Effects of Petasitin as Natural Extract on Proliferation and Pathological Changes of Pediatric Neuroblastoma SK-N-SH Cells

Abstract

Scalp alopecia areata (SAA) is a common non-scarring hair loss condition, associated with factors such as autoimmune responses, genetics, emotional stress, and endocrine imbalances. Current treatments for SAA included minoxidil, topical steroid creams, biologics, and plant extracts. Tea tree oil (TTO), a natural plant extract, is known for its antibacterial, anti-inflammatory, and acaricidal properties, and it also provides nourishment for hair. In this research, a natural extract of TTO, was prepared to analyze its antibacterial properties. The hair follicle stem cells (HFSCs) of patients with primary SAA were analyzed to understand the influences of TTO on migration of HFSCs. TTO was extracted from fresh tea tree leaves using steam distillation. Quantitative analysis of gas chromatography-mass spectrometry (GC/MS) qualitative analysis and its total ion chromatogram using area normalization method were conducted. Meanwhile, its antibacterial activity was tested against five common pathogens (Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Staphylococcus albus (S. albus), Pseudomonas aeruginosa (P. aeruginosa), and Candida albicans (C. albicans)) by measuring the diameter of inhibition zones (DIZ), minimal inhibitory concentration (MIC), and minimum bactericidal concentration (MBC). HFSCs were isolated from patients with SAA and cultured in vitro, with cell identification performed through cytokeratin 15 (K15) immunofluorescent staining. The HFSCs were then exposed to varying concentrations (0.0, 0.5, 2.0, 5.0, 10.0, and 25.0 mmol/L) of TTO for culture, and cell proliferation activity (CPA) was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay, while migration of HFSCs was evaluated using the Transwell chamber assay. Results demonstrated that the extracted TTO had a content of 0.69 g and an extraction rate of 2.32%. 36 components were identified, constituting 98.67% of the total, with 4-terpineol reaching a high concentration of 48.35%. It exhibited a DIZ of less than 25 mm against all tested pathogens, with MIC values lower than 29 mg/mL and MBC values below 38 mg/mL. Patients with SAA displayed yellow and black dots, broken hair, malnourished and exclamation mark hairs, with few flag hairs observed in skin microscope examination. Isolated and cultured HFSCs expressed K15 primarily in the cytoplasm and exhibited favorable growth dynamics. Treatment with various concentrations of TTO greatly increased CPA and migrated cell numbers in HFSCs, with the optimal effect observed at 5.0 mmol/L concentration of TTO. In conclusion, the plant extract TTO possessed significant antibacterial activity and can promote proliferation and migration in vitro of HFSCs, suggesting its potential therapeutic application for SAA.