Fig. 4

Preconditioning with LL-37 stimulates hair regeneration in vivo and production of regenerative factors in vitro. a VEGF, (b) TB4, (c) SDF-1α, and (d) MCP-1 mRNA expression was determined by real-time PCR. e VEGF, (f) TB4, (g) SDF-1α, and (h) MCP-1 proteins were detected using the supernatants of confluent cell cultures. ASCs were treated with 20 μg/mL LL-37 for 48 hr, and then the protein levels of VEGF, TB4, SDF-1α, and MCP-1 were analyzed using specific ELISAs. A representative experiment from three independent experiments is shown. Bars represent the mean ± SD. *, P < 0.05 vs. control. i Hair was removed from the backs of C57BL/6 mice and the hair growth rate was monitored for 3 weeks. CM of human ASCs pretreated with or without LL-37 (20 μg/mL) was topically applied daily for up to 18 days to mice with hair loss. Gross views observed by photographs. j Hair growth was scored as described in the Materials and methods section. *, P < 0.05 for control vs. group treated with ASC CM. ^§, P < 0.05 for group treated with ASC CM vs. group treated with CM of ASCs pre-activated with LL-37. VEGF vascular endothelial growth factor, TB4 thymosin beta-4, SDF-1α stromal cell-derived factor-1α, MCP-1 monocyte chemoattractant protein-1, ASC adipose-derived stromal/stem cell, ELISA enzyme-linked immunosorbent assay, CM conditioned medium