A NaDES extract of “Jardin de Granville” rose flower displays pro-resolving and epidermal strengthening properties
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Presented by: Virginie Pecher
Introduction
Inflammation is a local reaction towards a disturbance of tissue homeostasis caused by damage to tissue structure and infection. It is normally self-limited and ends with its complete resolution, which involves the production of a class of lipid mediators capable of counter-acting inflammation, namely Specialized Pro-resolving Mediators (SPMs). Derived from poly-unsaturated fatty acids, such as arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid and, their biosynthesis involves different members of the cyclooxygenase and lipoxygenase families. Increasing attention has been paid lately to skin-relevant SPMs such as Resolvin D1 or Lipoxins A4, that have been reported to alleviate skin conditions such as psoriasis or contact dermatitis. Comparing the SPM patterns in old vs. young biopsies, we previously observed an altered responsiveness to pro-inflammatory challenge by Phorbol 12-myristate 13-acetate (PMA), with a switch towards LOX-5 mediated SPM syntheses.
A natural deep eutectic solvent (NaDES) extract of Rose flowers was developed and evaluated for its ability to increase biosynthesis of SPMs both in old and young skin biopsies and to improve epidermal structure, cohesion and differentiation.
Materials and methods
“Jardin de Granville” (“Evanrat”) is a variety of garden rose created in France by crossing between the ‘Auscent’ and ‘Dorblan’ cultivars. “Jardin de Granville” rose flower extract was obtained by solid-liquid extraction of freeze-dried rose flowers in a natural deep eutectic solvent (NaDES) mixture comprised of fructose, propanediol and water, in a 1/1/3 molar ratio, followed but solid-liquid separation and successive filtration steps. The phytochemical profile was studied using UHPLC-UV methods.
Healthy normal human skin biopsies were obtained after informed consent of patients undergoing surgical discard, in accordance with French national ethical guidelines. 10-mm punches were sampled from skin explants, transferred to Snapwell™ culture inserts (Corning) and cultured at the air-liquid interface at 37 °C in 5 % CO2 in Prime-3D medium (CnT-PR-3D, CELLnTEC) supplemented with normocin™ (Invivogen). Skin explants were pre-treated for 16h by topical application of 1% NaDES Rose extract in a gel formulation, before being challenged by PMA at 1.5% in the same gel formulation. Skin biopsies were then sampled after 2, 4, 8, 24 and 48h. Untreated skin explants were used as control. Explants topically treated by gel formulation without PMA were used as placebo reference.
Lipidomic analysis was carried out using previously described LC-MS/MS methods. Briefly, tissues were lyzed and solid phase extraction was performed to obtain bioactive lipids. LC-MS/MS analysis was performed on UHPLC system (EXION LCAD, SCIEX) coupled to QTRAP6500+ (SCIEX) equipped with electro-spray ionization operating in negative mode. A profile of 24 molecules including resolvins, SPMs and precursors was analyzed.
A TLDA (TaqMan Low Density Array) microfluidic card was designed to include a list of selected genes known to code for proteins involved in keratinocyte metabolism. Keratinocytes were pre-treated for 24h.
Results
Seven different flavonoids could be identified amongst which astragalin was the most abundant. Significant amounts of free amino acids could also be detected, amongst which asparagine accounted for 40%.
Lipidomic analysis showed increased production of D-series resolvins in both young and old skin biopsies further to PMA challenge. D-series resolvins have been reported to be involved in wound-healing, re-epithelialization and tissue repair. In particular, RvD1 has been shown to improve skin resistance to UV-induced oxidative stress and inflammation. Additionally, LXA4 and LXB4, both of which are early resolution mediators, were significantly increased in young skin biopsies thus demonstrating an ability of the NaDES extract of “Jardin de Granville” rose to facilitate inflammation resolution at all age.
Moreover, this rose flower extract induced interesting epidermal biological activities such as positive effect on genes relevant to skin barrier functions especially Filaggrin, Transglutaminase, Cornifin, Cytokeratin 10, Connexin 43, Desmoglein, Occludin, Claudin and Aquaporins.
Conclusion
Taken together, these results highlight this NaDES extract of “Jardin de Granville” rose as a potent ingredient capable of enhancing inflammation resolution in both young and old skins, while improving epidermal structure and homeostasis.
Inflammation is a local reaction towards a disturbance of tissue homeostasis caused by damage to tissue structure and infection. It is normally self-limited and ends with its complete resolution, which involves the production of a class of lipid mediators capable of counter-acting inflammation, namely Specialized Pro-resolving Mediators (SPMs). Derived from poly-unsaturated fatty acids, such as arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid and, their biosynthesis involves different members of the cyclooxygenase and lipoxygenase families. Increasing attention has been paid lately to skin-relevant SPMs such as Resolvin D1 or Lipoxins A4, that have been reported to alleviate skin conditions such as psoriasis or contact dermatitis. Comparing the SPM patterns in old vs. young biopsies, we previously observed an altered responsiveness to pro-inflammatory challenge by Phorbol 12-myristate 13-acetate (PMA), with a switch towards LOX-5 mediated SPM syntheses.
A natural deep eutectic solvent (NaDES) extract of Rose flowers was developed and evaluated for its ability to increase biosynthesis of SPMs both in old and young skin biopsies and to improve epidermal structure, cohesion and differentiation.
Materials and methods
“Jardin de Granville” (“Evanrat”) is a variety of garden rose created in France by crossing between the ‘Auscent’ and ‘Dorblan’ cultivars. “Jardin de Granville” rose flower extract was obtained by solid-liquid extraction of freeze-dried rose flowers in a natural deep eutectic solvent (NaDES) mixture comprised of fructose, propanediol and water, in a 1/1/3 molar ratio, followed but solid-liquid separation and successive filtration steps. The phytochemical profile was studied using UHPLC-UV methods.
Healthy normal human skin biopsies were obtained after informed consent of patients undergoing surgical discard, in accordance with French national ethical guidelines. 10-mm punches were sampled from skin explants, transferred to Snapwell™ culture inserts (Corning) and cultured at the air-liquid interface at 37 °C in 5 % CO2 in Prime-3D medium (CnT-PR-3D, CELLnTEC) supplemented with normocin™ (Invivogen). Skin explants were pre-treated for 16h by topical application of 1% NaDES Rose extract in a gel formulation, before being challenged by PMA at 1.5% in the same gel formulation. Skin biopsies were then sampled after 2, 4, 8, 24 and 48h. Untreated skin explants were used as control. Explants topically treated by gel formulation without PMA were used as placebo reference.
Lipidomic analysis was carried out using previously described LC-MS/MS methods. Briefly, tissues were lyzed and solid phase extraction was performed to obtain bioactive lipids. LC-MS/MS analysis was performed on UHPLC system (EXION LCAD, SCIEX) coupled to QTRAP6500+ (SCIEX) equipped with electro-spray ionization operating in negative mode. A profile of 24 molecules including resolvins, SPMs and precursors was analyzed.
A TLDA (TaqMan Low Density Array) microfluidic card was designed to include a list of selected genes known to code for proteins involved in keratinocyte metabolism. Keratinocytes were pre-treated for 24h.
Results
Seven different flavonoids could be identified amongst which astragalin was the most abundant. Significant amounts of free amino acids could also be detected, amongst which asparagine accounted for 40%.
Lipidomic analysis showed increased production of D-series resolvins in both young and old skin biopsies further to PMA challenge. D-series resolvins have been reported to be involved in wound-healing, re-epithelialization and tissue repair. In particular, RvD1 has been shown to improve skin resistance to UV-induced oxidative stress and inflammation. Additionally, LXA4 and LXB4, both of which are early resolution mediators, were significantly increased in young skin biopsies thus demonstrating an ability of the NaDES extract of “Jardin de Granville” rose to facilitate inflammation resolution at all age.
Moreover, this rose flower extract induced interesting epidermal biological activities such as positive effect on genes relevant to skin barrier functions especially Filaggrin, Transglutaminase, Cornifin, Cytokeratin 10, Connexin 43, Desmoglein, Occludin, Claudin and Aquaporins.
Conclusion
Taken together, these results highlight this NaDES extract of “Jardin de Granville” rose as a potent ingredient capable of enhancing inflammation resolution in both young and old skins, while improving epidermal structure and homeostasis.