J. App/. Cosmetol. 27, 13-25 (January/March 2009)

EFFECTS OF ORAL SUPPLEMENTATION ANO TOPICAL APPLICATION OF SUPERCRITICAL C02 EXTRACTED SEA BUCKTHORN OIL ON SKIN AGEING OF FEMALE SUBJECTS

Baoru Yang•·, Adriana Bonfìgli', Velerie Pagani', Tiina lsohanni3, Asa von-Knorring, Arimatti Jutila', Vesa-Pekka Judin'

' Aromtech Ltd, Tornio, Finland ' lnstitute of Skin and Product Evaluation (ISPE Sri), Milan, ltaly 3 LUMENE group, Finland

Received: November, 2008.

Key words: rhamnoides; Sea buckthom oil; Ora/ supplementation; Topica/ appli­

cation; Supercritica/ C02 extraction; Skin ageing; Skin hydration; Skin elasticity; Skin surface roughness; Cutaneous thickness; Beauty from within;

Summary

Sea buckthom () seed and pulp o il is known to promote skin regeneration, speed up wound healing and ease skin inflammation . The present study was carried out to investiga­ te the anti -ageing effects of ora! supplementation of a standardized supercritica! C02-extracted sea buckthorn oi l and topica! skin application of supercriti ca! COrextracted sea buckthorn seed oil. Sixty femaJe subjects of age 50-70 years were randomly di vided into two groups, thirty subjects in each group. In one group, the subjects took sea buckthorn oil capsules, four capsules (4 x 0.5 g oil) per day, for three months. In the other group, the subjects applied topically sea buckthom seed oil rej u­ venating night cream on the face, twice per day, for three months. Skin hydration status, elasticity, surface roughness, lurninosity and cutaneous thickness were determined using non-invasive instru­ me ntal measurements before, after one month and at the end of treatments. Both treatments signifi­ cantly improved the skin hydration status and the overall skin elasticity of the subjects (P < 0.001). Oral supplementation with the sea buckthom oil capsules resulted in decreases in the mean roughness (Ra) and maximum roughness (Rz) of the skin surface, indicating anti-wrinkle efficacy of the pro­ duct. Topica! application of the sea buckthorn oil cream increased cutaneous thickness, suggesting

13 Effects of Ora/ Supplementation and Topica/ Application of Supercritica/ Co2 Extracted Sea Buckthorn

positive structural changes and improvement in collagen synthesis in the skin. This is the first study showing beneficiai effects of orally used sea buckthorn oil in reducing the signs of skjn ageing. This is also an important breakthrough demonstrating the great potential of sea buckthorn oil in suppor­ ting the health and well being of healthy sbn through internal use. Riassunto

E' noto come l'olio ricavato dai semi e dalla polpa dell'olivello spinoso promuova la rigenerazione della pelle, acceleri la sua cicatrizzazione riducendo i processi infiammatori. Il presente studio è stato condotto per verificare gli effetti anti-età della supplementazione orale del­

l'olio di olivello spinoso estratto dai suoi semi con il metodo di C02 ad alta pressione. Sessanta donne di età compresa tra i 50 e i 70 anni sono state suddivise, in modo casuale, in due grup­ pi da 30 individui. In un gruppo i soggetti assumevano 4 capsule al giorno dell'olio (4x0, 5 g) per tre mesi. Nell'altro gruppo i soggetti applicavano topicamente due volte al giorno, sempre per tre mesi, una crema da notte contenente lo stesso olio. Dopo il primo mese ed alla fine del trattamento sono state controllate con metodologia non invasi­ va l'idratazione, l'elasticità, la rugosità, la luminosità e lo spessore della cute. Entrambi i trattamenti hanno migli orato significativamente lo stato di idratazione e l'elasticità (p<0,001). La supplementazione orale con le capsule dell'olio di oli vello spinoso poneva in risalto sia una ridu­ zione della rugosità media (Ra) che di quell a massima (Rz) riscontrata a livello della superficie cuta­ nea, ponendo in risalto l'efficacia anti-rughe del prodotto. L'applicazione topica dello stesso oli o incrementava lo spessore medio della pelle raggiungendo un positivo incremento della produzione di collageno. Questo è il primo studio che, ponendo in evidenza l'effetto benefico che l'olio di olivello spinoso svolge nel ridurre i segni dell 'età , dimostra la grande potenzialità insita nel suo uso per migliorare il benessere generale e l'aspetto della cute.

14 B. Yang, A. Bonfigli, V. Pagani. T. lsohanni, A. von-Knorring, A. Jutila, V. Judin

INTRODUCTION oil are rich sources of , , plant sterols and natural . The carote­ Lipids are important components of the diet for noid content can be as high as 0.3-0.5% in the maintaining the health and the functions of the pulp oil (5). Sea buckthorn seed oil and pulp oil skin. Triacylglycerols, free fatty acids, sterols, have been shown to inhibit lipid oxidation, pro­ squalene, ceramides and some polar lipids are tect celi membranes from oxidation damage, essential components of the stratum comeum, reduce inflammation and promote tissue regene­ forming the barrier structure of the skin ( I). ration (6; 7; 8; 9; 10; 11). Phospholi pids and sterols are components of celi Sea buckthom pulp oil and seed oil from diffe­ membranes responsible for both the transduction rent sources have been shown to be effective in of signals and the transportation of substances. treating skin wounds, burns and scalds (12; 13 ; Polyunsaturated fa tty acids are precursors of 14) as well as irradiation derrnatitis (15; 16). locai hormones regulating the inflammation and Topica! application of test creams containing 3- proliferation of skin cell s. lnsufficient or unba­ 5% C02 extracted sea buckthom seed oil or pulp lanced dietary intake of lipids results in dry, oil effectively scavenged free radicals formed by scaly and inflammatory skin (2). UVA radiation (by 40-50%) and protected skin Polyunsaturated fatty acids have been shown to lipids from UVA-induced photo-oxidation by have potential in inhibiting melanin synthesis 60-80%. The test creams also reduced skin and reducing UV-induced hyperpigmentation inflammation and speeded up the restoration of (3). In topica] applications, COi-extracted lin­ skin barrier function after repeated tape-strip­ gonberry seed oil rich in linoleic ( 18:2n-6) and ping (17). Ora! supplementation of capsules of a-linolenic (18:3n-3) acids increased skin moi­ supercritica! COrextracted sea buckthorn seed sture and lightened the skin tone of normai skin oil and pulp oil improved the conditions of ato­ and age spots (4). pic skin. The symptom improvement in the seed Oxidation damages of cellular components are oil group was positively correlateci with the often the primary cause of aging and onset of incorporation of sea buckthorn fatty acids into diseases. Environmental factors such as UV­ the plasma lipids ( 18). So far, no scientific radiation, pollution and temperature change pose reports have been published on the effects of sea constant oxidative stress to the skin. A proper buckthorn oil on healthy skin . supply of through diet and dietary supplementation forrns an indispensible element MATERIALS ANO METHODS of modem skin care. Sea buckthorn ( H ippophae· rhamnoides) is a The ai m of the study is to investigate the effects Eurasian plant that has been used in traditional of ora! supplementation and topica) application health care since ancient times. The berry of sea of supercritica) COrextracted sea buckthorn oil buckthorn is a rich source of a wide range of on healthy mature skin using non-invasive skin lipophilic and hydrophilic bioactive compounds. measurement methods ( 19; 20). Changes in skin Sea buckthom seed oil is rich in the two essen­ hydration, elasticity, colorimetry, skin roughness tial fatty acids, linoleic and a-linolenic acids. Oil and cutaneous thickness were investigateci isolateci from the pulp/peel fraction of sea buck­ during and after the treatments. thorn berries contains high levels of monounsa­ turated fatty acids, ( 16: I n-7) and (18: ln-9). Both seed oil and pulp

15 Effects of Ora/ Suppfementation and Topica/ Appfication of Supercritica/ Co2 Extracted Sea Buckthorn

SUBJECTS oil cream (LUMENE Group, Espoo, Finland) was a rejuvenating night cream containing I% Sixty Caucasian females of age 50-70 (mean age sea buckthorn seed oil manufactured by 61 years) were recruited. The subjects did not Aromtech using supercritica] C02 extraction have skin diseases or other pathological events technology. The cream was an oil-in-water­ during the period immediately before or during emulsion with an oil/aqueous-phase ratio of the study period. Other exclusion criteria approx. 27/73. The active ingredients of the included topica] application and systematic use cream include sea buckthorn seed oil, acetyl of drugs, pregnancy, breast-feeding, and history hexapeptide-8, palmitoyl pentapeptide-4, glyce­ of intolerance of drugs and cosmetic products. rin, biosaccharide gum-1, saccharide isomerate, Ali the subjects were informed of the purpose sodium hyaluronate. The composition of and the procedures of the study and signed writ­ SBA24* Sea Buckthorn Oil and supercritica! ten consents before starting the trial. The sub­ COrextracted sea buckthom seed oil is presen­ jects were allowed to interrupt the treatments ted in Table I. based on their own will or medicai reasons rela­ ted or not related to the treatments. Details of STUDY DESIGN any cases of dropping out were reported. The study was canied out at the Institute of Skin TEST PRODUCTS and Product Evaluation (Milan, Italy) in com­ pliance with the principles of Good Laboratory The sea buckthorn oil capsule (Aromtech Ltd, Practice and Good Clinica] Practice as well as Kiviranta/Tornio, Finland) was a vegetable the principles established by the World Medicai based product, with fi ll ing materiai consisting of Association in the Declaration of Helsinki. The l00% SBA24* Sea Buckthorn Oil, a standardi­ study was approved by the Ethical Committee of ISPE srl. sed composition containing C02 extracted sea buckthorn seed and pulp oil. The sea buckthorn

TABLEI Composition ofS BA 24 sea buckthorn oil and sea buckthorn seed oil Fatty acids (weight % ) 16:0 16: ln-7 18:0 18:ln-9 18:ln-7 18:2n-6 18:3n-3 Seed oil 7 I 2 18 3 37 31 SBA24 24 24 1 14 5 18 13 Carotenoids Plant sterols Tocopherols and tocotrienols (mg/100 g) (wei2ht% of oil) (m2/100 2) Seed oil 30 0.5 100 SBA24 180 1.2 400

16 B. Yang, A. Bonfigli, V. Pagani, T. /sohanni, A. von-Knorring, A. Jutila, V. Judin

The subjects were randornly divided into two Elasticity groups, thirty subjects in each group. In group I, the subjects took 4 capsules per day for three The elasticity of the skin was rneasured using a rnonths. The subjects were advised not to apply Cutometer SEM 575 (Courage + Khazaka any skin care products on the forehead. In group Electronic GmbH, Koln, Germany). The instru­ Il, the subjects applied the sea buckthorn test rnent rneasured the vertical deformation of the cream on the face. Ali the subjects were advised skin sucked into the opening of a rneasuring to avoid UV radiation on the skin of the face and probe by a constant negati ve pressure of 350 forehead throughout the duration of the study. At rnbars lasting for one second. The negative pres­ the beginning, after 1 rnonth and 3 rnonths of tre­ sure was then annulled , and the skin was allowed atrnent, non invasive instrurnental rneasurernents to relax for two second . An optical systern rnea­ of the selected pararneters were perforrned on sured the variations in electrical capacitance, the forehead of the subjects of group I and on the which were proportional to the rise (in millime­ peri ocular area of the subjects of group II. ters) of the skin surface being measured. Three Instrurnental rneasurernents included skin hydra­ rneasurernent cycles ( l cycle: suction/release) tion, skin elasticity, skin surface roughness, skin were performed for each rneasurernent. The luminosity, and cutaneous thickness. The instru­ three suction/release cycles represented three rnental rneasurernents were carried out in a bio­ successive curves providing the deformation cl imatic roorn (temperature: 24 ± 2°C; relative parameters relating to the elastic features of the humidity: 50 ± IO %). skin (22; 23). Figure 1 presents the demonstra­ tion of the deforrnation curve of one rneasure­ lnstrumental measurements rnent cycle. The skin rises are shown on Cartesian axes, where the deformation of the Hydration status skin (expressed in mm) is a function of time (expressed in seconds). The effects of the sup­ The skin hydration status was rneasured using a plernentation on skin elastici ty were investigated Corneorneter CM 825 (Courage + Khazaka by rnonitoring the changes in rnaxirnal deforrna­ Electronic GmbH, Koln, Germany). The physi­ tion (RO= UF), overall elasticity (R2 = UA I UF), cal principle the instrurnent is based on the rnea­ and viscoelastic ratio (R6 = Uv I UE) during and surement of capacitance (21 ). The device con­ after the treatrnents. UA, Uv, and UE represent 2 sists of a square-shaped sensor (area: 49 rnrn ) on tota] deforrnation recovery at the end of the a mobile axis connected to the base unit by a spi­ stress-off period, visco-elastic creep occurring rai cable. When the front surface of the sensor is after the elastic deforrnation, and elastic defor­ pressed against the skin, a nurnber appears on mation of the skin due to the application of stress the liquid crystal display. This reading, directly delivered by the instrurnent, respectively. proportional to the amount of water contai ned in straturn corneum and the hydration leve) of cuta­ Luminosity neous surface, is expressed in corneometric unit (c.u.), an arbitrary unit of the instrument. The Skin lurninosity (L* value) was rneasured using measurement range of the corneometer was 0- a Chrorna Meter CR-300 (Minolta Carnera Co. 150 c.u. Ltd, Osaka, Japan). L * values ranged frorn O to 100, where O corresponded to black colour and 100 to white (24).

17 Effects of Ora/ Supplementation and Topica/ Application of Supercritica/ Co2 Extracted Sea Buckthorn

skin deformation

Uv f I ~ I f UR t LIA UF t + l Lh

Time ! ' t =o

Fig. 1 Skin deformarion curve of 011e measuremenr cyc/e 11si11g a curomer. UE, elasric deformation of the skin. due to rhe application of stress by the i11srrn111e111; UV, viscoelastic creep occurring afrer the elastic deformation; UR, elastic defor111atio11 recovery due to stress removal; VA, tota/ deformatio n recovery at the end of the stress-off period; UF, maxima/ defonnation of rhe skin.

Surface roughness

To obtain negative imprints of the skin surface, max imum roughness were calculated and used skin replicas were prepared using adhesive discs for evaluation of the effects of sea buckthorn oil with an internal diameter of24 mm and an exter­ treatment on skin surface roughness (27). An nal diameter of 40 mm (3M, St. Paul, image covering an area of 12 x 9 mm 2 of the sur­ Minnesota) and fast hardening synthetic poly­ face of each skin replica was acquired through a mer SILFLO (Flexico Ltd, Potters Bar, Herts, High Performance CCD video-camera (Cohu, United Kingdom). The adhesive disc was put Inc., San Diego, CA, USA). onto the subject's skin in order to delimit the in vestigated area and to avoid skin stretching Cutaneous thickness during the polymer application. A small amount of polymer was then spread onto the internal The thickness of the skin was measured using a area of the disc and left in situ for a few minutes Dermascan C®, high resolution ultrasound scan­ unti! it hardened. The di sc was then removed, ner (Cortex Technology, Danmark) operated in and a replica with negative imprints of surface A-scan (amplitude-scan) mode (28; 29; 30). structure of the skin obtained. The skin replicas were then analysed by an Data analysis image analysis system Quantilines (Monaderm, Monaco) allowing analysis of a range of relief For each instrumental measurement, mean parameters (25; 26). The parameters Ra repre­ values and standard deviations were calculated senting mean roughness and Rz representing for results at each of the three time points, base-

18 B. Yang. A. Bonfigli. V. Pagani. T. lsohanni. A. von-Knorring. A. Jutila. V. Judin

line, after one and three months of supplementa­ Skin elasticity tion. The values obtained at the three check points were compared by Analysis of Variance Topica! treatment with sea buckthorn cream sli­ and Tukey's test. Differences reaching a confi­ ghtly increased the maximal deformation of the dence level of 95% (P < 0.05) were considered skin , whereas no clear effect was observed from statistically significant. the supplementation of SBA24 sea buckthorn oil (Figure 3). Both treatments significantly increa­ RESULTS sed the overall elasticity of the skin (Figure 4). After one and three months of treatment, overall Skin hydration status elasticity was increased from the baseline by 16.3% and 25.8%, respecti vely, in the capsule Skin capacitance measured as corneometric group, 18.6% and 21.9%, respectively, in the units significantly increased in both groups cream group. At the same time some decrease in during and at the end of the study (Figure 2). skin visco-elastic ratio was detected in both After one month of supplementation with groups during and after the treatments, although SBA24 sea buckthorn oil , the capacitance was the changes did not reach a statistically signifi­ increased by 33.6%, after three months of treat­ cant leve! (Figure 5). ment by 48.6%, from the baseline value (P < The clear increase in the overall elasticity and 0.001). Correspondingly, one-month application the decreasing trend in the visco-elastic ratio of sea buckthom oil cream increased the capaci­ demonstrated clear positi ve effects of the test tance of the skin by 30.5% (P < 0.001). This products in improving the elasticity of the skin. increase was maintained during the rest of the study period. No difference in skin hydration status was fou nd between the group receiving the sea buckthorn oil capsules and the group using the sea buckthorn oil cream.

p < 0.001 p < 0.001 capsule, capsule. 80 0.28 i K -.,.. vX .rie.v..,, p > IO<0.05 ; compared compared o o Capsule Cream, Cream, :s with w·lh o Capsule Q) 70 a Cream E 0.23 p <0.05;1 T p E 1 baseline bas ~line

Fig. 2 Capaciumce expressed as comeometric 1mirs be/o­ Fig. 3 Maximum deformatio11 of rhe skin before, during re, d11ri11g and after supple111ema1io11 of sea buck­ and after supplememarion of sea buckrhom oil cap­ thom oi/ capsule and topica/ applicarion of sea sule and topica/ applicarion of sea buckthom oil buckrhom oil cream. cream.

19 Effects of Oro/ Supplementotion ond Topico/ Applicotion of Supercritico/ Co2 Extrocted Seo Buckthorn

p < 0.001 p < 0.001 decreasing big wrinkles and fine lines of the 0.7 compared compared skin. The cream treatment did not show clear o Capsule I witt 0.6 wi'1 effects on the skin surface roughness in the study oCream I base I ne base ine >. >- peri od. ·E o.5 - u; -- >->- -- ~ 0.4 ---- ro 35 i o Capsule o Cream~ P > 0.0 ID 0.3 5 p > 0.05 >o com "ared comuared 0.2 Cl. 30 '" wth (/) eline (/) bas bas ~ l i ne -- 0.1 ~ 25 - ~ Base line 1 month 3 month O> ::J time points e 20 - e _,_ Q) ,_ -- Fig. 4 Overall elasticity of the skin before, during and "'E 15 - - after supplementation of sea buckthorn oil capsule and topica! applicarion of sea buckthorn oil cream. 10 ~ Base line 1 month 3 month time points 0.35 l oCaosule Fig. 6 Mean roughness (Ra) ofrhe skin before, during and l o cream p > 0.05 p > 0.05 0.30 after supplementation of sea buckthorn oil capsule comi ared compared ... b and topica/ applicarion of sea buckrhom oil cream. ~ 0.25 with '1) bas line bas li ne ~ 0.20 "' -- ->- ~ 0.15 -- ,_ -- - p > 0.05 p > 0.05 (..) 1 (/) 180 rnmn?Sran ,....,...mn-::::r.r.c.~

- ~ ·o;: 0.10 - - - [;_(J) 160 ...... 0.05 -- - - - ~ 140 e O.OD ~ Base line 1 month 3 month g> 120 time points e E ::J Fig. 5 Viscoe/asricity of rhe skin before, during and after ; 80 - X suppleme/1/ation of sea buckrhorn oil capsule and ~ 60 topica/ app/ication ofsea buckrhorn oil cream. 40 I I I I I I I I I I I Base line 1 month 3 month Surface roughness time points

Fig. 7 Maximum roughness (Rz) ofthe skin before, during The mean roughness (Ra) of the skin surface was and after supp/ementation ofsea buckrhorn oil cap­ decreased by 3.0% after one month of supple­ sule and ropical application of sea buckthom oil mentati on and by 7.4% after three months of cream. supplementation of SBA24 from the baseline value (Figure 6). The maximum roughness para­ Luminosify and fhickness meter Rz was decreased from the baseline value by 3.6% and 9.2%, respectively, after one and Throughout the study skin luminosity remained three months of the supplementation (Figure 7). rather constant in both groups (Figure 8), indica­ The results indicate potential of ora! supplemen­ ting no clear effects of the two treatments on the tation SBA24 sea buckthom oil capsules in skin tone of the subjects. The cream treatment

20 B. Yang. A. Bonfigli, V Pagani. T. lsohanni, A. von-Knorring, A. Jutila. V Judin

significantly increased the cutaneous thickness DISCUSSION (P < 0.05 after one month of treatment, P < 0.001 after three months of treatment) (Figure 9). With an increasing awareness of the importance Increase in skin thickness may have resulted of diet and nutrition in health management, from improvement of hydration leve! and increa­ "beauty from within" is becoming an important se of collagen synthesis and are often associated part of the strategy for comprehensive skin care. with increased elasticity of the skin. Despite the Dietary supplementations and functional foods deviation, the average cutaneous thickness have great potential in promoting the health and remained rather constant in the capsule group well-being of the skin through different biologi­ during the study (Figure 9). ca! activities such as enhancing collagen synthe­ sis, scavenging free radicals and reducing the 70 p > o 05 p > 0.05 tendency of inflammation. Beneficiai effects of sea buckthorn oil on skin 65 I d-...... :::L .... :.·~ I I have been widely reported. However, most of _J ~ 60 the previous studies have investigated the effica­ ·u; o cy of sea buckthorn oils in treating skin pro­ -~ 55 ·1 blems such as wounds, burns, skalds, and diffe­ ~ 50 ·1 rent types of dermatitis. To our knowledge, this 45 paper is the first report on the positive effects of dietary supplementation of sea buckthorn oil in 40 I I I I I I I I I I I improving hydration status and reducing the Base fine 1 month 3 month time points ageing of healthy and mature skin. SBA24 sea buckthorn oil and the sea buckthorn seed oil Fig. 8 Lumi11osity of1he ski11 before, during a11d after s11p­ used in the present study are standardised natu­ ple111e111a1ion of sea buck1horn oil capsule and topi­ ca/ applicario11 ofsea buck1hom oil cream. ral products manufactured by environmental

friendly supercritica! C02 extraction technology. By avoiding thermal and oxidative damage during the manufacturing process, the bioactive lipids were kept in the natural form in these pro­ Capsule, Capsule, p > 0.05; p > 0.05; ducts. _ Cream Cream 1 80 - p < 0.05; p < 0.01; Essential fatty acids, long chain alcohols, and E 1 .7 5 -t---t----<::em §, o Capsule sterols in the products are essential nutrients gi 1.70 o Cream supporting the regeneration of skin cells and ~ 1.65 -"' restoration of skin bruTier structure. Both SBA24 ~ 1.60 sea buckthorn oil and the sea buckthorn seed oil ~ 1.55 are rich in natural a- and y-tocopherols. It is fil 1.50 -I e: known that natural a- has a better :;"' 1.45 u bioavailability and is more effective as an antio­ 1.40 I I I I i I I I I I I I xidant and in internal applications Base line 1 monlh 3 month time points compared with the synthetic forms (31). Recent Fig. 9 Cutaneous rhickness of the skin before, during and studies suggest that y-tocopherol is not only an after supple111entatio11 of sea buckthorn oil capsule effective but also a potent anti­ and 1opical application of sea buckrhorn oil cream. inflammatory compound (32). Tocopherols,

21 Effects of Ora/ Supplementation and Topica/ Application of Supercritica/ Co,, Extracted Sea Buckthorn

tocotrienols, and carotenoids are natural antioxi­ dants working synergisticall y, protecting skin from the oxidative damage caused by stress, ageing, and the unfavourable factors in the envi­ ronment (33; 34). It is also important to notice that ora! supplemen­ tation with sea buckthorn oil resulted in impro­ vement in the skin conditions of the subjects comparable to the effects shown by the topica! treatment with the sea buckthorn oil cream. While effects of topica! treatments tend to be restricted to the skin, dietary supplementation of sea buckthorn oil positively influences the well being of the whole body. The beneficia! effects of ingested SBA24 sea buckthorn oil on the skin, mucous membranes, the heart and the vascul ar system have been demonstrated by earlier stu­ dies (18; 35; 36).

ACKNOWLEDGEMENTS

The authors thank the Finnish Funding Agency for Technology and lnnovationfor financial sup­ port for the study.

22 8. Yang. A Bonfigli. V Pagani. T. lsohanni. A. von-Knorring. A Jutila. V Judin

References

1) Moller H. (2002) The chemistry of natural and syntheti c skin barrier lipids. In: Cosmetic lipids and the skin barrier. Marce! Dekker, Inc., New York, p. 1-36. 2) Ziboh VA, Miller CC, Cho Y (2000) Metabolism of polyunsaturated fatty acids by skin epider­ mal enzymes: generation of antiinflammatory and antiproliferative metabolites. Am. J. Clin. Nutr., 71(suppl):361- 366. 3) Ando H, Ryu A. and Hashimoto A. (1998) and a -linolenic acid lightens ul travio­ let-induced hyperpigmentation of skin. Arch. Dermatol. Res ., 290:375-381.

4) Yang B. and Judin V-P. (2004) Supercritica! C02 extracts from peat and lingonberry: innovati­ ve natural anti-ageing ingredients for skincare. and Toiletries Manufacture Worldwide. Quest Magazines & Events Wilmington, UK. p. 7-12. 5) Yang B. and Kallio H. (2002) Lipophilic components in seeds and berries of sea buckthorn and physiological effects of sea buckthorn oils. Trends Food Sci. Technol., 13: J 60-J 67. 6) Ji YB. and Gao Y. (1991) Effect of feeding sea buckthorn seed oil and sea buckthorn seed oil supplemented with sodium selenite in vivo on Na-K-ATPase acti vity in erythrocyte ghost in rats. Acta Nutrimenta Sinica, 13 (1):20-24 . 7) Ji YB. and Gao Y. (1991) Effect of feeding sea buckthorn seed oil and sea buckthorn seed oil supplemented with sodium selenite in vivo on structural stability of erythrocyte ghosts in rats. Chinese Biochemical Journal, 7(4):441 -446. 8) Song ZH. and Gao Y. (1995) Effect of sea buckthorn oil and vi tamin E on the lipid peroxida­ tion of rats after cold exposure. Acta Mutrimenta Sinica, 17 (1):27-3 1. 9) Shi HL, Cai H J , Chen XY. and Yang CM. (1994) Studies on the antioxidative effects of Hippophae rhamnoides L. seed oil. Acta Nutrimenta Sinica, 16 (3):292-295. 10) Yang B. and Kallio H. (2003) Effects of sea buckthorn (Hippophae rhamnoides) oil on skin: Eastern tradition and modem research. Asia Pacific Persona! Care, 4 (5): 47-49. 11) Yang B. and Kallio H. (2005) Physiological effects of seabuckthorn (Hippophae rhamnoides) fruit pulp and seed oils). In: Seabuckthorn (Hippophae L.). A Multipurpose Wonder Plant. Voi. II: Biochernisty and Pharmacology. Dya Publishing House, New Delhi, India, p. 363-389. 12) Vlasov VV. (1970) Hippophae oi l in the treatment of superficial burns of the skin. Vestn. Dermatol. Venero!, 44 (6):69-72. 13) Zhao YS. (1994) A preliminary report on treatment of 32 cases of burns with sea buckthorn seed oil. Hoppophae, 7 (3):36-37. 14) Mekhtiev NKh, Azizov FSh, Salamov AA, Nasudari AA, Guseinova SYu. and Agaev EM. (1991) Chemical, technological and pharmacological studies of naturally growing sea buckthorn (Hippophaif rhamnoides) of Azerbaijan. In : Nov. v Biol. Khimii i Farmakol. Oblepikhi ., Novosibirsk, SO AN SSSR, p. 166-170. 15) Zhang WL, Zhang ZF, Fan JJ, Yang SY, Li ZM, Deng ZC, Wang GL. and Zhang FS. (1988) Experimental observation and clinica! investigation effect of sea buckthorn oil on acute radio dermatitis. Hippophaif, 1 (1):27-30. 16) Quirin KW. and Gerard D. (1993) Sanddomlipide - interessante Wirkstoffe fur die Kosmetik. Parfamerie Kosmetik, 10:6 18-625.

23 Effects of Ora/ Supplementation and Topica/ Application of Supercritica/ Co" Extracted Sea Buckthorn

17) Derma Consultant (1998) Protective effects oftopically applied sea buckthorn seed oil and pulp oil on skin against free radical s and lipid oxidation induced by UVA radiation and mechanically induced skin irritation. Unpublished data, 1998. 18) Yang B, Kalimo K, Mattila L, Kallio S, Katajisto J, Peltola O. and Kallio H. (1999) Effects of dietary supplementation wi th sea buckthorn (Hippophae rhamnoides) seed and pulp oils on atopic dermatitis. J. Nutr. Biochem., 10:622-630. 19) Seidenari S. (1998) Diagnostica non invasiva in dermatologia. EDRA, Milano. 20) Serup J, Jemec GBE. (1995) Handbook of non - invasive methods and the skin. CRC Press, Florida. 21) Berardesca E., EEMCO group (1997) EEMCO guidance for the assessment of stratum cor­ neum hydration: electrical methods. Skin Res . Technol., 3:126-132. 22) Pierard GE., EEMCO group (1999) EEMCO guidance for the in-vivo assessment of tensile functional properties of the skin - part l. Skin Pharmacol. Appl. Skin Physiol., 12(6):352-362. 23) Rodrigues L., EEMCO group (2001) EEMCO guidance for the in-vivo assessment of tensile functional properties of the skin - part 2. Skin Pharmacol, Appl, Skin Physiol., 14(1):52-67. 24) Pierard GE. (1998) "EEMCO guidance for the assessment of skin colour. J. Eur. Acad. Dermatol. Venereo!., 10(1): 1- 11. 25) Corcuff P, Chatenay F. and Brun A. (1985) Evaluation of anti-wrinkle effects on humans; Int. J. Cosm. Sci., 7:117-126. 26) Corcuff P. and Leveque JL. (1995) Skin surface replica image analysis of furrows and wrinkles in Handbook of non-invasive methods and the skin , Serup and Jermec (eds.) CRC Press. 27) Leveque JL. (1999) EEMCO guidance for the assessment of skin topography. J. Eur. Acad. Dermatol. Venereo I ., 12(2): I 03- 1 14. 28) Kirsch JM, Hanson ME. and Gibson JR. (1984) The deterrnination of the skin thickness using conventional diagnostic ultrasound equipment. Clin. Exp. Dermato/., 9:280-285. 29) Sondergaard J, Serup J. and Tikjob G. (1985) Ultrasonic A and B scanning in clinica! and experimental dermatology. Acta Dermatovener (Stock) Suppi., 120:76-82. 30) Serup J. (1992) Ten years experience with high-frequency ultrasound examination of the skin: development and refinement of the technique and equipment. In: Ultrasound in Dermatology, Springer-Yerlag Berlin Heidelberg, Berlin , p.41-54. 31) Traber MG, Elsner A. and Brigelius-Flohé R.(1998) Synthetic as compared with natural vita­ min E is preferentially excreted as a-CEHC in human urine: studies using deuterated a-tocophe­ ryl acetates. FEBS Letters, 437:145- 148. 32) Jiang Q, Elson-Schwab I. and Courtemanche C. (2000) y-tocopherol and its major metaboli­ te, in contrast to a-tocopherol , inhibit cyclooxygenase activity in macrophages and epithelial cells. Proc Nati Acad Sci USA, 97: 11494-11499. 33) Serbinova E, Kagan V. and Han D. (1991) Free radical recycling and intramembrane mobility in the antioxidant properties of alpha-tocopherol and alpha-. Free Rodie. Biol. Med., 10:263-275. 34) Stone W. and Papas A. (2003) In: Gunstone (eds.) Lipids for Functional Foods and . The Oily Press, Bridgwater, England, p. 53-72.

24 B. Yang, A. Bonfigli, V Pagani, T. /sohanni, A. von-Knorring. A. Jutila, V Judin

35) Yang B. and Erkkola R. (2007) Sea buckthom oil s, mucous membranes and Sjogren's syndro­ me - with special reference to latest studies. In: Seabuckthom (Hippophae L.). A Multipurpose Wonder Plant. Voi. III: Advances in Research and Development, Daya Publishing House, Delhi, India, p. 254-267. 36) Johansson A, Korte H, Yang B, Stanley J. and Kallio H. (2000) Sea buckthorn berry oil inhi­ bits platelet aggregation. J. Nutr. Biochem., 11,491-495.

Author Address:

Dr Baoru Yang Aromtech Ltd. Veturitallintie l FIN-9541 O Tornio - Finland Fax: +358 2 4101710 E-mail: [email protected]

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