研究目的
To demonstrate a direct link between vascular function/dysfunction, the state of lipids and the cortical stiffness of endothelial cells, by detecting all parameters within the same murine ex vivo aorta preparation.
研究成果
The study demonstrated a direct link between the phenotype of endothelial dysfunction, increased intracellular lipids and elevated cortical stiffness in the ApoE/LDLR-/- mice. The combined Raman/AFM setup provided detailed information related to the outer endothelial layer of the aorta, highlighting the interdependency between intracellular lipid contents and cortical nanomechanics contributing to the development of endothelial dysfunction and cardiovascular diseases.
研究不足
The study was limited to ex vivo preparations, and the exact localization of the probed cell area in AFM measurements was challenging. The study also did not analyze the cholesterol content due to the weak specific cholesterol marker bands in Raman spectroscopy.
1:Experimental Design and Method Selection:
Combined Raman spectroscopy and Atomic Force Microscopy (AFM) was used to analyze the biochemical and nanomechanical properties of living endothelial cells within the same area of the ex vivo aorta preparation.
2:Sample Selection and Data Sources:
Aortas from ApoE/LDLR-/- mice and wild type (WT) mice at the age of 6 months were used.
3:List of Experimental Equipment and Materials:
Confocal Raman Imaging system (WITec alpha 300, Germany), AFM (WITec alpha 300), soft triangular cantilevers with gold coated backs and a polystyrene sphere with 10 μm diameter as a tip (Novascan, USA).
4:Experimental Procedures and Operational Workflow:
Raman imaging and AFM-measurements were performed in fluid. Nanoindentation measurements on in situ endothelial cells of ex vivo aortic explants were performed.
5:Data Analysis Methods:
Raman integral intensities were calculated in OPUS 5, whilst all other values were calculated using OriginPro. The two sample t-test or Tukey test (Raman), the two sample t-test or Mann-Whitney test (AFM) or the unpaired, one-tailed t-test (EPR) was performed.
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