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

Ebrahim Mirzadegan Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Hannaneh Golshahi Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Zahra Saffarian Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Haleh Edalatkhah Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Maryam Darzi Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Somayeh Khorasani Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Kioomars Saliminejad Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
Somaieh Kazemnejad Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

DOI: https://doi.org/10.18502/ajmb.v15i3.12923

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

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 nanofibers 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.