Abstract
The contractile behavior of smooth muscle cells (SMCs) in the aorta is an important determinant of growth, remodeling, and homeostasis. However, quantitative values of SMC basal tone have never been characterized precisely on individual SMCs. Therefore, to address this lack, we developed an in vitro technique based on Traction Force Microscopy (TFM). Aortic SMCs from a human lineage at low passages (4-7) were cultured 2 days in conditions promoting the development of their contractile apparatus and seeded on hydrogels of varying elastic modulus (1, 4, 12 and 25 kPa) with embedded fluorescent microspheres. After complete adhesion, SMCs were artificially detached from the gel by trypsin treatment. The microbeads movement was tracked and the deformation fields were processed with a mechanical model, assuming linear elasticity, isotropic material, plane strain, to extract the traction forces formerly applied by individual SMCs on the gel. Two major interesting and original observations about SMC traction forces were deduced from the obtained results: 1. they are variable but driven by cell dynamics and show an exponential distribution, with 40% to 80% of traction forces in the range 0-10 µN. 2. They depend on the substrate stiffness: the fraction of adhesion forces below 10 µN tend to decrease when the substrate stiffness increases, whereas the fraction of higher adhesion forces increases. As these two aspects of cell adhesion (variability and stiffness dependence) and the distribution of their traction forces can be predicted by the probabilistic motor-clutch model, we conclude that this model could be applied to SMCs. Further studies will consider stimulated contractility and primary culture of cells extracted from aneurysmal human aortic tissue.
Highlights
IntroductionIt is widely acknowledged that the biomechanical behavior of smooth muscle cells (SMCs) in the aorta, through mechanosensitivity and mechanotransduction, is an important determinant of growth, remodeling, and homeostasis [1,2,3,4,5,6,7].Aortic SMCs are highly sensitive to:-biochemical [8,9,10,11,12,13].-and mechanical stimuli from the surrounding extracellular matrix (ECM) [2,14,15,16,17,18,19].In pathologies such as aortic aneurysms, the function of aortic SMCs may be altered by:-genetics ([20,21,22,23,24,25,26,27,28]),-hemodynamics ([29,30]),-or biomechanics ([3,21,31]), DOI:10.32604/mcb.2019.06415 www.techscience.comMCB, 2019, vol., no.2This may induce manifestations such as-SMC apoptosis [16,32,33]
We have found a significant effect of the substrate stiffness on the adhesion forces generated by cells and measured using Traction Force Microscopy (TFM)
The contractile behavior of SMCs being an important determinant of growth, remodeling, and homeostasis in the aorta, we applied a TFM technique to measure traction forces of individual human aortic SMCs
Summary
It is widely acknowledged that the biomechanical behavior of smooth muscle cells (SMCs) in the aorta, through mechanosensitivity and mechanotransduction, is an important determinant of growth, remodeling, and homeostasis [1,2,3,4,5,6,7].Aortic SMCs are highly sensitive to:-biochemical [8,9,10,11,12,13].-and mechanical stimuli from the surrounding extracellular matrix (ECM) [2,14,15,16,17,18,19].In pathologies such as aortic aneurysms, the function of aortic SMCs may be altered by:-genetics ([20,21,22,23,24,25,26,27,28]),-hemodynamics ([29,30]),-or biomechanics ([3,21,31]), DOI:10.32604/mcb.2019.06415 www.techscience.comMCB, 2019, vol., no.2This may induce manifestations such as-SMC apoptosis [16,32,33]. -and mechanical stimuli from the surrounding extracellular matrix (ECM) [2,14,15,16,17,18,19]. In pathologies such as aortic aneurysms, the function of aortic SMCs may be altered by:. -or biomechanics ([3,21,31]), DOI:10.32604/mcb.2019.06415 www.techscience.com.
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