Figure 4.

Ex vivo validation of cp-asiAR therapeutic efficacy in human hair follicles. Hair follicles were isolated from the human scalp and incubated with DHT (100 nM) to mimic AGA-like hair loss conditions. (A) Each group of hair follicles was co-treated with flutamide or cp-asiAR at the indicated doses and observed on days three and six after incubation (n = seven-nine/group). (Top) Images were obtained on day six, and telogen hair was identified based on morphological characteristics as described previously.⁴ Tapered, elongated bulb shape, and dissociation of very compact papilla were considered as the key features of telogen hair. (Bottom) Hair follicles from each group were isolated on day two after incubation. Frozen sections of the tissue were hybridized with the AR riboprobe to visualize the mRNA levels of AR in the hair follicle. (B) Fluorescence intensity in the DP area was analyzed using ImageJ, and the AR levels relative to the untreated control are presented as mean and standard deviation. Statistical significance was calculated using a t-test with the vehicle control (**, p < 0.01; ***, p < 0.001). (C) Telogen ratio in each treatment group was quantified based on the number of anagen and telogen hair follicles. Statistical significance was calculated using the Chi-square test with the vehicle control. (D) Effect of cp-asiAR on the hair follicle diameter following DHT treatment. A total of 130 hair follicles from 13 patients (8 with AGA and 5 without AGA) were collected, and the hair follicle diameter was measured on days three and six after incubation with the indicated treatment. Statistical significance was calculated using the t-test with the vehicle control of the same day (*, p < 0.05, **, p < 0.01, ***, p < 0.001).