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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

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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

Cannabidiol alleviates LPS-inhibited odonto/osteogenic differentiation in human dental pulp stem cells in vitro

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“Aim: Cannabidiol (CBD), derived from the Cannabis sativa plant, exhibits benefits in potentially alleviating a number of oral and dental pathoses, including pulpitis and periodontal diseases. This study aimed to explore the impact of CBD on several traits of human dental pulp stem cells (hDPSC), such as their proliferation, apoptosis, migration and odonto/osteogenic differentiation.

Methodology: hDPSCs were harvested from human dental pulp tissues. The cells were treated with CBD at concentrations of 1.25, 2.5, 5, 10, 25 and 50 μg/mL. Cell responses in terms of cell proliferation, colony-forming unit, cell cycle progression, cell migration, apoptosis and odonto/osteogenic differentiation of hDPSCs were assessed in the normal culture condition and P. gingivalis lipopolysaccharide (LPS)-induced ‘inflammatory’ milieus. RNA sequencing and proteomic analysis were performed to predict target pathways impacted by CBD.

Results: CBD minimally affects hDPSCs’ behaviour under normal culture growth milieu in normal conditions. However, an optimal concentration of 1.25 μg/mL CBD significantly countered the harmful effects of LPS, indicated by the promoting cell proliferation and restoring the odonto/osteogenic differentiation potential of hDPSCs under LPS-treated conditions. The proteomic analysis demonstrated that several proteins involved in cell proliferation and differentiation were upregulated following CBD exposure, including CCL8, CDC42 and KFL5. RNA sequencing data indicated that CBD upregulated the Notch signalling pathway. In an inhibitory experiment, DAPT, a Notch inhibitor, reduced the effect of CBD-rescued LPS-attenuated mineralization in hDPSCs, suggesting that CBD potentially mediates Notch activation to exert its impact on odonto/osteogenic differentiation of hDPSCs.

Conclusions: CBD recovers the proliferation and survival of hDPSCs following exposure to LPS. Additionally, we report that CBD-mediated Notch activation effectively restores the odonto/osteogenic differentiation ability of hDPSCs under inflamed conditions. These results underscore the potential role of CBD as a therapeutic option to enhance dentine regeneration.”

https://pubmed.ncbi.nlm.nih.gov/39697062/

https://onlinelibrary.wiley.com/doi/10.1111/iej.14183

Deciphering the Phytochemical Potential of Hemp Hairy Roots: A Promising Source of Cannabisins and Triterpenes as Bioactive Compounds

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“Cannabis sativa L., specifically hemp, is a traditional herbaceous plant with industrial and medicinal uses.

While much research has focused on cannabinoids and terpenes, the potential of hemp roots is less explored due to bioproduction challenges. Still, this material is rich in bioactive compounds and demonstrates promising anti-inflammatory, antimicrobial, and antioxidant properties. Biotechnological methods, such as hairy root cultures, enable the efficient production of specialized metabolites while avoiding the issues of outdoors cultures. Despite these benefits, the chemical diversity understanding of hemp hairy roots remains limited.

In this study, we conducted an extensive NMR and LC/MS chemical profiling of hemp hairy roots to determine their chemical composition, revealing the presence of cannabisins for the first time. We then investigated the accumulation of cannabisins and triterpenes in both hemp hairy roots and hemp aeroponic roots.

Our findings reveal that hairy roots produce 12 times more cannabisins and 6 times more triterpenes than aeroponic roots, respectively, in addition to yielding 3 times more biomass in bioreactors. Preliminary bioassays also suggest antioxidant and antifungal properties. This research underscores the potential of hemp hairy roots as a valuable source of specialized metabolites and calls for further exploration into their bioactive compounds and applications.”

https://pubmed.ncbi.nlm.nih.gov/39683949/

“This study highlights the unique phytochemical profile of hemp hairy roots and underscores their potential for various applications. The advantages offered by hairy root cultures, such as improved productivity of biomass and metabolites, better reliability due to in vitro controlled culture and genetic consistency, and water- and energy-saving potential, make them a promising avenue for further exploration and utilization in industrial and medicinal contexts.”

https://www.mdpi.com/1420-3049/29/23/5792

Exploring the Biological Activity of Phytocannabinoid Formulations for Skin Health Care: A Special Focus on Molecular Pathways

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“Recent advancements have highlighted the potential of cannabis and its phytocannabinoids (pCBs) in skin health applications.

These compounds, through their interaction with the endocannabinoid system (ECS), show promise for skin health products. Their ability to regulate inflammation, oxidative stress and cell proliferation makes them useful in addressing skin problems such as inflammation, scarring, healing, acne and aging, positioning them as valuable tools for innovative skincare solutions.

In the present work, the cellular and molecular effects of proprietary pCB-based formulations on ECS modulation, inflammation and skin regeneration were investigated.

Using human dermal fibroblasts (HDF) and keratinocytes (HaCaT), the effect of formulations in both pre-treatment and treatment scenarios following exposure to stress-inducing agents was assessed. Key molecular markers were analyzed to tackle their efficacy in mitigating inflammation and promoting structural integrity and regeneration.

In vitro results showed that these formulations significantly reduced inflammation, promoted skin regeneration and improved structural functions. In vivo studies confirmed that the formulations were well-tolerated and led to noticeable improvements in skin health, including enhanced barrier function.

This study demonstrates the safety and efficacy of pCB-based formulations for cosmeceutical applications. By combining molecular analysis with in vivo testing, this research provides new insights into the therapeutic potential of pCBs for managing various skin conditions.”

https://pubmed.ncbi.nlm.nih.gov/39684852/

“This study confirms the safety and efficacy of pCB-based formulations for skin applications, highlighting their potential to enhance regeneration and structural processes. The findings underscore the promise of cannabis-based products in cosmetics and dermatology, meeting the rising demand for natural, effective skincare solutions and shaping the future of modern skincare and therapeutic approaches.”

https://www.mdpi.com/1422-0067/25/23/13142

Anti-staphylococcal activity of soilless cultivated cannabis across the whole vegetation cycle under various nutritional treatments in relation to cannabinoid content

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Scientific Reports

“Antibiotic resistance in staphylococcal strains and its impact on public health and agriculture are global problems. The development of new anti-staphylococcal agents is an effective strategy for addressing the increasing incidence of bacterial resistance.

In this study, ethanolic extracts of Cannabis sativa L. made from plant parts harvested during the whole vegetation cycle under various nutritional treatments were assessed for in vitro anti-staphylococcal effects.

The results showed that all the cannabis extracts tested exhibited a certain degree of growth inhibition against bacterial strains of Staphylococcus aureus, including antibiotic-resistant and antibiotic-sensitive forms. The highest antibacterial activity of the extracts was observed from the 5th to the 13th week of plant growth across all the nutritional treatments tested, with minimum inhibitory concentrations ranging from 32 to 64 µg/mL. Using HPLC, Δ9-tetrahydrocannabinolic acid (THCA) was identified as the most abundant cannabinoid in the ethanolic extracts. A homolog of THCA, tetrahydrocannabivarinic acid (THCVA), reduced bacterial growth by 74%.

These findings suggest that the cannabis extracts tested in this study can be used for the development of new anti-staphylococcal compounds with improved efficacy.”

“In summary, the present study demonstrated the antistaphylococcal activity of ethanolic extracts of C. sativa L. against both of the bacterial strains tested, MSSA and MRSA, across all the vegetation stages, especially from the 5th to the 13th week. The various nutritional treatments had no impact on the resulting antibacterial effect.”

https://www.nature.com/articles/s41598-024-54805-3