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Optical Measurement of Micromechanic...

Kotlarchyk, Maxwell Aaron.

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Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix./
Author:	Kotlarchyk, Maxwell Aaron.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2011,
Description:	174 p.
Notes:	Source: Dissertations Abstracts International, Volume: 73-06, Section: B.
Contained By:	Dissertations Abstracts International73-06B.
Subject:	Biomedical engineering. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3482434
ISBN:	9781267037985

Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix.
Kotlarchyk, Maxwell Aaron.

Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix. - Ann Arbor : ProQuest Dissertations & Theses, 2011 - 174 p.

Source: Dissertations Abstracts International, Volume: 73-06, Section: B.

Thesis (Ph.D.)--University of California, Irvine, 2011.

In recent years, a significant number of studies have focused on linking substrate mechanics to cell function using standard methodologies to characterize the bulk properties of the hydrogel substrates. However, current understanding of the correlations between the microstructural mechanical properties of hydrogels and cell function in 3D is poor, in part because of a lack of appropriate techniques. Methods for tuning extracellular matrix (ECM) mechanics in 3D cell culture that rely on increasing the concentration of either protein or cross-linking molecules fail to control important parameters such as pore size, ligand density, and molecular diffusivity. Alternatively, ECM stiffness can be modulated independently from protein concentration by mechanically loading the ECM. We have developed an optical tweezers-based microrheology system to investigate the fundamental role of ECM mechanical properties in determining cellular behavior. Further, this thesis outlines the development of a novel device for generating stiffness gradients in naturally derived ECMs, where stiffness is tuned by inducing strain, while local structure and mechanical properties are directly determined by laser tweezers-based passive and active microrheology respectively. Hydrogel substrates polymerized within 35 mm diameter Petri dishes are strained non-uniformly by the precise rotation of an embedded cylindrical post, and exhibit a position-dependent stiffness with little to no modulation of local mesh geometry. Here we present microrheological studies in the context of fibrin hydrogels. Microrheology and confocal imaging were used to directly measure local changes in micromechanics and structure respectively in unstrained hydrogels of increasing fibrinogen concentration, as well as in our strain gradient device, in which the concentration of fibrinogen is held constant. Orbital particle tracking, and raster image correlation analysis are used to quantify changes in fibrin mechanics on the single fiber level. Furthermore, we show that aortic smooth muscle cells cultured within our device are morphologically sensitive to the induced mechanical gradient, and strain drives cell populations from a proliferative to a contractile-like phenotype. This powerful platform-independent cell culture tool will enable a more complete understanding of mechanical effects on cellular physiology in naturally derived 3D ECM tissues.

ISBN: 9781267037985Subjects--Topical Terms:

535387
Biomedical engineering.
Subjects--Index Terms:

Extracellular matrix

Optical Measurement of Micromechanics and Structure in a 3D Fibrin Extracellular Matrix.
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520 $a In recent years, a significant number of studies have focused on linking substrate mechanics to cell function using standard methodologies to characterize the bulk properties of the hydrogel substrates. However, current understanding of the correlations between the microstructural mechanical properties of hydrogels and cell function in 3D is poor, in part because of a lack of appropriate techniques. Methods for tuning extracellular matrix (ECM) mechanics in 3D cell culture that rely on increasing the concentration of either protein or cross-linking molecules fail to control important parameters such as pore size, ligand density, and molecular diffusivity. Alternatively, ECM stiffness can be modulated independently from protein concentration by mechanically loading the ECM. We have developed an optical tweezers-based microrheology system to investigate the fundamental role of ECM mechanical properties in determining cellular behavior. Further, this thesis outlines the development of a novel device for generating stiffness gradients in naturally derived ECMs, where stiffness is tuned by inducing strain, while local structure and mechanical properties are directly determined by laser tweezers-based passive and active microrheology respectively. Hydrogel substrates polymerized within 35 mm diameter Petri dishes are strained non-uniformly by the precise rotation of an embedded cylindrical post, and exhibit a position-dependent stiffness with little to no modulation of local mesh geometry. Here we present microrheological studies in the context of fibrin hydrogels. Microrheology and confocal imaging were used to directly measure local changes in micromechanics and structure respectively in unstrained hydrogels of increasing fibrinogen concentration, as well as in our strain gradient device, in which the concentration of fibrinogen is held constant. Orbital particle tracking, and raster image correlation analysis are used to quantify changes in fibrin mechanics on the single fiber level. Furthermore, we show that aortic smooth muscle cells cultured within our device are morphologically sensitive to the induced mechanical gradient, and strain drives cell populations from a proliferative to a contractile-like phenotype. This powerful platform-independent cell culture tool will enable a more complete understanding of mechanical effects on cellular physiology in naturally derived 3D ECM tissues.
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