibomian lipids Lipid Cory (1973) (%) Tiffany (1978) (%) Nicolaides (1981) (%) McCulley (1997) (%) Mathers (1998) (%) Range (%) HC WE 64 a SE See a Diest ers TG Alcs FFA Ch Polar Butovich et al. Curr Eye Res May;33(5):405-20
19 TABLE 1 Comparison of the major lipid composition of normal human meibomian lipids Lipid Cory (1973) (%) Tiffany (1978) (%) Nicolaides (1981) (%) McCulley (1997) (%) Mathers (1998) (%) Range (%) HC WE 64 a SE See a Diest ers TG Alcs FFA Ch Polar Butovich et al. Curr Eye Res May;33(5):405-20
20 Composition Alkanes compose 0 % to 36 % of the meibum
21 Composition Esters compose between 20-80% of the meibum Wax Ester 13-32%
22 Composition Cholesterol esters make up 7-50% of the meibum
23 Is the large variation in lipid classes due to experimental variation, or variation from person to person, time, sex, age or Meibomian Gland dysfunction?
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25 Cholesterol Esters TG DG Wax Esters RCH OH 2 CH 2 R-C=O H CH=CH CH 3 Parts per Million
26 100 % of Measured Lipids Normal Dry Eye 0 Wax Esters Chol. Esters Triglycerides Diglycerides
27 Fourier Transform Infrared and Raman Spectroscopies
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29 Wax Ester Meibum CHOLESTEROL SM FA DPPC
30 Meibum Lipid Composition is Different from Tear Film Lipid Composition
31 Raman Spectra of Human Meibum and Standards Human Meibum Wax Frequency (cm -1 )
32 Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF/MS)
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34 Lipids were measured with a sensitivity of 9 pmoles. In spectra where the wax was made to be invisible, tear fluid lipid composition was much more complex than for meibum lipid. The opposite was true for spectra where the wax was made visible.
35 Composition Structure Function
36 What is Known about Tear Lipid Molecular Structure/ conformation?
37 Nothing
38 Tear Lipid Hydrocarbon Chain Conformation Using FTIR
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41 ν sym = P 1 + (P 2 /((1+P 3 /T) P 4 )) 100 P 4 l l 50 P 2 P 1 0 P3 Temperature ν sym (a measure of lipid disorder) gauche/(gauche + trans) X100
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43 . As the temperature increased from 25 to 40 o C, hydrocarbon chains became disordered (20% to 62%). These structural changes may contribute to the therapeutic spreading of meibum with heat.
44 Crystal Field Splitting
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47 Does Meibum Lipid Compositon/Molecular-Structure Change With Age or Meibomian Gland Disfunction?
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49 Meibum from Normal Donors 84 years old 3 years old
50 Meibum from Normal Donors
51 From Raman Spectra of Human Meibum Age (y) Conjugated C=C bonds from carotinoids I1520 / I 1303
52 cicatricial disease Foulks. Survey of Ophthalmology Volume 52, Issue 4, July-August 2007, Pages
53 60 Phase Transition Temperature ( o C) Lipid Order at 34 o C (% trans rotomers) Dry Eye 30 Normal 20 Dry Eye Normal
54 From: Nagymihalyi, A.; Dikstein, S.; Tiffany, J. M Exp Eye Res 2004, 78,
55
56 Conclusions Meibum lipid composition changes with age and dry-eye symptoms. These compositional changes are enough to change lipid hydrocarbon conformation and lipid-lipid interactions. Wax, a major component of meibum, binds to proteins found in tears. Wax bound to proteins in solution does not influence the rate of evaporation in vitro. However, lipid on the surface strongly lowers the rate of evaporation. This work also highlights the power of spectroscopic methods to characterize tear film lipid composition-structure-function relationships and lipid-protein interactions that will be applied in future studies in relation to age, sex and dry eye symptoms.
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58 The End
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