effects of oral supplementation and topical application of ...


effects of oral supplementation and topical application of ...

B. Yang, A. Bonfigli, V. Pagani, T. Isohanni, Å. von-Knorring, A. Jutila, V. JudinINTRODUCTIONLipids are important components of the diet formaintaining the health and the functions of theskin. Triacylglycerols, free fatty acids, sterols,squalene, ceramides and some polar lipids areessential components of the stratum corneum,forming the barrier structure of the skin (1).Phospholipids and sterols are components of cellmembranes responsible for both the transductionof signals and the transportation of substances.Polyunsaturated fatty acids are precursors oflocal hormones regulating the inflammation andproliferation of skin cells. Insufficient or unbalanceddietary intake of lipids results in dry,scaly and inflammatory skin (2).Polyunsaturated fatty acids have been shown tohave potential in inhibiting melanin synthesisand reducing UV-induced hyperpigmentation(3). In topical applications, CO 2 -extracted lingonberryseed oil rich in linoleic (18:2n-6) andα-linolenic (18:3n-3) acids increased skin moistureand lightened the skin tone of normal skinand age spots (4).Oxidation damages of cellular components areoften the primary cause of aging and onset ofdiseases. Environmental factors such as UVradiation,pollution and temperature change poseconstant oxidative stress to the skin. A propersupply of antioxidants through diet and dietarysupplementation forms an indispensible elementof modern skin care.Sea buckthorn (Hippophaë rhamnoides) is aEurasian plant that has been used in traditionalhealth care since ancient times. The berry of seabuckthorn is a rich source of a wide range oflipophilic and hydrophilic bioactive compounds.Sea buckthorn seed oil is rich in the two essentialfatty acids, linoleic and α-linolenic acids. Oilisolated from the pulp/peel fraction of sea buckthornberries contains high levels of monounsaturatedfatty acids, palmitoleic acid (16:1n-7)and oleic acid (18:1n-9). Both seed oil and pulpoil are rich sources of tocopherols, tocotrienols,plant sterols and natural carotenoids. The carotenoidcontent can be as high as 0.3-0.5% in thepulp oil (5). Sea buckthorn seed oil and pulp oilhave been shown to inhibit lipid oxidation, protectcell membranes from oxidation damage,reduce inflammation and promote tissue regeneration(6; 7; 8; 9; 10; 11).Sea buckthorn pulp oil and seed oil from differentsources have been shown to be effective intreating skin wounds, burns and scalds (12; 13;14) as well as irradiation dermatitis (15; 16).Topical application of test creams containing 3-5% CO 2 extracted sea buckthorn seed oil or pulpoil effectively scavenged free radicals formed byUVA radiation (by 40-50%) and protected skinlipids from UVA-induced photo-oxidation by60-80%. The test creams also reduced skininflammation and speeded up the restoration ofskin barrier function after repeated tape-stripping(17). Oral supplementation of capsules ofsupercritical CO 2 -extracted sea buckthorn seedoil and pulp oil improved the conditions of atopicskin. The symptom improvement in the seedoil group was positively correlated with theincorporation of sea buckthorn fatty acids intothe plasma lipids (18). So far, no scientificreports have been published on the effects of seabuckthorn oil on healthy skin.MATERIALS AND METHODSThe aim of the study is to investigate the effectsof oral supplementation and topical applicationof supercritical CO 2 -extracted sea buckthorn oilon healthy mature skin using non-invasive skinmeasurement methods (19; 20). Changes in skinhydration, elasticity, colorimetry, skin roughnessand cutaneous thickness were investigatedduring and after the treatments.3

Effects of Oral Supplementation and Topical Application of Supercritical Co 2 Extracted Sea BuckthornSUBJECTSSixty Caucasian females of age 50-70 (mean age61 years) were recruited. The subjects did nothave skin diseases or other pathological eventsduring the period immediately before or duringthe study period. Other exclusion criteriaincluded topical application and systematic useof drugs, pregnancy, breast-feeding, and historyof intolerance of drugs and cosmetic products.All the subjects were informed of the purposeand the procedures of the study and signed writtenconsents before starting the trial. The subjectswere allowed to interrupt the treatmentsbased on their own will or medical reasons relatedor not related to the treatments. Details ofany cases of dropping out were reported.TEST PRODUCTSThe sea buckthorn oil capsule (Aromtech Ltd,Kiviranta/Tornio, Finland) was a vegetablebased product, with filling material consisting of100% SBA24 ® Sea Buckthorn Oil, a standardisedcomposition containing CO 2 extracted seabuckthorn seed and pulp oil. The sea buckthornoil cream (LUMENE Group, Espoo, Finland)was a rejuvenating night cream containing 1%sea buckthorn seed oil manufactured byAromtech using supercritical CO 2 extractiontechnology. The cream was an oil-in-wateremulsionwith an oil/aqueous-phase ratio ofapprox. 27/73. The active ingredients of thecream include sea buckthorn seed oil, acetylhexapeptide-8, palmitoyl pentapeptide-4, glycerin,biosaccharide gum-1, saccharide isomerate,sodium hyaluronate. The composition ofSBA24 ® Sea Buckthorn Oil and supercriticalCO 2 -extracted sea buckthorn seed oil is presentedin Table I.STUDY DESIGNThe study was carried out at the Institute of Skinand Product Evaluation (Milan, Italy) in compliancewith the principles of Good LaboratoryPractice and Good Clinical Practice as well asthe principles established by the World MedicalAssociation in the Declaration of Helsinki. Thestudy was approved by the Ethical Committee ofISPE srl.TABLE IComposition of SBA 24 sea buckthorn oil and sea buckthorn seed oilFatty acids (weight %)16:0 16:1n-7 18:0 18:1n-9 18:1n-7 18:2n-6 18:3n-3Seed oil 7 1 2 18 3 37 31SBA 24 24 24 1 14 5 18 13Carotenoids(mg/100 g)Plant sterols(weight% of oil)Seed oil 30 0.5 100SBA 24 180 1.2 400Tocopherols and tocotrienols(mg/100 g)4

B. Yang, A. Bonfigli, V. Pagani, T. Isohanni, Å. von-Knorring, A. Jutila, V. JudinThe subjects were randomly divided into twogroups, thirty subjects in each group. In group I,the subjects took 4 capsules per day for threemonths. The subjects were advised not to applyany skin care products on the forehead. In groupII, the subjects applied the sea buckthorn testcream on the face. All the subjects were advisedto avoid UV radiation on the skin of the face andforehead throughout the duration of the study. Atthe beginning, after 1 month and 3 months of treatment,non invasive instrumental measurementsof the selected parameters were performed onthe forehead of the subjects of group I and on theperiocular area of the subjects of group II.Instrumental measurements included skin hydration,skin elasticity, skin surface roughness, skinluminosity, and cutaneous thickness. The instrumentalmeasurements were carried out in a bioclimaticroom (temperature: 24 ± 2°C; relativehumidity: 50 ± 10 %).Instrumental measurementsHydration statusThe skin hydration status was measured using aCorneometer CM 825 (Courage + KhazakaElectronic GmbH, Köln, Germany). The physicalprinciple the instrument is based on the measurementof capacitance (21). The device consistsof a square-shaped sensor (area: 49 mm 2 ) ona mobile axis connected to the base unit by a spiralcable. When the front surface of the sensor ispressed against the skin, a number appears onthe liquid crystal display. This reading, directlyproportional to the amount of water contained instratum corneum and the hydration level of cutaneoussurface, is expressed in corneometric unit(c.u.), an arbitrary unit of the instrument. Themeasurement range of the corneometer was 0-150 c.u.ElasticityThe elasticity of the skin was measured using aCutometer SEM 575 (Courage + KhazakaElectronic GmbH, Köln, Germany). The instrumentmeasured the vertical deformation of theskin sucked into the opening of a measuringprobe by a constant negative pressure of 350mbars lasting for one second. The negative pressurewas then annulled, and the skin was allowedto relax for two second. An optical system measuredthe variations in electrical capacitance,which were proportional to the rise (in millimeters)of the skin surface being measured. Threemeasurement cycles (1 cycle: suction/release)were performed for each measurement. Thethree suction/release cycles represented threesuccessive curves providing the deformationparameters relating to the elastic features of theskin (22; 23). Figure 1 presents the demonstrationof the deformation curve of one measurementcycle. The skin rises are shown onCartesian axes, where the deformation of theskin (expressed in mm) is a function of time(expressed in seconds). The effects of the supplementationon skin elasticity were investigatedby monitoring the changes in maximal deformation(R0= U F ), overall elasticity (R2 = U A / U F ),and viscoelastic ratio (R6 = U V / U E ) during andafter the treatments. U A , U V , and U E representtotal deformation recovery at the end of thestress-off period, visco-elastic creep occurringafter the elastic deformation, and elastic deformationof the skin due to the application of stressdelivered by the instrument, respectively.LuminositySkin luminosity (L* value) was measured usinga Chroma Meter CR-300 (Minolta Camera Co.Ltd, Osaka, Japan). L* values ranged from 0 to100, where 0 corresponded to black colour and100 to white (24).5

Effects of Oral Supplementation and Topical Application of Supercritical Co 2 Extracted Sea BuckthornFig. 1 Skin deformation curve of one measurement cycle using a cutomer. UE, elastic deformation of the skin due to theapplication of stress by the instrument; UV, viscoelastic creep occurring after the elastic deformation; UR, elasticdeformation recovery due to stress removal; UA, total deformation recovery at the end of the stress-off period; UF,maximal deformation of the skin.Surface roughnessTo obtain negative imprints of the skin surface,skin replicas were prepared using adhesive discswith an internal diameter of 24 mm and an externaldiameter of 40 mm (3M, St. Paul,Minnesota) and fast hardening synthetic polymerSILFLO (Flexico Ltd, Potters Bar, Herts,United Kingdom). The adhesive disc was putonto the subject’s skin in order to delimit theinvestigated area and to avoid skin stretchingduring the polymer application. A small amountof polymer was then spread onto the internalarea of the disc and left in situ for a few minutesuntil it hardened. The disc was then removed,and a replica with negative imprints of surfacestructure of the skin obtained.The skin replicas were then analysed by animage analysis system Quantilines (Monaderm,Monaco) allowing analysis of a range of reliefparameters (25; 26). The parameters Ra representingmean roughness and Rz representingmaximum roughness were calculated and usedfor evaluation of the effects of sea buckthorn oiltreatment on skin surface roughness (27). Animage covering an area of 12 x 9 mm 2 of the surfaceof each skin replica was acquired through aHigh Performance CCD video-camera (Cohu,Inc., San Diego, CA, USA).Cutaneous thicknessThe thickness of the skin was measured using aDermascan C ® ‚ high resolution ultrasound scanner(Cortex Technology, Danmark) operated inA–scan (amplitude-scan) mode (28; 29; 30).Data analysisFor each instrumental measurement, meanvalues and standard deviations were calculatedfor results at each of the three time points, base-6

B. Yang, A. Bonfigli, V. Pagani, T. Isohanni, Å. von-Knorring, A. Jutila, V. Judinsignificantly increased the cutaneous thickness(P < 0.05 after one month of treatment, P < 0.001after three months of treatment) (Figure 9).Increase in skin thickness may have resultedfrom improvement of hydration level and increaseof collagen synthesis and are often associatedwith increased elasticity of the skin. Despite thedeviation, the average cutaneous thicknessremained rather constant in the capsule groupduring the study (Figure 9).L*luminositySkin(mm)thicknesscutaneous70656055504540Fig. 8 Luminosity of the skin before, during and after supplementationof sea buckthorn oil capsule and topicalapplication of sea buckthorn oil cream.1.801.751.701.651.601.551.501.451.40P > 0.05comparedwithbaselineCapsule,P > 0.05;Cream,P < 0.05;comparedwithbaselineCapsule,P> 0.05;Cream,P< 0.01;comparedwithbaselineP > 0.05comparedwithbaselineBase line 1 month 3 monthtime pointsBase line 1 month 3 monthtime pointsCapsuleCreamFig. 9 Cutaneous thickness of the skin before, during andafter supplementation of sea buckthorn oil capsuleand topical application of sea buckthorn oil cream.DISCUSSIONWith an increasing awareness of the importanceof diet and nutrition in health management,“beauty from within” is becoming an importantpart of the strategy for comprehensive skin care.Dietary supplementations and functional foodshave great potential in promoting the health andwell-being of the skin through different biologicalactivities such as enhancing collagen synthesis,scavenging free radicals and reducing thetendency of inflammation.Beneficial effects of sea buckthorn oil on skinhave been widely reported. However, most ofthe previous studies have investigated the efficacyof sea buckthorn oils in treating skin problemssuch as wounds, burns, skalds, and differenttypes of dermatitis. To our knowledge, thispaper is the first report on the positive effects ofdietary supplementation of sea buckthorn oil inimproving hydration status and reducing theageing of healthy and mature skin. SBA24 seabuckthorn oil and the sea buckthorn seed oilused in the present study are standardised naturalproducts manufactured by environmentalfriendly supercritical CO 2 extraction technology.By avoiding thermal and oxidative damageduring the manufacturing process, the bioactivelipids were kept in the natural form in these products.Essential fatty acids, long chain alcohols, andsterols in the products are essential nutrientssupporting the regeneration of skin cells andrestoration of skin barrier structure. Both SBA24sea buckthorn oil and the sea buckthorn seed oilare rich in natural α- and γ-tocopherols. It isknown that natural α-tocopherol has a betterbioavailability and is more effective as an antioxidantand vitamin E in internal applicationscompared with the synthetic forms (31). Recentstudies suggest that γ-tocopherol is not only aneffective antioxidant but also a potent antiinflammatorycompound (32). Tocopherols,9

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