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British loiirna! of !}ert}uitoloi]ij ( 1 9 9 2 ) 1 2 7 . 2 4 7 - 2 5 J.

Topical vitamin C protects porcine skin from ultraviolet

radiation-induced damage
Du!(e University Medical Center. Division of Dennatology. Durham. NC. U.S.A.
'NCSU School of Veterinary Medicine, Raleigh. NC. U.S.A.
Accepted for publication 8 April 1992

Summary Ultraviolet radiation damage to the skin is due. in part, to the generation of reactive oxygen species.
Vitamin C (i.-ascorbic acid) functions as a biological co-factor and antioxidant due to its reducing
properties. Topical application of vitamin C has been shown to elevate significantly cutaneous levels of
this vitamin in pigs, and this correlates with protection ofthe skin from UVB damage as measured by
erythema and sunburn cell formation. This protection is biological and due to the reducing properties
of the molecule. Further, we provide evidence that the vitamin C levels of the skin can be severely
depleted after UV irradiation, which would lower this organ's innate protective mechanism as well as
leaving it at risk of impaired healing after photoinduced damage. In addition, vitamin C protects
porcine skin from UVA-mediated phototoxic reactions (PUVA) and therefore shows promise as a
broad-spectrum photoprotectant.

In a spate of research activity over the past few years, the oedema)." Finally, a recent report has shown topically
conclusion that oxygen free radicals are involved in applied vitamin E or vitamin C effective in moderating
cutaneous sun damage has become well accepted.'"* low level, chronic UVB (but not UVA) damage to mouse
The proposal that the skin's native antioxidant protec- skin.'"*
tion system breaks down under this 'photo-onslaught" Vitamin C (L-ascorbic acid) has received renewed
has been suggested: for example, it has been shown that attention as an antioxidant of biological importance in
in response to acute UV exposure, epidermal superoxide systems varying from blood ('. . . an outstanding anti-
dismutase activity declines.^ ^ Glutathione reductase. oxidant in human blood plasma'' "^) to skin."' On the skin
catalase and reduced glutathione levels in skin have also surface, it and thioredoxin reductase may be very
been shown to decline after only one exposure to UVB important in eliminating reasonably long-lived free
radiation:'^ except for glutathione. the same results hold radicals.'^'' Although vitamin C is valuable as a free-
for UVA-visible irradiation of skin.*^ Loss of other skin radical quencher, it may itself be susceptible to UV-
antioxidants (ubiquiones. vitamins E and C. etc.) ranged mediated destruction. As such, we investigated the
from minimal to significant. Prophylactic antioxidant ability of pharmacological amounts of topically applied
therapy has been investigated in so far as it has been vitamin C to increase cutaneous concentration of this
shown that increased dietary antioxidants (including vitamin and to lessen UV damage to porcine skin.
vitamin C) lessen UV-induced skin lesions.'"" that
topically applied, liposomally encapsulated superoxide
dismutase reduces a IlV-medlated Ioss of the native Methods
enzyme" and that free-radical scavengers inhibit ultra-
i,-Ascorbic acid (A.C.S. reagent). D-isoascorbic acid. 1.2
violet radiation's formation of 'sunburn cells' in excised
propanediol (propylene glycol) and hydroxypropylcellu-
skin.'^ In addition, topical application of several free-
lose (average MW^ 500.000} were purchased from
radical scavengers on guinea-pig skin lessened sub-
Aldrich. Experiments utilized a 10% L-ascorbic acid
sequent photosensitized reactions (erythema and
(w/v) solution in 20% propylene glycol (v/v) with 0-5%
hydroxypropylcellulose (w/v) incorporated as thickener.
Correspondence: Dr D.Darr, Box J I J T , Duke University Medical Stored at 4°C. this solution was stable for 1 week as
Center, D u r h a m , NC 2 7 7 1 0 , U.S.A. measured spectrophotometrically.

248 D.DARR et al.

Animals tion of the the two values gave an estimate of the

penetration of the ascorbic acid into and through the
Domestic Yorkshire pigs (males weighing 32-45 kg)
were used in this study because of the similarity of their
skin to humans. They were housed in barns at the North
Carolina State Veterinary School and fed a standard diet. Histology
Prior to experiments, animals were anaesthetized with
The experimental sites were biopsied with a 4-mm
ketamine/xylazine (2 mg/kg) and their backs clipped
punch at 24 h for UVB experiments. For UVA experi-
with animal shears. During irradiation or before biopsy,
ments, biopsies were taken at 48 h. Like humans.' '* these
all animals were additionally placed under halothane
Yorkshire pigs showed variable, delayed responses to
anaesthesia. Typically four animals were used per
PUVA treatment. The biopsies were tixed in formalin and
processed for routine histology. For "sunburn cell'
analysis, duplicate haematoxylin and cosin-stained sec-
tions of triplicate biopsies for each experimental site were
UV source analysed for sunburn cells (basal keratinocytes having
A bankof two WestinghouseFS4() lamps was used in the pyknotic nuclei as well as eosinophilic cytoplasm) and
bulk of the studies. They were placed above a restrained normalized to the 4-mm punch diameter. The number of
animal at a distance of approximately 10 cm. The sunburn cells per given condition was calculated in this
intensity at that distance measured ()'45-()-5 mW/cm- way from four animals per experiment.
using a National Biological Corporation model UVR
LMH06C photodetector. Alternatively, specially made
HPLC analysis of vitamin C levels in skin
Philips TL 4()W/01 fluorescent bulbs (four housed in a
planar array) were used experimentally. They were To determine vitamin C concentrations in the skin.
placed above the restrained animals to achieve an HPLC with electrochemical detection was used.''* Briefly,
intensity of ^ 0 - 8 mW/cm^. In the PUVA experiments, after extensive washing, skin was excised at the subcuta-
four GE F4() BL fluorescent bulbs were used. These were neous boundary from two or more animals using a 4-
placed above the animals so that the measured intensity mm punch, and immediately frozen in liquid nitrogen.
was 2 mW/cm^ (using an International Light Model IL Weighed skin samples were placed on a glass dish in cold
440 detector). In these experiments. 8-methoxypsoraIen 5% metaphosphoric acid (MPA) and finely minced with
was formulated at a 0 1 % (w/v) concentration in 90% surgical scalpels. Following a wash with additional MPA
ethanol. The solution was applied to the sites at 10 /ig/ the minced skin sample was subjected to three cycles of
cm". Additional experimental details are given in the freeze-thawing ( — 70°C). Samples were spun down and
Figure and Table legends. the supernatant filtered through a 0 22 ^m filter. All
samples were frozen at — 70°C until analysis.

Percutaneotis absorption
Laser-Doppler measurement of cutaneous blood flow
Skin was removed from a non-treated animal at the time
Laser-Doppler velocimetry (LDV) is an accepted tion-
of killing to a depth of 500 /im with a Padgett Model B
invasive technique for monitoring relative blood fiow
dermatome. The skin was cut into 1 5-cm squares and
and has been used in quantifying erythema caused by
placed between the upper and lower chambers of a
UV irradiation.-" Cutaneous blood flow was estimated by
modiiied Franz cell. Vitamin C samples (20 //I) contain-
LDV using a LaserFlo™ BPM model 403 24 h after UV
ing 1 ^Ci '•^C-i.-ascorbic acid (Amersham) were pipetted
irradiation of vitamin C or vehicle-treated sites on four
on to skin in multiple cells. At 24 and 48 h. 100-/(1
aliquots were removed from the buffered saline reservoir,
mixed with scintiliant and counted on an LKB model
1219 scintillation counter. The skin sample itself was Results
then washed by dipping for 10 s into three consecutive
water baths (this was shown to remove virtually all Skin levels of vitamin C after topical treatment with this
surface counts) and burned in a stream of oxygen in a vitamin
Packard model 306 tissue oxidizer. The '"^CO^ released Using modified Franz cells, percutaneous absorption
was trapped in scintiliant and also counted. Combina- studies with '^C-labelled ascorbic acid showed that

Table 1. Increase in cutaneuus vitamin C levels by topical treatment 'I'able 2. Reduction in UVB-mediated sunburn cell formation by topical
with this vitamin. Four-millimetre biopsies from each treatment site on vitamin C, Paired sites on the backs ofdomestic pigs were pretreated for
each of three animals were weighed, pooled and extracted tis in ? days, and 30 min before irradiation, with a 10% vitamin C solution
Methods. Samples were analysed in duplicate by HPLC with electro- or the vehicle control 1100 fii/U) cm'). The sites were then irradiated
chemical detection and peak areas compared to known standards. with the indicated dose of UVB from a bank of Philips Tl, 4(1W/1)I
Values are mean ± SI! oi" duplicate analyses of two pooled samples. bulbs. At 24 h. three biopsies were taken from each site. Duplicate
sections from each biopsy were stained with hacmatoxyiin and eosin
and the number uf 'sunburn cells' counted. Cell counts from the six
Vitamin C concentration histological sections per condition were normalized to the 4-mm biopsy
Site ;jg/g wet weight diameter. Values are the mean±SE, Numbers in parentheses equal
animal number. From this DVsourcein these animals l()()mj/cm- =: I
Vehicle-treated MVD
Vitamin C-treated 542-6±5O

No. of sunburn ceUs/4-nim biopsy

* P<0-005, Student's Mest.
Condition (dose 400 ml/cm-')

0 - 7 ± 0 - l % (meaniSEM) passed through and Control (121 331±S 1

8-2 ± 1 •l%(mean±SEM) ofthe applied dose (2 mg) was Vitamin C-treated (12)
present within a 500-/mi thick section of porcine skin 48
h after topical application. However, this technique only •P<0-0()5: paired 1-test.
follows the movement of the radioactive label and as
vitamin C is notoriously unstable, we also measured was a bank of four Philips TL 40W/01 bulbs whose
cutaneous levels of vitamin C proper by HPLC. Table 1 is emission centres around 3 11 nm with little or no output
derived from an experiment in which three animals were below 305 nm (Fig. I), The L-ascorbic acid in this
treated topically with vitamin C. then thoroughly formulation has virtually no absorbance above 300 nm.
washed, and full thickness skin samples excised and Indeed, if this solution is applied to a piece of quartz glass
assayed for ascorbic acid levels. Additional experiments in a similar manner to that employed experimentally
with different animals have confirmed that skin levels of (^H)O /(I/IO cm-) and allowed to air dry for JO-fiO
vitamin C increase 4-40-foId or more following multiple min, virtually 100% of the energy at wavelengths
treatments with this vitamin. Assuming porcine skin to greater than 300 nm is transmitted through the quartz
be approximately 75% water (a fair approximation), cuvette. Therefore, the protection seen against UVB is
both methods give values of vitamin C concentration in biological, not physical. Responses to these Philips bulbs
the skin of approximately 3-4 mM after topical appli- (sunburn cell formation and erythema) were indis-
cation. Because of the obvious gradient, even higher tinguishable from those seen with more typical UVB
levels of this vitamin can be assumed in the upper levels bulbs, e.g. Westingbouse FS40,
ofthe epidermis and stratum corneum while somewhat
lower in the reticular dermis.
100 400

Protection of swine skin from UVB by topical vitamin C

Topical application of vitamin C protects porcine skin
from UVB damage. Using sunburn cell (dyskeratotic
basal epidermal ceils) formation as a marker for UV-
induced damage. Table 2 shows a significant reduction
in the UV-mediated formation of this cell type by topical
Eipplication of vitamin C. Although a single application
(approximately 15-30 min prior to exposure) is gener- 220 240 260 280 300 320 340 360 380
ally sufficient to give UV protection, some animals Wavelength (nm)
showed better results if the vitamin C was applied more
Figure 1. Absorption spectrum of L-ascorbic acid versus emission
than once, and as such, a typical protocol called for spectrum of Philips Tl, 4()W/01 fluorescent bulbs. The absorbence of
several applications ofthe vitamin to the skin prior to the vitamin C in the test solution was measured through quartz
irradiation. cuvettes using a Shimadzu Model 2M) spectrophotometer. The
emission spectral data was obtained from the manufacturer. I—)
The protection noted in Table 2 is not, however, due to Relative absorbence of r,-ascorbic acid in experimental formulation.
a 'sunscreen effect'. The UV source in those experiments ( ) Emission spectrum ofT!, 4()W/0] fluorescent bulb.
250 D.DARR et al.

Table i. Reduction in lIVB-elicited erythema by topical vitamin C. be mimicked by its isomer isoascorbic acid, assuming
Paired sites on the backs of domestic pigs were prelreated for 1 week, cellular uptake and/or in vivo penetration rates are
and i() min before irradiation, with a 10% vitamin C solution or the
vehicle control, Thesites were then irradiated wilh approximately 2-J
similar for the two molecules (not an absolute, e.g. Kipp
MFD of UVB radiation ( ^ 1 50 m|/cm- from two Westinghouse FS40 and Schwartz^'). Table 4 shows that both ascorbic acid
bulbs}. At 24 h. triplicate measures ofcutaneous blood flow from each and isoascorbic acid exhibit identical protective abilities
site were taken using a TSI Laserl'lo''" model 403 laser-Doppler against UVB damage. In a separate experiment, the two-
velocimeter. Adjacent non-irradiated skin was used for 'background'
blood tiow determination. Values are mean ±S[i of arbitrary bUxxJ flow
electron oxidation product of ascorbic acid, dehydro-
values after subtraction of'background' flow. Numbers in parentheses ascorbate (DHA). provided little or no visual protection
equal animal number against UV erythema when compared with its parent
molecule (data not shown).
Condition b!o(jd flow
Prnteclion of piij skin frow UVA-mediated phototoxicily bij
Control (4) topical vitamin C
Vitamin C-treated (4)
Although nowhere near as crythemogenic or carcino-
genic as UVB. UVA is responsible for many biological
• ('<t)-()t)l; paired t-test.
effects, at least partially because the relative intensity of
UVA in solar UV is many times that of UVB. Because of
the aforementioned ineffectiveness of low-dose UVA to
A second hallmark of UVB-induced sunburn is ery- elicit measurable biological responses in skin, we chose
thema. One quantitative tneasure of erythema is provided to use 8-niethoxypsoralen plus UVA (PUVA) to increase
by laser-Doppler velocimetry. Using this technique, skin damage, i.e. sunburn cell formation, and allow us to
increased skin blood flow occurred 24 h after a 2- 3 MHD quantify possible protection by topical vitamin C. This
dose of TlVB was measured. In this experiment, blood method has been used to test UVA sunscreen efficacy,
tlow approximately doubled compared with adjacent and to assess the role of reactive oxygen in PUVA
non-irradiated control sites in response to the radiation. damage in vivo."-'^-'' Table 5 shows that topical
Pretreatment with topical vitamin C halved this increase application of vitamin C does inhibit the PUVA-mediated
(Table i). production of this cell type, indicating that vitamin C is
capable of ameliorating a UVA-mediated phototoxic
response in skin. Indeed, in experiments using UVA
Protection against UVB damage by reducing and non- doses greater than 500 mj/cm-. gross pathological
reducing analogues of vitamin C changes (leucocytic infiltrates, blistering, etc.) are seen
The antioxidant protection of skin by vitamin C should
Table S. Reduction in psoralen-LIVA-mediated sunburn cell formation
by topical vitamin C, Paired sites on the backs of domestic pigs were
Table 4. Comparative effectiveness of ascorbic acid vs. isoascorbic acid pretreated for i days with a 1(1% vitamin C solution or the vehicle
in preventing UVB-induced 'sunburn cell' formation. Solutions were control. One hour prior to irradiation. S-methoxypsoralen |()-l% w/v
10% Iw/v} ir( 20% propylene glycol: 1()% hydroxy-propylcellulose. in 90%) ethanoH was evenly applied to all sites (10 /ig/cm-). Thirty
Paired sites on the backs of domestic pigs were treated daily for 1 week, minutes later a final treatment of vitamin C or vehicle (iOO ;/i| was
and JO min before irradiation, with 2-3 MHU of UVB (Westinghonse applied. The sites were then irradiated with a bank of OIE 1 40 BL
FS4() biiibsl. After 24 h. 4 mm biopsies (three per site) were taken, lluorescent tubes (emission peak ^ ibU nm) at the indicaled dose. In
processed for histology, stained with haematoxylin and eosin and these animals, this is approximately equal to three times the minimum
'sunburn cells' counted as in Table 2. Numbers in parentheses equal phototoxic dose (MPD). Biopsies were taken at 48 h. Sunburn cell
animal number. analysis was performed as in Table 2. Numbers in parentheses equal
animal number

Treatment No. of sunburn cells/4 mm % Reduction

No, ofsunburn cells/4-mm biopsy
Vehicle (8) 172±6O Condition (dose 500 m|/cm')
L-ascorbic acid (8) 10'0±3-7* 42
Vehicle (8) ]8-3±4-5 Control (10) tl4-5±17'8
isoascorbic acid (8) 1()'8±2'5* 41 Vitamin C-treated (10) 48'4±]()-r

• P < 0 - 0 2 ; paired I-test. ' P<0-002; Paired 1-test.


Table 6. UV-mediated loss of cutaneous vitamin C. Sites on three Discussion

animals were treated with topical vitamin C Idaily for 5 days). One site
was then covered and the other exposed to approximately 4 MKD of
UVB. Immediately after the exposure, the sites were thoroughly Although the meehanism(s) of ultraviolet damage to
washed. Multipie biopsies were taken from the two sites on each skin is certain to be complex, reactive oxygen species are
animal, frozen in liquid nitrogen and stored at 7 0 ^ until analysed. likely to be a major contributor to this damage. Unlike
In two separate analyses, biopsies from the irradiated or non-irradiated model systems, UV probably generates the entire spec-
sites from the three animals were pooled, weighed and extracted for
HFLC quantification of vitamin C content. Vitamin C-treated, non- trum of oxygen radicals, radical-derived species and
irradiated values were treated as 100% and irradiated values com- other reactive molecules. In this regard, ascorbic acid
pared with them. Numbers represent the average of tive HPLC should be particularly effective in interfering with the
determinations per experimental condition. UV-mediated generation and/or propagation of the
reactive oxygen species, as it reacts with or quenches
Treatment % Vitamin C remaining the superoxide anion.^^ the hydroxyl radical,-'' singlet
oxygen^^ and hypochlorous acid.^*^ In addition, vitamin
Vitamin C-treated C is the primary replenisher of vitamin E. the pre-
Non-irradiated 100
eminent inhibitor of lipid peroxidation.-'*^" Durham et al.
showed that increases in dietary vitamin C led to a
statistically significant reduction in subsequent UV-
elicited tumours.'" We believed that topical application
would achieve a much higher tissue concentration of
in vehicle-treated sites, whereas vitamin C pretreated this vitamin compared with oral intake. Experiments are
sites maintained substantially normal histology {unpub- in progress to specifically test this. However, conserva-
lished data). No evidence was found lor any interaction tive calculations based on the percutaneous absorption
between vitamin C and the ground-state psoralen and HPLC experiments give ascorbic acid concentrations
molecule (measured spectrophotometritally) so the in the millimolar range in full or partial thickness skin
protection is presumed to be due to either quenching of after several days of topical application. These levels
the excited state psoralen molecule by vitamin C or its would be expected to be even higher in the superficial
scavenging of reactive oxygen species known to be layers ofthe skin. i.e. the epidermis and upper papillary
generated during PUVA therapy.' * dermis. In vitro evidence suggests that at these concen-
trations ascorbic acid strictly acts as an antioxidant.''
Experiments with the isoascorbate and dehydroascor-
Loss of ciitaneotts vitamin C bij UV radiation bate also show the effects to be biological and dependent
on the reducing property of this vitamin. It is of note that,
Vitamin C is essential to tissue repair, a tact noted many although it is known that dehydroascorbate can be
years ago. We, and others.^' hypothesized that UV metabolically converted back to the parent ascorbic acid
damage may in part be due to a depletion of the skin's molecule within cells, this apparently does not occur
natural defences by the radiation. In fact. Fuchs et al.~ under our experimental conditions, or more likely, the
have shown UV-mediated losses of several key antioxi- bulk of the protection occurs in the metabolically
dants in mice exposed to UVB. Vitamin C levels (mea- inactive cell layers, i.e. the stratum corneum. In defense
sured spectrophotometrically) declined, although not of this thesis, using vitamin C solutions at either pH 2 • 5
significantly. We found it difficult to docutiient ultra- (totally un-ionized) or 5-0 (totally ionized) makes no
violet radiation-induced changes in vitamin C concen- difference in protection against UVH damage even
tration in lull thickness skin: vasodilatation and skin though the un-ionized form traverses the skin to a much
inflammation may lead to variable levels. To circumvent greater degree (data not shown). In addition, as pre-
this problem, we used several days of topical vitamin C viously mentioned, protection is noted after only one
treatment to raise skin levels ofthe vitamin. The treated application of the vitamin. 15-JO min before UV
areas were then UVB-irradiated. washed thoroughly to exposure, presumably long before dehydroascorbate
remove any free vitamin C. and the cutaneous vitamin C could reach the metabolically active layers of the
concentration measured by HPT.C, 'I'able (i shows a epidermis. Thus, it is not unexpected for DHA to be
dramatic UV-mediated diminution ofthe tissue vitamin inactive in this model and reasserts the importance ofthe
C. It thus appears likely that the natural vitamin C levels reductant nature of vitamin C in the lessening of damage
ofthe skin are also subjected to this loss, perhaps to near nt)ted.
depletion in areas.
252 D.DARR ct aL

These studies in part employed fluorescent UVB- those leading to chronic UVA damage is a subject for
emitting bulbs, wavelengths long known to be respon- debate. As there are no good models for UVA damage
sible for eliciting skin cancer. Multiple exposures to alone, PUVA has become an acceptable experimental
significantly lower doses than used in this study has been model for predicting product efficacy in providing protec-
shown to be carcinogenic.'- Additionally, reactive oxy- tion against longer wavelength ultraviolet radiation
gen species/oxygen radicals have been implicated in the (UVA).^''^'* Topical vitamin C treatment may thus prove
tumour promotion phase.'-' " One study has shown a to be a particularly effective broad spectrum photopro-
positive effect of topical vitamin C in preventing the onset tectant.
of tumourogenesis in mice.'^ This is an ongoing avenue Participation of reactive oxygen species in. and
of study. antioxidant prevention of. UV damage is now well
Quantitative measures (sunburn cell counts and laser- documented.' ^ i*''^^-' We report that topically applied
Doppler measurements) show that topical vitamin C L-ascorbic acid is effective in preventing UVB- and
treatment can reduce UVB-mediated increases in these psoralen-UVA-mediated damage to pig skin. The mech-
parameters ^ 4 0 % at moderate UV doses ( i - 4 MED). anism(s) of this protection is presumed to be due to its
Tissue hypoxia has also been shown to reduce UVB- 'antioxidant' status. The protection may be due to
induced formation of suburn cells, again by approxima- vitamin C directly reacting with, or quenching certain
tely 40-50%.*^ These data suggest that, although reactive species, or by regenerating vitamin E and thus
important, reactive oxygen species cannot be considered participating in the inhibition of UV-induced iipid peroxi-
the only source of UVB damage. Certainly, historical dation. As significant free-radical reducing potential is
experiments showing direct (non-oxygen-dependent) found in the extracellular spaces,"' topical vitamin C
interaction between UV (particularly UVB) and impor- may significantly bolster this free-radical reducing capa-
tant biological targets, e.g. DNA. are well known. city and thus beneiit the skin. The skin's innate antioxi-
Vitamin C obviously need not interfere with these other dant defense system is complex. We believe that vitamin
reactions. C is an important part of the UV defense system of the
skin, although its exact role has not yet been fully
Recent studies hint at a potential drawback to many
ordinary sunscreens. They absorb the sunburning rays
of the sun so effectively that they allow the user to stay Finally, vitamin C is essential to tissue and/or wound
outdoors much longer than would ordinarily be possible. repair. Perhaps UV damages skin not only directly, but
Unfortunately, this allows considerable amounts ofthe indirectly via inactivation of protective agents such as
longer wavelength, non-absorbed UVA to enter the skin, superoxide dismutase. catalase. etc. We provide evidence
which may be related to an increased risk of certain skin that additionally, skin vitamin C levels can be dramati-
cancers or ageing changes. In addition, there appears to cally reduced in response to UV radiation. Thus, in
be a stronger correlation between UVA-induced skin addition to a loss of antioxidant status, it may be
damage and oxygen radical formation than with UVB. '^ hypothesized that other tissue damage may be less
As we have now shown, topical vitamin C is also effective effectively repaired in this 'vitamin deficient' state.
in tnoderating UVA-mediated phototoxic reactions in Therefore, replenishment of skin vitamin C would be an
skin, apparently due to its antioxidant status. Earlier in important pharmacological intervention against sun
vitro work showed that ascorbic acid and anaerobiosis damage.
were equally effective in inhibiting PUVA-mediated
intermolecular cross-link formation of a-crystallin. pro-
viding further evidence for the participation of reactive Acknowledgments
oxygen species in at least some psoralen-UVA photo-^
sensitized reactions. ^^ These results are in contrast with This work was supported in part by the Dermatology
investigations performed in mice. i.e. vitamin C was Foundation (Dermik. Inc.). the Skin Cancer Foundation
ineffective in preventing []VA-induced skin wrinkling.'^ and NIH Grants 5R37 AR17128 and 5R01 AR28 3O4.
Considerable differences, e.g. long-term UVA exposure
vs. acute PUVA treatment, mouse vs. pig skin, as well as
measured endpoints. exist between these two studies. References
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