International Journal of Cosmetic Science 21: 437447 (1999)

Horse chestnut Aesculus hippocastanum: potential applications in cosmetic skin-care products

J . A . W I L K I N S O N * and A . M . G . B R O W N
Herbal Research Laboratories, School of Health, Biological and Environmental Sciences, Middlesex University, Queensway, Eneld, Middlesex EN3 4SF, UK

Received 12 March 1999 Accepted 22 April 1999

Keywords: anti-inammatory, cosmoceutical, herbal, phytochemistry, saponin Synopsis In addition to the well reported benecial effects of horse chestnut (Aesculus hippocastanum) extracts on venous insufciency and associated conditions, such preparations also have many potential positive pharmacological effects on the skin. Extracts from this species, and in particular, those based on horse chestnut seeds, contain saponins, known collectively as aescin, which have a gentle soapy feel, and are potent anti-inammatory compounds. Saponins, such as escin, also reduce capillary fragility, and therefore help to prevent leakage of uids into surrounding tissues, which can cause swelling. An extract of horse chestnut has recently been shown to have one of the highest active-oxygen scavenging abilities of 65 different plant extracts tested. Such extracts are more powerful anti-oxidants than vitamin E, and also exhibit potent cell-protective effects, which are linked to the well-known anti-ageing properties of anti-oxidants. The extract is also rich in a number of avonoids, such as derivatives of quercetin and kaempferol. Flavonoids also have protective effects on blood vessels, and are well-known, powerful anti-oxidants. R sum e e En plus des effets b n ques bien connus des extraits de marron dInde (Aesculus hippocastanum) sur e e linsufsance veineuse et les troubles associ s, de telles pr parations ont aussi de nombreux effets e e pharmacologiques potentiels positifs sur la peau. Des extraits de cette esp` ce, et en particulier, ceux a base de e ` graines de marron dInde, contiennent des saponines, collectivement connues comme aescine, qui ont un toucher savonneux doux, et sont des compos s anti-inammatoires puissants. Les saponines, telles que e laescine, r duisent aussi la fragilit capillaire, et aident par cons quent a emp cher la fuite des uides dans les e e e ` e tissus environnants, ce qui peut causer un gonement. Un extrait de marron dInde sest r cemment av r avoir e ee une capacit de capture de loxyg` ne actif parmi les plus elev es de 65 extraits de plantes diff rentes test s. e e e e e De tels extraits sont des anti-oxydants plus puissants que la vitamine E, et pr sentent aussi des effets potentiels e de protection de la cellule, qui sont li s aux propri t s anti-vieillissement bien connues des anti-oxydants. e ee Lextrait est aussi riche en un certain nombre de avonodes, tels que des d riv s de la querc tine et du e e e kaempf rol. Les avonodes ont aussi des effets protecteurs sur les vaisseaux sanguins, et sont des anti-oxydants e puissants bien connus.

Introduction
In recent years, the cosmetic industry has become increasingly interested in using plantderived natural products in cosmetic formulations. This is not merely a marketing tool, but
* To whom correspondence should be addressed: (T) J.Wilkinson@mdx.ac.uk 44 181 362 6425; (F) 44 181 805 0702. E-mail:

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a result of the plant material itself having biological activity that is benecial to skin [1]. The use of horse chestnut preparations as herbal medicines has been adequately described elsewhere, so this paper therefore focuses on the biological potential of extracts from this species as cosmoceuticals and for skin-care products.

Existing uses of Aesculus hippocastanum

Extracts of the seeds of Aesculus hippocastanum (horse chestnut) are a well established treatment for conditions such as varicose veins, haemorrhoids, phlebitis (inammation of the veins), diarrhoea, fever, enlargement of the prostate gland [2], rheumatism, neuralgia and rectal complaints [3]. Such treatments are usually administered orally, at a dose of 0.51.2 ml of liquid seed extract per day. In addition to these uses for seed extracts, preparations from both the bark and the leaves of A. hippocastanum are also used as therapeutic agents. The bark is no longer in common use, but has a folk medicinal application as a febrifuge, and as an astringent in cases of diarrhoea and haemorrhoids [4]. Decoctions of the bark are also used, albeit rarely, for the topical treatment of skin disorders, such as sores, lupus and ulcers [4]. The bark has also previously been used as an anti-malarial agent, as a cinchona substitute, but this practice is no longer continued [3]. Horse chestnut leaf preparations are used in folk medicine to treat coughs, rheumatism and arthritis, although the underlying phytochemical basis for these applications has not been determined [3].

Chemical constituents of Aesculus hippocastanum seeds

Saponins The total saponin content of seeds, often expressed as (a)escin (Figure 1), actually consists of -escin and -escin, the latter of which is, in turn, composed of more than 30 derivatives of the triterpenoids, protoaescigenin and barringtogenol C. These compounds are primarily found in the seed cotyledons (they can constitute up to 28% of the weight of the dry seeds), but have also been detected in the seed integument, the bark, buds, leaves and the immature fruit pericarp of A. hippocastanum [5]. Flavonoids A number of avonoids (mainly glycoside derivatives of quercetin and kaempferol) have been detected in A. hippocastanum (Figure 2), including astragalin (kaempferol 3-Oglucoside), isoquercitrin (quercetin-3 -glucoside), leucocyanidin (3,3',4,4',5,7 hexahydroxyavone) and rutin (quercetin 3-rutinoside) [3]. Most of these compounds are found in the seeds (not including the integument), but have also been found in the fruit pericarp, bark, leaves and buds. Results concerning quercitrin are contradictory, with different authors reporting that this compound is present either in the leaves or in the seeds of A. hippocastanum. [5] In addition to these avones, epicatechin and its dimer proanthocyanidin A2 (Figure 3) have been reported in A. hippocastanum. These compounds are primarily found in the bark, leaves buds and fruit pericarp, but have also been found in the seed integument. Previously, this latter tissue has been eliminated from extracts used in the preparation of drugs [6], but, if included, it would provide a valuable additional source of bioactive compounds.

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escin Ia escin Ib desacylescin I escin IIa escin IIb desacylescin II

R1 Tig Ang H Tig Ang H

R2 Ac Ac H Ac Ac H

R3 CH2OH CH2OH CH2OH H H H

Figure 1. Selected components of the saponin mixture escin.

R R R R

Rutinoside: Rutin Rhamnoside: Quercitrin Glucose: Isoquercitrin Arabinoside: Quercetin 3-arabinoside

R1 R1 R1 R1

Rutinoside: Kaempferol 3-O-rutinoside Rhamnoside: Kaempferol 3-O-rhamnoside Glucose: Astragalin Arabinoside: Kaempferol 3-arabinoside

Figure 2. Selected avonoids found in Aesculus hippocastanum.

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Figure 3. Structure of proanthocyanidin A2.

Other constituents Seeds of A. hippocastanum have also been described as containing starch (4050% weight) [6], sugars, proteins (specically the globulin, hippocastanine, containing L-( )lysine and L-( )-tryptophan), a fatty oil (containing oleic, linoleic, linolenic, stearic and palmitic acids) and purines (adenosine, adenine and guanine). [5]

Bark
The bark of Aesculus hippocastanum, in common with the seeds, has been described as containing the saponin mixture escin [3]. Coumarin glycosides, including esculin, scopolin and fraxin, and their respective aglycones, esculetin, scopoletin and fraxetin, are also present in the bark of A. hippocastanum (Figure 4), in contrast to seed tissues, in which these compounds have not been detected. The avonoid glycoside quercitrin (Figure 2), and its corresponding aglycone have also been detected in bark tissues. Additional compounds, including allantoin, sterols, leucocyanidin, leucodelphinidin, catechol tannins and alkanes have also been described as occurring in bark tissues. [3]

R1 R2 R3

H: Aesculetin, R H: Scopoletin, R H: Fraxetin, R

Glucose: Aesculin Glucose: Scopolin Glucose: Fraxin

Figure 4. Coumarins and coumarin glycosides present in the leaves and bark of Aesculus hippocastanum.

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Leaves
In common with the bark of A. hippocastanum, leaf tissues contain the coumarin glycosides scopolin, fraxin and esculin (Figure 4) [3]. A range of avonoid glycosides of quercetin (e.g. quercitrin, rutin, isoquercitrin and quercetin 3-arabinoside) and the corresponding glycosides of kaemperfol have also been detected in leaf tissues (Figure 2). In addition to these glycosides, escin has been detected (but only in trace amounts), as well as leucanthocyans, cis,trans-polyprenols, amino acids, fatty acids and sterols (sitosterol, stigmasterol and campesterol).

Biological activities of horse chestnut phytochemicals

Saponins Isolated components of the escin complex have been demonstrated to have potent antiinammatory effects, especially on the early stages of induced inammation [7, 8] and can also reduce ethanol absorption and display hypoglycaemic actions when taken orally [9]. Escin also has positive therapeutic effects on the microvasculature and its surrounding connective tissues. Such tissues are composed of cellular and brous components in a uid ground substance or extracellular matrix, which consists of proteoglycans and glucosaminoglycans, and provides support and cushioning to blood vessels and brous tissues. One of the key components of this ground substance is hyaluronic acid, a proteoglycan, which gives the extracellular matrix its viscosity. The activity of hyaluronidase, the enzyme which breaks down hyaluronic acid, reduces the viscosity of the ground substance [10], thus reducing its supporting and cushioning capacity. An additional effect of this activity is that there is reduced resistance to uid leakage from capillary vessels, as a result of the compromised integrity of the extracellular matrix, and hence there is an increased exchange of uids across the microvasculature membranes (Figure 5) [11]. Whilst this activity can be associated with pathogenic infections (e.g. Staphylococcus aureus secretes hyaluronidase, allowing the bacterium to move more freely through the connective tissues [10]), it has also been exploited for benecial effect, as injection of hyaluronidase prior to drug administration can be used to deliberately increase capillary permeability, thus facilitating easier access of the therapeutic agent to the blood stream. One hypothesis for the oedematous symptoms associated with venous insufciency is that hyaluronidase activity results in increased vascular leakage from the capillaries [11]. This hypothesis could certainly be used to explain the benecial effects of horse chestnut extracts on conditions resulting from venous insufciency, as more than one component of these extracts have been demonstrated to display anti-hyaluronidase activity [11, 5]. The saponin mixture, escin, is one component of horse chestnut extracts capable of inhibiting hyaluronidase activity (another is proanthocyanidin A2; see later). In the case of escin, however, inhibition of hyaluronidase has been determined to be insufcient to account for the powerful venotonic properties of A. hippocastanum extracts on the microvasculature, and it has been suggested that inhibition of other enzymes, including collagenase, elastase and -glucuronidase, all of which are involved in dening the integrity of the extravascular matrix, might explain why extracts of A. hippocastanum were markedly more effective than escin alone [11].

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1 Ground tissue consists of proteoglycans and glucosaminoglycans, including hyaluronic acid. Cellular components (broblasts, macrophages, plasma cells, adipocytes and mast cells) and tissues (collagen, elastic and reticular bres) omitted for clarity. 2 Fluid leakage due to capillary permeability and density of surrounding ground substance. Increased capillary permeability can result in oedema typical of diseases resulting from venous insufciency.

Figure 5. Simplied representations of capillary vessel permeability in connective tissues. Left: Permeability in tissue containing normal amounts of hyaluronic acid in ground substance. Right: Increased permeability, as a result of reduced hyaluronic acid content (due to hyaluronidase activity).

Flavonoids As a class of phytochemicals, avonoids are widely accepted as possessing a range of biological activities [12] which may be of relevance in the context of topical application of avonoid-containing preparations. Some of these activities are listed in Table I. In the specic case of horse chestnut avonoids, rutin has been described as possessing radical scavenging, anti-bacterial and anti-viral activities, and is also used medicinally in the treatment of varicosis and capillary fragility [13]. Quercitrin and isoquercitrin are also antibacterial (e.g. against Pseudomonas maltophilia), and the former additionally possesses anti-haemorrhagic activity [13]. Proanthocyanidin A2 has been demonstrated to possess a range of biological activities, similar to those of escin, which would be benecial in cosmetic applications [5, and refs.
Table I. General biological activities of flavonoids Activity Reduction of capillary fragility and permeability Anti-inammatory Anti-allergenic Anti-bacterial Anti-viral Enzyme inhibitory
Activities as described in Bruneton (1995) [6]

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therein]. These activities are primarily venotonic and vasoprotective, such as depermeabilisation of capillaries, but also include wound healing, powerful antioxidant, antiinammatory, and anti-enzymatic properties. In a similar manner to escin, proanthocyanidin A2 has been shown to be able to inhibit lipid peroxidation, which is indicative of cellularprotective effects of this compound. Similarly, proteolytic enzymes in the extravascular matrix are inhibited by proanthocyanidin A2, with -glucuronidase, elastase and collagenase all being more strongly inhibited than hyaluronidase. Compounds such as proanthocyanidin A2 may well be at least partly responsible for the differences between the venotonic effects of escin (which inhibits hyaluronidase [11]), and the more powerful effects of crude A. hippocastanum extracts.

Existing cosmetic applications

In 1980, the applications of horse chestnut and escin to cosmetic products were described for treating the following areas: scalp, oral cavity, face, body, hands, legs and feet, body hygiene, foot hygiene, intimate hygiene and prevention of haemorrhoids [14]. Recently, it has been reported that extracts of horse chestnut [or escin at 0.250.5% (w/v)] have been included in a range of commercial products which apply to these areas, including shampoos, shower foams, foam baths, creams, lotions and toothpastes [15]. Horse chestnut seeds are on the Medicines Control Agencys General Sales List in the UK [2] for external use only. In Germany, they are the subject of a positive therapeutic monograph, indicated for venous insufciency, although in the United States, they are of undetermined regulatory status [15].

Additional cosmetic applications

Recent pharmacological studies suggest that there is, in addition to the uses described above, considerable potential for use of these extracts in cosmetic preparations in the following categories: Anti-ageing effects Many reports suggest a link between skin ageing and active oxygen species, especially when combined with ultra-violet radiation. Recently, an extract of A. hippocastanum has been shown to be highly effective in reducing the production and damaging effects of active oxygen species in vitro, using both biochemical and biological assays [16]. In vitro active-oxygen-scavenging properties of A. hippocastanum. An extract of A. hippocastanum has been demonstrated to be a potent scavenger of active oxygen, being almost 20 times more effective at absorbing superoxide anions than ascorbic acid, a recognised anti-oxidant [16; Table II]. Similarly, the extract of A. hippocastanum reduced the release of hydroxyl radicals and singlet oxygens in vitro, demonstrating a broad spectrum of active oxygen scavenging properties (Table II). As these oxygen species are associated with cellular damage and inammation, their absorption by cosmetic preparations containing horse chestnut phytochemicals has great potential to alleviate damage to the skin, which could ultimately be manifested as an anti-ageing effect. In vitro cell-protective effects of A. hippocastanum. Lipid peroxidation, a process indicative of damage to cellular membranes, was induced in vitro, under controlled conditions, and

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Table II. Superoxide-anion, hydroxyl radical and singlet oxygen scavenging activities of Aesculus hippocastanum and antioxidants IC50 Treatment Aesculus hippocastanum L-Ascorbic acid dl- -Tocopherol DMSO Sodium azide g (dry material) ml 0.01 0.39 38.70
1

Superoxide anions 0.24 4.10 576.20

Hydroxyl radicals 7.79 3.30 253.70 (2.55 0.26 0.03 38.70 0.1) 105

Singlet oxygens 72.98 21.18 43.75 6.20 0.39

8.99

Data represent mean standard deviation (n 3). Data using L-Ascorbic acid, dl- -Tocopherol, DMSO and Sodium azide are included as reference values, IC50 values for these compounds are expresed as gml 1 Modied from Masaki et al., 1995 [16].

was found to be signicantly reduced in experiments using A. hippocastanum extracts [16]. Such inhibition was comparable to that obtained by including the anti-oxidant -tocopherol in the assay. Cultured broblasts, which are cells usually found in the connective tissue secreting a matrix associated with the repair and isolation of wounded tissues, were also strongly protected by the extract of A. hippocastanum from damage caused by active oxygen species in vitro. Almost four times the number of these cells survived damage induced by oxygen species, generated using the hypoxanthine/xanthineoxidase system, in the presence of the A. hippocastanum extract than when the extract was absent. Solar protective effects Closely linked to the anti-ageing effects of A. hippocastanum are its potential applications in cosmetics with sun-screen properties. A recent study [17] has investigated the potential use of plant extracts, including A. hippocastanum seed extract, as novel anti-solar agents, based on their ability to absorb light. Such absorption is of particular interest in the ultraviolet region of the spectrum, as UV radiation has been linked with a number of cutaneous conditions, including sunburn, photosensitivity, actinic elastosis, cutaneous degeneration and perhaps most importantly, skin cancer [10, 17]. Although an extract of A. hippocastanum did not display signicant UV-absorbing properties when studied in isolation [in which it had a sun protection factor (SPF) of 0], when combined with a synthetic sunscreen, octylmethoxyciunamate, signicant increases in the effectiveness of the synthetic compound were demonstrated (SPF rose from 4 to 6 on the addition of the A. hippocastanum extract; Table III). Whilst the precise reasons for this improvement of SPF remain unresolved, the authors speculated that such an increase may be as a result of synergistic interactions between components of the A. hippocastanum extract and octylmethoxycinnamate [17]. Venous effects One of the most widely used applications of A. hippocastanum is in the treatment of poor circulation, and its associated conditions. Whilst many of these studies rely on oral delivery of the extracts to improve circulation, conditions such as varicose veins, phlebitis

A. hippocastanum: potential applications in cosmetic skin-care products

Table III. Sun protection factors and UV absorptions of dry extracts of Aesculus hippocastanum (prepared by maceration) alone, and in combination with 2% solution of octylmethoxycinnamate. Absorbance Wavelength (nm) 290 295 300 305 310 315 320 SPFa
a b

445

10% dry extract 0.091 0.079 0.065 0.053 0.045 0.038 0.034 0

10% dry extract 2% octylmethoxycinnamate 0.528 0.569 0.598 0.625 0.651 0.625 0.553 6b

SPF Sun protection factor 2% octylmethoxycinnamate has an SPF rating of 4 Modied from Ramos et al., 1996 [17].

and post-thrombotic syndrome have all been treated using gels containing escin [5, and refs. therein], suggesting that the topical administration of extracts, as would be the case in the application of cosmetics, is potentially an equally valid delivery system. The positive effects of A. hippocastanum extracts on circulation are due to a range of complex interactions with veins, the lymphatic system, capillaries and connective tissue. Extracts of A. hippocastanum have been demonstrated to possess venotonic effects in vitro and in vivo, in both animal and human systems [5, and refs. therein]. For example, A. hippocastanum extract led to an increase in pressure of ow through both normal and constricted isolated veins, and an increased effect of noradrenaline on increasing the blood pressure (and consequently ow) through such vessels [18]. Similarly, capillaries have been shown to be strengthened by administration of A. hippocastanum extract. In addition to the direct effects of A. hippocastanum extract on blood vessels, benet may also be gained from the protective effects of the extract on connective tissues which surround the capillary vessels. In chronic venous insufciency, the capillaries become highly permeable, resulting in water and proteins leaving the vascular system, which in turn cause swelling; this may be as a result of the degradation of the extracellular matrix surrounding the microvasculature (Figure 5). The main component of this extracellular matrix is hyaluronic acid, a simple glucosaminoglycan consisting of up to several thousand sugar residues; levels of this compound are usually regulated by the enzyme hyaluronidase, which promotes the degradation of hyaluronic acid. Recently, escin and its aglycone, escinol, have been demonstrated to possess non-competitive antihyaluronidase activity (Table IV), even at concentrations well below that expected as a result of topical application of these compounds [11]. This inhibition of hyaluronidase should lead to the recovery of the integrity of hyaluronic acid, and consequently the extracellular matrix surrounding the microvasculature, and may therefore be responsible for some of the benecial effects of A. hippocastanum extracts.

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Table IV. Anti-hyaluronidase activity of selected sapogenins and saponins IC50 (Hyaluronidase) Sapogenins Hederagenina (from Hedera helixc) Oleanolic acida (from H. helixc) Ruscogeninb (from Ruscus aesculeatusd) Escinola (from Aesculus hippocastanume) Saponins Hederacoside C (from H. helixc) -hederin (from H. helixc) Escin (from A. hippocastanume)
a

280.4 2.7 M 300.2 3.1 M 5 mM 1.65 0.03 mM 5 mM 5 mM 149.9 2.6

Triterpene sapogenin Steroidal sapogenin c Obtained from leaf tissues Modied from Facino et al., 1995 [11].
b

d e

Obtained from rhizome tissues Obtained from seed tissues

One of the conditions associated with venous insufciency is the development of oedematous panniculopathies (inammation of the sub-cutaneous layer of fat, leading to tender nodules, typically on the thighs and breasts), and this condition has recently been treated with phytochemicals from horse chestnut. Application of an emulsion containing escin and biological carriers, such as -sitosterol and phosphatidylcholine, over a 30 day treatment period, led to signicant improvements to both thigh and breast pauniculopathies in human volunteers. These improvements were ascribed to improvements in microcirculation, as recorded by Laser Doppler Flowmetry, resulting in reduced swelling and normalisation of skin temperature [5, and refs. therein].

Conclusions
In addition to the existing uses of extracts of horse chestnut as treatments for venous insufciency, preparations containing phytochemicals from horse chestnut have considerable potential for inclusion in a range of cosmetic formulations. Inclusion of horse chestnut extracts (especially those from the seeds of the plant, rather than the more poorlycharacterised extracts from leaf and bark tissue) could result in a number of benecial actions on the skin after topical administration. Such effects would include potent antioxidative activity, which could help decrease the visible signs of skin ageing, anti-bacterial and anti-viral properties (largely due to avonoids), reduction in skin reddening and cutaneous swelling as a result of effects on capillary vessels and their surrounding connective tissues (due to saponins and the epicatechin dimer proanthocyanidin A2) and also the potential synergistic enhancement of the efcacy (SPF) of sun screens.

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