Evaluation of Hypoxia-Conditioned MSC Exosomes' Effects on GM-CSF and IL-1α Expression in a Rat Model of Androgenic Alopecia.

Abstract

Objective: This study aims to evaluate the effects of hypoxia-conditioned mesenchymal stem cell exosomes (EH-MSCs) on granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-1α (IL-1α) expression in a dihydrotestosterone (DHT)-induced murine model of androgenetic alopecia.

Methods: This experimental study employed a randomized posttest-only control-group with 34 male C57BL/6 mice. These mice were divided into five groups: healthy control Group 1 (G1); DHT-induced control receiving saline (G2); DHT-induced mice treated with topical 5% minoxidil (G3); DHT-induced mice treated with EH-MSCs at 100 μg/kgBW (G4); and DHT-induced mice treated with EH-MSCs at 200 μg/kgBW (G5). The MSCs were isolated from umbilical cords, validated based on morphology, surface marker expression, and differentiation assays, and the exosomes were characterized by the presence of CD63 and CD9 markers. Gene expression of GM-CSF and IL-1α was quantified by real-time polymerase chain reaction. Statistical analyses included ANOVA or non-parametric equivalents, with p<0.05 being considered significant.

Results: EH-MSC treatment significantly modulated cytokine expression compared with controls. GM-CSF expression was upregulated in EH-MSC groups, with the highest level being found in G5 (3.42±0.31). This showed a dose-dependent effect (p<0.001). Conversely, IL-1α expression was downregulated, with the lowest level being found in G5 (0.95±0.33). This indicated potent anti-inflammatory activity (p=0.001). Finally, minoxidil did not significantly reduce IL-1α levels compared with saline.

Conclusions: EH-MSCs increased GM-CSF and decreased IL-1α expression in a dose-dependent manner, demonstrating dual regenerative and anti-inflammatory effects. These findings support the idea that EH-MSCs can serve as a promising therapeutic candidate for androgenetic alopecia.