Amelioration of Diabetic Wound Using Menstrual Blood Stem Cells Seeded on Bilayer Scaffold Composed of Amniotic Membrane and Silk Fibroin in Mouse Model

Objective: Recently, we fabricated a bilayer scaffold composed of amniotic membrane and silk fibroin in aim for skin regen-eration which was characterized physically, biochemically and biologically in vitro and in vivo. In this study, we evaluated the efficiency of menstrual blood stem cells (MenSCs) seeded on the fabricated scaffold in healing of diabetic wound in animal model.Materials and Methods: The ۷۵ male diabetic C۵۷/BL۶ mice were equally divided into five groups including amniotic mem-brane group, no treatment group, scaffold group, MenSCs-seed-ed scaffold group and group constituted of foreskin- derived ke-ratinocytes/fibroblasts seeded on scaffold. All animals received humane care in according to the criteria outlined in the Declara-tion of Helsinki. The study was approved by bioethics commit-tee of Avicenna Research Institute. The excisional bilateral full thickness circular wounds were created in the dorsal dermis of diabetic mice. Donut-shaped silicon splints were utilized to pre-vent wound contraction in mouse skin and simulate re-epithe-lialization. The healing of diabetic wounds in each group was evaluated at ۳, ۷ and ۱۴ days after their treatment. The healing progression measurements, quantification of dermal and epi-dermal hypertrophy and quantification of immunofluorescence staining assays were processed in ImageJ software.Results: The gross morphology, epithelization and histologic changes in regenerated tissue in MenSCs-seeded bilayer nanofi-brous scaffold group showed the most similarity to intact tissue.Moreover, the wound healing was typically faster in MenSCs-seeded scaffold group compared to other diabetic groups. Im-munofluorescence staining of mouse skin depicted higher levels of CD۳۱ and von Willebrand factor (VWF), higher ratio of M۲/ M۱ macrophages, higher expression of involucrin as a keratino-cyte marker and higher levels of neural marker in the MenSCs-seeded bilayer nanofibrous scaffold group in comparison with other treatment groups.Conclusion: The dramatic effects of MenSCs as a promising adult stem cell population accompanied by significant impact of fabricated bilayer scaffold sounds to mediate a promoting and appropriate circumstance on wound healing and skin regenera-tion.