Bioengineered Human Ovarian ECM Reconstructs GDF-۹ Positive Follicles

Background: Since access to a practical way for achievingthe mature follicles from cryopreserved or native ovarian tissuesspecial in huge animals is difficult, tissue engineering isvery promising to provide artificial folliculogenesis. Decellularizationis a process to remove cellular materials except theorgan skeleton and extracellular elements (ECE) to make a bioscaffolds.In the present study, the effect of sodium hydroxide(NaOH) has been investigated in decellularization of humanovarian tissue applied to follicular reconstruct.Materials and Methods: Human ovarian pieces were distributedin two groups consisted of control and NaOH treated (۰.۵M). Qualitative histological evaluations, quantitative assessments(DNA, total collagen and glycosaminoglycan contents),immunohistochemistry (IHC) staining for Laminin, Fibronectinand Collagen I, cell viability and electron microscopic assay were performed. Finally, human NaOH treated scaffolds alongwith mouse ovarian cells as hybrid artificial ovary (hAO) wereallo-transplanted to ovariectomized mice. H & E technique andIHC for GDF-۹ was performed on ovarian tissue decellularizedone month after transplantation.Results: Histological studies and quantitative evaluations confirmedthe successful decellularization and presence of keyfactors in ovarian scaffolds. Also, toxicity test showed NaOHtreated scaffolds well maintained the survivability of fibroblastcells. Moreover, spherical associations with cuboidal cells intransplanted scaffolds were observed which GDF-۹ expressionwas confirmed follicular reconstruction.Conclusion: NaOH is an appropriate material for eliminatingthe ovarian cells and supporting the new implanted ovariancells to follicular reconstruction which can be a valuablefinding in tissue bioengineering research to provide a hybridartificial ovary.

کلیدواژه ها:

Ovary ، Decellularization ، NaOH ، Bioscaffold ، Follicular Reconstruction