Ziziphora tenuior L. essential oil from Dana Biosphere Reserve (Southern Jordan); Chemical characterization and assessment of biological activities.

ETHNOPHARMACOLOGIC RELEVANCE: Ziziphora tenuior L. (Lamiaceae) is a medicinal plant in Jordan, which is included in various antimicrobial, antiseptic, expectorant and wound healing preparations. It is used for the treatment of cough, stomach ache, dysentery, fever, uterus infection, gut inflammation and painful menstruation. AIM OF THE STUDY: The aim of this study was to assess, for the first time, the chemical composition of the essential oil of Z. tenuior originated from southern Jordan and its antifungal effects against several yeasts. Concomitantly, the mechanisms behind the anti-fungal activity against Candida albicans were also disclosed. Since the Z. tenuior traditional uses are related with inflammatory-associated conditions, the putative anti-inflammatory activity of the oil was also unveiled. Importantly, the potential toxicity of pharmacologically active concentrations was screened in different types of mammalian cells. MATERIALS AND METHODS: Z. tenuior essential oil, isolated by hydrodistillation, was analyzed by gas chromatography, gas chromatography-mass spectrometry and 13C nuclear magnetic resonance spectroscopy. Antifungal activity was evaluated against yeasts, dermatophytes and Aspergillus strains. Germ tube inhibition and biofilm formation assays were evaluated using C. albicans. Assessment of cell viability was made by the MTT assay using different types of mammalian cells, including hepatocytes, keratinocytes and macrophages. The in vitro anti-inflammatory potential of the oil was evaluated by measuring nitric oxide production using lipopolysaccharide-stimulated mouse macrophages. RESULTS: Oxygen-containing monoterpenes are the main oil compounds: pulegone (46.8%), p-menth-3-en-8-ol (12.5%), isomenthone (6.6%) and 8-hydroxymenthone (6.2%). The highest antifungal activity was against Cryptococcus neoformans, with a MIC value of 0.16µL/mL. The oil revealed an important inhibitory effect on germ tube formation with a filamentation inhibition rate higher than 80% at 0.16µL/mL. The amount of the attached biomass was reduced. Importantly, concentrations devoid of toxicity on several mammalian cell types still displayed anti-inflammatory activity (0.16 and 0.32µL/mL). CONCLUSIONS: These findings add significant information to the pharmacological activity of Z. tenuior, thus justifying and reinforcing the use of this plant in traditional medicine. Additionally, the antifungal and anti-inflammatory potential of the oil at non-toxic concentrations, opens new avenues for its further exploitation, for instance in health-care product development.

Chemical composition and biological activities of Artemisia judaica essential oil from southern desert of Jordan.

ETHNOPHARMACOLOGIC RELEVANCE: Artemisia judaica L. (Arabic name: Beithran), is a medicinal and aromatic plant growing in the valley bottoms of desert areas, particularly in the southern desert of Jordan nearest to the Jordan-Saudi Arabia borders and in Wadi Araba in the Southern Badia. In Jordan, A. judaica is widely used in traditional medicine being recommended by aboriginal Bedouins in the North Badia region of Jordan as calmative. Furthermore, it is used for the treatment of stomach ache, heart diseases, sexual weakness, diabetes, gastro-intestinal disorders and external wounding. Additionally, other folk medicines of the Arabic region commonly use this aromatic plant for the treatment of inflammatory-related diseases, for instance fungal infections, diabetes, atherosclerosis, cancer and arthritis. AIM OF THE STUDY: Considering the traditional medicinal uses and the lack of scientific studies addressing the cellular and molecular mechanisms behind A. judaica claimed activities, the present study was designed to validate some of the traditional uses ascribed to this species, specifically the antifungal and anti-inflammatory activities of A. judaica essential oil at doses devoid of cytotoxicity to mammalian cells. MATERIALS AND METHODS: Chemical analysis of A. judaica essential oil isolated by hydrodistillation from aerial parts was carried out by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antifungal activity (minimal inhibitory concentrations and minimal lethal concentrations) was evaluated against yeasts, dermatophyte and Aspergillus strains. In order to deeply explore the mechanisms behind the anti-fungal effect of the essential oil, the germ tube inhibition assay and the biofilms formation assay were evaluated using Candida albicans. The assessment of cell viability was accomplished using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both hepatocytes and macrophages. Furthermore, the in vitro anti-inflammatory potential of A. judaica oil was evaluated by measuring nitric oxide (NO) production using lipopolysaccharide (LPS)-stimulated mouse macrophages. RESULTS: Oxygen containing monoterpenes are a representative group of constituents (68.7%) with piperitone (30.4%), camphor (16.1%) and ethyl cinnamate (11.0%) as main compounds. The highest antifungal activity of the oil was observed against Cryptococcus neoformans, with a MIC value of 0.16µL/mL. The oil revealed an important inhibitory effect on germ tube formation in C. albicans with 80% inhibition of filamentation at a concentration of 0.16µL/mL. Importantly, the oil also interfered with pre-formed biofilms by reducing the amount of the attached biomass. Furthermore, the essential oil significantly inhibited NO production evoked by LPS on macrophages at concentrations with very low toxicity (0.32µL/mL) or without toxicity (0.16µL/mL) to both macrophages and hepatocytes. CONCLUSIONS: The present study revealed that A. judaica essential oil from Jordan significantly inhibited germ tube formation and disrupted preformed biofilms of C. albicans, emphasizing the therapeutic potential for the treatment of disseminated candidiasis. Additionally, safe concentrations of this essential oil significantly inhibited NO production elicited by LPS in macrophages, highlighting its potential anti-inflammatory activity. Overall, A. judaica bears promising therapeutic potential for further drug development. Importantly, this work also validates some of the traditional uses of A. judaica.

Artemisia herba-alba essential oil from Buseirah (South Jordan): Chemical characterization and assessment of safe antifungal and anti-inflammatory doses.

ETHNOPHARMACOLOGIC RELEVANCE: Artemisia herba-alba Asso ("desert wormwood" in English; "armoise blanche" in French; "shaih" in Arabic), is a medicinal and strongly aromatic plant widely used in traditional medicine by many cultures since ancient times. It is used to treat inflammatory disorders (colds, coughing, bronchitis, diarrhea), infectious diseases (skin diseases, scabies, syphilis) and others (diabetes, neuralgias). In Jordanian traditional medicine, this plant is used as antiseptic and against skin diseases, scabies, syphilis, fever as well as menstrual and nervous disorders. AIM OF THE STUDY: Considering the traditional medicinal uses and the lack of scientific studies addressing the cellular and molecular players involved in these biological activities, the present study was designed to unveil the antifungal and anti-inflammatory activities of A. herba-alba Asso essential oil at doses devoid of toxicity to mammalian cells. MATERIALS AND METHODS: Chemical analysis of A. herba-alba essential oil isolated by hydrodistillation from aerial parts was carried out by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antifungal activity (minimal inhibitory concentrations and minimal lethal concentrations) was evaluated against yeasts, dermatophyte and Aspergillus strains. In order to explore the mechanisms behind the anti-fungal effect of the essential oil, the germ tube inhibition assay was evaluated using Candida albicans. The assessment of cell viability was accomplished using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the in vitro anti-inflammatory potential of A. herba-alba oil at the periphery and central nervous system was evaluated by measuring nitric oxide (NO) production using lipopolysaccharide (LPS)-stimulated mouse macrophages and microglia, respectively. RESULTS: Oxygen-containing monoterpenes are the main compounds of the oil, namely 1,8-cineole (20.1%), ß-thujone (25.1%), α-thujone (22.9%) and camphor (10.5%). Among the fungal strains tested, the oil demonstrated potential against Trichophyton rubrum and Epidermophyton floccosum, with minimal inhibitory concentration (MIC) and minimal lethal concentration (MCL) values of 0.32 mg/mL and Cryptococcus neoformans with MIC of 0.64 mg/mL. The oil revealed a strong inhibitory effect on germ tube formation in C. albicans with inhibition of filamentation around 90% at a concentration 0.16 mg/mL. Importantly, the essential oil significantly inhibited NO production evoked by LPS without cytotoxicity at concentrations up to 1.25 µL/mL in macrophages and up to 0.32 µL/mL in microglia. Furthermore, evaluation of cell viability in RAW 264.7 macrophages, BW2 microgliacells and HaCaT keratinocytes showed no cytotoxicity at concentrations up to 0.32 µL/mL. CONCLUSIONS: It was possible to find appropriate doses of A. herba-alba oil with both antifungal and anti-inflammatory activities and without detrimental effects towards several mammalian cell types. These findings add significant information to the pharmacological activity of A. herba-alba essential oil, specifically to its antifungal and anti-inflammatory therapeutic value, thus justifying and reinforcing the use of this plant in traditional medicine.

Essential oil of common sage (Salvia officinalis L.) from Jordan: assessment of safety in mammalian cells and its antifungal and anti-inflammatory potential.

Salvia officinalis L. (Lamiaceae) is a Mediterranean species, naturalized in many countries. In Jordan, it is used in traditional medicine as antiseptic, antiscabies, antisyphilitic, and anti-inflammatory, being frequently used against skin diseases. This study aimed the assessment of the antifungal and anti-inflammatory potential of its essential oils, and their cytotoxicity on macrophages and keratinocytes. The oils were investigated by gas chromatography and gas chromatography-mass spectrometry and the antifungal activity was evaluated against yeasts, dermatophyte and Aspergillus strains. Assessment of cell viability was made by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and the in vitro anti-inflammatory potential was evaluated by measuring nitric oxide production using lipopolysaccharide-stimulated mouse macrophages. The main compounds of S. officinalis oils were 1,8-cineole (39.5-50.3%) and camphor (8.8-25.0%). The oils revealed antifungal activity against dermatophyte strains and significantly inhibited NO production stimulated by LPS in macrophages, without affecting cell viability, in concentrations up to 0.64 µL/mL. This is the first report addressing the in vitro anti-inflammatory potential of S. officinalis oil. These findings demonstrated that bioactive concentrations of S. officinalis oils do not affect mammalian macrophages and keratinocytes viability making them suitable to be incorporated in skin care formulations for cosmetic and pharmaceutical purposes.