Helium–Neon Laser Irradiation Stimulates Migration and Proliferation in Melanocytes and Induces Repigmentation in Segmental-Type Vitiligo Hsin-Su Yu,

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Helium–Neon Laser Irradiation Stimulates Migration and Proliferation in Melanocytes and Induces Repigmentation in Segmental-Type Vitiligo Hsin-Su Yu, Chieh-Shan Wu, Ying-Hsien Kao, Min-Hsi Chiou Journal of Investigative Dermatology Volume 120, Issue 1, Pages 56-64 (January 2003) DOI: 10.1046/j.1523-1747.2003.12011.x Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Effects of He–Ne laser irradiation on bFGF release from cultured human keratinocytes and fibroblasts. Cultured human keratinocytes and fibroblasts were irradiated with 0, 0.5. 1.0, and 1.5 J per cm2 of He–Ne laser radiation, respectively. The total amounts of bFGF released in the supernatants (including during irradiation and 30 min after irradiation) were measured with a commercially available bFGF ELISA kit. The stimulatory effect on bFGF release from both keratinocytes (open bar) and fibroblasts (solid bar) by He–Ne laser irradiation showed a dose-responsive effect. Journal of Investigative Dermatology 2003 120, 56-64DOI: (10.1046/j.1523-1747.2003.12011.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Dose-dependent effect of He–Ne laser irradiation on NGF release from cultured human keratinocytes. Dishes of cultured human keratinocytes were irradiated with 0, 0.5, 1.0, and 1.5 J per cm2 of He–Ne laser radiation, respectively. At 24 h after irradiation, NGF concentrations of the collected supernatants were measured with an NGF ELISA kit. The pattern of NGF release by keratinocytes showed a dose-dependent effect produced by He–Ne laser irradiation. Journal of Investigative Dermatology 2003 120, 56-64DOI: (10.1046/j.1523-1747.2003.12011.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Stimulatory effect of He–Ne laser irradiation on NGF gene expression in cultured human keratinocytes. Dishes of cultured human keratinocytes were irradiated with an He–Ne laser as previously described. At 4 h after irradiation, the cells were collected for total RNA extraction and RT-PCR as described in Materials and Methods. The expression of NGF mRNA by keratinocytes was semiquantitatively analyzed by normalization with β-actin mRNA expression. A stimulatory effect was observed in laser-irradiated groups. The ratios of NGF to β-actin in integrated density value were 0.075 (0 J per cm2), 0.123 (0.5 J per cm2), 0.222 (1.0 J per cm2), and 0.378 (1.5 J per cm2), respectively. PCR product sizes of β-actin and NGF were 314 bp and 302 bp. M, 100 bp ladder DNA marker. Journal of Investigative Dermatology 2003 120, 56-64DOI: (10.1046/j.1523-1747.2003.12011.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Direct and indirect effects of He–Ne laser irradiation on migration of cultured human melanocytes. Cultured human melanocytes were seeded on collagen type I coated Cellocate slips at 1×104 cells per dish. Time-lapse photographs were taken at the indicated time points and the migration distance (μm) was measured. (A) Melanocytes were treated with either mock irradiation or 1 J per cm2 He–Ne laser irradiation. (B) Melanocytes were treated with cultured medium or 10% (vol/vol) supernatants (derived from keratinocytes treated with 1 J per cm2 He–Ne laser irradiation) with or without 1 μg per ml of neutralizing antibody (Ab) against bFGF or NGF. *p<0.05, **p<0.01 compared to mock treatment or control medium. Bars: 1 SD; n=20. Journal of Investigative Dermatology 2003 120, 56-64DOI: (10.1046/j.1523-1747.2003.12011.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 He–Ne laser treatment induced repigmentation in three typical patients with segmental-type vitiligo. Figure 5,1(AA) A 39-y-old man had vitiligo on right forehead and paranasal area for 1.5 y. Figure 5,1B After 142 sessions of He–Ne laser irradiation, 90% repigmentation was observed. Figure 5,2(AA) A 13-y-old boy had vitiligo on left eyebrow and cheek areas for 5 y. Leukotrichia was noticed on the left eyebrow for about 4 y. Figure 5,2(B) 78% repigmentation was observed after 132 sessions of treatment. Figure 5,3(AA) A 12-y-old boy had depigmented lesions on right chin and upper neck for 2 y. Figure 5,3(B) 65% repigmentation was found after 139 sessions of treatment. Journal of Investigative Dermatology 2003 120, 56-64DOI: (10.1046/j.1523-1747.2003.12011.x) Copyright © 2003 The Society for Investigative Dermatology, Inc Terms and Conditions