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

H E G PH E G PInserm

AAA: which target ?

• Loss of smooth muscle cells

• Destruction of elastin network

• Increased activity of MMP-9

• Inflammatory reaction

AAA: why cell therapy ?

• To restore cells

• To restore elastin network

• To control MMP-9 activity

AAA: which cell ?

• Gingiva: embryo-like repair– A model for arterial repair

– No scar/no inflammation

• Gingival fibroblast:– Responsible for gingival tissue healing

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

METHODOLOGY

• Cell Culture

• Artery Culture

• Animal model– 1- Healthy arteries– 2- Stenosis model (Lafont A, 1995) – 3- Aneurism model (Anidjar S, 1992, 1994).

Cell Coculture SMC/HGF3D Coculture Artery/HGF

• MMP9

• TIMP1

• Complex MMP9/TIMP1

Human gingival fibroblasts

inhibit MMP-9 activity

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

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

TIMP-1 translation is also increased:

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

D3

D 21

Control

Elastic network is protected in presence of human gingival fibroblasts

Artery + HGF

X20

Orcéine

HGF tranfer in healthy arteries:

(1 million de fibroblastes gingivaux injectés)

X 20

X 10

X 100

X 40

Perls

HLA detection

Elastic network in elastase model

Elastase model Control

Orceine X 20

In vivo: Aneurysm model

Control Elastase model

HGF injection in aneurysmal arteries:

Perls X 5 Perls X 10

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

ACKNOWLEDGEMENTS:

Bernard Coulomb, CR1Bruno Gogly, PU-PH

Camille Brasselet, PHU, post-docEric Durand, PHU , post-doc

Sylvie Séguier, MCU-PH

Ludovic Couty, ingénieur de rechercheMathilde Lemitre, ITA

Dominique Urbain, ITAKarima Ichegour,

Agnès Bodineau, AHU, doctorante 2ème année

Marco Giacci, DEABenjamin Fournier, doctorant 2ème année

Adrien Naveau, doctorant 3ème annéeNicoletta Reynald, doctorante 2ème année