Encapsulation of Hemp (Cannabis sativa L.) Essential Oils into Nanoemulsions for Potential Therapeutic Applications: Assessment of Cytotoxicological Profiles
Abstract
:1. Introduction
2. Results
2.1. EO Chemical Composition
2.2. Characterization and Physical Stability over Time of C. sativa EO-Based NEs
2.3. Evaluation of C. sativa EO-Based NEs’ Safety Profile in Human Cell Lines
2.4. Evaluation of C. sativa EO-Based NEs’ Effect on Inflammatory Condition of Human Cell Lines
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Microwave-Assisted Extraction (MAE)
4.3. EO GC-MS Characterization
4.4. C. sativa EO-Based Nanoemulsions (NEs) Formulation and Characterization
4.5. Cell Lines
4.6. Cell Viability Assay
4.7. RNA Isolation, Reverse Transcription and Quantitative Real-Time PCR, and TaqMan Array
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
- Fike, J. Industrial Hemp: Renewed Opportunities for an Ancient Crop. Crit. Rev. Plant Sci. 2017, 35, 406–424. [Google Scholar] [CrossRef]
- Zheljazkov, V.D.; Sikora, V.; Dincheva, I.; Kačániová, M.; Astatkie, T.; Semerdjieva, I.B.; Latkovic, D. Industrial, CBD, and Wild Hemp: How Different Are Their Essential Oil Profile and Antimicrobial Activity? Molecules 2020, 25, 4631. [Google Scholar] [CrossRef] [PubMed]
- Pieracci, Y.; Ascrizzi, R.; Terreni, V.; Pistelli, L.; Flamini, G.; Bassolino, L.; Fulvio, F.; Montanari, M.; Paris, R. Essential Oil of Cannabis sativa L.: Comparison of Yield and Chemical Composition of 11 Hemp Genotypes. Molecules 2021, 26, 4080. [Google Scholar] [CrossRef] [PubMed]
- Zengin, G.; Menghini, L.; Di Sotto, A.; Mancinelli, R.; Sisto, F.; Carradori, S.; Cesa, S.; Fraschetti, C.; Filippi, A.; Angiolella, L.; et al. Chromatographic Analyses, In Vitro Biological Activities, and Cytotoxicity of Cannabis sativa L. Essential Oil: A Multidisciplinary Study. Molecules 2018, 23, 3266. [Google Scholar] [CrossRef]
- Benelli, G.; Pavela, R.; Lupidi, G.; Nabissi, M.; Petrelli, R.; Ngahang Kamte, S.L.; Cappellacci, L.; Fiorini, D.; Sut, S.; Dall’Acqua, S.; et al. The crop-residue of fiber hemp cv. Futura 75: From a waste product to a source of botanical insecticides. Environ. Sci. Pollut. Res. 2018, 25, 10515–10525. [Google Scholar] [CrossRef]
- Benelli, G.; Pavela, R.; Petrelli, R.; Cappellacci, L.; Santini, G.; Fiorini, D.; Sut, S.; Dall’Acqua, S.; Canale, A.; Maggi, F. The essential oil from industrial hemp (Cannabis sativa L.) by-products as an effective tool for insect pest management in organic crops. Ind. Crops Prod. 2018, 122, 308–315. [Google Scholar] [CrossRef]
- Tabari, M.A.; Khodashenas, A.; Jafari, M.; Petrelli, R.; Cappellacci, L.; Nabissi, M.; Maggi, F.; Pavela, R.; Youssefi, M.R. Acaricidal properties of hemp (Cannabis sativa L.) essential oil against Dermanyssus gallinae and Hyalomma dromedarii. Ind. Crops Prod. 2020, 147, 112238. [Google Scholar] [CrossRef]
- Pagano, S.; Coniglio, M.; Valenti, C.; Federici, M.I.; Lombardo, G.; Cianetti, S.; Marinucci, L. Biological effects of Cannabidiol on normal human healthy cell populations: Systematic review of the literature. Biomed. Pharmacother. 2020, 132, 110728. [Google Scholar] [CrossRef]
- Cerino, P.; Buonerba, C.; Cannazza, G.; D’Auria, J.; Ottoni, E.; Fulgione, A.; Di Stasio, A.; Pierri, B.; Gallo, A. A Review of Hemp as Food and Nutritional Supplement. Cannabis Cannabinoid Res. 2021, 6, 19. [Google Scholar] [CrossRef]
- Petrovici, A.R.; Simionescu, N.; Sandu, A.I.; Paraschiv, V.; Silion, M.; Pinteala, M. New Insights on Hemp Oil Enriched in Cannabidiol: Decarboxylation, Antioxidant Properties and In Vitro Anticancer Effect. Antioxidants 2021, 10, 738. [Google Scholar] [CrossRef]
- Mikulcová, V.; Kašpárková, V.; Humpolíček, P.; Buňková, L. Formulation, Characterization and Properties of Hemp Seed Oil and Its Emulsions. Molecules 2017, 22, 700. [Google Scholar] [CrossRef] [PubMed]
- Martins, A.M.; Gomes, A.L.; Boas, I.V.; Marto, J.; Ribeiro, H.M. Cannabis-Based Products for the Treatment of Skin Inflammatory Diseases: A Timely Review. Pharmaceuticals 2022, 15, 210. [Google Scholar] [CrossRef] [PubMed]
- Baswan, S.M.; Klosner, A.E.; Glynn, K.; Rajgopal, A.; Malik, K.; Yim, S.; Stern, N. Therapeutic Potential of Cannabidiol (CBD) for Skin Health and Disorders. Clin. Cosmet. Investig. Dermatol. 2020, 13, 927. [Google Scholar] [CrossRef] [PubMed]
- Baroni, A.; Buommino, E.; De Gregorio, V.; Ruocco, E.; Ruocco, V.; Wolf, R. Structure and function of the epidermis related to barrier properties. Clin. Dermatol. 2012, 30, 257–262. [Google Scholar] [CrossRef]
- Hänel, K.H.; Cornelissen, C.; Lüscher, B.; Baron, J.M. Cytokines and the skin barrier. Int. J. Mol. Sci. 2013, 14, 6720–6745. [Google Scholar] [CrossRef]
- Pastar, I.; Stojadinovic, O.; Yin, N.C.; Ramirez, H.; Nusbaum, A.G.; Sawaya, A.; Patel, S.B.; Khalid, L.; Isseroff, R.R.; Tomic-Canic, M. Epithelialization in Wound Healing: A Comprehensive Review. Adv. Wound Care 2014, 3, 445–464. [Google Scholar] [CrossRef] [PubMed]
- Kowalczyk, A.; Przychodna, M.; Sopata, S.; Bodalska, A.; Fecka, I. Thymol and Thyme Essential Oil-New Insights into Selected Therapeutic Applications. Molecules 2020, 25, 4125. [Google Scholar] [CrossRef]
- Alotaibi, B.; Negm, W.A.; Elekhnawy, E.; El-Masry, T.A.; Elseady, W.S.; Saleh, A.; Alotaibi, K.N.; El-Sherbeni, S.A. Antibacterial, Immunomodulatory, and Lung Protective Effects of Boswelliadalzielii Oleoresin Ethanol Extract in Pulmonary Diseases: In Vitro and In Vivo Studies. Antibiotics 2021, 10, 1444. [Google Scholar] [CrossRef]
- Mamber, S.W.; Gurel, V.; Lins, J.; Ferri, F.; Beseme, S.; McMichael, J. Effects of cannabis oil extract on immune response gene expression in human small airway epithelial cells (HSAEpC): Implications for chronic obstructive pulmonary disease (COPD). J. Cannabis Res. 2020, 2, 5. [Google Scholar] [CrossRef]
- Aswad, M.; Hamza, H.; Pechkovsky, A.; Zikrach, A.; Popov, T.; Zohar, Y.; Shahar, E.; Louria-Hayon, I. High-CBD Extract (CBD-X) Downregulates Cytokine Storm Systemically and Locally in Inflamed Lungs. Front. Immunol. 2022, 13, 877546. [Google Scholar] [CrossRef]
- Rossi, P.; Cappelli, A.; Marinelli, O.; Valzano, M.; Pavoni, L.; Bonacucina, G.; Petrelli, R.; Pompei, P.; Mazzara, E.; Ricci, I.; et al. Mosquitocidal and Anti-Inflammatory Properties of The Essential Oils Obtained from Monoecious, Male, and Female Inflorescences of Hemp (Cannabis sativa L.) and Their Encapsulation in Nanoemulsions. Molecules 2020, 25, 3451. [Google Scholar] [CrossRef]
- Pavoni, L.; Perinelli, D.R.; Ciacciarelli, A.; Quassinti, L.; Bramucci, M.; Miano, A.; Casettari, L.; Cespi, M.; Bonacucina, G.; Palmieri, G.F. Properties and stability of nanoemulsions: How relevant is the type of surfactant? J. Drug Deliv. Sci. Technol. 2020, 58, 101772. [Google Scholar] [CrossRef]
- Hanuš, L.O.; Hod, Y. Terpenes/Terpenoids in Cannabis: Are They Important? Med. Cannabis Cannabinoids 2020, 3, 25–60. [Google Scholar] [CrossRef] [PubMed]
- Fiorini, D.; Molle, A.; Nabissi, M.; Santini, G.; Benelli, G.; Maggi, F. Valorizing industrial hemp (Cannabis sativa L.) by-products: Cannabidiol enrichment in the inflorescence essential oil optimizing sample pre-treatment prior to distillation. Ind. Crops Prod. 2019, 128, 581–589. [Google Scholar] [CrossRef]
- Ascrizzi, R.; Iannone, M.; Cinque, G.; Marianelli, A.; Pistelli, L.; Flamini, G. “Hemping” the drinks: Aromatizing alcoholic beverages with a blend of Cannabis sativa L. flowers. Food Chem. 2020, 325, 126909. [Google Scholar] [CrossRef]
- Pavela, R.; Pavoni, L.; Bonacucina, G.; Cespi, M.; Cappellacci, L.; Petrelli, R.; Spinozzi, E.; Aguzzi, C.; Zeppa, L.; Ubaldi, M.; et al. Encapsulation of Carlina acaulis essential oil and carlina oxide to develop long-lasting mosquito larvicides: Microemulsions versus nanoemulsions. J. Pest Sci. 2021, 94, 899–915. [Google Scholar] [CrossRef]
- Mazzara, E.; Torresi, J.; Fico, G.; Papini, A.; Kulbaka, N.; Dall’acqua, S.; Sut, S.; Garzoli, S.; Mustafa, A.M.; Cappellacci, L.; et al. A Comprehensive Phytochemical Analysis of Terpenes, Polyphenols and Cannabinoids, and Micromorphological Characterization of 9 Commercial Varieties of Cannabis sativa L. Plants 2022, 11, 891. [Google Scholar] [CrossRef]
- Sparkman, O.D. Identification of essential oil components by gas chromatography/mass spectrometry Robert P. Adams. J. Am. Soc. Mass Spectrom. 2005, 16, 11. [Google Scholar] [CrossRef]
- NIST 17 Mass Spectral Library (NIST/EPA/NIH); National Institute of Standards and Technology: Gaithersburg, MD, USA, 2017.
- Mondello, L. FFNSC 3 Mass Spectra of Flavors and Fragrances of Natural and Synthetic Compounds, 3rd ed.; John Wiley & Sons, Inc.: Hoboken, NJ, USA, 2015; ISBN 978-1-119-06984-3. [Google Scholar]
- Procaccia, S.; Lewitus, G.M.; Lipson Feder, C.; Shapira, A.; Berman, P.; Meiri, D. Cannabis for Medical Use: Versatile Plant Rather Than a Single Drug. Front. Pharmacol. 2022, 13, 1308. [Google Scholar] [CrossRef]
- Bukke, V.N.; Archana, M.; Villani, R.; Serviddio, G.; Cassano, T. Pharmacological and Toxicological Effects of Phytocannabinoids and Recreational Synthetic Cannabinoids: Increasing Risk of Public Health. Pharmaceuticals 2021, 14, 965. [Google Scholar] [CrossRef]
Component a | RI Calc. b | RI Lit c | % Uso 31 | % Carmagnola CS |
---|---|---|---|---|
α-pinene | 926 | 932 | 11.07 | 13.45 |
β-pinene | 968 | 974 | 3.59 | 5.45 |
myrcene | 989 | 988 | 15.28 | 37.57 |
limonene | 1025 | 1024 | 1.59 | 5.37 |
(E)-β-ocimene | 1047 | 1044 | 6.71 | 1.79 |
terpinolene | 1085 | 1086 | 5.46 | 2.97 |
(E)-caryophyllene | 1409 | 1417 | 25.93 | 16.99 |
α-humulene | 1443 | 1452 | 8.92 | 6.11 |
caryophyllene oxide | 1571 | 1582 | 7.23 | 2.95 |
Total identified (%) | 99.03 | 97.72 | ||
Monoterpene hydrocarbons (%) | 44.24 | 67.27 | ||
Oxygenated monoterpenes (%) | 0.43 | |||
Sesquiterpene hydrocarbons (%) | 45.31 | 26.18 | ||
Oxygenated sesquiterpenes (%) | 9.04 | 3.58 | ||
Cannabinoids (%) | 0.43 | 0.23 |
IC50 mg mL−1 | |||
---|---|---|---|
HaCaT | NHF A12 | HBEpC | |
Carmagnola CS EO | 0.052 ± 0.002 | 0.119 ± 0.009 | 0.034 ± 0.002 |
Carmagnola CS NE | 0.354 ± 0.018 (0.021 ± 0.001) | 0.448 ± 0.020 (0.023 ± 0.001) | 0.154 ± 0.010 (0.009 ± 0.001) |
CTRL NE | 2.606 ± 0.050 | 9.666 ± 0.600 | 1.221 ± 0.500 |
Uso 31 EO | 0.049 ± 0.002 | 0.103 ± 0.008 | 0.041 ± 0.003 |
Uso 31 NE | 0.857 ± 0.030 (0.043 ± 0.002) | 1.505 ± 0.058 (0.075 ± 0.003) | 0.245 ± 0.010 (0.012 ± 0.001) |
CTRL NE | 5.388 ± 0.100 | >10.04 | 3.255 ± 0.090 |
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Aguzzi, C.; Perinelli, D.R.; Cespi, M.; Zeppa, L.; Mazzara, E.; Maggi, F.; Petrelli, R.; Bonacucina, G.; Nabissi, M. Encapsulation of Hemp (Cannabis sativa L.) Essential Oils into Nanoemulsions for Potential Therapeutic Applications: Assessment of Cytotoxicological Profiles. Molecules 2023, 28, 6479. https://doi.org/10.3390/molecules28186479
Aguzzi C, Perinelli DR, Cespi M, Zeppa L, Mazzara E, Maggi F, Petrelli R, Bonacucina G, Nabissi M. Encapsulation of Hemp (Cannabis sativa L.) Essential Oils into Nanoemulsions for Potential Therapeutic Applications: Assessment of Cytotoxicological Profiles. Molecules. 2023; 28(18):6479. https://doi.org/10.3390/molecules28186479
Chicago/Turabian StyleAguzzi, Cristina, Diego Romano Perinelli, Marco Cespi, Laura Zeppa, Eugenia Mazzara, Filippo Maggi, Riccardo Petrelli, Giulia Bonacucina, and Massimo Nabissi. 2023. "Encapsulation of Hemp (Cannabis sativa L.) Essential Oils into Nanoemulsions for Potential Therapeutic Applications: Assessment of Cytotoxicological Profiles" Molecules 28, no. 18: 6479. https://doi.org/10.3390/molecules28186479