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Titre de la communication :
Autologous cell-laden hydrogel sheet for prevention of post-surgical abdominal adhesion (PAA)
Auteurs (et leurs adresses) de la communication :
L Bresson1,2, E Leblanc1, Teru Okitsu3, F Chai2 1 Centre Oscar Lambret, Gynecology and Oncology, 59045 Lille, France 2 INSERM 1008, Biomatériaux, University Lille 2, 59045 Lille, France 3 Takeuchi Lab, Institut of Industrial Science, Tokyo, Japan
Résumé de la communication :
Introduction: Postoperative abdominal adhesions (PAA) are a major complication leading to medical and economical problems. Replace injured peritoneum, which is composed of a monolayer of mesothelial cells (MC), using cell-therapy or cell-laden scaffold are two strategies to prevent PAA. MCs are crucial and adipose derived stem cells (ASCs) could replace thanks to their potential of differentiation. Hydrogel polymers are attractive scaffolds mimicking the properties of the native extracellularmatrix. Milestones of our project of regenerative medicine are (1) comparison of cell-laden scaffolds and cell-therapy, (2) choice of cell source and (3) biomaterial, and finally (4) transplantation of the cell-laden hydrogel scaffold, (5) in a pre-established rat model of PAA.
Materiel and Method:
Firstly, model of PAA has been validating comparing two peritoneal injuries (5). Fours groups were compared (1): Peritoneal graft with MC exposed to the abdominal cavity (group MC), Peritoneal graft with sub serosa containing fibroblasts (FB) exposed (group FB), MC cell-therapy using cell-sheet technology (group MC-FB sheet) and shame group (group control). After 14 days, PAA were assessed by macroscopic observation and histology. MCs and ASCs have been compared for isolation, culture, characterization, and differentiation (2). Hydrogels (3) have been compared for their mechanical properties and their biocompatibility. Cell-laden sheets (4) have been implanted and assessed on the capacity to prevent PAA.
Results: The animal model of PAA was validated and both models were effectives and clinically relevants. In group MC, no PAA were recorded on the surface of the graft. In the other groups, all rats were diagnosed with PAA. Extent of the adhesions and qualitative assessment revealed a significantly inferior score in group MC than in the other groups. Histological observation confirmed the respect of MCs only in group MC. MCs, with typical cobblestone morphology and bright edges, have been stained for Vimentin and Cytokeratin. Senescence arrived after only three passages for these adult well-differentiated cells. Spindle-shaped ASCs had a good capacity of expansion, were able to differentiate in osteocytes and adipocytes, and to form colonies as expected characteristics of stem cells. BD-Purastat® peptide has been tested in collaboration with 3D Matrix firm. MCs and cell lines were able to survive into the gel. Purastat® alone reduced significantly the severity and the extent of PAA in comparison with the group control. But, no significant difference were found between MCs in Purastat® and Purastat® or group control for the prevention of PAA.
Conclusion and perspectives: Our study supported that MCs and scaffold are both needed to succeed in peritoneum's engineering to prevent PAA. Characterization of MCs and ASCs will be detailed. ASCs differentiation into MCS phenotype will be assessed. Purastat® is promising but cell laden grafts need to be imrpoved.