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Related Concept Videos

Studying the Cytoskeleton01:17

Studying the Cytoskeleton

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The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
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Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy
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Atomic Force Microscopy Stiffness Mapping in Human Aortic Smooth Muscle Cells.

Claudie Petit1, Ali-Akbar Karkhaneh Yousefi1, Marine Guilbot1

  • 1Mines Saint-Etienne, Université de Lyon, INSERM, U 1059 SAINBIOSE, Saint-Etienne F - 42023, France.

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|January 26, 2022
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Aortic smooth muscle cells (SMCs) from aneurysms do not show significantly altered cytoskeleton stiffness compared to healthy cells. This suggests other factors contribute to increased traction forces in aneurysmal aortas.

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ascendant thoracic aortic aneurysm (ATAA)atomic force microscopy (AFM)cell biomechanicsfluorescent microscopy (FM)mechanotransductionnanoindentationsmooth muscle cells (SMC)

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Cardiovascular Research

Background:

  • Aortic smooth muscle cells (SMCs) are crucial for aortic mechanical homeostasis.
  • Aneurysmal SMCs exhibit increased traction forces, linked to higher abundance of hypertrophic SMCs.
  • Previous findings suggest potential alterations in SMCs within aneurysmal aortas.

Purpose of the Study:

  • To investigate alterations in cytoskeleton stiffness of SMCs from aneurysmal aortas.
  • To determine if SMC cytoskeleton stiffness differs between healthy and aneurysmal aortic tissues.
  • To correlate cytoskeleton stiffness with cellular mechanical properties in aortic aneurysms.

Main Methods:

  • Utilized atomic force microscopy (AFM) nano-indentation for subcellular stiffness mapping.
  • Cultured human aortic SMCs (AoSMCs) and primary aneurysmal SMCs (AnevSMCs).
  • Seeded cells on hydrogels with 12 kPa and 25 kPa stiffness to mimic physiological conditions.

Main Results:

  • SMCs displayed a lognormal stiffness distribution, with medians between 10-30 kPa.
  • Mean stiffness was 16 kPa for aneurysmal SMCs and 12 kPa for healthy SMCs.
  • Statistical analysis revealed no significant difference in stiffness due to high dispersion in AFM measurements.

Conclusions:

  • Cytoskeleton stiffness of SMCs is not significantly altered in aneurysmal aortas.
  • The increased traction forces observed in aneurysmal SMCs are likely not due to changes in cytoskeleton stiffness.
  • Further research is needed to identify the specific mechanisms behind altered SMC mechanical properties in aortic aneurysms.