{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T07:26:16Z","timestamp":1783581976659,"version":"3.55.0"},"reference-count":50,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T00:00:00Z","timestamp":1763510400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004704","name":"National Research Council of Thailand","doi-asserted-by":"publisher","award":["N41A650171"],"award-info":[{"award-number":["N41A650171"]}],"id":[{"id":"10.13039\/501100004704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002873","name":"Chulalongkorn University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100002873","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sore throat, commonly associated with pharyngitis and tonsillitis, is primarily caused by bacterial pathogens. Conventional therapies rely on antibiotics and anti-inflammatory drugs; however, concerns about adverse effects, antibiotic resistance, and drug interactions have encouraged the search for alternative remedies. Cannabis sativa L. (CS) has demonstrated potential in relieving sore throat and inflammation, while propolis, a bee-derived natural product, exhibits notable antibacterial and anti-inflammatory activities. This study aimed to investigate the enhanced antibacterial and anti-inflammatory effects of combining CS and propolis extracts (PE). Results found that CS and PE exhibited antibacterial effects against Streptococcus pyogenes DMST 4369, Staphylococcus aureus ATCC 25923, and Pseudomonas aeruginosa ATCC 9027. Their combination produced additive antibacterial effects against S. pyogenes and S. aureus. Cannabidiol (CBD) was identified as an active antibacterial constituent against S. pyogenes. Additionally, the PE-CBD in CS solution combination at concentration 625:0.125 \u00b5g\/mL significantly reduced NO production and suppressed proinflammatory cytokines in LPS-stimulated macrophages. This study highlights the enhanced antibacterial and anti-inflammatory potential of the C. sativa and propolis combination, emphasizing the need to verify synergistic effects and determine the appropriate ratio for rational product development. Further research is needed to investigate the underlying mechanisms of action, particularly the anti-inflammatory pathways, in animal models. In addition, studies on hepatotoxicity should be conducted to ensure safety.<\/jats:p>","DOI":"10.3390\/ijms262211181","type":"journal-article","created":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T12:55:31Z","timestamp":1763556931000},"page":"11181","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Enhanced Antibacterial and Anti-Inflammatory Activities of the Combination of Cannabis sativa and Propolis Extracts: An In Vitro Study"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-5203-7187","authenticated-orcid":false,"given":"Naruemon","family":"Perstwong","sequence":"first","affiliation":[{"name":"Graduate Program in Pharmaceutical Science and Technology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wongsakorn","family":"Phongsopitanun","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3582-5980","authenticated-orcid":false,"given":"Visarut","family":"Buranasudja","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1705-6217","authenticated-orcid":false,"given":"Sornkanok","family":"Vimolmangkang","sequence":"additional","affiliation":[{"name":"Research Cluster for Cannabis and Its Natural Substances, Chulalongkorn University, Bangkok 10330, Thailand"},{"name":"Center of Excellence in Plant-Produced Pharmaceuticals, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"},{"name":"Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S55","DOI":"10.1097\/00006454-199808001-00002","article-title":"Microbiology of bacterial respiratory infections","volume":"17","author":"Cappelletty","year":"1998","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_2","unstructured":"Ghlichloo, I., and Gerriets, V. (2019). Nonsteroidal Anti-Inflammatory Drugs (NSAIDs), StatPearls Publishing."},{"key":"ref_3","unstructured":"Department of Thai Traditional and Alternative Medicine, Ministry of Public Health of Thailand (2024). Thai Herbal Medicine in Thailand National List of Essential Medicines Replacing Modern Medicine, Ministry of Public Health of Thailand."},{"key":"ref_4","unstructured":"Herbal Product Division (2023). National List of Essential Medicines for Herbs 2023 Manual, Department of Food and Drug Administration (FDA), Ministry of Public Health."},{"key":"ref_5","unstructured":"Maneekul, U., and Samranjit, K. (1998). Textbook of General Thai Traditional Medicine, Branch of Thai Pharmacy, Division of Healing Arts Department of Health Service Support, Thai Phum Publishing. [2nd ed.]."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Odieka, A.E., Obuzor, G.U., Oyedeji, O.O., Gondwe, M., Hosu, Y.S., and Oyedeji, A.O. (2022). The medicinal natural products of Cannabis sativa Linn.: A review. Molecules, 27.","DOI":"10.3390\/molecules27051689"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1111\/j.1365-2044.2004.03674.x","article-title":"Initial experiences with medicinal extracts of cannabis for chronic pain: Results from 34 \u2018N of 1\u2019studies","volume":"59","author":"Notcutt","year":"2004","journal-title":"Anaesthesia"},{"key":"ref_8","first-page":"219","article-title":"The effectiveness of cannabis used in Thai Traditional way on the palliative care patients in Thailand","volume":"26","author":"Narongvit","year":"2021","journal-title":"J. Assoc. Res."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Mlost, J., Bryk, M., and Starowicz, K. (2020). Cannabidiol for pain treatment: Focus on pharmacology and mechanism of action. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21228870"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1055\/a-1581-3707","article-title":"Activity of THC, CBD, and CBN on human ACE2 and SARS-COV1\/2 main protease to understand antiviral defense mechanism","volume":"88","author":"Pitakbut","year":"2022","journal-title":"Planta Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"720","DOI":"10.56808\/3027-7922.2658","article-title":"Vegetable Oil-Based Cannabis: Its Cannabinoid Profiling and Photoprotective Effect on UVA-Irradiated Human Skin Keratinocytes","volume":"46","author":"Kobtrakul","year":"2023","journal-title":"Thai J. Pharm. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.4314\/tjpr.v16i7.13","article-title":"Anti-inflammatory effect of methanol extracts of hemp leaf in IL-1\u03b2-induced synovitis","volume":"16","author":"Duangnin","year":"2017","journal-title":"Trop. J. Pharm. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Niyangoda, D., Aung, M.L., Qader, M., Tesfaye, W., Bushell, M., Chiong, F., Tsai, D., Ahmad, D., Samarawickrema, I., and Sinnollareddy, M. (2024). Cannabinoids as Antibacterial Agents: A Systematic and Critical Review of In Vitro Efficacy Against Streptococcus and Staphylococcus. Antibiotics, 13.","DOI":"10.3390\/antibiotics13111023"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.fitote.2016.05.008","article-title":"Anti-inflammatory and antioxidant effects of a combination of cannabidiol and moringin in LPS-stimulated macrophages","volume":"112","author":"Rajan","year":"2016","journal-title":"Fitoterapia"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Ayad, A.S., Benchaabane, S., Daas, T., Smagghe, G., and Loucif-Ayad, W. (2025). Propolis Stands out as a Multifaceted Natural Product: Meta-Analysis on Its Sources, Bioactivities, Applications, and Future Perspectives. Life, 15.","DOI":"10.3390\/life15050764"},{"key":"ref_16","first-page":"206439","article-title":"Propolis: A Complex Natural Product with a Plethora of Biological Activities That Can Be Explored for Drug Development","volume":"2015","author":"Baltazar","year":"2015","journal-title":"Evid. Based Complement. Altern. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11065","DOI":"10.1021\/jf303004n","article-title":"Ethanol extract of Brazilian red propolis induces apoptosis in human breast cancer MCF-7 cells through endoplasmic reticulum stress","volume":"60","author":"Kamiya","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"12403","DOI":"10.1021\/jf3042775","article-title":"Identification and quantification of phytochemical composition and anti-inflammatory and radical scavenging properties of methanolic extracts of chinese propolis","volume":"60","author":"Shi","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11765","DOI":"10.1021\/jf303516w","article-title":"Solophenols b\u2013d and solomonin: New prenylated polyphenols isolated from propolis collected from the Solomon islands and their antibacterial activity","volume":"60","author":"Inui","year":"2012","journal-title":"J. Agric. Food Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"793","DOI":"10.7150\/ijms.4722","article-title":"Synergistic effects of honey and propolis toward drug multi-resistant Staphylococcus aureus, Escherichia coli and Candida albicans isolates in single and polymicrobial cultures","volume":"9","author":"Ansari","year":"2012","journal-title":"Int. J. Med. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2175","DOI":"10.1007\/s00217-023-04284-z","article-title":"Palynological, chemical, antimicrobial, and enzyme inhibition properties of Cannabis sativa L. propolis","volume":"249","author":"Keskin","year":"2023","journal-title":"Eur. Food Res. Technol."},{"key":"ref_22","first-page":"1086","article-title":"HPTLC\u2019an important tool in standardization of herbal medical product: A review","volume":"2","author":"Shivatare","year":"2013","journal-title":"J. Sci. Innov. Res."},{"key":"ref_23","unstructured":"Department of Medical Sciences, Ministry of Public Health (2020). Cannabis Extract (TP Monograph). Thai Pharmacopoeia II SUPPLEMENT 2020, The Agricultural Co-operative Federation of Thailand, Ltd.. Part 1."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1093\/chromsci\/bmw024","article-title":"TLC fingerprinting and pattern recognition methods in the assessment of authenticity of poplar-type propolis","volume":"54","author":"Vovk","year":"2016","journal-title":"J. Chromatogr. Sci."},{"key":"ref_25","unstructured":"United Nations Office on Drugs and Crime (2009). Recommended Methods for the Identification and Analysis of Cannabis and Cannabis Products: Manual for Use by National Drug Testing Laboratories."},{"key":"ref_26","unstructured":"The International Association for the Advancement of High Performance Thin Layer Chromatography (HPTLC Association) (2017). Cannabis (Cannabis sativa)\u2014Normal Phase Separation, The International Association for the Advancement of HPTLC."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"EUCAST (2000). Terminology relating to methods for the determination of susceptibility of bacteria to antimicrobial agents. Clin. Microbiol. Infect., 6, 503\u2013508.","DOI":"10.1046\/j.1469-0691.2000.00149.x"},{"key":"ref_28","unstructured":"Department of Medical Sciences, Ministry of Public Health (2021). KANCHA, RUEAN CHO DOK PHET MIA (Cannabis sativa L.). Thai Herbal Pharmacopoeia, The Agricultural Co-operative Federation of Thailand, Ltd.. The Thai Herbal Pharmacopoeia Committee."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Ristivojevi\u0107, P., Dimki\u0107, I., Trifkovi\u0107, J., Beri\u0107, T., Vovk, I., Milojkovi\u0107-Opsenica, D., and Stankovi\u0107, S. (2016). Antimicrobial Activity of Serbian Propolis Evaluated by Means of MIC, HPTLC, Bioautography and Chemometrics. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0157097"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"12826","DOI":"10.15835\/nbha51212826","article-title":"Comparative study on the antimicrobial activity of propolis, catechin, quercetin and gallic acid","volume":"51","author":"Borozan","year":"2023","journal-title":"Not. Bot. Horti Agrobot. Cluj-Napoca"},{"key":"ref_31","unstructured":"Department of Medical Sciences, Ministry of Public Health (2021). KOT PHUNG PLA (Terminalia chebula Retz.). Thai Herbal Pharmacopoeia, The Agricultural Co-operative Federation of Thailand, Ltd.. The Thai Herbal Pharmacopoeia Committee."},{"key":"ref_32","unstructured":"Department of Medical Sciences, Ministry of Public Health (2021). MAKHAM POM (Phyllanthus emblica L.). Thai Herbal Pharmacopoeia, The Agricultural Co-operative Federation of Thailand, Ltd.. The Thai Herbal Pharmacopoeia Committee."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Achenbach, J., Deyerling, N., Mello dos Santos, M., Sultana, S., Islam, M.K., and Locher, C. (2024). Physicochemical Properties, Antioxidant Activity, and High-Performance Thin-Layer Chromatography Profiling of Propolis Samples from Western Australia. Plants, 13.","DOI":"10.3390\/plants13141919"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e28621","DOI":"10.1016\/j.heliyon.2024.e28621","article-title":"Insight into the influence of natural deep eutectic solvents on the extraction of phenolic compounds from poplar type propolis: Composition and in vitro biological activity","volume":"10","author":"Trusheva","year":"2024","journal-title":"Heliyon"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"134801","DOI":"10.1016\/j.foodchem.2022.134801","article-title":"Almond gum-sodium caseinate complexes for loading propolis extract: Characterization, antibacterial activity, release, and in-vitro cytotoxicity","volume":"405","author":"Salehi","year":"2023","journal-title":"Food Chem."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Joki\u0107, S., Jerkovi\u0107, I., Pavi\u0107, V., Aladi\u0107, K., Molnar, M., Kova\u010d, M.J., and Vladimir-Kne\u017eevi\u0107, S. (2022). Terpenes and Cannabinoids in Supercritical CO2 Extracts of Industrial Hemp Inflorescences: Optimization of Extraction, Antiradical and Antibacterial Activity. Pharmaceuticals, 15.","DOI":"10.3390\/ph15091117"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1016\/j.fct.2019.01.035","article-title":"Multiple pharmacognostic characterization on hemp commercial cultivars: Focus on inflorescence water extract activity","volume":"125","author":"Ferrante","year":"2019","journal-title":"Food Chem. Toxicol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Martinenghi, L.D., J\u00f8nsson, R., Lund, T., and Jenssen, H. (2020). Isolation, Purification, and Antimicrobial Characterization of Cannabidiolic Acid and Cannabidiol from Cannabis sativa L. Biomolecules, 10.","DOI":"10.3390\/biom10060900"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Blaskovich, M.A.T., Kavanagh, A.M., Elliott, A.G., Zhang, B., Ramu, S., Amado, M., Lowe, G.J., Hinton, A.O., Pham, D.M.T., and Zuegg, J. (2021). The antimicrobial potential of cannabidiol. Commun. Biol., 4.","DOI":"10.1038\/s42003-020-01530-y"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.phytol.2014.08.017","article-title":"HPTLC-MS as an efficient hyphenated technique for the rapid identification of antimicrobial compounds from propolis","volume":"11","author":"Kasote","year":"2015","journal-title":"Phytochem. Lett."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Kongkadee, K., Wisuitiprot, W., Ingkaninan, K., and Waranuch, N. (2022). Anti-inflammation and gingival wound healing activities of Cannabis sativa L. subsp. sativa (hemp) extract and cannabidiol: An in vitro study. Arch. Oral Biol., 140.","DOI":"10.1016\/j.archoralbio.2022.105464"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Zhang, W., Cai, Y., Chen, X., Ji, T., and Sun, L. (2020). Optimized extraction based on the terpenoids of Heterotrigona itama propolis and their antioxidative and anti-inflammatory activities. J. Food Biochem., 44.","DOI":"10.1111\/jfbc.13296"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Tristao Santini, A., Otac\u00edlio Pinto, R.A., Goldoni Lazarini, J., Vieira de Morais, D., Alves de Piloto Fernandes, A.M., Franchin, M., Lunardelli Negreiros de Carvalho, P., Girotto Pressete, C., Rosalen, P.L., and Matias de Alencar, S. (2024). Bioactives of Melipona rufiventris propolis: Exploring its antimicrobial, anti-inflammatory, and antioxidant activities. Chem. Biodivers., 21.","DOI":"10.1002\/cbdv.202302084"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Zulhendri, F., Lesmana, R., Tandean, S., Christoper, A., Chandrasekaran, K., Irsyam, I., Suwantika, A.A., Abdulah, R., and Wathoni, N. (2022). Recent Update on the Anti-Inflammatory Activities of Propolis. Molecules, 27.","DOI":"10.3390\/molecules27238473"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1002\/cem.2592","article-title":"Pattern recognition methods and multivariate image analysis in HPTLC fingerprinting of propolis extracts","volume":"28","author":"Vovk","year":"2014","journal-title":"J. Chemom."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e01864-01819","DOI":"10.1128\/JCM.01864-19","article-title":"The Clinical and Laboratory Standards Institute Subcommittee on Antimicrobial Susceptibility Testing: Background, Organization, Functions, and Processes","volume":"58","author":"Weinstein","year":"2020","journal-title":"J. Clin. Microbiol."},{"key":"ref_47","unstructured":"EUCAST (2024, June 03). Determination of Minimum Inhibitory Concentrations (MICs) of Antibacterial Agents by Broth Dilution, Available online: https:\/\/www.clinicalmicrobiologyandinfection.org\/article\/S1198-743X(14)64063-5\/fulltext."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Buranasudja, V., Vimolmangkang, S., Sanookpan, K., Binalee, A., Bao Nguyen, A., Dong Thach, U., Hang Do, B., Son Dang, V., Minh Do, K., and Minh Nguyen, H. (2023). Antioxidant, Anti-Skin-Aging, Anti-Inflammatory, and Anti-Acetylcholinesterase Activities of Rourea oligophlebia Extracts. Chem. Biodivers., 20.","DOI":"10.1002\/cbdv.202201096"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Perstwong, N., Binalee, A., Kobtrakul, K., Phongsopitanun, W., Sanookpan, K., Areecheewakul, S., Buranasudja, V., and Vimolmangkang, S. (2025). Qualitative analysis and exploration of anti-inflammatory and antibacterial effects of a Thai traditional medicine formula from Wat Pho beyond its use for COVID-19 treatment. BMC Complement. Med. Ther., 25.","DOI":"10.1186\/s12906-025-04873-3"},{"key":"ref_50","unstructured":"Promega Corporation (2009). Techinical Bulletin: Griess Reagent System Instructions for Use of Product G2930, Promega Corporation. Available online: https:\/\/worldwide.promega.com\/resources\/protocols\/technical bulletins\/0\/griess-reagent-system-protocol\/."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/11181\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T05:27:17Z","timestamp":1763702837000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/22\/11181"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,19]]},"references-count":50,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262211181"],"URL":"https:\/\/doi.org\/10.3390\/ijms262211181","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,19]]}}}