Transcript Valvular Interstitial Cell Calcification Liwen Gu July 5, 2005

Valvular Interstitial
Cell Calcification
Liwen Gu
July 5, 2005
Prof C. Simmons
Background
20 000 deaths caused by valvular diseases in 2002 (US)
Aortic sclerosis: thickening of the valve leaftlets
calcification
Endothelial
Cells
Interstitial
Cells
Calcification process
http://www.ctsnet.org/edmunds/Chapter28section2.html
(Fig 28-4)
The American Journal of Cardiology Vol.94; 2004
Objectives
• How calcification occurs in
valvular interstitial cells (VIC)
• Effects of endothelial cell (EC)
paracrine factors on interstitial
cells - signaling process
Short-term/immediate goal:
induce calcification in VIC culture
1st Experiment
Purpose: to induce calcification in VIC culture by providing
an osteogenic supplemented (OS) media
Method:
-Porcine Aortic Valvular Interstitial Cells (PAVIC) – 14 days
-added 10 mmol/L βGP to VIC media
Hypothesis:
-more nodule formation compared to
control
-alkaline phosphatase activity
-mineralization
Ann Thorac Surg
2003; 75:457-66
st
1
Experiment (cont’d)
Results:
+OS
Explanations:
- βGP is not enough
-ALP protocol-no positive control
control (-OS)
2nd Experiment
Method:
-3 time points: 0, 14, 21 days
-10 mmol/L βGP + 50μg/mL ascorbic acid + 50ng RANKL
(J.J. Kaden et al. Journal of Molecular and Cellular Cardiology 36(2004)57-66)
Future Experiments
VIC alone
VIC + EC
1. VIC + osteogenic 4. VIC + EC
supplement
conditioned media
(βGP)
5. VIC-EC co-culture
2. VIC + OS (βGP + 6. VIC collagen gel +
ascorbic acid)
EC
3. Gene
expression
Future Experiments
VIC alone
VIC + EC
1. VIC + osteogenic 4. VIC + EC
supplement
conditioned media
(βGP)
5. VIC-EC co-culture
2. VIC + OS (βGP + 6. VIC collagen gel +
ascorbic acid)
EC
3. Gene expression
Future Experiments
VIC alone
VIC + EC
1. VIC + osteogenic 4. VIC + EC
supplement (βGP)
conditioned media
2. VIC + OS (βGP + 5. VIC-EC co-culture
ascorbic acid)
6. VIC collagen gel +
3. Gene expression
EC
Co-Culture
Shin et al. “Endothelial Cells Modulate Osteogenesis in Calcifying Vascular
Cells”. J Vasc Res 2004;41: 193-201
Method:
Results:
Future Experiments
VIC alone
VIC + EC
1. VIC + osteogenic 4. VIC + EC
supplement (βGP)
conditioned media
2. VIC + OS (βGP + 5. VIC-EC co-culture
ascorbic acid)
6. VIC collagen gel
3. Gene expression
+ EC
Acknowledgements
• Prof C. Simmons
• Simmons Group:
Alice Ho
Yifei Wang
Samir Raza
Jan Chen
Joel Rebello
• Davies lab
Thank You!
Contact: [email protected]