Shikonin was provided by Pharmacrea Kobe Co., Ltd. (Hyogo, Japan) and β-1,3-1,6 glucan was provided by Osaka Soda Co., Ltd. (Osaka, Japan). Inclusion dispersion of shikonin in β-1,3-1,6 glucan was performed using a wet-milling method by Fuji Pigment Co., Ltd. (Hyogo, Japan). Briefly, β-1,3-1,6 glucan (3.0 g) and arginine (3.0 g) were mixed with 94 mL of demineralized water and stirred using a homo disper (PRIMIX Corporation, Hyogo, Japan) at 100 g for 3 h at 85 °C to obtain a 3.0% (w/v) of β-1,3-1,6 glucan solution. The glucan solution (0.52 g) was diluted with 9.47 mL deionized water and mixed with 0.015 g shikonin extract (powder). The solution was dispersed using a paint conditioner (Red Devil Inc., Pryor, OK, USA) with zirconia beads as the medium for 2 h to give a shikonin dispersion.
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