Hyperforin/HP-β-Cyclodextrin Enhances Mechanosensitive Ca²⁺ Signaling in HaCaT Keratinocytes and in Atopic Skin Ex Vivo Which Accelerates Wound Healing

Figure 2

Stretch-induced Ca²⁺ wave propagation from the wound edge and ATP release from the leading cells. Hyperforin/HP-β-CD-treated HaCaT cells at 3 h after scratching were subjected to a single stretch (20% for 1 s), which was applied perpendicular to the linear gap. (a) The intracellular Ca²⁺ responses were measured using the Ca²⁺ fluorescence indicator, Fluo-8. In response to the stretch, the cells at the leading edge exhibited a remarkably long-lasting increase in intracellular Ca²⁺, and the Ca²⁺ increase subsequently propagated to the cells located behind the edge (Movie S1 online). (b) The release of ATP was visualized using a real-time luciferin-luciferase bioluminescence imaging system. Representative overlay images of the ATP-dependent luminescence (red) and infrared DIC images (green) are shown. Following stretching, the release of ATP was only observed in the cells at the leading edge. The released ATP diffused into the entire area and remained at a high concentration for several minutes (Movie S2 online).

(a)

(b)