Technical Brief

Effect of Osmotic Pressure on Cellular Stiffness as Evaluated Through Force Mapping Measurements

[+] Author and Article Information
Hsien-Shun Liao

Department of Mechanical Engineering,
National Taiwan University,
Taipei 10617, Taiwan
e-mail: liaohs@ntu.edu.tw

Peter J. Wen, Ling-Gang Wu

National Institute of Neurological Disorders and Stroke,
Bethesda, MD 20892

Albert J. Jin

National Institute of Biomedical Imaging
and Bioengineering (NIBIB),
Bethesda, MD 20892

1Corresponding author.

Manuscript received August 23, 2017; final manuscript received January 28, 2018; published online March 16, 2018. Assoc. Editor: Nathan Sniadecki.

J Biomech Eng 140(5), 054502 (Mar 16, 2018) (5 pages) Paper No: BIO-17-1381; doi: 10.1115/1.4039378 History: Received August 23, 2017; Revised January 28, 2018

Atomic force microscopy (AFM) has been used to measure cellular stiffness at different osmolarities to investigate the effect of osmotic pressure on cells. However, substantial direct evidence is essential to clarify the phenomena derived from the experimental results. This study used both the single-point and force mapping methods to measure the effective Young's modulus of the cell by using temporal and spatial information. The single-point force measurements confirmed the positive correlation between cellular stiffness and osmolarity. The force mapping measurements provided local stiffness on the cellular surface and identified the cytoskeleton distribution underneath the plasma membrane. At hyper-osmolarity, the cytoskeleton was observed to cover most of the area underneath the plasma membrane, and the effective Young's modulus on the area with cytoskeleton support was determined to be higher than that at iso-osmolarity. The overall increase in cellular Young's modulus confirmed the occurrence of cytoskeleton compression at hyper-osmolarity. On the other hand, although the average Young's modulus at hypo-osmolarity was lower than that at iso-osmolarity, we observed that the local Young's modulus measured on the areas with cytoskeleton support remained similar from iso-osmolarity to hypo-osmolarity. The reduction of the average Young's modulus at hypo-osmolarity was attributed to reduced cytoskeleton coverage underneath the plasma membrane.

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Grahic Jump Location
Fig. 1

Single-point measurements from iso-osmolarity (305 mOsm/kg) to (a) hyper-osmolarity (650 mOsm/kg) and (b) hypo-osmolarity (165 mOsm/kg). (c) Young's modulus versus osmolarity (*p = 0.0554, **p = 0.0095, two-tailed t-test).

Grahic Jump Location
Fig. 2

(a)–(c) Cell height images and (d)–(f) Young's modulus mapping at hyper-osmolarity (650 mOsm/kg) (a) and (d), iso-osmolarity (305 mOsm/kg) (b) and (e), and hypo-osmolarity (165 mOsm/kg) (c) and (f). (g) Force curves at marked positions in (e) and (f).

Grahic Jump Location
Fig. 3

Histograms of Young's modulus (a)–(c) and correlations with local cell heights (d)–(f). Mapping at hyper-osmolarity (a) and (d), iso-osmolarity (b) and (e), and hypo-osmolarity (c) and (f).



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