Bjarne Thorsted1, Maria Bloksgaard2, Alexandra Groza3, Lars Peter Schousboe4, Nils J Færgeman3, Jens A Sørensen5, Viggo Svane-Knudsen, Jonathan R Brewer6. 1. MEMPHYS-Centre for Biomembrane Physics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark. 2. Department of Cardiovascular and Renal Research, Institute of Molecular Medicine, University of Southern, Odense C, Denmark. 3. Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark. 4. Department of ENT Surgery, Vejle Hospital, Vejle, Denmark. 5. Department of Reconstructive Surgery, Odense University Hospital, Odense C, Denmark. 6. MEMPHYS-Centre for Biomembrane Physics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark brewer@bmb.sdu.dk.
Abstract
OBJECTIVES: To quantify the barrier sterols and image the lipid structures in the matrix of acquired cholesteatoma and compare the distribution with that found in stratum corneum from normal skin, with the goal to resolve their potential influence on cholesteatoma growth. METHODS: High-performance thin-layer chromatography (HPTLC) was used to achieve a quantitative biochemical determination of the sterols. The intercellular lipids were visualized by Coherent Anti-Stokes Raman scattering (CARS) microscopy, which enables label-free imaging of the lipids in intact tissue samples. RESULTS: The results show that the total lipid content of the cholesteatoma matrix is similar to that of stratum corneum from skin and that the cholesteatoma matrix unquestionably contains cholesterol. The cholesterol content in the cholesteatoma matrix is increased by over 30% (w/w dry weight) compared to the control. The cholesterol sulfate content is below 1% of the total lipids in both the cholesteatoma and the control. Cholesterol ester was reduced by over 30% when compared to the control. CONCLUSIONS: The content of cholesterol in the cholesteatoma matrix is significantly different from that in stratum corneum from skin, and we confirm that the main structure of the cholesteatoma resembles very thick stratum corneum.
OBJECTIVES: To quantify the barrier sterols and image the lipid structures in the matrix of acquired cholesteatoma and compare the distribution with that found in stratum corneum from normal skin, with the goal to resolve their potential influence on cholesteatoma growth. METHODS: High-performance thin-layer chromatography (HPTLC) was used to achieve a quantitative biochemical determination of the sterols. The intercellular lipids were visualized by Coherent Anti-Stokes Raman scattering (CARS) microscopy, which enables label-free imaging of the lipids in intact tissue samples. RESULTS: The results show that the total lipid content of the cholesteatoma matrix is similar to that of stratum corneum from skin and that the cholesteatoma matrix unquestionably contains cholesterol. The cholesterol content in the cholesteatoma matrix is increased by over 30% (w/w dry weight) compared to the control. The cholesterol sulfate content is below 1% of the total lipids in both the cholesteatoma and the control. Cholesterol ester was reduced by over 30% when compared to the control. CONCLUSIONS: The content of cholesterol in the cholesteatoma matrix is significantly different from that in stratum corneum from skin, and we confirm that the main structure of the cholesteatoma resembles very thick stratum corneum.
Authors: Vinzent Kevin Ortner; Nhi Nguyen; Jonathan R Brewer; Vita Solovyeva; Merete Haedersdal; Peter Alshede Philipsen Journal: Lasers Surg Med Date: 2022-04-22