Protective Action of Cannabidiol on Tiamulin Toxicity in Humans-In Vitro Study

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“The growing awareness and need to protect public health, including food safety, require a thorough study of the mechanism of action of veterinary drugs in consumers to reduce their negative impact on humans. Inappropriate use of veterinary drugs in animal husbandry, such as tiamulin, leads to the appearance of residues in edible animal tissues.

The use of natural substances of plant origin, extracted from hemp (Cannabis sativa L.), such as cannabidiol (CBD), is one of the solutions to minimize the negative effects of tiamulin.

This study aimed to determine the effect of CBD on the cytotoxicity of tiamulin in humans.

The cytotoxic activity of tiamulin and the effect of its mixtures with CBD were tested after 72 h exposure to three human cell lines: SH-SY5Y, HepG2 and HEK-293. Cytotoxic concentrations (IC50) of the tested drug and in combination with CBD were assessed using five biochemical endpoints: mitochondrial and lysosomal activity, proliferation, cell membrane integrity and effects on DNA synthesis. Oxidative stress, cell death and cellular morphology were also assessed. The nature of the interaction between the veterinary drug and CBD was assessed using the combination index. The long-term effect of tiamulin inhibited lysosomal (SH-SY5SY) and mitochondrial (HepG2) activity and DNA synthesis (HEK-293). IC50 values for tiamulin ranged from 2.1 to >200 µg/mL (SH-SY5SY), 13.9 to 39.5 µg/mL (HepG2) and 8.5 to 76.9 µg/mL (HEK-293). IC50 values for the drug/CBD mixtures were higher.

Reduced levels of oxidative stress, apoptosis and changes in cell morphology were demonstrated after exposure to the mixtures. Interactions between the veterinary drug and CBD showed a concentration-dependent nature of tiamulin in cell culture, ranging from antagonistic (low concentrations) to synergistic effects at high drug concentrations.

The increased risk to human health associated with the presence of the veterinary drug in food products and the protective nature of CBD use underline the importance of these studies in food toxicology and require further investigation.”

https://pubmed.ncbi.nlm.nih.gov/39769305/

https://www.mdpi.com/1422-0067/25/24/13542

Cannabichromene as a Novel Inhibitor of Th2 Cytokine and JAK/STAT Pathway Activation in Atopic Dermatitis Models

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“Cannabichromene (CBC) is one of the main cannabinoids found in the cannabis plant, and although less well known than tetrahydrocannabinol (THC) and cannabidiol (CBD), it is gaining attention for its potential therapeutic benefits.

To date, CBC’s known mechanisms of action include anti-inflammatory, analgesic, antidepressant, antimicrobial, neuroprotective, and anti-acne effects through TRP channel activation and the inhibition of inflammatory pathways, suggesting that it may have therapeutic potential in the treatment of inflammatory skin diseases, such as atopic dermatitis (AD), but its exact mechanism of action remains unclear. Therefore, in this study, we investigated the effects of CBC on Th2 cytokines along with the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathways involved in AD pathogenesis. We used a 2,4-Dinitrochlorobenzene (DNCB)-induced BALB/c mouse model to topically administer CBC (0.1 mg/kg or 1 mg/kg).

The results showed that skin lesion severity, ear thickness, epithelial thickness of dorsal and ear skin, and mast cell infiltration were significantly reduced in the 0.1 mg/kg CBC-treated group compared with the DNCB-treated group (p < 0.001). In addition, real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed a significant decrease in the mRNA expression of Th2 cytokines (TSLPIL-4IL-13) and inflammatory mediators (IFN-γIL-1βIL-6IL-17IL-18, and IL-33) (p < 0.05). Western blot analysis also revealed a significant decrease in JAK1, JAK2, STAT1, STAT2, STAT3, and STAT6 protein expression (p < 0.05).

These results suggest that CBC is a promising candidate for the treatment of AD and demonstrates the potential to alleviate AD symptoms by suppressing the Th2 immune response.”

https://pubmed.ncbi.nlm.nih.gov/39769302/

https://www.mdpi.com/1422-0067/25/24/13539

An In Vitro Evaluation of Industrial Hemp Extracts Against the Phytopathogenic Bacteria Erwinia carotovora, Pseudomonas syringae pv. tomato, and Pseudomonas syringae pv. tabaci

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“Pests and diseases have caused significant problems since the domestication of crops, resulting in economic loss and hunger. To overcome these problems, synthetic pesticides were developed to control pests; however, there are significant detrimental side effects of synthetic pesticides on the environment and human health. There is an urgent need to develop safer and more sustainable pesticides.

Industrial hemp is a reservoir of compounds that could potentially replace some synthetic bactericides, fungicides, and insecticides.

We determined the efficacy of industrial hemp extracts against Pseudomonas syringae pv. tabaci (PSTA), Pseudomonas syringae pv. tomato (PSTO), and Erwinia carotovora (EC).

The study revealed a minimum inhibitory concentration (MIC) of 2.05 mg/mL and a non-inhibitory concentration (NIC) of 1.2 mg/mL for PSTA, an MIC of 5.7 mg/mL and NIC of 0.66 mg/mL for PSTO, and an MIC of 12.04 mg/mL and NIC of 5.4 mg/mL for EC. Time-kill assays indicated the regrowth of E. carotovora at 4 × MIC after 15 h and P. syringae pv. tomato at 2 × MIC after 20 h; however, P. syringae pv. tabaci had no regrowth. The susceptibility of test bacteria to hemp extract can be ordered from the most susceptible to the least susceptible, as follows: P. syringae pv. tabaci > P. syringae pv. tomato > E. carotovora.

Overall, the data indicate hemp extract is a potential source of sustainable and safe biopesticides against these major plant pathogens.”

https://pubmed.ncbi.nlm.nih.gov/39769990/

“The data show the susceptibility of the test bacteria to hemp extract, ordered from the most susceptible to the least susceptible, as follows: P. syringae pv. tabaci > P. syringae pv. tomato > E. carotovora.

This study indicates that hemp extract is effective in controlling E. carotovoraP. syringae pv. tabaci, and P. syringae pv. tomato. However, the hemp extract is more effective against both Pseudomonas spp. than E. carotovora. The difference could be due to their difference in cell wall structure, resistance mechanisms, and metabolic pathways.

More studies are needed to determine how hemp extract causes stress to bacteria such as interference with quorum sensing, biofilm formation, and oxidative stress. Moreover, to ensure sustainable agricultural practices that are safe and affordable for low-income farmers, synergistic effect studies are needed between hemp compounds in the extract and, more importantly, between hemp extracts and other plant extracts.”

https://www.mdpi.com/1420-3049/29/24/5902

Three new α-glucosidase inhibitors from aqueous extract of Cannabis sativa leaves: isolation, characterisation, and kinetic study

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“α-Glucosidase inhibitory assay-guided isolation of the aqueous extract from Cannabis sativa leaves afforded three new compounds named cannabisaldehyde (8), cannacone A (9), and canniprene C (10), along with eight previously known compounds (1711). The structures of new compounds were determined through extensive analysis of various spectroscopic data. Of isolated compounds, cannacone A (9) demonstrated most potent inhibition against maltase and sucrase with IC50 values of 80.0 and 82.9 μM, respectively. Cannacone A (9) inhibited both maltase and sucrase by competitive mechanism.”

https://pubmed.ncbi.nlm.nih.gov/39756038/

https://www.tandfonline.com/doi/full/10.1080/14786419.2024.2448841

“Alpha-glucosidase inhibitors (AGIs) are used to treat type 2 diabetes and to prevent or delay the development of type 2 diabetes in people at risk.”

“Alpha-glucosidase inhibitors are antihyperglycemic agents that lower blood glucose by delaying the digestion and absorption of complex carbohydrates.”

The antibacterial and antibiofilm role of cannabidiol against periodontopathogenic bacteria

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“Aims: Bacterial resistance and systemic risks associated with periodontitis underscore the need for novel antimicrobial agents. Cannabis sativa is a promising source of antimicrobial molecules, and cannabidiol (CBD) attracts significant interest. This study evaluated the antibacterial and antibiofilm activity of CBD against periodontopathogens, and assessed its toxicity in vivo model.

Methods and results: Antibacterial activity was determined by the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Biofilm inhibition was determined the Minimum Inhibitory Concentration of Biofilm (MICB50). Toxicity was assessed using Caeonorhabditis elegans. The periodontopathogens tested were Actinomyces naeslundii (ATCC 19039), Peptostreptococcus anaerobius (ATCC 27337), Veillonella parvula (ATCC 17745), Fusobacterium nucleatum (ATCC 10953) and Aggregatibacter actinomycetemcomitans (ATCC 43717). CBD exhibited antibacterial effects with MICs of 0.39 to 3.12 μg ml-1 and MICB50 of 0.39 μg ml-1 to 1.56 μg ml-1 against biofilms, without toxicity below 375 μg ml1.

Conclusion: The results suggest that CBD is a non-toxic product with antibacterial and antibiofilm potential, exhibiting promise as a therapeutic alternative for oral diseases.”

https://pubmed.ncbi.nlm.nih.gov/39737707/

https://academic.oup.com/jambio/advance-article-abstract/doi/10.1093/jambio/lxae316/7934962?redirectedFrom=fulltext&login=false

Genotoxicity study of Cannabis sativa L. extract

Toxicology Reports

“Cannabis sativa L., a member of the Cannabaceae family, has been thoroughly investigated for its diverse therapeutic properties, primarily attributed to cannabinoids such as delta-9-tetrahydrocannabinol (THC) and cannabidiol (CBD). Secondary, metabolites like terpenes also exhibit pharmacological effects.

This study examined the genotoxicity of a whole Cannabis sativa flower extract 160.32 mg/mL using three OECD-recommended protocols: the Ames test, micronucleus test, and comet assay. Five groups of six Wistar rats were used. Three doses of the extract (500, 1000, and 2000 mg/kgbw) or negative control (placebo) were administered orally, while cyclophosphamide monohydrate (20 mg/kgbw) was used as a positive control via intraperitoneal injection. Blood was collected for the comet test, and the animals were euthanized for bone marrow collection for the micronucleus test.

The Cannabis extract did not increase the number of revertant bacterial colonies at (375, 250, 125, and 62.5 μg/plate) in TA100 or TA98, nor did it affect the number of micronucleated polychromatic erythrocytes (MNPCEs) or the ratio of polychromatic to normochromatic erythrocytes (PCEs/NCEs). It also did not alter the index or frequency of DNA damage in hematopoietic cells.

These results suggest no genotoxic effects, supporting its potential therapeutic use.”

“Cannabis sativa extract shows no significant genotoxic effects in tested models.”

“Study supports therapeutic use of whole Cannabis sativa extract.”

https://www.sciencedirect.com/science/article/pii/S2214750024002476

https://pubmed.ncbi.nlm.nih.gov/39816045/

https://www.forbes.com/sites/emilyearlenbaugh/2024/12/30/cannabis-study-finds-no-genotoxic-effects/

Cannabinoids as cytotoxic agents and potential modulators of the human parasite Trichomonas vaginalis

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“Trichomoniasis, a globally prevalent sexually transmitted infection caused by Trichomonas vaginalis, affects approximately 278 million people each year. It presents a challenge due to resistance to the current treatment, Metronidazole (MTZ), which is also associated with side effects.

Cannabis sativa, with more than 100 phytocannabinoids and numerous studies for therapeutic applications, including parasitic infections, has undergone a significant shift in acceptance worldwide, highlighted by legalizations and substantial revenue projections.

In this context, the present study delves into the effects of cannabinoids, specifically WIN 55,212-2 (WIN), Cannabivarin (CBV) showcasing their anti-parasitic actions that influence the growth and morphology of T. vaginalis. The analysis extends to encompass the pharmacokinetic properties of these cannabinoids.

Among the analyzed cannabinoids, CBV stands out for adhering to Lipinski’s rules, indicating its potential suitability for oral drug delivery. They also demonstrated inhibitory effects on the growth of T. vaginalis trophozoites and a reduction in the parasite’s adhesion to host cells. Several morphological alterations were observed, such as membrane projections, blebbing, autophagosomes and damaged hydrogenosomes.

These results highlight the need for further research to explore the therapeutic potential of cannabinoids and understand their mechanisms of action in T. vaginalis.”

https://pubmed.ncbi.nlm.nih.gov/39724679/

“The treatment of trichomoniasis faces significant challenges, primarily due to the limited options and drug resistance issues associated with nitroimidazole derivatives like Metronidazole. However, exploring alternative therapeutic approaches is crucial. One promising avenue is the use of C. sativa and its compounds which have demonstrated anti-parasitic properties. In conclusion, cannabinoids inhibit T. vaginalis proliferation and alter its morphology, warranting further research into their therapeutic potential and mechanisms of action. Such exploration could revolutionize the current understanding and treatment of parasitic infections, offering new hope for combating these persistent pathogens.”

https://www.sciencedirect.com/science/article/pii/S0753332224016809?via%3Dihub

Biopeptide-rich fermented hemp seeds: Boosting anti-inflammatory and immune responses through Lactiplantibacillus plantarum probiotic fermentation

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“Cannabis sativa L. (hemp) seeds are increasingly recognized as a promising food source rich in phytochemicals that support inflammatory and immunological reactions.

This study investigates whether fermentation with Lactiplantibacillus plantarum can further enhance these functional properties, paving the way for hemp seeds to be developed into potent functional food ingredients.

Aqueous, 70 % ethanol, and ethyl acetate extracts from both L. plantarum-fermented (FHS) and unfermented hemp seeds (HS) were evaluated for their anti-inflammatory activities using cell-based assays.

The 70 % ethanol extract of FHS exhibited marked inhibitory effects on cytokines, including TNF-α, IL-1β, and IL-10, with fermentation significantly enhancing these effects by 25 %, 39.3 %, and 29.6 %, respectively, compared to the unfermented extracts. Additionally, mRNA expression analysis confirmed the strong immunomodulatory potential of the fermented extracts. Intracellular metabolomic analysis revealed that the ‘antifolate resistance’, ‘nicotine addiction’, ‘aminoacyl-tRNA biosynthesis’, and ‘D-amino acid metabolism’ are highlighted in the reasons for this enhancement. Furthermore, FHS significantly prolonged the survival of C. elegans exposed to pathogens, with gene expression analysis indicating modulation of the innate immune system via regulation of genes such as gcs-1, lys-1, dbl-1, pmk-1, elt-2, and dod-22. A comprehensive metabolomic and correlation analysis identified five novel bioactive peptides (AAELIGVP, AAVPYPQ, VFPEVAP, DVIGVPLG, PVPKVL) and bioactive acids (indoleacetic acid and homovanillic acid) that were enriched during fermentation, which are strongly linked to the enhanced anti-inflammatory and immunomodulatory effects observed.

These findings suggest that L. plantarum-fermented hemp seeds hold significant promise as functional ingredients in anti-inflammatory and immunomodulatory food products, with potential applications in health and wellness industries.”

https://pubmed.ncbi.nlm.nih.gov/39706455/

“L. plantarum fermentation amplified the anti-inflammatory properties of hemp seeds.”

https://www.sciencedirect.com/science/article/abs/pii/S014181302409593X?via%3Dihub

Cannabidiol induces autophagy via CB1 receptor and reduces α-synuclein cytosolic levels

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“Numerous studies have explored the role of cannabinoids in neurological conditions, chronic pain and neurodegenerative diseases. Restoring autophagy has been proposed as a potential target for the treatment of neurodegenerative diseases.

In our study, we used a neuroblastoma cell line that overexpresses wild-type α-synuclein to investigate the effects of cannabidiol on autophagy modulation and reduction in the level of cytosolic α-synuclein.

Our results demonstrated that cannabidiol enhances the accumulation of LC3-II- and GFP-LC3-positive vesicles, which indicates an increase in autophagic flux. In addition, cannabidiol-treated cells showed a reduction in cytosolic α-synuclein levels. These effects were inhibited when the cells were treated with a CB1 receptor-selective antagonist, which indicates that the biological effects of cannabidiol are mediated via its interaction with CB1 receptor. Additionally, we also observed that cannabinoid compounds induce autophagy and α-synuclein degradation after they interact with the CB1 receptor.

In summary, our data suggest that cannabidiol induces autophagy and reduces cytosolic α-synuclein levels. These biological effects are mediated preferentially through the interaction of cannabidiol with CB1 receptors, and therefore, cannabinoid compounds that act selectively on this receptor could represent a new approach for autophagy modulation and degradation of protein aggregates.”

https://pubmed.ncbi.nlm.nih.gov/39710053/

https://www.sciencedirect.com/science/article/pii/S0006899324006693?via%3Dihub

Effects of Cannabidiol on Biomineralization and Inflammatory Mediators Expression in Immortalized Murine Dental Pulp Cells and Macrophages under Pro-Inflammatory Conditions

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“Objectives: This study investigated the in vitro effects of cannabidiol (CBD) on dental pulp cells and macrophages under pro-inflammatory conditions.

Materials and methods: Mouse dental pulp undifferentiated cells (OD-21) were pre-stimulated with tumor necrosis factor alpha (10 ng/mL) or left untreated, then exposed to CBD at concentrations of 0.01 µM, 0.1 µM, 1 µM, and 10 µM for 24 hours and 7 days. Cell viability was assessed using the MTT assay, while gene expression related to mineralization-Dentin Sialophosphoprotein (Dspp), Dentin Matrix Protein 1 (Dmp1), Runt-related transcription factor 2 (Runx2), TNF-α (Tnf), and prostaglandin-endoperoxide synthase 2 (Ptgs2) were analyzed via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Mineralization nodule formation was evaluated using alizarin red staining. Macrophages (RAW 264.7) were stimulated with lipopolysaccharide (LPS) for 2 hours before exposure to the same CBD concentrations. Data analysis included the Shapiro-Wilk normality test and comparisons using ANOVA and Tukey’s post-hoc test (α = 0.05).

Results: The findings indicated that CBD did not significantly affect OD-21 cell viability, except for the 10 µM concentration after 7 days (p < 0.05). CBD treatment promoted mineralization, with significant differences observed among groups (p < 0.05). Notably, Ptgs2 expression varied between time points, while Runx2 expression was significantly reduced at 24 hours (p < 0.05). In macrophages, Ptgs2 expression was low, and TNF-α levels were downregulated across all tested CBD concentrations (p < 0.05).

Conclusion: These results suggest that cannabidiol may positively influence the biomineralization process and modulate inflammatory mediator expression.”

https://pubmed.ncbi.nlm.nih.gov/39706322/

“Cannabidiol is a compelling candidate for innovative dental therapies aimed at both reparative and preventive care.”

https://www.sciencedirect.com/science/article/abs/pii/S0300571224007048?via%3Dihub