Quantitative analysis of cytoskeletal remodeling in vascular smooth muscle cells during phenotypic modulation

The authors developed a quantitative image processing technique to investigate cytoskeletal remodeling of the vascular smooth muscle cells (VSMCs). Cells were harvested from 1-day neonatal mice. Two different culture substrates, laminin and fibronectin, were used to modulate VSMCs phenotype. Contractile and synthetic. The authors also applied cyclic stretch to VSMCs, which increases contractile phenotype. Cytoskeletal fibers were stained using immunofluorescent techniques. The software was designed based on spatial Fourier transform to quantitatively analyzes cytoskeletal fiber orientation within the cell. Quantitative fiber orientation index (OI) was defined for individual cells. OI of VSMCs grown on laminin and fibronectin were 33±14% and 21±11%, respectively. OI of VSMCs grown with and without cyclic stretch were 40±12% and 28±10%, respectively. Thus, cytoskeletnl remodeling of VSMCs during phenotypic modulation can be studied quantitatively