Malus pumila Mill. cv Annurca apple extract might be therapeutically useful against oxidative stress and patterned hair loss

Abstract

Patterned hair loss (PHL) or androgenetic alopecia is a condition affecting about 50% of people worldwide. Several pharmacological medications have been developed over the years, but few studies have investigated their effectiveness. Therefore, new, safer and more effective strategies are required. Recent investigations showed that Annurca apple extract application could induce keratin production and promote hair growth thanks to the high amount of procyanidin B2 contained in. Hence, this study aimed to investigate the role of an Annurca apple extract in preventing PHL by testing it on human follicle dermal papilla cells (HFDPCs) for the first time. Treatment of HFDPCs with Annurca apple extract counteracted intracellular reactive oxygen species accumulation by increasing the activity of antioxidant enzymes such as superoxide dismutase 2 and catalase. Furthermore, treatment with Annurca apple extract increased β-catenin and fibroblast growth factor 2, which are involved in hair growth stimulation. These data suggest that Annurca apple extract may be a potential therapeutically useful nutraceutical product for preventing or treating hair loss by reducing oxidative stress and inducing the expression of hair growth-related factors.

Keywords: Annurca apple extract; androgenetic alopecia; antioxidant effect; nutraceutical products; patterned hair loss.

Fig. 1

Representative chromatogram of AT HAIR‐FUL AA®. (1) Chlorogenic acid; (2) procyanidin B2. The analysis was performed using a wavelength of 280 nm.

Fig. 2

Cell viability of human hair follicle dermal papilla cells (HFDPCs) treated with H₂O₂. Cells were treated with different concentrations (100–700 μm) of H₂O₂ for 2 h. Cell viability was evaluated by MTT assay. Data are expressed as the mean ± SD of three independent experiments (n = 3). **P < 0.01, ****P < 0.0001.

Fig. 3

Evaluation of the protective effect of AT HAIR‐FUL AA® against H₂O₂‐induced oxidative stress. Human hair follicle dermal papilla cells (HFDPC) were (A) preincubated with different concentrations of AT HAIR‐FUL AA® (0.25–2 mg·mL⁻¹) for 24 h and then treated with H₂O₂ 400 μm for 2 h. (B) Cells were preincubated with H₂O₂ 400 μm for 2 h and then treated with different concentrations of AT HAIR‐FUL AA® (0.25–2 mg·mL⁻¹). Cell viability was evaluated by MTT assay. Data are expressed as mean ± SD of three independent experiments (n = 3). ^#### P < 0.0001 vs CTRL. **P < 0.01, ***P < 0.001, vs H₂O₂‐treated group.

Fig. 4

Evaluation of the protective effect of procyanidin B2 and chlorogenic acid against H₂O₂‐induced oxidative stress. Human hair follicle dermal papilla cells (HFDPC) were preincubated with procyanidin B2 (PB2, 0.16 μg·mL⁻¹) and chlorogenic acid (CGA, 1.05 μg·mL⁻¹) for 24 h and then treated with H₂O₂ 400 μm for 2 h. Cell viability was evaluated by 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide dye (MTT) assay. Data are expressed as mean ± SD of three independent experiments (n = 3). ^#### P < 0.001 vs CTRL group and ****P < 0.0001 vs H₂O₂‐treated group.

Fig. 5

Human hair follicle dermal papilla cells (HFDPC) were pretreated with AT HAIR‐FUL AA® (2 mg·mL⁻¹), procyanidin B2 (PB2, 0.16 μg·mL⁻¹) and chlorogenic acid (CGA, 1.05 μg·mL⁻¹), and N‐acetyl cysteine (NAC) for 24 h, and then treated with or without H₂O₂ 400 μm for 2 h. ROS generation was measured by flow cytometry using 2′,7′‐dichlorodihydrofluorescein diacetate (DCFH‐DA) staining. Data are expressed as the mean ± SD of three independent experiments (n = 3). ^{# ###} P < 0.0001 vs CTRL and ****P < 0.0001 vs H₂O₂‐treated cells.

Fig. 6

Effect of AT HAIR‐FUL AA® on antioxidant enzymes. Human hair follicle dermal papilla cells (HFDPCs) treated with AT HAIR‐FUL AA® (2 mg·mL⁻¹), procyanidin B2 (PB2 0.16 μg·mL⁻¹) and chlorogenic acid (CGA 1.05 μg·mL⁻¹) (A–C); HFDPCs were stressed with H₂O₂ 400 μm for 2 h and then treated with AT HAIR‐FUL AA® (2 mg·mL⁻¹), procyanidin B2 (PB2, 0.16 μg·mL⁻¹) and CGA (1.05 μg·mL⁻¹) (B–D) The protein levels were normalised with β‐actin content. Data were normalised to control cells set to 100% and expressed as the mean ± SD of three independent experiments (n = 3). *P < 0.05, **P < 0.01 vs CTRL group and ^# P < 0.05, ^## P < 0.01 vs H₂O₂‐treated group.

Fig. 7

Effect of AT HAIR‐FUL AA® on hair growth factors. Human hair follicle dermal papilla cells (HFDPCs) were treated with AT HAIR‐FUL AA® (2 mg·mL⁻¹) or procyanidin B2 (PB2, 0.16 μg·mL⁻¹) or chlorogenic acid (CGA, 1.05 μg·mL⁻¹) for 4 h and 24 h. The protein levels were normalised with β‐actin content. Data were normalised to CTRL group, which was set to 100% and expressed as the mean ± SD of three independent experiments (n = 3). *P < 0.05, ***P < 0.001.