Synergistic Effects of Mesenchymal Stem Cell Secretome and Wnt10b on Hair Follicle Regeneration in a 3D Bioengineering Model: A Preclinical Study Publisher Pubmed



Bajouri A ; Aghdami N ; Nilforoushzadeh MA
Source: Stem Cell Research and Therapy Published:2025

Abstract

Background: Despite all advances, constructing functional hair follicles remains a significant clinical challenge. Building on recent advancements in mesenchymal stem cells secretome and Wnt signaling, we aimed to fabricate hair follicles in a 3D hydrogel scaffold and assess the effects of placenta-derived MSCs (PMSC) secretome and Wnt10b on follicle formation in ex vivo and in vivo models. Methods: A gelatin methacryloyl (GelMA) hydrogel scaffold was engineered via a 3D-printed We engineered a gelatin methacryloyl (GelMA) hydrogel scaffold using a 3D-printed polydimethylsiloxane mold to create a co-culture system for hair follicle biofabrication. Dermal papilla cells (DPCs) and epithelial cells from a donor were isolated and seeded into the scaffold to form microfollicles. We treated hair microfollicles with PMSC secretome, Wnt10b protein, or combination of both. The generated hair microfollicles were assessed using specific molecular and protein markers and then implanted into nude mice skin to investigate the hair growth in vivo. Results: After three weeks of co-culturing, the group receiving both secretome and Wnt10b showed 90% cell aggregation and highest hair follicle formation rate at 28% (p < 0.0001). Immunofluorescent analysis revealed elevated expression levels of markers for DPCs and epithelial cells in formed hair follicles. Real-time PCR analysis indicated significantly enhanced expression of Wnt10b, Bmp6, Notch1 and Notch2 genes, particularly in the group receiving secretome and Wnt10b. Transplantation of GelMA hydrogels containing hair microfollicles into nude mice resulted in successful hair formation along with immunofluorescence and gene expression analyses showing marked increases in hair follicle growth markers. Conclusion: Our study demonstrates that the combination of PMSC-secretome and Wnt10b in a GelMA hydrogel scaffold effectively promotes hair follicle biofabrication and successful hair formation in vivo. However, the in vivo portion was limited by a small sample size (n = 3 per group), underscoring the need for larger-scale studies to confirm these findings. © The Author(s) 2025.