origin may be separated. The first subgroup is made up of cells from large blood
vessels, among which the best-defined and the most commonly used in endothelial
research is the EA.hy926 cell line [68]. The second subgroup is composed of
endothelial cell lines for the study of microvasculature with the first characterized
HMEC-1 (human microvascular endothelial cells) line from the foreskin microcirculation [69]. Compared with other culture lines, they proved to be the best model
for endothelial research, because they most closely reflected the processes observed
in primary cells [70].
6.2.2 In Vitro Cell Models of Pathophysiology
of the Endothelium
The number of published papers on endothelial studies using microscopic and
spectroscopic methods is rather small, and the majority of these works deal with the
endothelium in the blood vessel wall [71]. Thematically, they can be divided into
works addressing the general characterization of the endothelium and referring to
the disease states, mainly in murine models [60, 71–76]. The Raman imaging
combined with atomic force microscopy (AFM) and immunohistochemical staining
was used to characterize the chemical composition and topography of the
endothelium [75]. In vivo Raman studies on the endothelium in the aorta wall of the
sheep and in vitro in the human aorta have also been published [77]. In addition,
spectroscopic methods were used in studies of endothelial pathologies:
atherosclerosis [76, 78], hypertension [60, 73], diabetes type 2 [74] and cancer
metastasis [72]. In these studies, changes in the protein and/or lipid content
observed in the endothelial layer were treated as a marker of endothelial condition
in diseases and, therefore, could be used in diagnostic context. Analysis of alterations arising from disease development requires knowledge about normal,
non-disease-affected samples. Raman microscopy was used for comparison of the
control EA.hy926 endothelial cells in reference to the endothelium of the murine
aortic wall [79]. This work involved Raman 3D profiling combined with the cluster
analysis (CA) to analyze the biochemical composition and endothelial morphology,
as well as to define the size, shape and composition of the main cell compartments
and organelles.
Many works on endothelial cells explore the topic of infecting the endothelium
with bacteria or parasites [80–82]. Studies on the interaction of Staphylococcus
aureus bacteria with EA.hy926 cells enabled to visualize the location and spectral
differences in the Raman spectral profile of cells after the bacteria invasion [81].
Presented research provide the chance to develop new preventive and therapeutic
strategies against intracellular bacteria. Other studies focus on the protozoan
Neospora caninum that attacks vertebrates and remains inside the brain and other
tissues for whole lifetime. In the study by Elsheikha and Kong [80, 82], Raman and
fluorescence imaging supported by biochemical tests were applied to analyze the
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K. Czamara et al.
vessels, among which the best-defined and the most commonly used in endothelial
research is the EA.hy926 cell line [68]. The second subgroup is composed of
endothelial cell lines for the study of microvasculature with the first characterized
HMEC-1 (human microvascular endothelial cells) line from the foreskin microcirculation [69]. Compared with other culture lines, they proved to be the best model
for endothelial research, because they most closely reflected the processes observed
in primary cells [70].
6.2.2 In Vitro Cell Models of Pathophysiology
of the Endothelium
The number of published papers on endothelial studies using microscopic and
spectroscopic methods is rather small, and the majority of these works deal with the
endothelium in the blood vessel wall [71]. Thematically, they can be divided into
works addressing the general characterization of the endothelium and referring to
the disease states, mainly in murine models [60, 71–76]. The Raman imaging
combined with atomic force microscopy (AFM) and immunohistochemical staining
was used to characterize the chemical composition and topography of the
endothelium [75]. In vivo Raman studies on the endothelium in the aorta wall of the
sheep and in vitro in the human aorta have also been published [77]. In addition,
spectroscopic methods were used in studies of endothelial pathologies:
atherosclerosis [76, 78], hypertension [60, 73], diabetes type 2 [74] and cancer
metastasis [72]. In these studies, changes in the protein and/or lipid content
observed in the endothelial layer were treated as a marker of endothelial condition
in diseases and, therefore, could be used in diagnostic context. Analysis of alterations arising from disease development requires knowledge about normal,
non-disease-affected samples. Raman microscopy was used for comparison of the
control EA.hy926 endothelial cells in reference to the endothelium of the murine
aortic wall [79]. This work involved Raman 3D profiling combined with the cluster
analysis (CA) to analyze the biochemical composition and endothelial morphology,
as well as to define the size, shape and composition of the main cell compartments
and organelles.
Many works on endothelial cells explore the topic of infecting the endothelium
with bacteria or parasites [80–82]. Studies on the interaction of Staphylococcus
aureus bacteria with EA.hy926 cells enabled to visualize the location and spectral
differences in the Raman spectral profile of cells after the bacteria invasion [81].
Presented research provide the chance to develop new preventive and therapeutic
strategies against intracellular bacteria. Other studies focus on the protozoan
Neospora caninum that attacks vertebrates and remains inside the brain and other
tissues for whole lifetime. In the study by Elsheikha and Kong [80, 82], Raman and
fluorescence imaging supported by biochemical tests were applied to analyze the
172
K. Czamara et al.