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Autologuous cell-ladened hydrogel sheet for prevention of post-surgical abdominal adhesion

Abstract : Introduction: Postoperative abdominal adhesions 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 promising strategies to prevent adhesions. MC as functional and differentiated cells are crucial. However, adipose stem cells (ASCs) could replace MCs thanks to their potential of differentiation. Furthermore, hydrogel polymers are attractive scaffolds mimicking the properties of the native ECM. So, milestones of our project of regenerative medicine are comparison of cell-laden scaffolds and cell-therapy, choice of cell source and biomaterial, and finally transplantation of the cell-laden hydrogel scaffold, in a pre-established rat model of adhesion.Materiel and Method: Firstly, models of adhesion were compared usingdifferent peritoneal injuries. Then, MCs and ASCs were compared for isolation, culture, characterization, and differentiation. Nextly, cell-therapy using MC intra peritoneal injection (IP) or cell-sheet technology was compared with tissue-therapy using peritoneal grafts through a proof of concept study. BD-Purastat® hydrogel was tested in collaboration with 3D Matrix firm. Cell-laden hydrogel gels were implanted and assessed on adhesion prevention.Results: Two animal models of adhesion were validated and both techniques were effective and clinically relevant. MCs and ASCs were isolated from respectively tunica vaginalis and subcutaneous inguinal fat pad. MCs, with typical cobble-stone morphology and bright edges, were positively 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 for stem cells. Autologuous peritoneal grafts prevented postoperative abdominal adhesions in the rat model. As the mechanism of this prevention, the MC survived and contributed to reperitonealization, only when they were transplanted as a part of the autologous peritoneal grafts and were located on the surface exposed to the abdomen. Cell sheet technology and MCs IP injection failed in the adhesion prevention. BD-Purastat® presented good mechanical properties and biocompatibility. Alone, it reduced significantly adhesion extent. But, cell encapsulation into BD-Purastat® did not improve this prevention.Conclusion and perspectives: Our study supported that MCs and scaffold are both needed to succeed in peritoneum’s engineering to prevent adhesion. ASCs differentiation into MCs phenotype has still to be shown. BD- Purastat® decreases adhesion extent and behavior of the cell into this scaffold needs to be studied to improve the effectiveness of the cell-laden hydrogel application.
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Submitted on : Wednesday, July 11, 2018 - 12:19:05 PM
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  • HAL Id : tel-01835340, version 1



Lucie Bresson. Autologuous cell-ladened hydrogel sheet for prevention of post-surgical abdominal adhesion. Human health and pathology. Université du Droit et de la Santé - Lille II, 2017. English. ⟨NNT : 2017LIL2S033⟩. ⟨tel-01835340⟩



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