ABSTRACT
Although medical cannabis was legalized in Czechia in 2013 and its use in topical treatments of skin disorders is now allowed, galenic formulations prepared from medical cannabis have not been widely implemented in the Czech healthcare system. One of the main reasons is the lack of a straightforward standardized protocol for their preparation. Cannabinoids, e.g., cannabidiol (CBD) and tetrahydrocannabinol (THC), have been shown to have therapeutic effects on various skin conditions, such as atopic dermatitis, psoriasis, scleroderma, acne and skin pigmentation. Recognizing the potential of dermatological treatment with medical cannabis, the present study aimed to evaluate the extraction capacity of various pharmaceutical bases for cannabinoids and the stability of prepared galenic formulations for dermatological applications with respect to cannabinoid content. The results showed that the stability of cannabinoids in formulations depended on the bases' physical and chemical properties. The highest THC decomposition was observed in cream bases and Vaseline, with estimated percentage loss of total content of up to 5.4% and 5.6% per week, respectively. In contrast, CBD was more stable than THC. Overall, the tested bases were comparably effective in extracting cannabinoids from plant material. However, olive oil and Synderman bases exhibited the highest cannabinoid extraction efficiencies (approximately 70%) and the best storage stabilities in terms of the content of monitored compounds. The proposed preparation protocol is fast and easily implementable in pharmacies and medical facilities.
ABSTRACT
Cannabis preparations are gaining popularity among patients with various skin diseases. Due to the lack of scientific evidence, dermatologists remain cautious about their prescriptions. So far, only a few studies have been published about the effects of high-potency cannabis extracts on microorganisms (especially dermatophytes) causing skin problems that affect more than 25% of the worldwide population. Even though, the high-potency cannabis extracts prepared by cold extraction are mostly composed of non-psychoactive tetrahydrocannabinolic acid (THCA) and only low amount of THC, their use in topical treatment can be stigmatized. The in vitro antimicrobial and antifungal activity of two high potent cannabis strains extracted by three solvents traditionally or currently used by cannabis users (ethanol; EtOH, butane; BUT, dimethyl ether; DME) was investigated by broth dilution method. The chemical profile of cannabis was determined by high-performance liquid chromatography with ultraviolet detection and gas chromatography with mass spectrometer and flame ionization detector. The extraction methods significantly influenced chemical profile of extracts. The yield of EtOH extracts contained less cannabinoids and terpenes compared to BUT and DME ones. Most of the extracts was predominantly (>60%) composed of various cannabinoids, especially THCA. All of them demonstrated activity against 18 of the 19 microorganisms tested. The minimal inhibitory concentrations (MICs) of the extracts ranged from 4 to 256 µg/mL. In general, the bacteria were more susceptible to the extracts than dermatophytes. Due to the lower content of biologically active substances, the EtOH extracts were less effective against microorganisms. Cannabis extracts may be of value to treat dermatophytosis and other skin diseases caused by various microorganisms. Therefore, they could serve as an alternative or supportive treatment to commonly used antibiotics.
ABSTRACT
There is growing evidence to support the involvement of nutrients and biostimulants in plant secondary metabolism. Therefore, this study evaluated the potential of amino acid-based supplements that can influence different hydroponic nutrient cycles (systems) to enhance the cannabinoid and terpene profiles of medical cannabis plants. The results demonstrate that amino acid biostimulation significantly affected ion levels in different plant tissues (the "ionome"), increasing nitrogen and sulfur content but reducing calcium and iron content in both nutrient cycles. A significantly higher accumulation of nitrogen and sulfur was observed during the recirculation cycle, but the calcium level was lower in the whole plant. Medical cannabis plants in the drain-to-waste cycle matured 4 weeks earlier, but at the expense of a 196% lower maximum tetrahydrocannabinolic acid yield from flowers and a significantly lower concentration of monoterpene compounds than in the recirculation cycle. The amino acid treatments reduced the cannabinolic acid content in flowers by 44% compared to control in both nutritional cycles and increased the monoterpene content (limonene) up to 81% in the recirculation cycle and up to 123% in the drain-to-waste cycle; ß-myrcene content was increased up to 139% in the recirculation cycle and up to 167% in the drain-to-waste cycle. Our results suggest that amino acid biostimulant supplements may help standardize the content of secondary metabolites in medical cannabis. Further experiments are needed to identify the optimal nutrient dosage and method of administration for various cannabis chemotypes grown in different media.
ABSTRACT
Cannabis (Cannabis sativa L.) for medical purposes has been legalized again in many countries in recent years. Currently, only two major cannabinoids (Δ9-THC and CBD) are considered in the legislation and medication, which is not sufficient in case of dried plant material or resulting extract. Other substances (mainly terpenes/terpenoids), or their specific combinations, could influence the resulting therapeutic effect for specific oncology diagnosis and specific patients. Six different genotypes (Conspiracy Kush, Jilly Bean, Jack Cleaner 2, Jack Skellington, Nordle and Nurse Jackie) were cultivated indoor at the Czech University of Life Sciences Prague. Ethanol extracts taken from the inflorescences were assayed for their content of main cannabinoids and terpenes/terpenoids. The extracts were used for in vitro cytotoxicity studies on hepatocarcinoma human cell lines Hep-G2 and colorectal carcinoma human cell lines Caco-2 and Ht-29. Healthy lung fibroblast MRC-5 and healthy intestinal cells FHs 74 Int were used to compare selectivity of cytotoxicity. The average content of Δ9-THC in extracts was 59.1 ± 2.43%, and of CBD 1.84 ± 0.17%. The content of main cannabinoids in the Nurse Jackie genotype extract was significantly greater than that of the other genotypes. Overall, more than 60 different terpenes/terpenoids were identified in the extracts. The major terpenes/terpenoids detected in most genotypes were limonene, linalool, α-terpineol, ß-caryophyllene, trans-α-bergamotene, α-humulene, ß-caryophyllene oxide, guaiol, γ-eudesmol, ß-eudesmol and α-bisabolol. Differences in the terpene composition of individual genotypes were caused by minor terpenoids, such as ß-ocimene, isopulegol acetate, ß-elemene, ß-selinene and spathulenol. All extracts were highly cytotoxic to Ht-29 colorectal carcinoma cells and showed positive selectivity compared to healthy FHs 74 Int colon cells. The Jack Cleaner 2 extract was cytotoxic to all cell lines tested at the lowest concentrations (8.48 ± 2.4-16.14 ± 0,07 µg/mL), but was positively selective only for colorectal cancer cells, especially Ht-29 and to a lesser extent for Caco-2. Similarly, the Nordle extract showed positive selectivity for Ht-29 and Caco-2 only. Jilly Bean was unique in this study, in that its extract functioned on all cell lines at the highest concentrations (20.13 ± 3.05-49.88 ± 1.5 µg/mL), whilst also being highly positively selective in all carcinoma lines (Ht-29, Caco-2 and Hep-G2 hepatocarcinoma) compared to healthy cell lines (FHs 74 Int and MRC-5). The results suggest that Δ9-THC and CBD are responsible for the in vitro cytotoxicity of the extracts, but observed differences in selectivity reveal their synergies with other substances. According to chemical analysis, higher concentrations of myrcene, ß-elemene, ß-selinene and α-bisabolol oxide found in the Jilly Bean genotype may positively affect the selectivity of cytotoxic activity. It is therefore vital that similar studies are performed on other cell lines, in order to be able to recommend these cannabis genotypes for preclinical and clinical studies, which are still lacking.
