Proliferation of Alveolar Type II Cells on Fibronectin Fragments
Claire Couch, Carrie Harris, Jeremy Martz, Paul Wach
The Wallace H. Coulter School of Biomedical Engineering
Introduction
Results
Pulmonary Fibrosis
The exact etiology of this disease is unknown, but is
theorized to be the result of a genetic predisposition,
viral infection, autoimmune disorder, or microscopic
injury to the alveolar cells of the lung. There are
currently no effective treatments or cures.
Alveolar Type II Cells (ATII)
Residual Plots for No Serum Media Cultures
99
500
90
250
50
10
1
-500
-250
250
- Showed similarity between F10 and F9-10
500
Source
Factor
Error
Total
2
0
-400
-200
0
Residual
600
700
Fitted Value
800
900
200
DF
SS
5 951868
30 1615026
35 2566894
MS
190374
53834
F
3.54
P
0.012
99
400
90
200
50
10
-200
0
Residual
200
Frequency
0
-200
Source
Factor
Error
Total
7.5
5.0
2.5
-160
0
Residual
1200
1350
1500
Fitted Value
1650
One-way ANOVA:
BSA, TCP, FN, F10, F9-10, F9*10
10.0
-320
Overall, the results show higher proliferation in cells
cultured with 0.2% FBS serum. Furthermore, the F10
and F9-10 fibronectin fragments illustrated greater
influence over cell proliferation than the other fragments.
-400
400
Histogram
0.0
Conclusion
Versus Fits
Residual
Percent
Normal Probability Plot
160
320
DF
SS
MS
F
5 1691034 338207 18.06
30 561719 18724
35 2252752
P
0.000
S = 136.8
R-Sq = 75.07%
R-Sq(adj) = 70.91%
Two-way ANOVA: C1 versus C3, C2
Figure 1: Fibronectin 3D structure.
Source
C3
C2
Interaction
Error
Total
Statistical Method
One-Way ANOVA Test
S = 189.0
• Comparison of each of the 6 cell cultures to test for
statistically significant differences in proliferation
DF
1
5
5
60
71
SS
7287948
251462
3034490
2144134
12718033
MS
7287948
50292
606898
35736
R-Sq = 83.14%
F
203.94
1.41
16.98
P
0.000
0.235
0.000
Therefore it is our recommendation that F9 and F9-10
fragments cultured in 0.2% FBS media should be used
to analyze ATII cells and their role in reducing the
severity of pulmonary fibrosis.
1. “What is Pulmonary Fibrosis?” Pulmonary Fibrosis Foundation.
Access Date: 1 December 2007.
<http://www.pulmonaryfibrosis.org/ipf.htm>.
R-Sq(adj) = 80.05%
2. “Respiratory System-Cells.” IUPS Physiome Project. Access Date: 1
December 2007. <http://www.bioeng.auckland.ac.nz/physiome/
ontologies/respiratory/cells.php>.
2000
3. “Fibronectin.” Wikipedia. Access Date: 1 December 2007.
<http://en.wikipedia.org/wiki/Fibronectin>.
1500
Cell Count
The presence of large amounts of fibronectin in lung
tissue is characteristic of pulmonary fibrosis. The
fragment of fibronectin that results in the highest number
of ATII cells present should be targeted clinically for
treatment pharmacology.
References
Boxplot of Two-Way ANOVA Data
• Test applied for cultures with 0.2% FBS Serum and with
No Serum
•Low p-value suggests that there is a significant
difference in cell proliferation under the two culture
conditions
•Boxplot shows overall higher cell count for 0.2% FBS
serum cultured cells
S = 232.0
R-Sq = 37.08%
R-Sq(adj) = 26.60%
400
- Proliferation was higher in 0.2% FBS Serum
Two-Way ANOVA Results
Residual Plots for 0.2% FBS Media Cultures
• A large dimeric extracellular matrix (ECM) protein
crucial in wound healing and binding of cells to the ECM
• Comparison of 2 groups of cell cultures, No Serum and
0.2% FBS Serum to test for statistically significant
difference in proliferation
-250
One-way ANOVA:
BSA, TCP, FN, F10, F9-10, F9*10
4
Fibronectin
Two-Way ANOVA Test
•Due to low p-values, Tukey’s Tests were performed
0
500
6
1
-400
• Abnormalities with its transcription or expression result
in fibrosis
0
Residual
8
• ATI: form blood-gas barrier to allow gas exchange
• ATII: secrete pulmonary surfactant that reduces
surface tension of alveolar lining to allow cells to
expand and contract without collapsing during
respiration
•The histograms for both residual plots show normal
distributions
-500
Histogram
ATII cells are present in the lining of the alveolar ducts
and alveoli of the lungs. They can differentiate into both
ATI and ATII cells.
One Way ANOVA Results
Versus Fits
Residual
Percent
Normal Probability Plot
Frequency
This disease is characterized by replacement of the
air sacs in the lungs by fibrotic tissue resulting in
irreversible tissue damage and function loss. The
damaged lung can no longer properly re-oxygenate
blood.
Discussion
Acknowledgements
1000
We would like to thank Ashley Carson, Graduate Research
Assistant, for providing us data for the project.
500
BSA
TCP
FN
F10
No_Serum
F9-10 F9*10
BSA
TCP
FN
F10
FBS_Media
F9-10 F9*10
We also would like to thank Dr. Brani Vidakovic, BMED 2803
Professor, for his aid in our statistical analysis.