The role of fibroblast in ectopic calcification
•
0 likes•1,074 views
This study investigated the role of dermal fibroblasts in ectopic calcification using a cell culture model. The results show that calcification occurs slowly over time in cultured fibroblasts. Genetic studies implicate inorganic pyrophosphate (PPi) metabolism in calcification, and the results support that the process involves alterations in local PPi levels. Normal fibroblasts express proteins involved in PPi production and resorption, like NPP1 and TNAP. However, in a calcifying environment TNAP is highly upregulated, abolishing the inhibitory effects of PPi. This suggests TNAP plays a critical role by increasing inorganic phosphate levels needed for mineral deposition. Restoring extracellular PPi pools through TNAP inhibition can counteract mineral formation
Recommended
More Related Content
What's hot
What's hot (18)
Similar to The role of fibroblast in ectopic calcification
Similar to The role of fibroblast in ectopic calcification (20)
More from Giulia Annovi
More from Giulia Annovi (14)
Recently uploaded
Recently uploaded (20)
The role of fibroblast in ectopic calcification
- 1. The Role of Dermal Fibroblasts in the Development of Ectopic Calcifications Giulia Annovi Department of Life Sciences University of Modena and Reggio Emilia
- 2. Calcification Biological mineralization Ectopic calcification Physiological process Bone - Teeth Cell-mediated process Regulated by proteins, ions, functional molecules Pathological process Soft connective tissues Cell-mediated process Induced by several mechanisms
- 3. 5. Loss of “inhibitors” 4. Presence of “Bone Proteins” 1. Circulating nucleational complexes (high Ca / P) 2. Cell death 3. Proteolysis and ECM degradation
- 4. Cell Inhibitors: ANKH PC-1 MGP Osteopontin PPi Stimulators: Matrix vesicles ANKH PC-1 Annexins Apoptotic bodies TNAP Type I, II, X collagen Pi Mineral formation (Kirsch, 2006)
- 5. Osteoarthritis Ankylosing Spondylitis Keutel Syndrome Aging Atherosclerosis Aging Chronic renal failure Diabetes Bone formation Osteopetrosis Aging Osteoblasts Vascular Smooth Muscle Cells Chondrocytes Fibroblasts? Mesenchymal cells which are resident in a tissue that is only rarely affected by mineralization process.
- 6. Angioid streaks Haemorragies Central vision loss Papules Skin laxity and redundancy Cardiovascular complications Pseudoxanthoma elasticum as soft connective tissue calcification model
- 7. SEM analysis
- 8. Calcifying Medium Ascorbic Acid β glicerophosphate Dexamethasone
- 10. Day20Day30Day40
- 11. The calcification process in a fibroblast cell culture system is a slowly progressive event. Fibroblast's phenotype could be different from that of other mesenchymal cells.
- 12. Control PXE CM P CM C CM 20GG 6HPXE CM 20GG 6H 0 1 2 3 4 5 6 7 Control PXE ANKHgeneexpression 6h 6h 20d DMEM CM CM Control6 PXE6 Control6cm PXE6cm C XXg CM P xxg CM 0.0 0.5 1.0 1.5 2.0 2.5 Control PXE ANKH (Relativebandintensity) DMEM CM CM 6h 6h 20d
- 13. Control PXE CM P CM C Cm 20 GG 6HPXE CM 6H 20GG 0 10 20 PXE Control NPP1geneexpression DMEM CM CM 6h 6h 20d c 6h P 6h C 6h cm P 6h CM C XXg cm P XXg CM 0 1 2 3 4 Control PXE PC1 (Relativebandintensity) DMEM CM CM 6h 6h 20d
- 14. C CM PXE CM 0 1 2 3 4 Control PXE 4 14 24 34 ALPactivity(arbitraryunit) DMEM CM CM CM 6h 6h 10d 20d TNAP determins higher Pi levels.
- 15. DMEM CM CM +100 uM Levamisole Control PXE
- 16. # Genetic studies have implicated PPi metabolism in the development of calcification, but this finding sustains the hypothesis that the calcification process could be due to alterations in local PPi metabolism. # Results indicate that NPP1, being capable to produce PPi , is normally expressed by human dermal fibroblasts and is significantly upregulated in a calcifying environment. # However, the potent inhibitory action of PPi is abolished by the contemporary hyperactivation of TNAP, which is consistent with its critical role in mineralization process. # The higher upregulation of TNAP in PXE could be a condition associated with ECM calcification. # Restoration of PPi extracellular pool, by inhibiting TNAP activity, is able to counteract mineral deposits formation. Conclusions
- 17. Federica Boraldi Roberta Tiozzo Ivonne Ronchetti Daniela Quaglino University of Modena Maria I. Garcia Fernandez University of Malaga Thanks to Anne de Paepe Olivier Vanakker University of Ghent Leon J.Schurgers Cees Vermeer University of Maastricht
- 18. Is it TNAP sufficient to induce ectopic calcification in PXE? No,it is NECESSARY but NOT SUFFICIENT 50kDa 37kDa Actin Control 3-10NL 50kDa 37kDa Actin PXE 3-10NL Gla-MGP Control 15kDa 10kDa Gla-MGP PXE 15kDa 10kDa MGP
- 19. Control PXE C 20GG 6h PXE 20GG 6H 0 1 2 3 4 Control PXE BMP2GENEexpression 6h 20d Control6 PXE6 0.0 0.5 1.0 1.5 Control PXE BMP2 (Relativebandintensity) 6h 20d BMP2