Evaluation Three Methods of Ovarian Decellulariza-tion by Triton, Ammonium Hydroxide and SDS

Background: Decellularization is a novel technique in regen-erative medicine. Recently decellularized ovary introduced as a scaffold in the field of human fertility preservation. So the aim of the present study was producing a decellularized ovarian scaffold for fertility preservation approaches.Materials and Methods: In the present study 2 mm pieces of bovine and human ovarian cortex were prepared. In the SDS group, ovarian cortex was decellularized with 0.1% SDS for 24 h, in the SDS-Triton-ammonium group these pieces were decellularized with 0.5% SDS for 2 hours, 1% Triton and 0.1% Ammonium hydroxide for 22 hours and in SDS-Triton group pieces were decellularized with 0.5% SDS for 3 h and 1% Tri-ton for 9 hours. Human ovarian cortex was decellularized in three steps. First, it was treated with 0.1% SDS for 24 hours then according to freeze and thaw, it was treated with 0.2% SDS for 10 hours. After decellularization all of the scaffolds were washed with deionized water for 24 hours. The intact ovarian cortex was used as a control group. H&E and DAPI staining were applied to prove decellularization. Elastin and Masson s trichrome staining was carried out to evaluate the presence of elastin and collagen respectively in decellularized tissue. Fur-thermore, MTT test was done to assess the in vitro scaffold’s cytocompatibility.Results: According to the H&E staining results, the bovine ovarian cortex was decellularized in all groups. No residual nu-clei were observed by DAPI staining. Preservation of the ECM content was evaluated by Masson Trichrome and Gomori s al-dehyde-fuchsin staining. Elastic and collagen fibers were kept after the decellularization process in all groups. OD values of eluted formazan of MTT test showed that fibroblasts on the SDS-Triton-Amonium decellularized scaffolds were more vi-able than other groups. Human ovarian cortex was decellular-ized completely with mentioned protocol. Furthermore elastic and collagen fibers were kept in decellularized human ovarian cortex too. There was no significant difference in the Prolifera-tion rate of the fibroblast cells in the human decellularized scaf-fold and two-dimensional conventional culture system.Conclusion: In conclusion, human ovarian cortex was decel-lularized with the combination of different SDS solution. Struc-ture of ECM was not damaged in any groups. However, MTT test results of SDS-Triton-Ammonium was better than other groups.

کلیدواژه ها:

Decellularization ، Ovary ، Scaffold