Pigmentation after single and multiple UV-exposures depending on UV-spectrum
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- Ravnbak, M.H. & Wulf, H.C. Arch Dermatol Res (2007) 299: 25. doi:10.1007/s00403-006-0728-3
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Minimal pigmentation dose (MMD) after a single UV-exposure is well investigated. Whereas only few studies have established MMD after multiple UV-exposures and mainly in fair-skinned persons. The purpose of this study was to establish MMD 1 week after, respectively, one and five UV-exposures in volunteers with a large variation in constitutive pigmentation. A total of 52 volunteers (skin Types II–V) had skin pigmentation quantified by reflectance spectroscopy. They were UV-exposed on the back for 1 and 5 days using a Solar Simulator, narrowband UVB, broadband UVA and UVA1. For all sources a higher dose was needed the more pigmented the skin, except for UVA1. After one UV-exposure, we found a significant positive linear correlation between UV-dose to one MMD, skin type and pre-exposure skin pigmentation. After five UV-exposures the positive linear correlation between UV-dose and MMD and skin type was only significant for narrow band UVB, pre-exposure skin pigmentation was significant also for Solar Simulator. For UVA and particularly UVA1 the MMD was independent of pre-exposure pigmentation. The number of SED to MMD is therefore almost the same for very fair-skinned and dark-skinned persons. Pre-exposure pigmentation was clearly more predictive of MMD than skin type. 50% of MMD equals a pigmentation increase of 1%. The shorter the wavelengths the higher the SED to produce MMD. Solar was the least melanogenic and UVA1 the most melanogenic. For the UVB-sources a higher dose was needed the more pigmented the skin. For UVA the MMD was independent of pre-exposure pigmentation.
KeywordsSingle and multiple UV-exposuresPigmentation
Minimal erythema dose, the UV dose to elicit just perceptible erythema 24 h after a single UV-exposure
Minimal melanogenesis dose, the UV dose to elicit just perceptible tanning 7 days after the last UV-exposure. MMD will be expressed in SEDs 
Standard erythema dose, the UV dose that elicits just perceptible erythema in the most sensitive people in a group of very sun-sensitive but otherwise healthy individuals. One SED is defined as 100 J/m2 at 298 nm using the CIE erythema action spectrum
Sun and solarium exposure with the purpose of tanning is almost always performed as repetitive behavior. Nevertheless only few studies have investigated the minimal pigmentation dose (MMD) after multiple UV-exposures and then mainly in fair-skinned persons [1, 5, 10].
After the same sub-erythemogenic doses from UV-sources with different emission of UVB and UVA in skin types II and III, it was found that UVA irradiation enhanced pigmentation, whereas UVB irradiation did not . After five repetitive UV-exposures to 0.25, 0.5, 0.75 and one MED Kaidbey and Kligman found melanogenesis 7 days after the fifth UVA-exposure to only 0.25 MED, whereas no pigmentation was evident with UVB . Parrish found as well, in skin types II and III, that UVA was more melanogenic than erythemogenic; evidenced by MMD/MED < 1, whereas the opposite was true for UVB (MMD > MED). This was true for both single and multiple UV-exposures .
We chose to include also skin types IV and V to have a broader skin type (pigmentation) spectrum. The aim of this study was to examine the ability to tan, assessed as MMD, after single and multiple UV-exposures in relation to skin type and pre-exposure skin pigmentation. Moreover, we investigated how the melanogenic potential depends on the UV-spectrum (light-source) and finally by skin reflectance we investigated the objectively measured pigmentation increase.
Materials and methods
The local ethics committee approved the study (KF01-020/99), which took place outside the summer period to avoid seasonal variation of skin pigmentation. Sixty-two healthy volunteers, 34 females and 28 males, aged between 19 and 44 years (mean age 25 years) were recruited by advertisements in local papers. The volunteers were enrolled in the study after giving written consent. Skin diseases, sunbathing or exposure to artificial tanning 3 months prior entering into the study excluded participation as did medication with glucocorticoids, non-steroidal anti-inflammatory drugs, anti-histamines or retinoids. Volunteers with different skin types  were selected to achieve a broad range of skin pigmentation. A total of 10 volunteers only completed the single UV-exposure and then dropped out. Seven volunteers had self-evaluated skin type II, 16 skin type III, 16 skin type IV and 13 skin type V. The ethnic backgrounds varied from Scandinavians (n = 16), Hispanics (n = 12), and Asians (n = 12) to Indians and Pakistanis (n = 12). The pre-exposure pigmentation on the back varied from 13 to 55% .
At entry, the lower back of each volunteer was sub-divided into four areas. One area was allocated to UV irradiation with narrow-band UVB (nUVB) (TL01) (Philips, Rosendaal, Holland), the second area to UV irradiation with Solar Simulator (Solar)(Solar Light Co., Philadelphia, PA, USA), the third area to UV irradiation with broad-band UVA (bUVA) (Cleo-performance) (Philips) and the fourth area to UV irradiation with UVA1 (TL10) (Philips). To avoid anatomical differences in skin UV-sensitivity, the allocation of the four UVR sources was carried out by randomization. The pre-exposure skin pigmentation in the test areas was measured by a reflectance meter. To calculate the objectively measured pigmentation increase, the skin pigmentation was measured again by the reflectance meter 7 days after, respectively, the single UV-exposure and the five UV-exposures.
To determine the MMD after a single UV-exposure six UV-doses with 25% increments were used. MMD was determined clinically 7 days after the UV-exposure. When the individual MMD was determined, the volunteers were exposed to the four UV-sources in four new areas for five consecutive days in 24 h intervals.
About 7 days after the fifth UV-exposure MMD was evaluated clinically. The daily dose to give the same measurable pigmentation increase after one and five UV-exposures was calculated.
Throughout the 5 days of exposure we used six doses with 25% increments for the Solar Simulator. For the other UV-sources we used 100% increments since we did not know what to expect. Maximum dose for bUVA and UVA1 was two MMD as found after the single exposure. For the Solar Simulator and the nUVB maximum dose was one MMD to avoid burns.
The skin reflectance was measured using an instrument (UV-Optimize 555, Matic, Nærum, Denmark) with peak wavelength at 555 and 660 nm. The reflectance of these wavelengths is giving the melanin content or pigmentation of the skin, when the skin redness is zero. In the most pale persons, when the arm was drained from blood, the reflection of 660 nm light was 70%, which was defined to be 0% pigmentation, and 100% pigmentation corresponds to 0% reflection of the 660 nm light .
Dosimetry and UVR sources
The emission spectra and intensity of the radiation sources were measured before the start and regularly during the study using a hand held electronic spectroradiometer (Sola Tell, Sola-Hazard, 4D Controls Ltd., Redruth, UK). The manufacturer calibrated the spectroradiometer. The erythema activity of a UV source was measured in standard erythema dose (SED) as proposed on the 12th International Congress on Photobiology in 1996 [13, 14] and accepted by the CIE . The physical dimension of one SED is 10 mJ/cm2 at 298 nm using the CIE action spectrum . UV doses were therefore expressed in SED.
The Solar Simulator is equipped with a filtered xenon lamp that emits 8.7% in the UVB range and 91.3% in the UVA range (one SED corresponds to 1,490 mJ/cm2). The Solar Simulator conducts UV irradiation through six individually adjustable liquid light-guides with 10 mm Ø. During the irradiation the distance from the skin to the end of the light-guides was about 1 mm. Due to some instability and fluctuations in the intensity of the xenon lamp the intensity was measured just before and just after the exposure, and the average UV dose was used for the statistical analyses.
UV-exposure was interrupted in a specific area at erythema grade +++ or complaints of burning sensation irrespective of the erythema grade.
Regression analyses were performed to find the relation between UV-dose to one MMD and skin type and pre-exposure skin pigmentation. Not all the data were normally distributed, therefore we performed paired non-parametric tests (Wilcoxon signed rank test) to calculate if the differences in SED to one MMD between the UV-sources were significant.
Correlation coefficient r and P-values from Spearman’s rank correlation test of the relation between MMD and skin type (a) and between MMD and pre-exposure pigmentation for different light sources (b)
Number of exposures
(a) MMD and skin type
(b) MMD and pre-exposure pigmentation
A paired t-test was used to test if the differences in objectively measured pigmentation increase between the UV-sources were significant, when data were normally distributed, otherwise Wilcoxon signed rank test was performed. Moreover, for each of the UV-sources we used the Wilcoxon signed rank test to test if the objectively measured pigmentation increase was significantly higher after five compared to one UV-exposure.
Covariance analysis was performed to test for gender differences.
We used the Wilcoxon signed rank test to test if the difference in SED dose to one MMD between Solar Simulator (the least melanogenic) and UVA1 (TL10) (the most melanogenic) was significantly different after a single UV-exposure compared to five UV-exposures. P < 0.05 were considered to be significant.
Four of the Scandinavian persons (skin Type II) did not tan after a single UV-exposure to narrowband UVB or Solar Simulator and could therefore not be exposed for these UV-sources for 5 days.
Average UV-dose (SED) to give a minimal pigmentation (MMD) after one and five UV-exposures (a) and average UV-dose (mJ/cm2) to give a minimal pigmentation (MMD) after a single UV-exposure (b) in skin types II–V
(a) Average UV-dose (SED)
(b) Average UV-dose (mJ/cm2)
The UV-dose (SED) needed to produce a minimal pigmentation (one MMD) after, respectively, one and five UV-exposures are listed in Table 2 (part a) for each of the four UV-sources. The physical UV-dose (mJ/cm2) to MMD after a single exposure equivalent to SED is calculated in Table 2 (part b). Table 2 (part a) shows that a much lower UV-dose (SED) is needed to produce a minimal pigmentation as the UVB-content of the light source declines and the UVA-content increases. Except for Solar Simulator compared to nUVB.
The Solar Simulator was the least melanogenic of the four UV-sources and UVA1 was the most melanogenic. This means that the melanogenic effect is highly dependent on wavelength. The differences between the UV-sources were highly significant (P < 0.0001).
Single versus multiple UV-exposures
When giving five UV-exposures the daily dose needed to produce a minimal pigmentation (MMD evaluated visually) was ∼50% lowered compared to a single UV-exposure [Table 2 (part a)]. Therefore the cumulative dose is still higher after five UV-exposures.
The daily MMD dose to give the same measurable pigmentation percentage increase after a single and five UV-exposures was also calculated. It was found to be only ∼39% of the daily dose needed after a single UV-exposure.
Objectively measured pigmentation increase
The dose to 100% of MMD and 50% of MMD corresponds to the indicated objectively measured pigmentation percentage increase (reflectance)
After five UV-exposures the measurable pigmentation increase is on average 45% higher than after a single UV-exposure (for 100% MMD).
Ideally they should have been equal if the objective (measured) and clinical (visual) evaluation were identical. The daily dose to give the same measurable pigmentation increase after a single and five UV-exposures was calculated. It was found to be only ∼39% of the dose needed after a single UV-exposure.
To determine which of the parameters; skin type and pre-exposure pigmentation, that was more predictive of dose to one MMD, we compared the r and P-values obtained from Spearman’s rank correlation [Table 1 (parts a and b)]. The table shows a remarkable difference, pre-exposure pigmentation clearly preferable to skin type.
No gender differences were found in UV-dose to one MMD, except for bUVA. The slope of the bUVA curve for MMD in relation to pre-exposure pigmentation after one UV-exposure was steeper for females compared to males (P = 0.001), but no significant difference in pigmentation was found.
The Solar Simulator emits shorter waved UVB compared to nUVB (TL01). The short-wave UVB counts heavily to the SED and therefore the UVA dose only contributes to the SED by 14%. This is the reason why the Solar was less melanogenic than nUVB despite its higher UVA-content. Moreover does the Solar Simulator emit short-wave UVA that is less melanogenic than UVA1. nUVB was the most erythemogenic and a lower SED was needed to produce MMD than for the Solar Simulator .
Parrish found, in skin types II and III, that UVA was more melanogenic than erythemogenic; evidenced by MMD/MED < 1, whereas the opposite was true for UVB. This was true for both single and multiple UV-exposures on the buttocks .
The difference in SED to one MMD between Solar Simulator (the least melanogenic) and UVA1 (the most melanogenic) is more pronounced after five UV-exposures compared to a single UV-exposure (P = 0.001). Conversely to Parrish’ study , we did not use (sub-) erythemogenic doses, but equal melanogenic doses (MMD doses). Therefore our UVA-doses were relatively smaller, whereas our UVB-doses were relatively larger. The skin on the back is more sensitive to UV-irradiation than the buttock skin . We exposed our volunteers on the back and therefore presume that a smaller UV-dose was needed than the UV-dose to give minimal pigmentation on the buttocks.
Sub-erythemogenic UVA-doses induce pigmentation, whereas tanning from UVB-irradiation only occurs when preceded by erythema . This is in accordance with the finding of Kollias et al. that MED > MMD at 365 nm, and that MED < MMD at 295 and 305 nm and for UVB irradiation at 315 nm MED = MMD after a single UV-exposure in skin types I, II and V .
The larger dose intervals (100% increments) for the nUVB, bUVA and UVA1 compared to the Solar Simulator (25% increments) might be expected to result in a larger spread in these data than for the Solar Simulator. This was not the case, in fact, the range in SED to MMD for the Solar Simulator data was larger than for the other UV-sources [Table 2 (part a)].
The relation between UV-dose to one MMD, skin type and pre-exposure pigmentation was stronger after a single UV-exposure than after five UV-exposures [Table 1 (parts a and b)]. The reason for this is probably different initiation of protective mechanisms such as pigmentation and thickening of stratum corneum, already during the exposures. The relation to the pre-exposure pigmentation therefore becomes weaker during repeated exposures as the pigmentation may evolve with different speed and degree in individuals. We do not know if the minimum daily dose to tan after five exposures is reached or may be even lower after more exposures.
After, respectively, one and five exposures the objective (measured) and the clinical (visual) evaluation of pigmentation was identical for the different UV-sources (Table 3).
After five UV-exposures the objectively measured increase in pigmentation percentage was higher than after one UV-exposure, but this was only statistically significant for some UV-sources, nUVB (P = 0.007), UVA1 (P = 0.03) and for a mean of all UV-sources (P = 0.003). If the objective and the clinical evaluation of pigmentation had been identical, this difference would not occur. This means that the reflectance instrument (UV-optimize) is more sensitive at distinguishing pigmentation than the eye. The reason why more pigmentation is needed after five compared to one UV-exposure to be perceived by the eye is unknown. Regarding nUVB, we speculate that thickening of stratum corneum induced by the UV-exposures changes the optics of the skin and thereby changes how pigmentation is perceived. As thickening of stratum corneum mainly is caused by UVB  another yet unknown phenomena may play a role for UVA1. However, it is worth noting, that this did not apply to bUVA and the Solar Simulator.
In summary, the melanogenic effect of the UV-sources was highly dependent on wavelength: a lower SED dose was needed to produce a minimal pigmentation the longer the wavelengths. Solar was the least melanogenic and UVA1 the most melanogenic.
The UV-dose to one MMD was well correlated to skin type and pre-exposure skin pigmentation for one UV-exposure, except for the UVA1-source. For five UV-exposures MMD was only well correlated to skin type for nUVB whereas the relation to pre-exposure skin pigmentation was significant for both nUVB and Solar. This study showed that pre-exposure pigmentation was clearly more predictive of induction of pigmentation than skin type [Table 1 (parts a and b)].
Generally, for nUVB and Solar: the more pigmented a person was the higher SED dose was needed to tan. For UVA and particularly UVA1 the dose to one MMD was independent of pre-exposure pigmentation. A total of 50% of MMD corresponded to an invisible pigmentation increase of ∼1%, and was thus just measurable.
We thank engineers Jacob Heydenreich and Peter Alshede Philipsen, technicians Benedicte Wulf and Trine Ravn and medical students Pernille Fog Svendsen and Malene Rahbek Holm for their help in the project. This work was supported by Unilever, NJ, USA.