In vivo and in silico studies of the effects of oil extracted from Cannabis sativa L. seeds on healing of burned skin wounds in rats

IntroductionThis study investigates the potential effects of cannabis seed oil (CSO) on the wound healing process. The aim was to assess the efficacy of CSO in treating skin wounds using an animal model and to explore its anti-inflammatory properties through in silico analysis.MethodsEighteen male albino Wistar rats, weighing between 200 and 250 g, were divided into three groups: an untreated negative control group, a group treated with the reference drug silver sulfadiazine (SSD) (0.01 g/mL), and a group treated topically with CSO (0.962 g/mL). The initial wound diameter for all groups was 1 cm. In silico studies were conducted using Maestro 11.5 to evaluate the anti-inflammatory effects of phytoconstituents against cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2).ResultsCSO and SSD treatments led to a significant reduction (p <0.05) in the size of burned skin wounds by day 5, with contraction rates of 53.95% and 45.94%, respectively, compared to the untreated negative control group. By day 15, wounds treated with CSO and SSD had nearly healed, showing contraction rates of 98.8% and 98.15%, respectively. By day 20, the wounds treated with CSO had fully healed (100%), while those treated with SSD had almost completely healed, with a contraction rate of 98.97%. Histological examination revealed granulated tissue, neo-blood vessels, fibroblasts, and collagen fibers in wounds treated with CSO. In silico studies identified arachidic acid, γ-linolenic acid, and linolenic acid as potent inhibitors of COX-1 and COX-2. Serum biochemical parameters indicated no significant changes (p > 0.05) in liver and kidney function in rats treated with CSO, whereas a significant increase (p < 0.01) in ALAT level was observed in rats treated with SSD.DiscussionThe findings demonstrate that CSO has a promising effect on wound healing. The CSO treatment resulted in significant wound contraction and histological improvements, with no adverse effects on liver and kidney function.However, the study's limitations, including the small sample size and the need for detailed elucidation of CSO's mechanism of action, suggest that further research is necessary. Future studies should focus on exploring the molecular pathways and signaling processes involved in CSO’s pharmacological effects

Non-aqueous capillary electrophoresis with diode array and electrospray mass spectrometric detection for the analysis of selected steroidal alkaloids in plant extracts

Nonaqueous capillary electrophoresis coupled to UV detection is described for the separation and determination of steroidal alkaloids. After optimization of electrophoretic parameters, including the electrolyte nature and the organic solvent composition, a reliable separation of solasodine and solanidine was achieved in a methanol-acetonitrile (20:80, v/v) mixture containing 25 mM ammonium acetate and 1 M acetic acid. For quantitative purpose, a fused-silica capillary with a bubble cell was used and detection was performed at low wavelength (195 nm). Method performances, including migration time and peak area reproducibility, linearity, sensitivity and accuracy, were also evaluated. The method was applied to determine solasodine in Solanum elaeagnifolium berries and Solanum sodomaeum leaves and seeds. To further improve sensitivity in the analysis of solasodine-related compounds, solanidine, demissidine and tomatidine, the developed method was interfaced with electrospray ionization mass spectrometry. In the case of solasodine, the detection limit was estimated at 3 μg/ml for NACE-UV and at 0.05 μg/ml for NACE-MS, in the selected ion-monituring mode.</p

Plant growth, mineral nutrition and volatile oil composition of Mentha suaveolens subsp. timija (Briq.) Harley cultivated under salt stress conditions

Mentha suaveolens subsp. timija is a strict Moroccan endemic species that is intensively exploited from the wild because of its wide ranging medicinal and culinary properties. This has resulted in the species becoming rare or threatened with risk of extinction in the wild. In order to ensure the conservation and sustainable utilization of this valuable medicinal and aromatic plant, its cultivation has been suggested as a promising solution. However, many uncontrolled factors can affect this cultivation including salinity. Thus, the present work aims to evaluate the effects of saline conditions simulated by different NaCl concentrations (0, 50, 100 and 150 mM) on growth (fresh and dry weights), mineral content, essential oil (EO) quantity and quality of M suaveolens subsp. timija. The experiment was conducted inside a shade house at the experimental garden at the Faculty of Sciences, Semlalia-Marrakech during the growing season (February–May) of the year 2010. The results showed that increasing NaCl levels to 150 mM reduced significantly the fresh and dry biomass production by 47.558% and 69.324%, respectively, in comparison to the control. Application of NaCl induced also an increase in Na+ content, while K+ content and K+/Na+ ratio decreased with increasing salt stress. The EO yield decreased by 38.9% and 50.6% at 100 and 150 mM NaCl, respectively, compared with the control (0.77 ± 0.14%). Pulegone, menthone and isomenthone were found to be the principal components of M suaveolens subsp. timija EO. The relative abundance of menthone in particular was affected by salt stress. Based on the present results, it can be concluded that mint timija is a salt sensitive crop and cannot be cultivated in salt-affected areas

Comparative evaluation of antioxidant and insecticidal properties of essential oils from five Moroccan aromatic herbs

This study describes the antioxidant and insecticidal activities of essential oils (EOs) of Mentha suaveolens subsp. timija, Thymus satureioides, Achillea ageratum, Cotula cinerea and Salvia officinalis widely used in Morocco as flavorings, food additives and preservatives. Sixty seven components were identified accounting for more than 95.0 % of the total oils. M. suaveolens subsp. timija oil had as main components menthone and pulegone. A. ageratum oil was particularly rich in artemisyl acetate and yomogi alcohol. The essential oil of T. satureioides was characterized by high contents of carvacrol and borneol. C. cinerea oil contained trans-thujone and cis-verbenyl acetate as major constituents, whereas S. officinalis oil was characterized by trans-thujone and camphor. Antioxidant activities were examined by means of 2,2-diphenyl-1- picrylhydrazyl radical (DPPH), reducing power, β- carotene/linoleic acid bleaching and ABTS radical tests. In all assays, the highest antioxidant potency was observed in T. satureioides EO with IC50 values ranging from 0.15±0.36 μg mL-1 to 0.23±0.67 μg mL−1 across the four assays. The in vitro evaluation of the insecticidal activity showed that M. suaveolens subsp. timija EO present the highest insecticidal efficiency against adults of Tribolium castaneum with LD50 and LD90 values of 0.17 μL cm−2 and 0.26 μL cm−2, respectively and LT50, LT90 values ranged from 44.19 h to 2.98 h and 98.14 h to 6.02 h, respectively. Our data support the possible use of T. satureioides oil as potential antioxidant agent, while M. suaveolens subsp. timija oil can be developed as a new natural bio-insecticide

Comparative evaluation of antioxidant and insecticidal properties of essential oils from five Moroccan aromatic herbs

This study describes the antioxidant and insecticidal activities of essential oils (EOs) of Mentha suaveolens subsp. timija, Thymus satureioides, Achillea ageratum, Cotula cinerea and Salvia officinalis widely used in Morocco as flavorings, food additives and preservatives. Sixty seven components were identified accounting for more than 95.0% of the total oils. M. suaveolens subsp. timija oil had as main components menthone and pulegone. A. ageratum oil was particularly rich in artemisyl acetate and yomogi alcohol. The essential oil of T. satureioides was characterized by high contents of carvacrol and borneol. C. cinerea oil contained trans-thujone and cis-verbenyl acetate as major constituents, whereas S. officinalis oil was characterized by trans-thujone and camphor. Antioxidant activities were examined by means of 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), reducing power, β-carotene/linoleic acid bleaching and ABTS radical tests. In all assays, the highest antioxidant potency was observed in T. satureioides EO with IC50 values ranging from 0.15 ± 0.36 μg mL-1 to 0.23 ± 0.67 μg mL−1 across the four assays. The in vitro evaluation of the insecticidal activity showed that M. suaveolens subsp. timija EO present the highest insecticidal efficiency against adults of Tribolium castaneum with LD50 and LD90 values of 0.17 μL cm−2 and 0.26 μL cm−2, respectively and LT50, LT90 values ranged from 44.19 h to 2.98 h and 98.14 h to 6.02 h, respectively. Our data support the possible use of T. satureioides oil as potential antioxidant agent, while M. suaveolens subsp. timija oil can be developed as a new natural bio-insecticide

Phenological changes to the chemical composition and biologicalactivity of the essential oil from Moroccan endemic thyme (Thymusmaroccanus Ball)

The chemical composition, antioxidant and antimicrobial activities of essential oils isolated by hydrodistillation from the aerial parts of wild endemic Moroccan Thymus maroccanus at different developmental stages (vegetative, flowering and post-flowering) were evaluated. The yield of essential oils (v/w%) based on dry weight at different stages were: vegetative (2.14%), flowering (leaves: 1.80%; flowers: 3.46%), and post-flowering (leaves: 0.98%; post-flowers: 2.46%). Analysis of the essential oils by GC/MS revealed the presence of 28 components represented mainly by monoterpenes, both oxygenated (59.4–79.4%) and hydrocarbons (14.3–34.2%). The ranges of major constituents were as follow: carvacrol (14.1–77.6%), p-cymene (3.5–7.9%), γ-terpinene (3.8–6.6%) and α-pinene (1.2–7.8%). Leaves at post-flowering stage also contained a significant amount of borneol (16.3%) and thymol (27.8%). Essential oils obtained from flowering stage showed the highest antioxidant activity as measured by DPPH free radical scavenging ability, reductive potential and β-carotene/linoleic acid assays with IC50 values ranging from 61.48 ± 1.58 μg/mL to 182.86 ± 2.84 μg/mL for leaves and from 63.81 ± 1.96 μg/mL to 129.93 ± 6.16 μg/mL for flowers. The essential oils were also screened for antimicrobial activity against seven bacteria of significant importance for food hygiene and four pathogenic fungal strains. The results indicated that Candida strains were the most sensitive among the microorganisms tested (MIC values of 0.12–0.25 mg/mL) followed by Gram positive and Gram negative bacterial strains, with the Gram-negative Pseudomonas aeruginosa being the most resistant strain. The maximum antimicrobial activity was observed with oils obtained from the vegetative and flowering stages, which therefore represent the optimal harvesting times of this plant for application in the pharmaceutical and food industries

Effects of cultivation and application of inorganic fertilizer on volatile oils constituents and biological properties of Thymus leptobotrys Murb. and T. maroccanus Ball. from Morocco

Thymus leptobotrys and T. maroccanus are endemic Moroccan medicinal species that are intensively exploited from the wild because of their wide ranging therapeutic properties. In order to preserve these valuable medicinal plants from risk of extinction, the cultivation (with and without N, P, and K fertilizer) and its consequent effect on oil chemical composition and biological activities were studied. Carvacrol (64.1–78.1%) was found as the major constituent in oil samples, followed by p-cymene (4.7–12.4%) and γ-terpinene (3.2–7.9%). Cultivation without using N, P, and K fertilizer caused a slight decrease in carvacrol content against an increase of its main precursor p-cymene. However, application of N, P, and K fertilizer increased the carvacrol content of 3.9% for T. maroccanus and 9.4% for T. leptobotrys oils compared with unfertilized plants. Regarding the antioxidant activity, wild thyme oils showed the highest potency in all assays, while oils from cultivated plants fertilized with N, P, and K had greater activity than oils from plants not receiving fertilizer. Concerning the antimicrobial activity, oils from fertilized plants exhibited the highest inhibitory effect against most tested strains with MIC values ranged from 0.03 to 31.68 mg/mL. From this work, it can be concluded that cultivation could be a promising solution to ensure the sustainable utilization of these endemic and threatened medicinal thyme species

Chemical composition, antioxidant and antimicrobial activities of essential oil obtained from wild and cultivated Moroccan Thymus species

Thymus broussonetii, Thymus maroccanus and Thymus satureioides, are endemic Moroccan species that are intensively utilized because of their wide ranging medicinal and culinary properties. In an effort to preserve these over-exploited species, the effect of cultivation on the essential oil chemical composition, antioxidant and antimicrobial activities were investigated. The hydrodistilled oils obtained from wild and cultivated thyme species were analyzed by GC–MS. In total, 41 components were identified representing more than 98% of the oils, with carvacrol (26.0–71.6%), borneol (5.0–20.1%), γ-terpinene (4.0–8.9%) and p-cymene (5.2–10.3%) as the main constituents. Similar oil profiles were obtained from wild and cultivated T. maroccanus, whereas some quantitative differences were noted between oils obtained from wild and cultivated T. broussonetii and T. satureioides. The antioxidant and antimicrobial assays revealed that all oils tested from wild and cultivated studied thyme species showed strong activities

Chemical composition, antioxidant, antimicrobial and antibiofilm activities of essential oil from the Moroccan endemic Plant, Calendula maroccana (Ball) B. D. Jacks

The chemical characterization of the essential oil of the endemic Moroccan species, Calendula maroccana, was achieved by GC and GC-MS methods. The antioxidant, antimicrobial and antibiofilm activities of this essential oil were also studied. Eighty-one compounds were identified in the essential oil, representing 94.82% of the total. Sesquiterpenes predominate in the essential oil; 2-Methoxy-4-vinylphenol (16.13%), delta-amorphene (13.56%), alpha-cedrene (6.23%), gamma-curcumene (5.10%) and alpha-bisabolol (4.94%) were the main constituents. The antioxidant potential was evaluated by DPPH, FRAP and ABTS assays: the essential oil exhibited antioxidant effectiveness with IC50 values of 0.92, 0.3 and 1.7 mg/mL, respectively. Moreover, the essential oil showed potent antimicrobial activity as revealed by inhibition zone values against selected clinically important pathogenic microorganisms. Inhibition zone values of 15.33 +/- 0.19, 23 +/- 0.05 and 15.33 +/- 0.33 mm were observed against Staphylococcus aureus, Candida albicans and C. tropicalis, respectively. The maximal and minimal inhibition concentration values were in the range of 0.25 +/- 0.17 - 4.68 +/- 0.10 mg/mL for bacteria and 0.06 +/- 0.03 - 0.12 +/- 0.14 mg/mL, for Candida. The essential oil showed potent anti-biofilm activity with dual actions, inhibiting (69 - 88%) and eradicating (75 - 92%) the biofilms formed by the three tested bacteria strains. Based on these results, this essential oil can be considered as a natural antioxidant, antibacterial and antibiofilm agent that could be applied in phytotherapy, pharmaceutical and food sectors

Chemical characterization and insecticidal properties of essential oils from different wild populations of Mentha suaveolens subsp. timija (BRIQ.) HARLEY from Morocco

The present study is the first investigation of the volatile-oil variability and insecticidal properties of the endemic Moroccan mint Mentha suaveolens subsp. timija (mint timija). The yield of essential oils (EOs) obtained from different wild mint timija populations ranged from 0.20±0.02 to 1.17±0.25% (v/w). GC/MS Analysis revealed the presence of 44 oil constituents, comprising 97.3-99.9% of the total oil compositions. The main constituents were found to be menthone (1.2-62.6%), pulegone (0.8-26.6%), cis-piperitone epoxide (2.9-25.5%), piperitone (0.3-35.5%), trans-piperitone epoxide (8.1-15.7%), piperitenone (0.2-9.6%), piperitenone oxide (0.5-28.6%), (E)-caryophyllene (1.5-11.0%), germacrene D (1.0-15.7%), isomenthone (0.3-7.7%), and borneol (0.2-7.3%). Hierarchical-cluster analysis allowed the classification of the EOs of the different mint timija populations into four main groups according to the contents of their major components. This variability within the species showed to be linked to the altitude variation of the mint timija growing sites. The results of the insecticidal tests showed that all samples exhibited interesting activity against adults of Tribolium castaneum, but with different degrees. The highest toxicity was observed for the EOs belonging to Group IV, which were rich in menthone and pulegone, with LC50 and LC90 values of 19.0-23.4 and 54.9-58.0 μl/l air in the fumigation assay and LC50 and LC90 values of 0.17-0.18 and 0.40-0.52 μl/cm2 in the contact assay