Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis

Objectives: The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Methods: Specific colorimetric assays were used to estimate the total phenolic and flavo...

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Main Authors: Elisabetta Miraldi, Alessandro Giordano, Giorgio Cappellucci, Federica Vaccaro, Marco Biagi, Giulia Baini
Format: Article
Language:English
Published: MDPI AG 2024-10-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/17/11/1412
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author Elisabetta Miraldi
Alessandro Giordano
Giorgio Cappellucci
Federica Vaccaro
Marco Biagi
Giulia Baini
author_facet Elisabetta Miraldi
Alessandro Giordano
Giorgio Cappellucci
Federica Vaccaro
Marco Biagi
Giulia Baini
author_sort Elisabetta Miraldi
collection DOAJ
description Objectives: The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Methods: Specific colorimetric assays were used to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of Eurasian propolis (pinocembrin, pinobanksin, CAPE, chrysin and galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Human keratinocyte, fibroblast, and monocytic cell lines were used for the biological in vitro analyses. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide-LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8, and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. Network Pharmacology analysis was employed to further explore possible mechanisms of the action of propolis on the healing process. Results: The analyses confirmed the high phenolic content of Eurasian propolis (142.50–211.30 mg GAE/g), dominated by flavonoids (95.50–196.80 mg Galangin Equivalents/g), and terpenes (431.50–650.00 mg β-sitosterol Equivalents/g), while also verifying the significant antioxidant (4.9–8.9 mM/g Trolox Equivalents) and antiradical (DPPH IC<sub>50</sub> 26.1–54.4 μg/mL) activities. The samples showed indirect wound healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Conclusions: Eurasian propolis may not directly stimulate cell proliferation during wound healing. Its anti-inflammatory and immunomodulatory properties could indicate an indirect contribution in helping the process.
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spelling doaj-art-c50fdd020bf4487da537e0c092d7fcbc2024-11-26T18:17:03ZengMDPI AGPharmaceuticals1424-82472024-10-011711141210.3390/ph17111412Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian PropolisElisabetta Miraldi0Alessandro Giordano1Giorgio Cappellucci2Federica Vaccaro3Marco Biagi4Giulia Baini5Department of Physical Sciences, Earth and Environment, University of Siena, 53100 Siena, ItalyDepartment of Physical Sciences, Earth and Environment, University of Siena, 53100 Siena, ItalyDepartment of Physical Sciences, Earth and Environment, University of Siena, 53100 Siena, ItalyDepartment of Physical Sciences, Earth and Environment, University of Siena, 53100 Siena, ItalyDepartment of Food and Drug, University of Parma, 43124 Parma, ItalyDepartment of Physical Sciences, Earth and Environment, University of Siena, 53100 Siena, ItalyObjectives: The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Methods: Specific colorimetric assays were used to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of Eurasian propolis (pinocembrin, pinobanksin, CAPE, chrysin and galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Human keratinocyte, fibroblast, and monocytic cell lines were used for the biological in vitro analyses. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide-LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8, and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. Network Pharmacology analysis was employed to further explore possible mechanisms of the action of propolis on the healing process. Results: The analyses confirmed the high phenolic content of Eurasian propolis (142.50–211.30 mg GAE/g), dominated by flavonoids (95.50–196.80 mg Galangin Equivalents/g), and terpenes (431.50–650.00 mg β-sitosterol Equivalents/g), while also verifying the significant antioxidant (4.9–8.9 mM/g Trolox Equivalents) and antiradical (DPPH IC<sub>50</sub> 26.1–54.4 μg/mL) activities. The samples showed indirect wound healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Conclusions: Eurasian propolis may not directly stimulate cell proliferation during wound healing. Its anti-inflammatory and immunomodulatory properties could indicate an indirect contribution in helping the process.https://www.mdpi.com/1424-8247/17/11/1412propolisEurasian propolischemical characteristicsanti-inflammatory propertiesimmunomodulatory properties
spellingShingle Elisabetta Miraldi
Alessandro Giordano
Giorgio Cappellucci
Federica Vaccaro
Marco Biagi
Giulia Baini
Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
Pharmaceuticals
propolis
Eurasian propolis
chemical characteristics
anti-inflammatory properties
immunomodulatory properties
title Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
title_full Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
title_fullStr Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
title_full_unstemmed Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
title_short Phytochemical Characterization and Assessment of the Wound Healing Properties of Three Eurasian Propolis
title_sort phytochemical characterization and assessment of the wound healing properties of three eurasian propolis
topic propolis
Eurasian propolis
chemical characteristics
anti-inflammatory properties
immunomodulatory properties
url https://www.mdpi.com/1424-8247/17/11/1412
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AT giorgiocappellucci phytochemicalcharacterizationandassessmentofthewoundhealingpropertiesofthreeeurasianpropolis
AT federicavaccaro phytochemicalcharacterizationandassessmentofthewoundhealingpropertiesofthreeeurasianpropolis
AT marcobiagi phytochemicalcharacterizationandassessmentofthewoundhealingpropertiesofthreeeurasianpropolis
AT giuliabaini phytochemicalcharacterizationandassessmentofthewoundhealingpropertiesofthreeeurasianpropolis