Category Archives: Uncategorized

Extraction, purification and in vitro assessment of the antioxidant and anti-inflammatory activity of policosanols from non-psychoactive Cannabis sativa L

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“Policosanols (PCs) are bioactive compounds extracted from different natural waxes.

In this work, the purification, characterization and assessment of the antioxidant and anti-inflammatory activity was carried out on PCs from an innovative source, i.e. a waxy material from supercritical-fluid extraction (SFE) of non-psychoactive Cannabis sativa L. (hemp) inflorescences.

Starting from this material, PCs were obtained by microwave-assisted trans-esterification and hydrolysis, followed by preparative liquid chromatography under normal phase conditions. The purified product was characterized using high-performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). In vitro cell-free and cell-based antioxidant and anti-inflammatory assays were then performed to assess their bioactivity. HPLC-ELSED analysis of the purified mixture from hemp wax revealed C26OH and C28OH as the main compounds. 

In vitro assays indicated an inhibition of intracellular reactive oxygen species (ROS) production, a reduction of nuclear factor kappa B (NF-κB) activation and of the activity of the neutrophil elastase. Immunoblotting assays allowed us to hypothesize the mechanism of action of the compounds of interest, given the higher levels of MAPK-activated protein kinase 2 (MK2) and heme oxygenase-1 (HO-1) protein expression in the PC pretreated HaCaT cells.

In conclusion, even if more research is needed to unveil other molecular mechanisms involved in hemp PC activity, the results of this work suggest that these compounds may have potential for use in oxinflammation processes.”

https://pubmed.ncbi.nlm.nih.gov/38737258/

“A new extraction method for policosanols from hemp wax was developed. This new product may be useful against oxinflammation processes.”

https://www.cell.com/heliyon/fulltext/S2405-8440(24)06322-9?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS2405844024063229%3Fshowall%3Dtrue

“Policosanol suppresses tumor progression in a gastric cancer xenograft model”

https://pubmed.ncbi.nlm.nih.gov/36277362/

Hemp Seeds (Cannabis sativa L.) as a Valuable Source of Natural Ingredients for Functional Foods-A Review

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“Hemp (Cannabis sativa L.) has experienced a significant resurgence in popularity, and global interest in diversifying its use in various industries, including the food industry, is growing. Therefore, due to their exceptional nutritional value, hemp seeds have recently gained increasing interest as a valuable ingredient for obtaining high-quality foods and dietary supplements.

Hemp seeds stand out for their remarkable content of quality proteins, including edestin and albumin, two distinct types of proteins that contribute to exceptional nutritional value. Hemp seeds are also rich in healthy lipids with a high content of polyunsaturated fatty acids, such as linoleic acid (omega-6), alpha-linolenic acid (omega-3), and some vitamins (vitamins E, D, and A). Polyphenols and terpenoids, in particular, present in hemp seeds, provide antimicrobial, antioxidant, and anti-inflammatory properties.

This review examines the scientific literature regarding hemp seeds’ physicochemical and nutritional characteristics. The focus is on those characteristics that allow for their use in the food industry, aiming to transform ordinary food products into functional foods, offering additional benefits for the body’s health. Innovating opportunities to develop healthy, nutritionally superior food products are explored by integrating hemp seeds into food processes, promoting a balanced and sustainable diet.”

https://pubmed.ncbi.nlm.nih.gov/38731588/

“Cannabis sativa L. hemp seeds represent a valuable resource for the food industry, offering versatility and nutritional quality.”

https://www.mdpi.com/1420-3049/29/9/2097

Cannabidiol-Loaded Solid Lipid Nanoparticles Ameliorate the Inhibition of Proinflammatory Cytokines and Free Radicals in an In Vitro Inflammation-Induced Cell Model

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“Cannabidiol (CBD) is a non-psychoactive compound derived from Cannabis sativa. It has demonstrated promising effects in combating inflammation and holds potential as a treatment for the progression of chronic inflammation. However, the clinical application of CBD is limited due to its poor solubility and bioavailability.

This study introduces an effective method for preparing CBD-loaded solid lipid nanoparticles (CBD-SLNs) using a combination of low-energy hot homogenization and ultrasonication. We enhanced this process by employing statistical optimization with response surface methodology (RSM). The optimized CBD-SLN formulation utilizes glyceryl monostearate as the primary lipid component of the nanocarrier. The CBD-SLN formulation is screened as a potential tool for managing chronic inflammation. Stable, uniformly dispersed spherical nanoparticles with a size of 123 nm, a surface charge of -32.1 mV, an encapsulation efficiency of 95.16%, and a drug loading of 2.36% were obtained.

The CBD-SLNs exhibited sustained release properties, ensuring prolonged and controlled CBD delivery, which could potentially amplify its therapeutic effects. Additionally, we observed that CBD-SLNs significantly reduced both reactive oxygen and nitrogen species and proinflammatory cytokines in chondrocyte and macrophage cell lines, with these inhibitory effects being more pronounced than those of free CBD.

In conclusion, CBD-SLNs demonstrated superiority over free CBD, highlighting its potential as an effective delivery system for CBD.”

https://pubmed.ncbi.nlm.nih.gov/38731964/

https://www.mdpi.com/1422-0067/25/9/4744

Therapeutic potential of cannabidiol (CBD) in the treatment of cardiovascular diseases

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“Introduction: Cannabidiol (CBD) is the primary non-psychoactive chemical derived from Cannabis Sativa, and its growing popularity is due to its potential therapeutic properties while avoiding the psychotropic effects of other phytocannabinoids, such as tetrahydrocannabinol (THC). Numerous pre-clinical studies in cellular and animal models and human clinical trials have demonstrated a positive impact of CBD on physiological and pathological processes. Recently, the FDA approved its use for the treatment of seizures, and clinical trials to test the efficacy of CBD in myocarditis and pericarditis are ongoing.

Areas covered: We herein reviewed the current literature on the reported effects of CBD in the cardiovascular system, highlighting the physiological effects and the outcomes of using CBD as a therapeutic tool in pathological conditions to address this significant global health concern.

Expert opinion: The comprehensive examination of the literature emphasizes the potential of CBD as a therapeutic option for treating cardiovascular diseases through its anti-inflammatory, vasodilatory, anti-fibrotic, and antioxidant properties in different conditions such as diabetic cardiomyopathy, myocarditis, doxorubicin-induced cardiotoxicity, and ischemia-reperfusion injury.”

https://pubmed.ncbi.nlm.nih.gov/38703078/

https://www.tandfonline.com/doi/full/10.1080/13543784.2024.2351513

The Effect of Cannabinoids on Single-level Lumbar Arthrodesis Outcomes in a Rat Model

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“Background context: The opioid epidemic is a public health crisis affecting spine care and pain management. Medical marijuana is a potential non-opioid analgesic yet to be studied in the surgical setting since its effects on bone healing are not fully understood. Studies have demonstrated analgesic and potentially osteoinductive properties of cannabinoids with endocannabinoid receptor expression in bone tissue.

Purpose: We hypothesize that tetrahydrocannabinol (THC) and cannabidiol (CBD) will not decrease bone healing in spinal fusion.

Study design: Seventy-eight adult Sprague-Dawley rats were used for this study. Utilizing allogenic bone grafts (6 donor rats), posterolateral inter-transverse lumbar fusion at the L4-L5 level was performed. The animals were equally divided into four treatment groups, each receiving 0.1ml intraperitoneal injections weekly as follows: placebo (saline), 5mg/kg THC, 5mg/kg CBD, and a combination of 5mg/kg THC and 5mg/kg CBD (Combo).

Methods: Callus tissue was harvested 2- and 8-weeks post-surgery for qPCR assessment to quantify changes in the expression of osteogenic genes. Manual palpation was done to assess the strength of the L4-L5 arthrodesis on all rats. μCT image-based callus analysis and histology were performed. One-way ANOVA followed by post hoc comparisons was performed.

Results: μCT demonstrated no significant differences. Treatment groups had slightly increased bone volume and density compared to control. qPCR at two weeks indicated downregulated RANKL/OPG ratios skewing towards osteogenesis in the CBD group, with the THC and CBD+THC groups demonstrating a downward trend (P>0.05). ALPL, BMP4, and SOST were significantly higher in the CBD group, with CTNNB1 and RUNX2 also showing an upregulating trend. The CBD group showed elevation in Col1A1 and MMP13. Data at eight weeks showed ALPL, RUNX2, BMP4, and SOST were downregulated for all treatment groups. In the CBD+THC group, RANK, RANKL, and OPG were downregulated. OPG downregulation reached significance for the THC and CBD+THC group compared to saline. Interestingly, the RANKL/OPG ratio showed upregulation in the CBD and CBD+THC groups. RANKL showed upregulation in the CBD group. At 2 and 8 weeks, the CBD treatment group showed superior histological progression, increasing between time points.

Conclusion: This study demonstrates that CBD and THC have no adverse effect on bone healing and the rate of spinal fusion in rats. Osteogenic factors were upregulated in the CBD-treated groups at two weeks, which indicates a potential for bone regeneration. In this group, compared to control, the RANKL/OPG ratio at the early healing phase demonstrates the inhibition of osteoclast differentiation, enhancing bone formation. Interestingly, it shows promoted osteoclast differentiation at the later healing phase, enhancing bone remodeling. This aligns with the physiological expectation of a lower ratio in the early phases and a higher ratio in the later remodeling phases.

Clinical significance: CBD and THC showed no inhibitory effects on bone healing in a spinal fusion model. Moreover, histologic and gene expression analysis demonstrated that CBD may, in fact, enhance bone healing. Further research is needed to confirm the safe usage of THC and CBD in the post-operative setting following spinal fusions.”

https://pubmed.ncbi.nlm.nih.gov/38704096/

https://www.thespinejournalonline.com/article/S1529-9430(24)00217-1/abstract

Cannabinoids and healthy ageing: the potential for extending healthspan and lifespan in preclinical models with an emphasis on Caenorhabditis elegans

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“There is a significant global upsurge in the number and proportion of older persons in the population. With this comes an increasing prevalence of age-related conditions which pose a major challenge to healthcare systems. The development of anti-ageing treatments may help meet this challenge by targeting the ageing process which is a common denominator to many health problems.

Cannabis-like compounds (cannabinoids) are reported to improve quality of life and general well-being in human trials, and there is increasing preclinical research highlighting that they have anti-ageing activity. Moreover, preclinical evidence suggests that endogenous cannabinoids regulate ageing processes.

Here, we review the anti-ageing effects of the cannabinoids in various model systems, including the most extensively studied nematode model, Caenorhabditis elegans.

These studies highlight that the cannabinoids lengthen healthspan and lifespan, with emerging evidence that they may also hinder the development of cellular senescence. The non-psychoactive cannabinoid cannabidiol (CBD) shows particular promise, with mechanistic studies demonstrating it may work through autophagy induction and activation of antioxidative systems. Furthermore, CBD improves healthspan parameters such as diminishing age-related behavioural dysfunction in models of both healthy and accelerated ageing. Translation into mammalian systems provides an important next step. Moreover, looking beyond CBD, future studies could probe the multitude of other cannabis constituents for their anti-ageing activity.”

https://pubmed.ncbi.nlm.nih.gov/38696056/

“Ageing is a complex and multifactorial process that occurs as a gradual accumulation of cellular damage in various tissues of the body, leading to a decline in physiological functions across all systems. One main aim of ageing research is to identify compounds that can postpone deteriorative changes linked to ageing. Finding interventions that promote healthy ageing may provide a paradigm shift in medicine, by targeting the common denominator of many diseases, that is, the ageing process. With the ongoing trend of cannabis legalisation globally, there is a demand for research that explores the impact of cannabis and cannabinoids on healthy ageing and diseases of ageing.

The current review highlights that cannabinoids, whether endogenous or exogenous, extend lifespan and healthspan in model systems.

However, more research is needed to observe whether these results translate in mammalian systems and ultimately in the clinic. The anti-ageing effects of cannabinoids have a number of different mechanisms, including the reduction of oxidative stress and the triggering of autophagy. More research is needed to further explore the anti-ageing mechanisms of the cannabinoids and to more comprehensively examine their impact on the hallmarks of ageing including cellular senescence. The development of anti-ageing agents that tone up endocannabinoid transmission could also be examined. Moreover, plant cannabinoids beyond CBD could be explored, as well as other cannabis constituents, alone and in combination. In addition, given the robust findings with CBD, chemical analogues of CBD might be developed. The current review underscores that there is much promise for the further development of cannabinoids as anti-ageing agents, to improve healthy ageing and general well-being.”

https://link.springer.com/article/10.1007/s11357-024-01162-8

Hexahydrocannabinol (HHC) and Δ9-tetrahydrocannabinol (Δ9-THC) driven activation of cannabinoid receptor 1 results in biased intracellular signaling

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“The Cannabis sativa plant has been used for centuries as a recreational drug and more recently in the treatment of patients with neurological or psychiatric disorders.

In many instances, treatment goals include relief from posttraumatic disorders, anxiety, or to support treatment of chronic pain. Ligands acting on cannabinoid receptor 1 (CB1R) are also potential targets for the treatment of other health conditions. Using an evidence-based approach, pharmacological investigation of CB1R agonists is timely, with the aim to provide chronically ill patients relief using well-defined and characterized compounds from cannabis.

Hexahydrocannabinol (HHC), currently available over the counter in many countries to adults and even children, is of great interests to policy makers, legal administrators, and healthcare regulators, as well as pharmacologists. Herein, we studied the pharmacodynamics of HHC epimers, which activate CB1R. We compared their key CB1R-mediated signaling pathway activities and compared them to the pathways activated by Δ9-tetrahydrocannabinol (Δ9-THC). We provide evidence that activation of CB1R by HHC ligands is only broadly comparable to those mediated by Δ9-THC, and that both HHC epimers have unique properties.

Together with the greater chemical stability of HHC compared to Δ9-THC, these molecules have a potential to become a part of modern medicine.”

https://pubmed.ncbi.nlm.nih.gov/38649680/

https://www.nature.com/articles/s41598-024-58845-7

In vitro Antibacterial Activity of Ethanolic Tanao Si Kan Dang RD1 (Cannabis sativa L.) Extracts Against Human Antibiotic-Resistant Bacteria

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“Background and Objective: A new strain of cannabis, Cannabis sativa L. Tanao Si Kan Dang RD1, has been approved and registered by the Rajamangala University of Technology Isan, Thailand. The C. sativa is acknowledged for its medicinal properties which demonstrated various therapeutic properties, such as anti-cancer and antibacterial activities. This study aimed to investigate the antibacterial activity of ethanolic extracts from the stems and leaves of the Tanao Si Kan Dang RD1 strain against seven antibiotic-resistant bacteria. 

Materials and Methods: The primary antibacterial activity of ethanolic Tanao Si Kan Dang RD1 extracts were determined using the disc diffusion method, while the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined using the broth microdilution method. 

Results: The largest inhibition zone, measuring 12 mm, was observed in leaf extracts against Pseudomonas aeruginosa 101. The lowest MIC, at 0.78 mg/mL, was obtained from stem extracts against Stenotrophomonas maltophilia. The lowest MBCs, at 12.5 mg/mL, were observed in leaf extracts against Enterococcus faecalis, Acinetobacter baumannii, multidrug-resistant Klebsiellapneumoniae, Stenotrophomonas maltophilia and Pseudomonas aeruginosa 101 and stem extracts against Acinetobacter baumannii, multidrug-resistant Klebsiella pneumoniae, Stenotrophomonas maltophilia and Pseudomonas aeruginosa 101. 

Conclusion: This study presents a novel finding regarding the antibacterial activity of ethanolic extracts from the leaves and stems of Tanao Si Kan Dang RD1 against antibiotic-resistant bacteria. The potential application of these cannabis plant extracts in the development of antibiotics capable of combating antibiotic-resistant pathogenic bacteria represents a promising strategy to address a significant global health concern.”

https://pubmed.ncbi.nlm.nih.gov/38686733/

https://scialert.net/abstract/?doi=pjbs.2024.119.124

Hemp (Cannabis sativa L.) protein: Impact of extraction method and cultivar on structure, function, and nutritional quality

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“Hemp (Cannabis sativa L.) is increasingly gaining traction as a novel and sustainable source of plant protein.

Accordingly, the aim of this study was to investigate the effectiveness of two protein extraction methods, alkaline extraction coupled with isoelectric precipitation (AE-IEP) and salt extraction coupled with ultrafiltration (SE-UF) in producing hemp protein isolates (pH-HPI and salt-HPI) with high purity and yield. Structural characterization as impacted by extraction method and cultivar was performed and related to functional performance and nutritional quality.

Both extraction methods, with carefully selected parameters, resulted in HPI with high purity (86.6-88.1% protein) and protein extraction yields (81.6-87.3%). All HPI samples had poor solubility (∼9-20%) at neutral pH compared to commercial soy protein and pea protein isolates (cSPI, cPPI). A relatively high surface hydrophobicity and low surface charge contributed to such poor solubility of HPI. However, HPI demonstrated similar solubility at acidic pH (50-67%) and comparable gel strength (up to 24 N) to cSPI. Comparing experimental amino acid composition to the theoretical amino acid distribution in hemp protein provided insights to the functional performance of the protein isolates. While pH-HPI demonstrated better functionality than salt-HPI, minimal structural, functional, and nutritional differences were noted among the pH-HPI samples extracted from four different cultivars.

Overall, results from this work could be used to guide future attempts to further develop successful protein extraction processes, and to provide valuable insights to propel breeding efforts that target enhanced hemp protein characteristics for food applications.”

https://pubmed.ncbi.nlm.nih.gov/38681526/

https://linkinghub.elsevier.com/retrieve/pii/S2665927124000728

Immunostimulatory and Antibacterial Effects of Cannabis sativa L. Leaves on Broilers

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“The aim of this study was to evaluate the effect of dried Cannabis sativa L. leaves as a phytogenic mixture added to broiler feed on CD4+ and CD8+ T lymphocyte subpopulations,

Newcastle disease virus (NDV) antibody titres, and the presence of E. coli in faecal samples. The study was conducted on 100 male Ross 308 broilers, divided into four groups of 25 broilers, for a 42-day research period. The groups were housed separately in boxes on a litter of softwood shavings and were fed starter mixture from day 1 to day 21 and finisher mixture from day 22 to day 42. Industrial hemp (C. sativa) was grown in the Crkvina area, Croatia (latitude: 45°18’46.8″ N; longitude: 15°31’30″ E). The hemp leaves were manually separated, sun-dried, and ground to a powder. The mixture offered to the control group did not contain cannabis leaves, whereas the three experimental groups received mixtures containing mixed cannabis leaves in a quantity of 10 g/kg, 20 g/kg, or 30 g/kg (E_10, E_20, and E_30, respectively). The mean NDV antibody level was uniform in all study groups until post-vaccination day 14 and increased comparably with time. The percentage of CD4+ and CD8+ lymphocytes in the peripheral blood subpopulation showed statistically significant differences (p < 0.001) in the E_20 group as compared with the control group and both the E_10 and E_30 groups throughout the study period. As the broiler age increased, the CD4+-to-CD8+ ratios also increased and were statistically significant (p < 0.0001) on day 42 in all experimental groups as compared to the control group. Comparing the control group with the experimental groups indicated that the bacterial count was lower in broiler groups having received feed with the addition of 20 g/kg and 30 g/kg C. sativa leaves.

In conclusion, the C. sativa leaves were found to elicit a favourable immunomodulatory effect on cell-mediated and humoral immune responses in broilers via increased CD4+ and CD8+ lymphocyte subpopulations and higher CD4+:CD8+ cell ratios, thus indicating enhanced immune function capacity. In addition, C. sativa leaves may have complementary effects on the broiler post-vaccination immune response, increase broilers’ resistance to infectious diseases, reduce the effect of stress associated with vaccination, and improve broiler health and welfare.”

https://pubmed.ncbi.nlm.nih.gov/38672306/

“Food safety, climate change, the emergence of infectious diseases, the ban on the use of antibiotics as growth promoters, and increasingly demanding intensive production are daily challenges for poultry production. A functional immune system is a prerequisite for animal health, and nutrition is one of the modulators of the immune system; therefore, the appropriate balance of nutrients is extremely important for the proper development and maintenance of the immune system of animals. The antimicrobial and immunomodulatory effects of phytobiotics are properties that make their use important as feed additives for poultry.

Cannabis sativa L. contains many different compounds such as flavonoids, terpenes, and cannabinoids, each with different properties and effects. The effects of C. sativa seeds, essential oils, and cakes as feed additives for poultry have already been investigated, but the effect of C. sativa L. leaves as a feed additive on immunostimulatory and antibacterial activity has not. The results of this study show that C. sativa as a phytogenic additive to animal feed has a favourable antimicrobial and immunomodulatory effect in the production of broiler chickens.”

https://www.mdpi.com/2076-2615/14/8/1159