1. Comparison of the impact of preservation methods on amniotic membrane properties for tissue engineering applications
- Author
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Samantha Delmond, Florelle Gindraux, Robin Siadous, Sylvain Catros, Jean-Christophe Fricain, Stéphanie Brun, Mathilde Fenelon, Agathe Grémare, Delphine B. Maurel, Nicolas L'Heureux, Marlène Durand, Bioingénierie tissulaire (BIOTIS), Université de Bordeaux (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Bordeaux (UB), Plateforme Technologique d'Innovation Biomédicale (PTIB), Université de Bordeaux (UB)-Hôpital Xavier Arnozan, CHU Bordeaux [Bordeaux], Ecole Dentaire, Université Bordeaux Segalen - Bordeaux 2, Nanomédecine, imagerie, thérapeutique - UFC (EA 4662) (NIT / NANOMEDECINE), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Nanomédecine, imagerie, thérapeutique - UFC (UR 4662) (NIT / NANOMEDECINE), and CCSD, Accord Elsevier
- Subjects
Materials science ,Biocompatibility ,[SDV]Life Sciences [q-bio] ,Amniotic membrane ,Biocompatible Materials ,Bioengineering ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,in vivo biocompatibility ,Cryopreservation ,Biomaterials ,Freeze-drying ,Subcutaneous Tissue ,Implants, Experimental ,Tissue engineering ,immune system diseases ,hemic and lymphatic diseases ,mental disorders ,Animals ,Humans ,Amnion ,Rats, Wistar ,Inflammation ,Decellularization ,Cell Death ,Tissue Engineering ,Mesenchymal stem cell ,food and beverages ,virus diseases ,DNA ,021001 nanoscience & nanotechnology ,Extracellular Matrix ,0104 chemical sciences ,Staining ,[SDV] Life Sciences [q-bio] ,Acellular scaffold ,Membrane ,Mechanics of Materials ,Rat ,Female ,Processed amnion ,0210 nano-technology ,Biomedical engineering - Abstract
International audience; Human amniotic membrane (hAM) is considered as an attractive biological scaffold for tissue engineering. For this application, hAM has been mainly processed using cryopreservation, lyophilization and/or decellularization. However, no study has formally compared the influence of these treatments on hAM properties. The aim of this study was to develop a new decellularization-preservation process of hAM, and to compare it with other conventional treatments (fresh, cryopreserved and lyophilized). The hAM was decellularized (D-hAM) using an enzymatic method followed by a detergent decellularization method, and was then lyophilized and gamma-sterilized. Decellularization was assessed using DNA staining and quantification. D-hAM was compared to fresh (F-hAM), cryopreserved (C-hAM) and lyophilized/gamma-sterilized (L-hAM) hAM. Their cytotoxicity on human bone marrow mesenchymal stem cells (hBMSCs) and their biocompatibility in a rat subcutaneous model were also evaluated. The protocol was effective as judged by the absence of nuclei staining and the residual DNA lower than 50 ng/mg. Histological staining showed a disruption of the D-hAM architecture, and its thickness was 84% lower than fresh hAM (p < 0.001). Despite this, the labeling of type IV and type V collagen, elastin and laminin were preserved on D-hAM. Maximal force before rupture of D-hAM was 92% higher than C-hAM and L-hAM (p < 0.01), and D-hAM was 37% more stretchable than F-hAM (p < 0.05). None of the four hAM were cytotoxic, and D-hAM was the most suitable scaffold for hBMSCs proliferation. Finally, D-hAM was well integrated in vivo. In conclusion, this new hAM decellularization process appears promising for tissue engineering applications.
- Published
- 2019