ABSTRACT
Aquaporin 3 (AQP3) channels are tetrameric membrane-bound channels that facilitate the transport of water and other small solutes across cell membranes in the skin. Decreased AQP3 expression is associated with skin dryness, skin aging, psoriasis, and delayed wound healing. Thus, our study focused on a novel combination based on Aloe barbadensis leaf extract and trimethylglycine for targeted AQP3 regulation in skin keratinocytes and deep skin moisturization. Firstly, a dose-finding cytotoxicity assay of the selected substances was performed with a 2,5-diphenyl-2H-tetrazolium bromide (MTT) indicator on HaCaT cells. The substances' ability to increase the amount of AQP3 in keratinocytes was evaluated in a keratinocyte cell culture by means of ELISA. Additionally, the deep skin hydration effect was confirmed in clinical research with healthy volunteers. According to the results, the maximum tolerated doses providing viability at 70% (MTDs) values for Aloe barbadensis leaf extract and trimethylglycine were 24.50% and 39.00%, respectively. Following the research and development, a complex based on Aloe barbadensis leaf extract and trimethylglycine in a 1:1 mass ratio exhibited a good cytotoxicity profile, with an MTDs value of 37.90%. Furthermore, it was shown that the combination had a clear synergetic effect and significantly increased AQP3 by up to 380% compared to the negative control and glyceryl glucoside (p < 0.001). It was clinically confirmed that the developed shower gel containing Aloe barbadensis leaf extract and trimethylglycine safely improved skin hydration after one use and over 28 days. Thus, this novel plant-based combination has promising potential for AQP3 regulation in the skin epidermis and a role in the development of dermatological drugs for the treatment of skin xerosis and atopic-related conditions.
Subject(s)
Aloe , Humans , Aquaporin 3 , Skin , Keratinocytes , Betaine , Plant Extracts/pharmacologyABSTRACT
The research into new pharmaceutical substances based on essential oils, individual biologically active phytochemicals, and plant extracts is a priority in field of pharmaceutical sciences. A novel multicomponent substance based on Melaleuca alternifolia (M. alternifolia) leaf oil (TTO), 1,8-cineole (eucalyptol), and (-)-α-bisabolol with potent synergetic antimicrobial activity was investigated and suggested for the treatment of seborrheic dermatitis (SD) and dandruff. The objective of this research was to establish and validate a specific, accurate, and precise gas chromatography-mass spectrometry (GC-MS) method for further quantitative and qualitative analysis in order to ensure quality control. The main parameters of validation were suitability, specificity, linearity, accuracy, and intermediate precision according to the European Pharmacopoeia (XI edition), Russian Pharmacopoeia (XIV edition), and some parameters of ICH requirements. The peaks of fifteen chemical phytoconstituents were identified in the test sample solution with the prevalence of (-)-α-bisabolol (27.67%), 1,8-cineole (25.63%), and terpinen-4-ol (16.98%). These phytochemicals in the novel substance were chosen for standardization and validation of the GC-MS method. The chosen chromatographic conditions were confirmed for testing of the plant-based substance in a suitability test. It was established that the GC-MS method provides a significant separation, symmetry of peaks and resolution between phytochemicals. The calibration curves of each phytochemical had good linearity (R2 > 0.999) in five concentrations. In the same concertation range, the accuracy of terpinen-4-ol, 1,8-cineol, and (-)-α-bisabolol determination using the method of additives was 98.3-101.60%; the relative standard deviation (RSD) ranged from 0.89% to 1.51% and corresponded to requirements. The intraday and interday precision was ≤2.56%. Thus, the GC-MS method was validated to be specific, sensitive, linear, accurate, and precise. This GC-MS method could be recommended as a routine analytic technique for multicomponent plant-based substances-enriched terpenes.
ABSTRACT
Seborrheic dermatitis (SD) is the most prevalent dermatological disease, occurring in up to 50% of newborns, children, and adults around the world. The antibacterial and antifungal resistance contributed to the search for new natural substances and the development of a novel substance based on Melaleuca alternifolia (M. alternifolia) leaf oil (TTO), 1,8-cineole (eucalyptol), and α-(-)-bisabolol. Thus, this work aimed to determine the chemical composition of the novel plant-based substance and to evaluate its antimicrobial activity against standard microorganisms involved in the pathogenesis of SD. Moreover, the chemical composition of the substance was analyzed by gas chromatography coupled with mass spectrometry (GC/MS). Staphylococcus epidermidis (S. epidermidis), Staphylococcus aureus (S. aureus), Micrococcus luteus (M. luteus), and Candida albicans (C. albicans) were used for antimicrobial and antifungal assays by means of the broth microdilution method to determine the minimal inhibitory concentration (MIC). Finally, the substance's ability to inhibit Malassezia furfur (M. furfur) was evaluated. Eighteen compounds from different chemical groups were identified by GC/MS. The major biologically active compounds of the substance were terpinen-4-ol (20.88%), 1,8-cineole (22.28%), (-)-α-bisabolol (25.73%), and o-cymene (8.16%). The results showed that the substance has a synergistic antimicrobial and antifungal activity, while S. epidermidis and C. albicans strains were the most susceptible. Furthermore, the substance inhibited M. furfur, which is a main pathogen involved in the pathogenesis of SD and clinical manifestations. It can be concluded that the novel plant-based substance has a promising potential against M. furfur and scalp commensal bacteria and may be helpful for the development of new drugs for treatment of dandruff and SD.
