Cardiac Cell Mechanics at the Single Cell Level

Front Cover
ProQuest, 2009 - 187 pages
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The analysis of the cellular microenvironment is an area that has received much attention recently in the study of the tractions cells use for locomotion. Specifically, the study of cardiac cell mechanics is of particular interest as cardiac dysfunction is one of the leading causes of death in America. This progression is fueled by the need to have a system capable of reproducing mechanical environments before, during and after a dysfunctional event in order to fully characterize the causes and prevent any permanent damage. The current study proposes a system capable of providing quantitative measurements of the cellular microenvironment while concurrently allowing the same environment to be spatially controlled through collagen deposition and alignment. In this study, we have developed a system capable of characterizing the mechanical environment of contractile cardiac myocytes and migratory cardiac fibroblasts while concurrently allowing for fine control over cell position and alignment using a combination of Traction Force Microscopy with a deformable substrate and Finite Element Analysis. The results of this study indicate that this system can be applied to the study of the mechanical nature of cardiac dysfunctions like hypertrophy, myocardial infarction and hypertension.
  

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Contents

Introduction
1
Cartoon of Contractile Proteins
6
Cartoon Diagram of Gap Junction Structure
8
Cartoon Diagram of Adherens Junction Structure
10
The ReninAngiotensin System RAS
14
Gap Junction Relocation
20
Cardiac Mechanical Studies
35
Cell Culture Stress Frame
36
40x Aligned Myocytes Undergoing Contraction
97
Stress Maps in the XDirection on Layer8
98
Stress Maps in the XDirection on Layer4
99
40x Aligned Myocytes Undergoing Contraction
100
Stress Maps in the XDirection on Layer8
101
Stress Maps in the XDirection on Layer4
102
40x Unaligned Myocytes Undergoing Contraction
103
Stress Maps in the XDirection on Layer8
104

Mechanical Damage of Perpendicular Stretch
38
Equibiaxial Stretch Device
40
Polycarbonate Indenter Ring
43
PDMS Stretch Cartoon
44
Fibroblasts on Elastic Substrate
46
Locomotion of Fish Keratocyte
47
Traction Map
49
Development of Traction Force Microscopy
50
Equations Page Equation 1 Correlation Coefficient
53
Stress Fields
54
Displacement Field
55
Probability of Correct Hypothesis
56
Myocytes on Aligned Collagen
59
Fluorescent Image with Outofplane Beads
61
Fluorescent Image with no Outofplane Beads
62
Cardiac Myocytes Patterned with Photolithographic Stencil
63
Cardiac Myocytes Aligned with Coverslip Edge
64
Newtons Second Law
65
Displacement Map Created with Optical Flow
66
Hookes Law Derivation
67
Experimental Procedure
69
GUI of Optical Flow Program
74
Three Dimensional Representation of Simulated Substrate
75
40x Aligned Myocytes Undergoing Contraction
78
Stress Maps in the XDirection on Layer8
79
Stress Maps in the YDirection on Layer4
80
40x Aligned Myocytes Undergoing Contraction
81
Stress Maps in the YDirection on Layer8
82
40x Aligned Myocytes Undergoing Contraction
83
Stress Maps in the XDirection on Layer8
84
Stress Maps in the YDirection on Layer4
85
Fluorescent Bead Image Masked with Displacement Data
86
Stress Maps in the XDirection on Layer4
87
40x Unaligned Myocytes Undergoing Contraction
88
Stress Maps in the XDirection on Layer8
89
Stress Maps in the YDirection on Layer4
90
40x Unaligned Myocytes Undergoing Contraction
91
Stress Maps in the XDirection on Layer8
92
Stress Maps in the YDirection on Layer4
93
Fluorescent Bead Image Masked with Displacement Data
94
Stress Maps in the YDirection on Layer8
95
Stress Maps in the YDirection on Layer4
96
Stress Maps in the XDirection on Layer4
105
40x Unaligned Myocytes Undergoing Contraction
106
Stress Maps in the XDirection on Layer8
107
Stress Maps in the XDirection on Layer4
108
40x Unaligned Myocytes Undergoing Contraction
109
Stress Maps in the XDirection on Layer8
110
Stress Maps in the XDirection on Layer4
111
40x Aligned Myocytes Undergoing Contraction
112
Fluorescent Bead Image Masked with Displacement Data
113
Stress Maps in the YDirection on Layer8
114
40x Aligned Myocytes Undergoing Contraction
115
Fluorescent Bead Image Masked with Displacement Data
116
Stress Maps in the YDirection on Layer8
117
40x Aligned Myocytes Undergoing Contraction
118
Fluorescent Bead Image Masked with Displacement Data
119
Stress Maps in the YDirection on Layer8
120
40x Unaligned Myocytes Undergoing Contraction
121
Fluorescent Bead Image Masked with Displacement Data
122
Stress Maps in the YDirection on Layer8
123
Stress Maps in the YDirection on Layer4
124
Fluorescent Bead Image Masked with Displacement Data
125
Stress Maps in the YDirection on Layer8
126
40x Unaligned Myocytes Undergoing Contraction
127
Stress Maps in the XDirection on Layer8
128
Stress Maps in the YDirection on Layer4
129
Random Myocytes Stained with AntiConnexin 43
130
Random Myocytes Stained with FITC Phalloidin
131
Discussion
132
Conclusions
138
Youngs Modulus
143
Sum Squared Difference
144
Correlation Coefficient
145
CorrelationBased Optical Flow
146
SubPixel Resolution
147
Strain Tensor
150
Midplane Deformations
151
Displacement Field
152
Total Mesh Area
153
Bayesian Hypothesis
154
Probability of Solution Given Correct Hypothesis
155
Traction Density
156
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