Cannabidiol-induced cellular and matrix-associated responses in primary equine sarcoid cells.
Abstract: Sarcoids are locally invasive skin tumors in equids, associated with bovine papillomavirus. Objective: Address potential applications of cannabidiol (CBD) in veterinary medicine. We evaluated the response of equine sarcoid cells to CBD in vitro, focusing on viability, invasiveness, and matrix remodeling. Methods: Three primary sarcoid cell lines. Methods: Cells were treated with CBD (20, 6.75, 2.25, 0.75 μM) and incubated for 6, 24, 48, 72 hours. Cell viability, cytotoxicity, and apoptosis were assessed using the ApoTox-Glo Assay. Based on these results, further analyses were performed for selected conditions only, including the assessment of cell invasiveness using the ECMatrix™ Cell Invasion Assay and the quantification of matrix metalloproteinase (MMP)-1, -2, and -9 in the culture medium by ELISA. Results: Treatment with CBD affected cell viability, cytotoxicity, and apoptosis. At 48 hours, apoptosis (measured as caspase 3/7 activity) reached 49.5% and further increased to 75% at 72 hours. Marked cytotoxicity (>96%) and decreased viability were observed at 72 hours. Cannabidiol also significantly decreased MMP-1 concentration by 48.9% at 24 hours and MMP-2 concentrations by 84% at 6 hours. Concentrations of MMP-9 also decreased by 37.2% and 45.3% at 6 and 48 hours, respectively, after treatment with 20 μM. Despite observed decreases in cell invasiveness ranging from 34% to 59% after 24 hours, these changes were not significant. Conclusions: Our findings support further investigation of CBD's role in extracellular matrix modulation in sarcoid tumors.
© The Author(s) 2026. Published by Oxford University Press on behalf of the American College of Veterinary Internal Medicine.
Publication Date: 2026-02-26 PubMed ID: 41742517PubMed Central: PMC12881954DOI: 10.1093/jvimsj/aalaf015Google Scholar: Lookup
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Summary
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Overview
- This research investigates the effects of cannabidiol (CBD) on primary equine sarcoid cells, particularly regarding cell viability, invasiveness, apoptosis, and matrix remodeling.
Background
- Equine sarcoids are locally invasive skin tumors found in horses and related species.
- These tumors are linked to infection by bovine papillomavirus (BPV).
- Current treatments for sarcoids are limited, and there is interest in exploring new therapeutic options.
- Cannabidiol (CBD), a non-psychoactive component of cannabis, has shown potential in modulating cancer cell behavior in various models, making it a candidate for veterinary oncology research.
Objectives
- The primary goal was to evaluate how CBD affects equine sarcoid cells in vitro, focusing on:
- Cell viability and cytotoxicity
- Apoptosis (programmed cell death)
- Cell invasiveness
- Extracellular matrix remodeling, particularly via matrix metalloproteinases (MMPs)
- These endpoints were chosen due to their relevance in tumor progression and metastasis.
Methods
- Three primary equine sarcoid cell lines were cultured and treated in vitro.
- Cells were exposed to varying concentrations of CBD: 0.75, 2.25, 6.75, and 20 μM.
- Incubation times included 6, 24, 48, and 72 hours to observe both short- and longer-term effects.
- Cell viability, cytotoxicity, and apoptosis were measured using the ApoTox-Glo Assay, which combines viability indicators and caspase 3/7 activity (a marker of apoptosis).
- Based on viability and apoptosis results, select conditions were further studied for:
- Cell invasiveness, using the ECMatrix™ Cell Invasion Assay – which measures the ability of cells to migrate through a matrix barrier, simulating invasion.
- Quantification of matrix metalloproteinases MMP-1, MMP-2, and MMP-9 in the culture medium by ELISA (enzyme-linked immunosorbent assay).
- MMPs are enzymes involved in degradation and remodeling of the extracellular matrix, which plays a key role in tumor invasiveness.
Results
- Apoptosis and Cytotoxicity:
- There was a significant increase in apoptotic activity over time, with caspase 3/7 activity reaching 49.5% at 48 hours and rising further to 75% after 72 hours of treatment.
- Cell cytotoxicity exceeded 96% at 72 hours, indicating substantial cell death.
- Correspondingly, cell viability decreased significantly by 72 hours.
- Matrix Metalloproteinase Levels:
- MMP-1 concentration decreased by 48.9% at 24 hours post-treatment with CBD.
- MMP-2 concentration was reduced by 84% as early as 6 hours post-treatment.
- MMP-9 levels were decreased by 37.2% at 6 hours and 45.3% at 48 hours after treatment with 20 μM CBD.
- Cell Invasiveness:
- Although CBD treatment led to decreases in invasiveness ranging from 34% to 59% after 24 hours, these reductions were not statistically significant.
Conclusions and Implications
- CBD exhibits cytotoxic and pro-apoptotic effects on equine sarcoid cells, significantly reducing their viability over time.
- The observed downregulation of MMP-1, MMP-2, and MMP-9 suggests that CBD influences extracellular matrix remodeling mechanisms, which could impact tumor progression and invasion capacity.
- Despite non-significant reductions in invasiveness in vitro, the trend suggests potential modulatory effects of CBD on tumor cell behavior related to metastasis.
- Overall, the findings support further research into CBD as a possible therapeutic agent for managing equine sarcoids, particularly by targeting the tumor microenvironment and matrix interactions.
- Further studies could explore optimization of dosing, mechanisms of action, and in vivo efficacy and safety.
Cite This Article
APA
Semik-Gurgul E, Ocłoń E, Zubel-Łojek J, Pędziwiatr R, Pawlina-Tyszko K.
(2026).
Cannabidiol-induced cellular and matrix-associated responses in primary equine sarcoid cells.
J Vet Intern Med, 40(1), aalaf015.
https://doi.org/10.1093/jvimsj/aalaf015 Publication
Researcher Affiliations
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
- Department of Infectious Diseases and Public Health Protection, University of Agriculture in Krakow, 1C Redzina Street, 30-248 Krakow, Poland.
- Laboratory of Recombinant Proteins Production, Faculty of Veterinary Medicine University of Agriculture in Krakow, 1C Redzina Street, 30-248 Krakow, Poland.
- Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Mickiewicza 24/28 Street, 30-059 Krakow, Poland.
- Department of Diagnostics and Clinical Sciences, Faculty of Veterinary Medicine, University of Agriculture in Krakow, Mickiewicza 24/28 Street, 30-059 Kraków, Poland.
- Equine Vet Clinic EQUI-VET, Proszowice 55A, 32-100 Stogniowice, Poland.
- Department of Animal Molecular Biology, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland.
MeSH Terms
- Animals
- Horses
- Cannabidiol / pharmacology
- Cannabidiol / therapeutic use
- Horse Diseases / drug therapy
- Horse Diseases / pathology
- Apoptosis / drug effects
- Cell Survival / drug effects
- Skin Neoplasms / veterinary
- Skin Neoplasms / drug therapy
- Skin Neoplasms / pathology
- Cell Line, Tumor
- Sarcoidosis / veterinary
- Sarcoidosis / drug therapy
- Matrix Metalloproteinases / metabolism
Conflict of Interest Statement
The authors declare no conflicts of interest.
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