The effect of keratinocyte-like cells differentiated from mouse embryonic stem cells on wound healing in a wound model created in mice under long-term steroid pressure
Ýlayda Gençer, Damla Akoğulları Çelik, Seda Hafize Vatansever, Hasan AydedeObjectives:
Wound healing is often impaired under long-term steroid treatment due to delayed keratinocyte migration and tissue repair. Cell-based therapies, including keratinocyte-like cells derived from embryonic stem cells, have shown potential to improve tissue regeneration in experimental models. This study aimed to investigate the effects of keratinocyte-like cells derived from mouse embryonic stem cells (mESCs) on wound healing in a mouse model under long-term steroid treatment.
Material and Methods:
An experimental, controlled animal study was conducted using a murine wound healing model under long-term steroid administration. Keratinocyte-like cells were cultured with mouse fibroblasts, expanded, and differentiated using bone morphogenetic protein-4. Their characterization was confirmed by assessing cytokeratin-8 and cytokeratin-14 levels through immunocytochemistry and flow cytometry. After creating wounds, tissue samples were collected on days 10 and 21, stained with hematoxylin–eosin, and evaluated immunohistochemically for cytokeratin-8, cytokeratin-14, macrophage chemoattractant protein (MCP-1), tumor necrosis factor-alpha, vascular endothelial growth factor (VEGF), interleukin-1 beta, and collagen type 1 alpha 1 chain (COL1A1) to compare healing processes across groups. Semiquantitative immunohistochemical scores were analyzed using the Kruskal–Wallis test followed by Dunn–Bonferroni post hoc comparisons. P < 0.05 was considered statistically significant.
Results:
A total of 48 wounds were analyzed. Statistical analysis demonstrated that keratinocyte-like cell treatment supported wound healing in mice receiving long-term steroid treatment on day 21 compared with the steroid-only group. These cells maintained cytokeratin-8 and -14 expression at the wound site and were associated with differences in MCP-1, VEGF, and COL1A1 immunoreactivity.
Conclusion:
This study offers preliminary evidence that keratinocyte-like cells derived from mESCs may enhance wound healing, particularly in patients with delayed healing, such as long-term steroid users.