1. New cells for new purposes: use of gingival fibroblast for arterial repair Antoine LAFONT, MD, PHD, FESC, Hopital Europeen Georges Pompidou INSERM U849 PARIS-DESCARTES University FRANCE E 00-16 Inserm

2. AAA: which target ? Loss of smooth muscle cells Destruction of elastin network Increased activity of MMP-9 Inflammatory reaction

3. AAA: why cell therapy ? To restore cells To restore elastin network To control MMP-9 activity

4. AAA: which cell ? Gingiva: embryo-like repair –A model for arterial repair –No scar/no inflammation Gingival fibroblast: –Responsible for gingival tissue healing

5. HYPOTHESIS Plasticity of HGF considered as embryo like cells Autologous HGF transferred in aneurysmal wall should modify the pathological remodeling : – enlarging remodeling: aneurysm –Contractil remodeling: stenosis

6. METHODOLOGY Cell Culture Artery Culture Animal model –1- Healthy arteries –2- Stenosis model (Lafont A, 1995) –3- Aneurism model (Anidjar S, 1992, 1994).

7. Cell Coculture SMC/HGF 3D Coculture Artery/HGF MMP9 TIMP1 Complex MMP9/TIMP1

8. Human gingival fibroblasts inhibit MMP-9 activity

9. Decrease of MMP-9 activity is not due to a translation modification

10. SMC/HGF coculture induce an increase (X7) of MMP-9 inhibitor: TIMP-1.

11. TIMP-1 translation is also increased:

12. Increase of MMP-9/TIMP-1 complex induces the inhibition of MMP-9:

13. D3 D 21 Control Elastic network is protected in presence of human gingival fibroblasts Artery + HGF X20 Orcéine

14. HGF tranfer in healthy arteries: (1 million de fibroblastes gingivaux injectés) X 20 X 10 X 100 X 40 Perls HLA detection

15. Elastic network in elastase model Elastase model Control Orceine X 20

16. In vivo: Aneurysm model Control Elastase model

17. HGF injection in aneurysmal arteries: Perls X 5 Perls X 10

18. Conclusions MMP-9: a real target to specifically treat AAA Gingival fibroblast: -able to inhibit MMP-9 expression via TIMP-1 stimulation -easy to collect/amplify Cell tansplantation: appropriate for AAA

19. ACKNOWLEDGEMENTS: Bernard Coulomb, CR1 Bruno Gogly, PU-PH Camille Brasselet, PHU, post-doc Eric Durand, PHU, post-doc Sylvie Séguier, MCU-PH Ludovic Couty, ingénieur de recherche Mathilde Lemitre, ITA Dominique Urbain, ITA Karima Ichegour, Agnès Bodineau, AHU, doctorante 2ème année Marco Giacci, DEA Benjamin Fournier, doctorant 2ème année Adrien Naveau, doctorant 3ème année Nicoletta Reynald, doctorante 2ème année