Development of ethosome comprising combined herbal extracts and its effect on hair growth

The petroleum ether (pet. ether) and ethanol extracts of leaves of Phyllanthus niruri ( P. niruri ), rhizomes of Zingiber officianale ( Z. officianale ) and Croton tiglium Linné ( C. tiglium Linné ) seeds were prepared and loaded in ethosomal formulations to evaluate the hair growth promoting activity along with safety. Phytochemical constituents in the extracts were determined. The heavy metal analysis, sterility tests for extracts, and high-performance thin layer chromatography (HPTLC) analysis of the petroleum ether, ethanol extracts, and formulations were carried out. HPTLC analysis confirmed prepared extracts and formulations contain flavonoids and phenols. The in-vitro evaluation of keratinocytes cell line studies with combined pet. ether extract formulation shows high cytotoxic activity IC 50 43 µg/mL. The hair follicular density, A/T ratio, and number of hair follicles are more in pet. ether extract of C. tigilum loaded ethosome formulation and combined pet. ether extract loaded ethosome formulation when compared to combined ethanol extract loaded ethosome formulation. Also, the combined ethanol extract loaded ethosome formulation showed more safety in topical application. Skin irritation studies showed no observed redness in combined ethanolic extract containing ethosome formulation. Histopathology report states that normal epidermis along with hair follicles and root of hair shaft on 21st day in combined extract containing ethosome formulation. Better outcome and safety were observed in the combined ethanolic extract of ethosomal formulations. The combined ethanol extracts loaded ethosome formulation was found to produce less cytotoxicity than combined pet. ether formulation and C. tiglium pet. ether extract loaded ethosome formulation.