OBJECTIVE: The aim of this study was to: (1) evaluate the anti-inflammatory effects of cannabidiol (CBD) on primary cultures of human gingival fibroblasts (HGFs) and (2) to clinically monitor the effect of CBD in subjects with periodontitis. BACKGROUND: The use of phytocannabinoids is a new approach in the treatment of widely prevalent periodontal disease. MATERIALS AND METHODS: Cannabinoid receptors were analyzed by western blot and interleukin production detected using enzyme immunoassay. Activation of the Nrf2 pathway was studied via monitoring the mRNA level of heme oxygenase-1. Antimicrobial effects were determined by standard microdilution and 16S rRNA screening. In the clinical part, a placebo-control double-blind randomized study was conducted (56 days) in three groups (n = 90) using dental gel without CBD (group A) and with 1% (w/w) CBD (group B) and corresponding toothpaste (group A - no CBD, group B - with CBD) for home use to maintain oral health. Group C used dental gel containing 1% chlorhexidine digluconate (active comparator) and toothpaste without CBD. RESULTS: Human gingival fibroblasts were confirmed to express the cannabinoid receptor CB2. Lipopolysaccharide-induced cells exhibited increased production of pro-inflammatory IL-6 and IL-8, with deceasing levels upon exposure to CBD. CBD also exhibited antimicrobial activities against Porphyromonas gingivalis, with an MIC of 1.5 μg/mL. Activation of the Nrf2 pathway was also demonstrated. In the clinical part, statistically significant improvement was found for the gingival, gingival bleeding, and modified gingival indices between placebo group A and CBD group B after 56 days. CONCLUSIONS: Cannabidiol reduced inflammation and the growth of selected periodontal pathogenic bacteria. The clinical trial demonstrated a statistically significant improvement after CBD application. No adverse effects of CBD were reported by patients or observed upon clinical examination during the study. The results are a promising basis for a more comprehensive investigation of the application of non-psychotropic cannabinoids in dentistry.
- MeSH
- antiflogistika terapeutické užití farmakologie MeSH
- chlorhexidin terapeutické užití farmakologie analogy a deriváty MeSH
- dospělí MeSH
- dvojitá slepá metoda MeSH
- faktor 2 související s NF-E2 MeSH
- fibroblasty * účinky léků MeSH
- gingiva * účinky léků MeSH
- gingivitida * farmakoterapie MeSH
- hemoxygenasa-1 MeSH
- interleukin-6 analýza MeSH
- interleukin-8 účinky léků MeSH
- kanabidiol * farmakologie terapeutické užití MeSH
- kultivované buňky MeSH
- lidé středního věku MeSH
- lidé MeSH
- parodontitida farmakoterapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- randomizované kontrolované studie MeSH
This study examined the biotransformation of phytocannabinoids in human hepatocytes. The susceptibility of the tested compounds to transformations in hepatocytes exhibited the following hierarchy: cannabinol (CBN) > cannabigerol (CBG) > cannabichromene (CBC) > cannabidiol (CBD). Biotransformation included hydroxylation, oxidation to a carboxylic acid, dehydrogenation, hydrogenation, dehydration, loss/shortening of alkyl, glucuronidation and sulfation. CBN was primarily metabolized by oxidation of a methyl to a carboxylic acid group, while CBD, CBG and CBC were preferentially metabolized by direct glucuronidation. The study also screened for the activity of recombinant human cytochromes P450 (CYPs) and UDP-glucuronosyltransferases (UGTs), which could catalyze the hydroxylation and glucuronidation of the tested compounds, respectively. We found that CBD was hydroxylated mainly by CYPs 2C8, 2C19, 2D6; CBN by 1A2, 2C9, 2C19 and 2D6; and CBG by 2B6, 2C9, 2C19 and 2D6. CBC exhibited higher susceptibility to CYP-mediated transformation than the other tested compounds, mainly with CYPs 1A2, 2B6, 2C8, 2C19, 2D6 and 3A4 being involved. Further, CBD was primarily glucuronidated by UGTs 1A3, 1A7, 1A8, 1A9 and 2B7; CBN by 1A7, 1A8, 1A9 and 2B7; CBG by 1A3, 1A7, 1A8, 1A9, 2B4, 2B7 and 2B17; and the glucuronidation of CBC was catalyzed by UGTs 1A1, 1A8, 1A9 and 2B7.
- MeSH
- biotransformace MeSH
- glukuronosyltransferasa metabolismus MeSH
- jaterní mikrozomy * metabolismus MeSH
- kyseliny karboxylové MeSH
- lidé MeSH
- systém (enzymů) cytochromů P-450 * metabolismus MeSH
- uridindifosfát metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvod a cíl: Zachování homeostázy v ústní dutině není spojeno pouze s imunitní reakcí a metabolismem měkkých tkání, ale také s komenzálními a patogenními bakteriemi. V rámci vývoje přípravků a prostředků ústní hygieny jsou hledána nová či alternativní antibiotická agens. Jednou z aktuálně studovaných skupin látek jsou fytokanabinoidy, především kanabidiol (CBD). Cílem předložené studie je zhodnotit antimikrobiální působení čtyř nepsychotropních fytokanabinoidů na vybrané orální bakterie. Metoda: Antimikrobiální účinky (MIC – minimální inhibiční koncentrace) fytokanabinoidů byly stanoveny in vitro standardní mikrodiluční technikou. Hodnocen byl účinek CBD, kanabigerolu (CBG), kanabichromenu (CBC), kanabinolu (CBN) a vybraných extraktů s obsahem 50 % CBD nebo CBG na Streptococcus mutans CCM 7409 a Porphyromonas gingivalis CCM 3985. Dále byl hodnocen účinek CBD na Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 1825, Aggregatibacter actinomycetemcomitans CCM 4688 a Eikenella corrodens CCM 5985. Jako aktivní komparátor byl použit chlorhexidin diglukonát (CHX). Výsledky: Hodnoty MIC pro CBD u P. gingivalis byly podobné hodnotám, kterých jsme dosáhli u CHX (1–2 μg/ml). V případě S. mutans byla nejnižší MIC (8 μg/ml) zjištěna pro CBG. U L. casei a L. acidophilus se MIC pro CBD blížily MIC pro CHX. Konkrétně pro L. casei byl CBD stejně účinný jako CHX (MIC 2–4 μg/ml). Pro L. acidophilus byla stanovena MIC pro CBD (4–8 μg/ml) a pro CHX (2–4 μg/ ml). U ostatních mikrobů byla účinnost CBD v porovnání s CHX nižší. A. actinomycetemcomitans je inhibován v růstu významně lépe CHX (MIC = 4 μg/ml) než v případě CBD (MIC > 128 μg/ml). Rozdíly také vykazovalo antimikrobiální působení CBD (MIC 16–32 μg/ml) a CHX (MIC 2–4 μg/ml) u E. corrodens. Testované komplexní směsi nevykazovaly významně vyšší antimikrobiální účinky ve srovnání s jednotlivými fytokanabinoidy. Závěr: Výsledky ukazují, že nepsychotropní fytokanabinoidy (převážně CBD) inhibují vybrané bakterie, které jsou součástí orální mikrobioty. Současně jsou schopné inhibovat růst periopatogenních bakterií, jako je P. gingivalis, a v případě CBD i E. corrodens, což naznačuje možnost dalšího výzkumu a využití v zubním lékařství.
Introduction, aim: Maintaining homeostasis of the oral cavity is associated not only with the immune response and soft tissue metabolism but also the action of commensal and pathogenic bacteria. In the development of dental remedies against pathogenic forms, new/alternative antibiotic agents are being sought. One of the currently studied groups of these substances are the phytocannabinoids, in particular cannabidiol (CBD). The aim of the presented study was to evaluate the antimicrobial effects of four nonpsychotropic phytocannabinoids on selected oral bacteria. Methods: The antimicrobial effects (MIC – minimal inhibition concentrations) of phytocannabinoids were determined in vitro using a standard microdilution technique. The effect of CBD, cannabigerol (CBG), cannabichromene (CBC), cannabinol (CBN) and selected phytocannabinoid extracts composed of 50% CBD or CBG was evaluated on Streptococcus mutans CCM 7409 and Porphyromonas gingivalis CCM 3985. The effect of CBD was further tested on Lactobacillus acidophilus CCM 4833, Lactobacillus casei CCM 1825, Aggregatibacter actinomycetemcomitans CCM 4688, and Eikenella corrodens CCM 5985. Chlorhexidine digluconate (CHX) was used as an active comparator. Results: The MIC values of CBD in P. gingivalis were similar to those for CHX (1–2 μg/ml). In case of S. mutans, the lowest MIC was found for CBG (8 μg/ml). For L. casei and L. acidophilus, MIC for CBD was close to MIC for CHX. Specifically for L. casei, CBD was as effective as CHX (MIC 2–4 μg/ml). In L. acidophilus, MICs were determined for CBD (4–8 μg/ml) and for CHX (2–4 μg/ml). For other microbes, the efficacy of CBD was lower than that for CHX. A. actinomycetemcomitans growth was significantly more inhibited by CHX (MIC = 4 μg/ml) than by CBD (MIC > 128 μg/ml). Similar effects were observed for E. corrodens with the following antimicrobial activity for CBD (MIC 16–32 μg/ml) and CHX (MIC 2–4 μg/ml). The tested complex mixtures showed no superior antimicrobial effects than individual phytocannabinoids. Conclusion: The results show that non-psychotropic phytocannabinoids (predominantly CBD) inhibit some oral bacteria. At the same time, they are able to inhibit the growth of periodontopathogens such as P. gingivalis, and in case of CBD also E. corrodens, which suggests the possibility of further research and application in dentistry.
Cannabidiol (CBD) and cannabigerol (CBG) are the two main non-psychotropic phytocannabinoids with high application potential in drug development. Both substances are redox-active and are intensively investigated for their cytoprotective and antioxidant action in vitro. In this study, we focused on an in vivo safety evaluation and the effect of CBD and CBG on the redox status in rats in a 90-d experiment. The substances were administered orogastrically in a dose of 0.66 mg synthetic CBD or 0.66 mg/1.33 mg CBG/kg/day. CBD produced no changes in the red or white blood count or biochemical blood parameters in comparison to the control. No deviations in the morphology or histology of the gastrointestinal tract and liver were observed. After 90 d of CBD exposure, a significant improvement in redox status was found in the blood plasma and liver. The concentration of malondialdehyde and carbonylated proteins was reduced compared to the control. In contrast to CBD, total oxidative stress was significantly increased and this was accompanied by an elevated level of malondialdehyde and carbonylated proteins in CBG-treated animals. Hepatotoxic (regressive changes) manifestations, disruption in white cell count, and alterations in the ALT activity, level of creatinine and ionized calcium were also found in CBG-treated animals. Based on liquid chromatography-mass spectrometry analysis, CBD/CBG accumulated in rat tissues (in the liver, brain, muscle, heart, kidney and skin) at a low ng level per gram. Both CBD and CBG molecular structures include a resorcinol moiety. In CBG, there is an extra dimethyloctadienyl structural pattern, which is most likely responsible for the disruption to the redox status and hepatic environment. The results are valuable to further investigation of the effects of CBD on redox status and should contribute towards opening up critical discussion on the applicability of other non-psychotropic cannabinoids.
- MeSH
- kanabidiol * toxicita MeSH
- kanabinoidy * toxicita MeSH
- krysa rodu rattus MeSH
- oxidace-redukce MeSH
- vápník MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cannabidiol (CBD), a non-psychotropic cannabinoid produced by the genus Cannabis, is a phytoceutical that activates the endocannabinoid system (ECS) through binding to CB1 and CB2 receptors. The ECS is involved in cellular homeostasis and regulates metabolic processes in virtually all mammalian tissues. Published studies on CBD focus, inter alia, on its use in prophylaxis and as an anti-inflammatory agent. Here the authors present a critical assessment of the effects of CBD on inflammatory periodontal diseases caused by bacterial virulence factors, and evaluate critically the possible benefits and drawbacks of CBD use in dentistry. Particular attention is paid to the interaction of CBD with microbially colonized oral tissues, the inflammatory response in relation to the immune response, and the destruction/regeneration of hard and soft tissues of the periodontium.
- MeSH
- analgetika MeSH
- kanabidiol * metabolismus farmakologie terapeutické užití MeSH
- kanabinoidy * MeSH
- lidé MeSH
- nemoci parodontu * farmakoterapie MeSH
- receptor kanabinoidní CB1 MeSH
- savci metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- MeSH
- kanabidiol * farmakologie MeSH
- krysa rodu rattus MeSH
- tetrahydrokanabinol * farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- dopisy MeSH
In this contribution, a comprehensive study of the redox transformation, electronic structure, stability and photoprotective properties of phytocannabinoids is presented. The non-psychotropic cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN), cannabichromene (CBC), and psychotropic tetrahydrocannabinol (THC) isomers and iso-THC were included in the study. The results show that under aqueous ambient conditions at pH 7.4, non-psychotropic cannabinoids are slight or moderate electron-donors and they are relatively stable, in the following order: CBD > CBG ≥ CBN > CBC. In contrast, psychotropic Δ9-THC degrades approximately one order of magnitude faster than CBD. The degradation (oxidation) is associated with the transformation of OH groups and changes in the double-bond system of the investigated molecules. The satisfactory stability of cannabinoids is associated with the fact that their OH groups are fully protonated at pH 7.4 (pKa is ≥ 9). The instability of CBN and CBC was accelerated after exposure to UVA radiation, with CBD (or CBG) being stable for up to 24 h. To support their topical applications, an in vitro dermatological comparative study of cytotoxic, phototoxic and UVA or UVB photoprotective effects using normal human dermal fibroblasts (NHDF) and keratinocytes (HaCaT) was done. NHDF are approx. twice as sensitive to the cannabinoids' toxicity as HaCaT. Specifically, toxicity IC50 values for CBD after 24 h of incubation are 7.1 and 12.8 μM for NHDF and HaCaT, respectively. None of the studied cannabinoids were phototoxic. Extensive testing has shown that CBD is the most effective protectant against UVA radiation of the studied cannabinoids. For UVB radiation, CBN was the most effective. The results acquired could be used for further redox biology studies on phytocannabinoids and evaluations of their mechanism of action at the molecular level. Furthermore, the UVA and UVB photoprotectivity of phytocannabinoids could also be utilized in the development of new cannabinoid-based topical preparations.
- MeSH
- antioxidancia * farmakologie MeSH
- kanabidiol * MeSH
- lidé MeSH
- tetrahydrokanabinol MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The oxidative photocyclization of aromatic Schiff bases was investigated as a potential method for synthesis of phenanthridine derivatives, biologically active compounds with medical applications. Although it is possible to prepare the desired phenanthridines using such an approach, the reaction has to be performed in the presence of acid and TEMPO to increase reaction rate and yield. The reaction kinetics was studied on a series of substituted imines covering the range from electron-withdrawing to electron-donating substituents. It was found that imines with electron-withdrawing substituents react one order of magnitude faster than imines bearing electron-donating groups. The 1H NMR monitoring of the reaction course showed that a significant part of the Z isomer in the reaction is transformed into E isomer which is more prone to photocyclization. The portion of the Z isomer transformed showed a linear correlation to the Hammett substituent constants. The reaction scope was expanded towards synthesis of larger aromatic systems, namely to the synthesis of strained aromatic systems, e.g., helicenes. In this respect, it was found that the scope of oxidative photocyclization of aromatic imines is limited to the formation of no more than five ortho-fused aromatic rings.
Carbohelicenes are a group of helical-shaped polycyclic aromatic hydrocarbons. This study examined the effect of hexahelicene (or [6]helicene) and of its imidazolium derivative, 1-butyl-3-(2-methyl[6]helicenyl)-imidazolium bromide (I[6]H), on the activity of the aryl hydrocarbon receptor (AhR) and expression of cytochrome P450 1A1 (CYP1A1) in human hepatoma HepG2 cells. An MTT viability assay showed that both [6]helicene and I[6]H were cytotoxic to HepG2 cells after 24 h of exposure, with IC50 values of 0.9 and 8.4 μM, respectively. Using a gene reporter assay performed in transiently transfected HepG2 cells, we found that 1 μM [6]helicene, unlike I[6]H, significantly increased the activity of AhR to 2.1-fold compared to the control after 24 h of exposure. Moreover, [6]helicene induced a small but significant increase in the level of CYP1A1 mRNA. On the other hand, neither the protein level nor activity of CYP1A1 were affected by [6]helicene in HepG2 cells. The effect of [6]helicene on the AhR pathway was thus much lower than that of 2,3,7,8-tetrachlorodibenzo-p-dioxin, a potent AhR activator. We conclude that [6]helicene is a poor activator of the AhR pathway in HepG2 cells, and that the possible activation of the AhR pathway in vivo remains to be investigated.
In this short communication we report optimized procedures for the chiral separation of non-charged [6]helicene (1) and cationic derivative 1-butyl-3-(2-methyl[6]helicenyl)-imidazolium bromide (2) using high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) methods. The possibility of using capillary electrophoresis (CE) was also tested. The satisfactory results were obtained with SFC, where the highly selective resolution of four enantiopure 1 and 2 helicenes was achieved in a single run within 5min. The semi-preparative procedure for the isolation of P and M enantiomers of compound 2, including circular dichroism data, is reported here for the first time. The results could be used in further separations and analytical applications targeting carbohelicenes vs. positively charged helicene derivatives.
