EM - IUEM - Técnicas Laboratoriais Forenses
Permanent URI for this collection
Browse
Browsing EM - IUEM - Técnicas Laboratoriais Forenses by advisor "Costa, Joana Couceiro da"
Now showing 1 - 3 of 3
Results Per Page
Sort Options
- Analysis of tobacco and cannabis smoke and its toxicological impact in neuroblastoma tumour cellsPublication . Grafinger, Katharina; Quintas, Alexandre; Costa, Joana Couceiro daThe dried products of Cannabis sativa have been consumed for their psychoactive effects for thousands of years. Almost five hundred different natural cannabinoids could be identified in Cannabis sativa with Δ9-THC being the most psychoactive one. The most effective way of consumption is by smoking, because it avoids digestion and the first pass in the liver. Usually so called "joints" are rolled in a 1:1 ratio of dried cannabis and tobacco filler. The reason for this is that cannabis has a lower burning efficiency and that tobacco is cheaper. Processed Nicotina tabaccum contains about 0.6- 9 % nicotine; an oily, volatile liquid alkaloid. Pyrolysis of tobacco and cannabis leads to the formation of new products whose cytotoxicity is not yet established. This study investigates the effect of smoke of tobacco, cannabis and their mixture in similar conditions as humans uptake it. For production of smoke an in-house developed smoking device was used which trapped the water-soluble and organic-soluble molecules separately. Qualitative analysis of the smoke was performed using GC-MS and measurement of cytotoxicity on neuroblastoma tumour cells SH-SY5Y was performed using the MTT assay. SH-SY5Y cell line selectively expresses Cannabinoid 1 (CB1) receptor and Nicotinic acetylcholine receptor (nAChR). The results allow us to identify several pyrolysated products which are hydroxylated forms of nicotine and Δ9-THC. Moreover, MTT analysis strongly suggests different toxicological impact between smoke of tobacco, cannabis and their mixture. In fact our results points smoke from the mixture of tobacco and cannabis having the higher impact on cell viability followed by the smoke of tobacco which in turn is more toxic than smoke of cannabis, the least toxic compound out of these three. This study shows that at 100 μM cannabis has a cell viability of 80 % while tobacco and the mixture have around 40 %.
- Assessment of neuronal cytotoxicity of JWH-073 and JWH-250Publication . Pereira, Carlos Victor Montefusco; Quintas, Alexandre; Costa, Joana Couceiro daSynthetic cannabinoids from marijuana herbal blends like ‘Spice’ and ‘K2’ are drawing the attention of drug of abuse organizations, including the UNODC1, the EMCCDA2 and emergency hospital all over the world. This concern rises from clinical episodes of psychotropic effects that go beyond the regular range of marijuana and THC – namely, panic attacks, psychosis, catatonia, addiction and withdrawal symptoms. Our study addressed two emergent synthetic cannabinoids (napthtoylindoles) denominated JWH-073 and JWH-250 that are currently detected on ‘Spice’-like products, in order to observe their cell toxicity profile on neuronal cells in vitro model (SH-SY5Y). Using 0.2% DMSO as negative control, MTT and LDH results revealed that within concentrations of 1, 5, 10, 25, 37.5 and 50 μM, JWH-250 is identified as ‘toxic’ in a statistically significant manner at higher concentrations. This work did not detect any statistically significant toxicity from JWH-073. This data suggests to extend these studies on new synthetic cannabinoids to neuronal cells with increased concentrations, as well as the application of assays assessing apoptosis (conditions and signalling), neuronal function and activity (as cell membrane potential assay) within differentiated cells as neurons and glia. At the same time, the evaluation of herbal mixtures of more than one cannabinoids and plant types is advisable in order to understand synergic effects.
- 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.