Human amniotic membrane successfully decellularization using of SDS

Background: Owing to its unique biological composition and regenerative properties, the human amniotic membrane (hAM) has been used effectively in the clinical grafting applications.Objective: In the present study, hAM was decellularized using sodium dodecyl sulfate (SDS) treatment to provide a three-dimensional (3D) bioscaffold.Materials and Methods: The qualitative histological evaluations including hematoxylin and eosin (H&E), Masson’s Trichrome (MT, for collagen assay), 4,6-diamidino-2- phenylindole (DAPI), and Alcian blue (AB) staining (for glycosaminoglycan (GAG) assessment), as well as the quantitative assessments including nuclear DNA contents and GAGs assays, were performed on 3D bioscaffolds to confirm the absence of cell nuclei in the decellularized hAM (dhAM) treated with SDS (1%) in comparison to the control group (intact human amniotic membrane (IAM)).Results: H&E and DAPI staining proved that hAM cells were removed from dhAM. MT and AB staining revealed that the GAGs and collagen fibers respectively were preserved in dhAM when compared to the IAM group. Quantitative assays showed that the DNA and GAGs content of dhAM was significantly reduced as compared to the IAM group (114 vs. 6207 ng/mg for DNA and 166 vs. 270 ng/mg for GAGs content; p<0.01).Conclusion: Decellularization of hAM by means of the SDS (1%) is an efficient method to provide a three-dimensional bioscaffold for using in tissue engineering.

Keywords:

Human amniotic membrane , Decellularization , SDS