valvular interstitial cell calcification
DESCRIPTION
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. - PowerPoint PPT PresentationTRANSCRIPT
Valvular Interstitial Valvular Interstitial Cell Calcification Cell Calcification
Liwen GuJuly 5, 2005
Prof C. Simmons
Background
Endothelial Cells
Interstitial Cells
http://www.ctsnet.org/edmunds/Chapter28section2.html (Fig 28-4)
20 000 deaths caused by valvular diseases in 2002 (US)
Aortic sclerosis: thickening of the valve leaftlets calcification
Calcification process
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 processShort-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
1st Experiment (cont’d)
Results:
+OS control (-OS)Explanations:
- βGP is not enough-ALP protocol-no positive control
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 supplement (βGP)
2. VIC + OS (βGP + ascorbic acid)
3. Gene expression
4. VIC + EC conditioned media
5. VIC-EC co-culture
6. VIC collagen gel + EC
Future Experiments
VIC alone VIC + EC
1. VIC + osteogenic supplement (βGP)
2. VIC + OS (βGP + ascorbic acid)
3. Gene expression
4. VIC + EC conditioned media
5. VIC-EC co-culture
6. VIC collagen gel + EC
Future Experiments
VIC alone VIC + EC
1. VIC + osteogenic supplement (βGP)
2. VIC + OS (βGP + ascorbic acid)
3. Gene expression
4. VIC + EC conditioned media
5. VIC-EC co-culture
6. VIC collagen gel + 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 supplement (βGP)
2. VIC + OS (βGP + ascorbic acid)
3. Gene expression
4. VIC + EC conditioned media
5. VIC-EC co-culture
6. VIC collagen gel + EC
Acknowledgements
• Prof C. Simmons
• Simmons Group:Alice HoYifei WangSamir RazaJan ChenJoel Rebello
• Davies lab