{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T03:04:33Z","timestamp":1776135873158,"version":"3.50.1"},"reference-count":32,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,4,28]],"date-time":"2025-04-28T00:00:00Z","timestamp":1745798400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"MIUR"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background:\u00a0Cannabis sativa L. (C. sativa) has a long history of medicinal use. Its inflorescences contain bioactive compounds like non-psychotropic cannabidiol (CBD), which is well known for its anti-inflammatory potential in skin conditions such as psoriasis, and psychotropic \u0394-9-tetrahydrocannabinol (THC). Keratinocytes, the main cells in the epidermis, are crucial for regulating skin inflammation by producing mediators like IL-8 when stimulated by agents like TNF\u03b1. Methods: This study explores the anti-inflammatory effects of a standardized C. sativa extract (CSE) with 5% CBD and less than 0.2% THC in human keratinocytes challenged by TNF\u03b1. The aim of this study is to analyze the specific contributions of the main constituents of CSE to inflammatory responses in human keratinocytes by fractionating the extract and examining the effects of its individual components. Results: MTT assays showed that CSE was non-toxic to HaCaT cells up to 50 \u03bcg\/mL. CSE inhibited NF-\u03baB activity and reduced IL-8 secretion in a concentration-dependent manner, with mean IC50 values of 28.94 \u00b1 10.40 \u03bcg\/mL and 20.06 \u00b1 2.78 \u03bcg\/mL (mean \u00b1 SEM), respectively. Fractionation of CSE into four subfractions revealed that the more lipophilic fractions (A and B) were the most effective in inhibiting NF-\u03baB, indicating that cannabinoids and cannflavins are key contributors. Pure CBD is one of the most active cannabinoids in reducing NF-\u03baB-driven transcription (together with THC and cannabigerol), and due to its abundance in CSE, it is primarily responsible for the anti-inflammatory activity. Conclusions: This study highlights CBD\u2019s significant role in reducing inflammation in human keratinocytes and underscores the need to consider the synergistic interactions of several molecules within C. sativa extracts for maximum efficacy. Standardized extracts are essential for reproducible results due to the variability in responses.<\/jats:p>","DOI":"10.3390\/ph18050647","type":"journal-article","created":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T11:35:13Z","timestamp":1746185713000},"page":"647","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Beyond Cannabidiol: The Contribution of Cannabis sativa Phytocomplex to Skin Anti-Inflammatory Activity in Human Skin Keratinocytes"],"prefix":"10.3390","volume":"18","author":[{"given":"Marco","family":"Fumagalli","sequence":"first","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0742-0521","authenticated-orcid":false,"given":"Giulia","family":"Martinelli","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giuseppe","family":"Paladino","sequence":"additional","affiliation":[{"name":"Linnea SA, 6595 Riazzino, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nora","family":"Rossini","sequence":"additional","affiliation":[{"name":"Linnea SA, 6595 Riazzino, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Umberto","family":"Ciriello","sequence":"additional","affiliation":[{"name":"Linnea SA, 6595 Riazzino, Switzerland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5752-1758","authenticated-orcid":false,"given":"Vincenzo","family":"Nicolaci","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-8416-1307","authenticated-orcid":false,"given":"Nicole","family":"Maranta","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2107-3033","authenticated-orcid":false,"given":"Carola","family":"Pozzoli","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Safwa Moheb","family":"El Haddad","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-0074-5770","authenticated-orcid":false,"given":"Elisa","family":"Sonzogni","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5378-402X","authenticated-orcid":false,"given":"Mario","family":"Dell\u2019Agli","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2470-6038","authenticated-orcid":false,"given":"Stefano","family":"Piazza","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2811-8628","authenticated-orcid":false,"given":"Enrico","family":"Sangiovanni","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences \u201cRodolfo Paoletti\u201d, Universit\u00e0 degli Studi di Milano, 20133 Milan, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.jep.2018.09.004","article-title":"Cannabis sativa: A comprehensive ethnopharmacological review of a medicinal plant with a long history","volume":"227","author":"Bonini","year":"2018","journal-title":"J. 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