Application of Menstrual Blood Derived Stromal (stem) Cells Exert Greater Regenerative Potency Than Fibroblasts/Keratinocytes in Chronic Wounds of Diabetic Mice

Ebrahim Mirzadegan; Hannaneh Golshahi; Zahra Saffarian; Haleh Edalatkhah; Maryam Darzi; Somayeh Khorasani; Kioomars Saliminejad; Somaieh Kazemnejad

doi:10.18502/ajmb.v15i3.12923

Application of Menstrual Blood Derived Stromal (stem) Cells Exert Greater Regenerative Potency Than Fibroblasts/Keratinocytes in Chronic Wounds of Diabetic Mice

Avicenna J Med Biotechnol. 2023 Jul-Sep;15(3):139-156. doi: 10.18502/ajmb.v15i3.12923.

Authors

Ebrahim Mirzadegan¹, Hannaneh Golshahi¹, Zahra Saffarian¹, Haleh Edalatkhah¹, Maryam Darzi¹, Somayeh Khorasani¹, Kioomars Saliminejad², Somaieh Kazemnejad¹

Affiliations

¹ Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
² Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Abstract

Background: In this study we differentially showed the effects of cell-seeded bilayer scaffold wound dressing in accelerating healing process in diabetic ulcers that still remains as a major clinical challenge. The aim of the study was to compare immunomodulatory and angiogenic activity, and regenerative effect differences between Menstrual blood-derived Stem Cells (MenSCs) and foreskin-derived keratinocytes/fibroblasts.

Methods: The streptozotocin-induced diabetic mice model was developed in male C57/BL6 mice. A bilayer scaffold was fabricated by electrospining silk fibroin nano-fibers on human Amniotic Membrane (AM). Dermal fibroblasts and keratinocyte isolated from neonatal foreskin and MenSCs were isolated from the menstrual blood of healthy women. The diabetic mice were randomly divided into three groups including no treatment group, fibroblast/keratinocyte-seeded bilayer scaffold group (bSC+FK), and MenSCs-seeded bilayer scaffold group. The healing of full-thickness excisional wounds evaluations in the diabetic mice model in each group were evaluated at 3, 7, and 14 days after treatment.

Results: The gross and histological data sets significantly showed wound healing promotion via re-epithelialization and wound contraction along with enhanced regeneration in MenSCs-seeded bilayer scaffold group with the most similarity to adjacent intact tissue. Immunofluorescence staining of mouse skin depicted a descending trend of type III collagen along with the higher expression of involucrin as keratinocyte marker in the MenSCs-seeded bilayer nanofibrous scaffold group in comparison with other treatment groups from day 7 to day 14. Moreover, higher levels of CD31 and von Willebrand factor (VWF), and also a higher ratio of M2/M1 macrophages in association with higher levels of the neural marker were observed in the bSC+MenSCs group in comparison with bSC+FK and no treatment groups.

Conclusion: Healing symptoms in wounds dressed with keratinocyte/fibroblast-seeded bilayer scaffold was significantly lower than MenSCs-seeded bilayer scaffold done on impaired diabetic wound chronicity.

Keywords: Bilayer scaffold; Diabetic wound; Fibroblasts; Keratinocyte; Menstrual blood stem cells.