EM - IUEM - Técnicas Laboratoriais Forenses
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- The cytotoxic effect of THJ-018 in human neuroblastoma SH-SY5Y cellsPublication . Khosasih, Vivia; Quintas, Alexandre; Costa, Joana Couceiro daSince its first appearing on 2004, synthetic cannabinoidsm the market of the so-called recreational drug, have regained popularity among adolescents and young adults. Current in vitro studies show that synthetic cannabinoids have stringer binding affinity to cannabinoids 1 (CB1) receptor than the Δ9-tetrahydrocannabinal. Therefore, it suggested that synthetic cannabinoids possess stronger psychotropic effect than cannabis. Albeit it has been increasingly abused as the legal alternative of cannabis, the pharmacological and toxicological profiles of synthetic cannabinoids remais poorly understood. One of the earliest synthetic cannabinoids to be identified was JWH-018. This study was performed to explore the toxicological effect of synthetic cannabinoids THJ-018 as the legal analog to JWH-018 on human cells model. SH-SY5Y cell lineage was used as the human cell model in this study since it has been reported to express cannabinoids 1 receptor (CB1). Seven designated concentrations of synthetic cannabinoids THJ-018, such as 1 μM, 5 μM, 10 μM, 25 μM, 50 μM, 75 μM and 100 μM were exposed for 24 hours to human neuroblastoma SH-SY5Y cell lineage. The cell viability was evaluated by performing the 3-(4,5-dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide (MTT) assay. The MTT results on SH-SY5Y cells show a statiscally significant increase on cell viability (* p<0.05, ** p<0.01 or *** p<0.001) for the concentration of 10, 25, 50, 75 and 100 μM of THJ-018. This results indicated that the THJ-018 is not toxic to the SH-SY5Y cell line. Furthermore, the cytotoxicity effect of THJ-018 combustion product, which is performed to evaluate the mimicry of the process of smoking as the common route of administration for synthetic cannabinoids, show a decrease of cell viability when SH-SY5Y cell exposed with THJ-018 combustion product.
- Toxicological profile of JWH-018 and JWH-018 N-(3-Hydroxypentyl) metabolite in the HEK 293 T cell linePublication . Sultan, Haider; Quintas, Alexandre; Dias, Mário JoãoThe rise in use of synthetic cannabinoids as herbal smoking mixtures has been attributed to a number of reasons; primary among them being their “legality” and their promotion as being safer than the natural alternatives. As their emergence has been quite recent, their pharmacological and toxicological profiles are not known completely. One of the earliest synthetic cannabinoid to be detected in these herbal mixtures was JWH-018. In the present study, the cell toxicity of JWH-018 and its N-(3-hydroxypentyl) metabolite was investigated using human cell line HEK-293 T. The cell viability was assessed using the MTT assay after treatment with standard JWH-018 and its N-hydroxymetabolite in a concentration range between 1ng/ml to 5000ng/ml of both respectively. The results show that the highest concentration of 5000ng/ml shows statistically significant decrease in cell viability for both the parent compound and the metabolite when compared to cell controls. In addition, the metabolite also showed a statistically significant decrease in cell viability at a concentration of 2500ng/ml. There were also significant differences observed between the smaller concentrations (1ng/ml and 10ng/ml) when compared to the highest concentration (5000ng/ml) for both the parent compound and metabolite. The cytotoxic effect was seen at levels which are above the expected tissue levels after use of JWH-018 either by ingestion or smoking. The cytotoxicity of both the parent compound and the metabolite highlights the adverse effects these drugs may have with consequent need for further research and regulation.