Title

Analyzing the Role of the Glycocalyx In Whole-Cell Mechanics Using Atomic Force Microscopy

Author(s)

Emily FastFollow

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Cell and Molecular Biology

Presentation Type

Mentored

Abstract

The glycocalyx is a layer of proteoglycans, glycolipids, integral proteins, etc. surrounding all animal cells. This layer plays a role in almost every cellular process and function, from cell communication to maintaining a viable concentration gradient. While its physical presence and function has been studied, its role in cell mechanics has yet to be established. The hypothesis of this research is that the absence of the glycocalyx can have a detrimental effect on cell mechanics. In this study, the apparent elastic modulus of endothelial cells (HUVEC), breast cancer cells (MCF-7), and vascular smooth muscle cells (VSMCs) was measured using Atomic Force Microscopy (AFM) before and after their glycocalyces were degraded. Elastic modulus is the resistance of cells to displacement, and this was calculated by recording the force that the cells exhibited when indented by the AFM cantilever mechanism. Following the control measurement, an enzyme solution comprised of chondroitinase and heparatinase at a concentration of 0.2 U/mL. was applied to the cells in order to degrade their glycocalyces, and the cells were indented once more. The results from the AFM were interpreted with a MATLAB program to determine the cells’ average apparent elastic modulus. The data showed that glycocalyx degradation decreased the elastic modulus measurement of all cell types by a range from 20-50% which confirmed the hypothesis. With this research, a more accurate model of the cell can be determined for future studies, as well as open doors for new solutions in tissue engineering and regenerative medicine.

Location

Founders Hall 114 A

Start Date

3-30-2019 1:45 PM

Presentation Format

Written Submission

Group Project

No

COinS
 
Mar 30th, 1:45 PM

Analyzing the Role of the Glycocalyx In Whole-Cell Mechanics Using Atomic Force Microscopy

Founders Hall 114 A

The glycocalyx is a layer of proteoglycans, glycolipids, integral proteins, etc. surrounding all animal cells. This layer plays a role in almost every cellular process and function, from cell communication to maintaining a viable concentration gradient. While its physical presence and function has been studied, its role in cell mechanics has yet to be established. The hypothesis of this research is that the absence of the glycocalyx can have a detrimental effect on cell mechanics. In this study, the apparent elastic modulus of endothelial cells (HUVEC), breast cancer cells (MCF-7), and vascular smooth muscle cells (VSMCs) was measured using Atomic Force Microscopy (AFM) before and after their glycocalyces were degraded. Elastic modulus is the resistance of cells to displacement, and this was calculated by recording the force that the cells exhibited when indented by the AFM cantilever mechanism. Following the control measurement, an enzyme solution comprised of chondroitinase and heparatinase at a concentration of 0.2 U/mL. was applied to the cells in order to degrade their glycocalyces, and the cells were indented once more. The results from the AFM were interpreted with a MATLAB program to determine the cells’ average apparent elastic modulus. The data showed that glycocalyx degradation decreased the elastic modulus measurement of all cell types by a range from 20-50% which confirmed the hypothesis. With this research, a more accurate model of the cell can be determined for future studies, as well as open doors for new solutions in tissue engineering and regenerative medicine.