Anti-Inflammatory and Antioxidant Effects of Diosmetin-3-O-ß-d-Glucuronide, the Main Metabolite of Diosmin: Evidence from Ex Vivo Human Skin Models.

Diosmin is used to relieve chronic venous disease (CVD) symptoms. This study aimed to investigate the anti-inflammatory and antioxidant effects of diosmetin-3-O-ß-d-glucuronide, the major metabolite of diosmin, using human skin explants. The explants were exposed to substance P (inflammation model) or UVB irradiation (oxidative model) and to five diosmetin-3-O-ß-d-glucuronide concentrations. Inflammation was evaluated through interleukin-8 (IL-8) secretion measurements and capillary dilation observation, and oxidation was evaluated by measuring the hydrogen peroxide levels and observing cyclobutane pyrimidine dimers (CPDs). In substance-P-exposed explants, diosmetin-3-O-ß-d-glucuronide induced a significant decrease in IL-8 secretions, with a maximal effect at 2700 pg/mL (-49.6%), and it reduced the proportion of dilated capillaries and the mean luminal cross-sectional area (p < 0.0001 at all tested concentrations), indicating a vasoconstrictive effect. In UVB-irradiated fragments, diosmetin-3-O-ß-d-glucuronide induced a significant decrease in hydrogen peroxide production and in the number of CPD-positive cells, reaching a maximal effect at the concentration of 2700 pg/mL (-48.6% and -52.0%, respectively). Diosmetin-3-O-ß-d-glucuronide induced anti-inflammatory and antioxidant responses, with the maximal effect being reached at 2700 pg/mL and corresponding to the peak plasma concentration estimated after the oral intake of 600 mg of diosmin, the daily dose usually recommended for the treatment of CVD. These ex vivo findings suggest a protective role of diosmetin-3-O-ß-d-glucuronide against inflammatory and oxidative stress affecting the vascular system in CVD pathophysiology.

A Harungana madagascariensis extract with retinol-like properties: Gene upregulations and protein expressions in human fibroblasts and skin explants.

OBJECTIVE: Because they limit, even reverse, age-induced skin alterations, retinoids became a staple in cosmetology. However, their use can result in undesired secondary effects and there is a demand for natural sources of compounds with retinoid-like effects. A preliminary screening identified a Harungana madagascariensis plant extract (HME) as possibly inducing genes stimulated by retinol. We analysed its effect on gene and protein expression, comparing it to retinoids. METHODS: Gene expression was analysed by real-time qPCR on RNA from isolated fibroblasts subjected to retinol or the plant extract for 6, 48 or 96 h. Skin markers were quantified in fibroblasts cultured with retinol or extract containing medium, and UV-aged skin explants subjected to topical applications of creams containing retinol, retinaldehyde or HME. RESULTS: Real-time qPCR shows that the extract induced all RARs and RXRs, even RXRγ that was not induced by retinol. Eighty-eight per cent of the 25 early retinoid reaction genes induced by a concentration of retinol are induced by the extract. In fibroblasts, only the extract increased collagen III labelling, while collagen I and fibronectin labelling are increased by retinol and the extract, with higher levels for the extract. When topically applied to UV-aged skin explants, only the cream containing the HME led to increased labelling of CRABP1 in the epidermis. CRABP2 and Ki67 are induced by all three creams and no effect was detected on RXRs. In the dermisthe extract containing cream increased CRABP2, total collagen, procollagen I and collagen I while creams with retinol or retinaldehyde only affected some of these proteins. CONCLUSIONS: The HME induces an overall retinol-like gene induction profile in isolated fibroblasts and retinoid-like stimulation of protein synthesis in both isolated fibroblasts and photoaged skin explants.

OBJECTIFS: Limitant, voire inversant les altérations cutanées induites par l'âge, les rétinoïdes sont devenus incontournables en cosmétologie. Cependant, leur application topique peut entraîner des effets secondaires indésirables et il existe une demande pour des composés naturels ayant des effets similaires à ceux des rétinoïdes. Un screening préliminaire nous avait permis d'identifier un extrait de la plante Harungana madagascariensis (HME) comme pouvant induire des gènes stimulés par le rétinol. Nous avons donc analysé son effet sur l'expression de gènes et de protéines induits par les rétinoïdes et comparé les résultats à ceux obtenus en présence de rétinoïdes. MÉTHODES: L'expression de gènes a été analysée par qPCR en temps réel réalisée sur l'ARN de fibroblastes isolés soumis au rétinol ou à l'extrait végétal pendant 6, 48 ou 96 heures. Différentes protéines cutanées ont été quantifiés dans des fibroblastes cultivés en présence de rétinol ou d'un milieu contenant l'extrait. Des quantifications ont également été faites sur des explants de peau vieillie par les UV et soumis à des applications topiques de crèmes contenant du rétinol, du rétinaldéhyde ou le HME. RESULTATS: La qPCR en temps réel montre que l'extrait induit tous les gènes RARs et RXRs, même RXRγ qui n'était pas induit par le rétinol. Quatre-vingt-huit pour cent des 25 gènes impliqués dans la réaction précoce aux rétinoïdes induits par une concentration de rétinol ont été induits par l'extrait. Dans les fibroblastes, seul l'extrait a augmenté le marquage du collagène III, tandis que le marquage du collagène I et de la fibronectine a été augmenté par le rétinol et l'extrait, avec des niveaux plus élevés pour l'extrait. En application topique sur des explants de peau vieillie par les UV, seule la crème contenant le HME a entraîné une augmentation du marquage de CRABP1 dans l'épiderme. CRABP2 et Ki67 ont été induits par les trois crèmes et aucun effet n'a été détecté sur les RXRs. Dans le derme, la crème contenant l'extrait a augmenté CRABP2, le collagène total, le procollagène I et le collagène I, tandis que les crèmes contenant du rétinol ou du rétinaldéhyde n'ont affecté que certaines de ces protéines. CONCLUSIONS: Chez les fibroblastes isolés, le HME induit un profil d'induction génique globalement similaire à celui du rétinol. Chez les fibroblastes isolés et des explants de peau photo-vieillie, il entraine une stimulation de la synthèse protéique similaire à celle des rétinoïdes.

Purple tulip extract improves signs of skin aging through dermal structural modulation as shown by genomic, protein expression and skin appearance of volunteers studied.

BACKGROUND: Purple tulip extract is a rich source of flavonoids which are powerful antioxidants and can hence be considered as an ideal candidate for use in skin care products. AIMS: We aimed to evaluate the effects of purple tulip extract on skin quality and to determine its molecular modes of interaction. METHODS: A pangenomic study on human skin fibroblasts was carried out to analyze multiple changes in gene expression. Ex vivo studies of human skin explants exposed to ultraviolet (UV) irradiation or H2 O2 were performed to assess modulations of protein expression. Finally, a clinical assay was carried out to evaluate the efficacy of purple tulip extract on skin appearance and condition of aged women. RESULTS: Genetic modulation analyses led us to infer the induction of many biological functions including cell differentiation, proliferation, migration, inflammatory responses, and matrix remodeling. The ex vivo studies revealed an enhancement of the collagen network and increased expression of glycosaminoglycans (GAG), fibronectin, and collagen VI. Finally, the clinical study highlighted the potential anti-aging properties of the purple tulip extract which decreased the relaxation of the oval face and improved skin elasticity after 28 days of treatment. Significant reductions of the length and depth of the nasolabial wrinkles were also observed. CONCLUSION: Our genomics data on the effect of purple tulip extract on the ex vivo UV-challenged skin showed that genes responsible for, among others, the upkeep of the skin, such as collagen induction, immune cell proliferation, and epidermal repair, were all up-regulated. More importantly, the clinical study corroborated these data by the visible and measurable effects of the topical purple tulip extract on the aged skin of 22 women, further demonstrating the beneficial impact of the extract on aged skin.

Tasmannia lanceolata leaf extract alleviates stretch mark appearance in a randomized, placebo-controlled clinical trial in women and stimulates extracellular matrix synthesis in ex vivo human skin explants.

BACKGROUND: The leaves of Tasmannia lanceolata mainly contain polygodial that is known to exhibit a range of biological functions including anti-inflammatory effects. AIMS: These studies aimed to assess the effects of Tasmannia lanceolata extract (TLE) on skin and more particularly on stretch marks in women. PATIENTS/METHODS: A double-blind, randomized, placebo-controlled clinical study was carried out on 29 women, aged from 25 to 60 years, to investigate the effects of TLE on stabilized stretch marks. TLE and placebo products were topically applied daily for 8 weeks. Skin roughness and firmness of stretch marks were assessed by 2D and 3D photograph processing and analyses. Dermal density and thickness were evaluated using ultrasound, while stretch mark conditions (length, color, and depth) were determined by clinical scoring. Matricial proteins (pro-collagen I and elastin) and pro-matricial factors, like TGF-ß concentrations, were quantified from cultures of human skin explants presenting stretch marks, treated with TLE or vehicle control. RESULTS: Skin roughness of stretch marks was significantly reduced in the TLE group after 8 weeks of treatment. Skin firmness of stretch marks was significantly increased in the TLE group after 4 weeks of treatment, and this improved effect was maintained until the end of the study. Dermal density and thickness were significantly increased in the TLE group compared to the placebo group. Furthermore, TLE restored the dermal condition of the stretch mark skin, up to normal skin levels. In addition, pro-collagen I and elastin concentrations were found to be higher in the TLE-treated stretch mark skin explants compared to the untreated ones, associated with higher quantities of TGF-ß production. CONCLUSION: These results revealed that TLE could help improve the aspect of stabilized stretch marks in women by restoring the matricial environment.

Polar lipids from wheat extract oil improve skin damages induced by aging: Evidence from a randomized, placebo-controlled clinical trial in women and an ex vivo study on human skin explant.

BACKGROUND: Polar lipids from wheat (Triticum vulgare/aestivum) extract oil (WEO) are known to improve skin hydration. AIMS: These studies aimed to assess WEO benefits on the skin appearance of middle-aged women. METHODS: A double-blind, randomized, placebo-controlled clinical study was carried out on 64 healthy women, aged from 45 to 60 years, to investigate antiaging effects and benefits for the skin. The study lasted 20 weeks including 12 weeks of oral supplementation with WEO or placebo and 8 weeks of follow-up. Wrinkles in the "crow's-feet" area were evaluated by the Lemperle score. Skin hydration was measured using a corneometer, while roughness and radiance were determined by clinical scoring. Collagen content was quantified in human skin explants exposed to ultraviolet (UV) irradiations and treated with WEO or vehicle control. RESULTS: Compared to the placebo group, the Lemperle score was significantly reduced in the WEO group between W0 and W8 to reach a clinically significant 1 grade at W12. Facial hydration was significantly improved in the WEO group from W0 to W12, whereas leg hydration was significantly increased after 4 weeks and lasted throughout the supplementation period. Skin roughness and radiance were also significantly improved from W0 to W8 in the WEO group compared to placebo group. A higher collagen content was measured in the UV-irradiated skin explants treated with WEO compared to the untreated ones. CONCLUSION: These results confirmed the moisturizing effect of WEO and, for the first time, revealed its potential antiaging properties.

Anti-inflammatory and antiradical effects of a 2% diosmin cream in a human skin organ culture as model.

BACKGROUND: Diosmin, a naturally occurring flavonoid, is considered as a vascular-protective agent and is used orally to treat chronic venous insufficiency. It exhibits anti-inflammatory and free radical scavenging properties, but, like many other flavonoids, it is poorly absorbed in the small intestine. OBJECTIVE: Our aim was to investigate the skin protective effects of a diosmin-based cream, using skin organ culture as model. METHODS: Fragments of human skin explants, cultured ex vivo, were allocated to four treatment groups: no cream, no cream + stress, placebo cream + stress, and 2% diosmin cream + stress. Stress was induced by exposure to either substance P (anti-inflammatory effects' assessment) or UVB irradiation (free radical scavenging effects' assessment). Vascular dilation and the pro-inflammatory mediator IL-8 release were determined in the first model, whereas hydrogen peroxide level and the number of cyclobutane pyrimidine-positive cells were evaluated in the second model. RESULTS: In the substance P-induced inflammation model, 2% diosmin cream exhibited significant vasoconstrictive (proportion of dilated capillaries: -29%, capillary luminal area: -49% vs no cream + stress) and anti-inflammatory (IL-8 release: -36% vs no cream + stress) effects. In the UVB irradiation model, 2% diosmin cream significantly reduced hydrogen peroxide production and cyclobutane pyrimidine dimer formation (-45% and -36% vs no cream + stress, respectively). These effects were not observed with placebo cream. CONCLUSION: Diosmin administered topically may protect skin against the biological effects of various exogenous or endogenous stresses, such as those involved in chronic venous disease.

Split-face histological and biochemical evaluation of tightening efficacy using temperature- and impedance-controlled continuous non-invasive radiofrequency energy.

BACKGROUND: Bipolar radiofrequency (RF) is capable of heating dermal collagen fibers and inducing skin tightening by collagen remodeling. OBJECTIVE: To substantiate safety and improvement of skin laxity following skin heating with a novel temperature- and impedance-controlled non-invasive radiofrequency (RF) device by histological and biochemical evaluations. METHODS: A split-face study was performed on 4 subjects who underwent 8 weekly RF sessions on one side of their face, leaving the other side an untreated control and then underwent facelift procedure. Clinical evaluation by photographs was done prior to the surgical procedure. Ex vivo fragments were harvested from both sides and compared. Morphometric analysis of dermal collagen fibers, collagen synthesis, and elastin synthesis evaluations were compared in triplicates. RESULTS: Facial skin tightening was apparent in split-face photographs. A significant increase of 7.9% in dermal collagen content, and a significant increase of 34.7% in collagen synthesis were demonstrated in the treated samples. No statistically significant effect on elastin synthesis was detected. CONCLUSIONS: Skin tightening following treatment with non-invasive RF has proven histologically and biochemically to derive from increase in dermal collagen synthesis and content.

Ex-vivo study of hybrid energy technology using a human skin model.

BACKGROUND: More aging adults and the social acceptance of aesthetic treatments have increased the demand for minimally invasive aesthetic treatments. Skin resurfacing is very effective at improving aging symptoms, including wrinkles and skin imperfections. Following the negative effects of full skin resurfacing, in addition to a very long downtime, fractional lasers and fractional radiofrequency (RF) technologies were introduced, since gaining popularity. Their efficacy, along with minimal downtime, has enabled an effective and safer treatment. A novel technology based on fractional Hybrid Energy™ (HE), combines RF and an additional electrical energy for enhancing the thermal effect. OBJECTIVE: This study evaluated the morphological and histological effects of the new HE technology on epidermal and dermal skin layers, using an ex-vivo human skin model. METHODS: Human skin samples were collected and treated ex-vivo with the HE applicator. The effect was evaluated by skin histology and quantitative analysis by assays of collagen fibers, elastin and glycosaminoglycan (GAGs) dosages, reflecting the hyaluronic acid content, in addition to epidermal mitotic index evaluation. RESULTS: Histology demonstrated immediate and long-term HE effects on both epidermal and dermal skin layers with a direct correlation between the treatment parameters and effects. Results demonstrated a significant increase of the epidermal mitotic index, significant dermal collagen remodeling and significant increases in both epidermal and dermal GAGs. CONCLUSIONS: HE technology significantly affected collagen remodeling and an increase in mid to deep dermis GAGs in addition to epidermal mitotic index, resulting in long term structural and biochemical dermal and epidermal improvement.

Anti-inflammatory and draining effect of the Deep Oscillation® device tested clinically and on a model of human skin maintained in survival condition.

The objective of this study was to evaluate the anti-inflammatory, toning and draining effect of the Deep Oscillation® device. The Deep Oscillation® device uses the forces of pulsed electrostatic attraction and friction to provoke oscillations that act on the epidermis, dermis, and sub-cutaneous layers of tissue. An ex-vivo study was first completed by using a model of skin maintained in survival condition. The draining and anti-inflammatory effects of the device were determined by pro-inflammatory cytokine assay and by histological analysis of capillary dilation. The analgesic effectiveness of the Deep Oscillation® was then evaluated by immunohistochemical analysis of TRPV1. To corroborate this data, a clinical study was conducted by selecting 20 subjects with periorbital bags or dark circles to undergo treatment with the device. Evaluations included photography, dermatological scores as well as ultrasound analysis. Using an ex-vivo model of human skin maintained in survival condition, the Deep Oscillation® device was effective in reducing inflammation with a significant reduction of dilated capillaries and IL8, while lowering sensory receptor levels. Clinically, the device was successful in reducing both dark circles and bags by an average of 40%.

Ex vivo study of the home-use TriPollar RF device using an experimental human skin model.

BACKGROUND: A wide variety of professional radio frequency (RF) aesthetic treatments for anti-aging are available aiming at skin tightening. A new home-use RF device for facial treatments has recently been developed based on TriPollar technology. OBJECTIVE: To evaluate the mechanism of the new home-use device, in the process of collagen remodeling, using an ex vivo skin model. METHODS: Human skin samples were collected in order to evaluate the anti-aging effect of a home-use device for facial treatments on an ex vivo human skin model. Skin tightening was evaluated by dermal histology, quantitative analysis of collagen fibers and dosage of collagen synthesis. RESULTS: Significant collagen remodeling following RF treatment with the device was found in the superficial and mid-deep dermis. Biochemical measurement of newly synthesized collagen showed an increase of 41% in the treated samples as compared to UV-aged control samples. CONCLUSIONS: The new home-use device has been demonstrated to affect significant collagen remodeling, in terms of the structural and biochemical improvement of dermal collagen on treated skin samples.

Clinical and histopathological study of the TriPollar home-use device for body treatments.

Professional non invasive treatments for body contouring based on radiofrequency (RF) became popular in aesthetic clinics due to proven efficacy and safety. A new home-use RF device for body treatments has been developed based on TriPollar technology. Our objective was to evaluate the TriPollar home-use device for circumference reduction, cellulite improvement and skin tightening using objective and subjective methods. An ex-vivo human skin model was used for histological and biochemical evaluations of the TriPollar clinical effect. Additionally, twenty four subjects used the new device on the abdomen and thigh areas and the circumference reduction was measured. Ex-vivo models indicated a significant increase of 82% in hypodermal glycerol release. Histology revealed a 34% alteration in adipocyte appearance. Collagen synthesis increased by 31% following TriPollar treatment. A significant average reduction of 2.4 cm was measured on the treated thighs. On the control thighs a lesser, non-significant reduction was found. Average abdominal laxity was reduced from 1.4 at baseline to 0.8 following treatments. A certain reduction was measured in the abdomen circumferences, although it was not significant. The reported results demonstrate the safety and efficacy of the new TriPollar home-use device for body contouring and skin tightening. Treatment may lead to discrete circumference reduction and moderate laxity improvement.

Ex vivo human skin evaluation of localized fat reduction and anti-aging effect by TriPollar radio frequency treatments.

BACKGROUND: A wide variety of radio frequency (RF) treatments for localized fat and cellulite reduction as well as anti-aging are available nowadays, but only a few have shown the biological mechanism responsible for the clinical results. OBJECTIVE: To determine the biological mechanism of the TriPollar RF device for localized fat and cellulite reduction as well as the collagen remodeling effect. METHODS: Human skin samples were collected from abdominoplasty surgery and facial lifts, in order to evaluate the lipolytic and anti-aging effects of the apollo device powered by TriPollar RF technology using an ex vivo human skin model. The anti-cellulite effect was evaluated by the dosage of released glycerol and histological analysis of the hypodermis. Skin tightening was evaluated by morphometric analysis of collagen fibers and the dosage of collagen synthesis. RESULTS: Following TriPollar treatment, a significant increase of glycerol release by skin samples was found. The structure of fat cells was altered in shape and a modification of the fibrous tract was also detected in the fat layer. Additional findings indicated stimulation of the dermal fibroblasts with increased collagen synthesis. CONCLUSIONS: The detected alteration in the hypodermal layer is manifested by fat and cellulite reduction accompanied by structural and biochemical improvement of dermal collagen, which result in overall skin tightening.