Eduardo Ruvolo

Irradiation of Skin with Visible Light Induces Reactive Oxygen Species and Matrix-Degrading Enzymes

Daily skin exposure to solar radiation causes cells to produce reactive oxygen species (ROS), which are a primary factor in skin damage. Although the contribution of the UV component to skin damage has been established, few studies have examined the effects of non-UV solar radiation on skin physiology. Solar radiation comprises <10% of UV, and thus the purpose of this study was to examine the physiological response of skin to visible light (400-700 nm). Irradiation of human skin equivalents with visible light induced production of ROS, proinflammatory cytokines, and matrix metalloproteinase (MMP)-1 expression. Commercially available sunscreens were found to have minimal effects on reducing visible light-induced ROS, suggesting that UVA/UVB sunscreens do not protect the skin from visible light-induced responses. Using clinical models to assess the generation of free radicals from oxidative stress, higher levels of free radical activity were found after visible light exposure. Pretreatment with a photostable UVA/UVB sunscreen containing an antioxidant combination significantly reduced the production of ROS, cytokines, and MMP expression in vitro, and decreased oxidative stress in human subjects after visible light irradiation. Taken together, these findings suggest that other portions of the solar spectrum aside from UV, particularly visible light, may also contribute to signs of premature photoaging in skin.

Impact of Long-Wavelength UVA and Visible Light on Melanocompetent Skin

The purpose of this study was to determine the effect of visible light on the immediate pigmentation and delayed tanning of melanocompetent skin; the results were compared with those induced by long-wavelength UVA (UVA1). Two electromagnetic radiation sources were used to irradiate the lower back of 20 volunteers with skin types IV-VI: UVA1 (340-400 nm) and visible light (400-700 nm). Pigmentation was assessed by visual examination, digital photography with a cross-polarized filter, and diffused reflectance spectroscopy at 7 time points over a 2-week period. Confocal microscopy and skin biopsies for histopathological examination using different stains were carried out. Irradiation was also carried out on skin type II. Results showed that although both UVA1 and visible light can induce pigmentation in skin types IV-VI, pigmentation induced by visible light was darker and more sustained. No pigmentation was observed in skin type II. The quality and quantity of pigment induced by visible light and UVA1 were different. These findings have potential implications on the management of photoaggravated pigmentary disorders, the proper use of sunscreens, and the treatment of depigmented lesions.

Visible Light Induces Melanogenesis in Human Skin through a Photoadaptive Response

Visible light (400–700 nm) lies outside of the spectral range of what photobiologists define as deleterious radiation and as a result few studies have studied the effects of visible light range of wavelengths on skin. This oversight is important considering that during outdoors activities skin is exposed to the full solar spectrum, including visible light, and to multiple exposures at different times and doses. Although the contribution of the UV component of sunlight to skin damage has been established, few studies have examined the effects of non-UV solar radiation on skin physiology in terms of inflammation, and limited information is available regarding the role of visible light on pigmentation. The purpose of this study was to determine the effect of visible light on the pro-pigmentation pathways and melanin formation in skin. Exposure to visible light in ex-vivo and clinical studies demonstrated an induction of pigmentation in skin by visible light. Results showed that a single exposure to visible light induced very little pigmentation whereas multiple exposures with visible light resulted in darker and sustained pigmentation. These findings have potential implications on the management of photo-aggravated pigmentary disorders, the proper use of sunscreens, and the treatment of depigmented lesions.

The value of the ratio of UVA to UVB in sunlight.

The objective of this communication is to present the calculated ratio between UVA and UVB irradiance from sunrise to sunset and under a number of weather conditions. UVA plays an important role in the sun spectrum and a lot of attention has been paid lately regarding the protection of people from UVA. Solar spectra were collected in Kuwait City located at 29.3oNorth latitude (similar to that of Houston, TX) over a period of 8 months and under various weather conditions. Spectra were collected from 260 nm to 400 nm in 2 nm increments for solar elevation angles from 10o to 90o using a calibrated Optronics Laboratories OL‐742 Spectroradiometer. The measurements reported in this study the ratio of UVA (320–400 nm) to UVB (280–320 nm) in solar terrestrial radiation remains essentially constant and equal to 20 for the part of the day when the solar elevation is greater than 60o. Consequently the value of the ratio of solar UVA/UVB should be considered as equal to 20 for studies in photobiology and photomedicine. When the wavelength limiting the range of UVA and UVB is 315 nm (i.e. UVB: 280–315 nm and UVA: 315–400 nm) the ratio of UVA to UVB becomes equal to 41.

Diffuse reflectance spectroscopy for ultraviolet A protection factor measurement: correlation studies between in vitro and in vivo measurements

Background/purpose: Assessing the ultraviolet (UVA) protection factor of sunscreen formulations has been discussed for the past 20 years. The purpose of this study is to correlate the measurements of the UVA protection factor value (PFA value) via in vivo diffuse reflectance spectroscopy (DRS) and to compare this method with the in vitro method of measuring the PFA value, as well as with the in vivo persistent pigment darkening (PPD) and PFA methodologies.

Assessment of Mechanical Properties of Skin by Shearwave Propagation and Acoustic Dispersion

In this chapter, we present a methodology of documenting the angular anisotropy of skin elasticity with high sensitivity and dynamic range using the Reviscometer® RVM 600 and its use to document changes in keratinocyte morphology elicited by topical application of a reversible antagonist to keratinocyte nicotinic acetylcholine receptors (nAChRs). The method is based on determining the directional dependence of the speed of an acoustic shear wave on the skin surface at intervals of 3°. Based on the angular distribution of the resonance running time, we define two parameters: the anisotropy and the width of the angular dispersion. The mechanical properties of the skin are not isotropic (uniform in all directions), and there is a need to assess this angular anisotropy. During development, from infancy to adolescence, skin is expected to respond to tension isotropically to accommodate for growth, while in adulthood this isotropic behavior regresses due to site-specific habituation to tension. We find that with increasing age the anisotropy increases while the angular dispersion width decreases. Changes in viable epidermis keratinocyte morphology and stratum corneum moisturization may be assessed and documented by these parameters. The ratio of these values provides a sensitive parameter for the assessment of the directional behavior of the skin mechanical properties. This parameter provides a large effective dynamic range capable of demonstrating close to an order of magnitude difference in skin viscoelasticity from infants up to 75 years of age and documenting efficacy of topical treatments. Furthermore, we show that the direction of the angular anisotropy relates to the direction of the dermal cleavage lines as defined by Langer, indicating that the anisotropy of the mechanical properties of skin stems from structural parameters. Based on these results, we conclude that the proposed methodology is able to capture accurately age-related and morphological changes in the skin of the mechanical properties and demonstrate a structure-function relationship.

Use of spectral imaging for documentation of skin parameters in face lift procedure

In rhytidectomy the postoperative edema (swelling) and ecchymosis (bruising) can influence the cosmetic results. Evaluation of edema has typically been performed by visual inspection by a trained physician using a fourlevel or, more commonly, a two-level grading(1). Few instruments exist capable of quantitatively assessing edema and ecchymosis in skin. Here we demonstrate that edema and ecchymosis can be objectively quantitated in vivo by a multispectral clinical imaging system (MSCIS). After a feasibility study of induced stasis to the forearms of volunteers and a benchtop study of an edema model, five subjects undergoing rhytidectomy were recruited for a clinical study and multispectral images were taken approximately at days 0, 1, 3, 6, 8, 10, 15, 22 and 29 (according with the day of their visit). Apparent concentrations of oxy-hemoglobin, deoxy-hemoglobin (ecchymosis), melanin, scattering and water (edema) were calculated for each pixel of a spectral image stack. From the blue channel on cross-polarized images bilirubin was extracted. These chromophore maps are two-dimensional quantitative representations of the involved skin areas that demonstrated characteristics of the recovery process of the patient after the procedure. We conclude that multispectral imaging can be a valuable noninvasive tool in the study of edema and ecchymosis and can be used to document these chromophores in vivo and determine the efficacy of treatments in a clinical setting.

Comparison of skin responses from macroscopic and microscopic UV challenges

The minimal erythema dose induced by solar-simulated radiation is a useful measure of UV sensitivity of skin. Most skin phototests have been conducted by projecting a flat field of UV radiation onto the skin in an area greater than 15 cm × 15 cm with an increment of radiation doses. In this study, we investigated the responses of human skin to solar-simulated radiation of different field sizes. Twelve human subjects of skin phototype I-IV were exposed to solar-simulated radiation (SSR) on their upper inner arm or on their lower back with a series of doses in increments of 20% in order to determine the threshold dose to induce a minimal perceptible erythema response (MED). Each dose was delivered with a liquid light guide (8 mm diameter on the back or 6 mm on the upper inner arm) and with quartz optical fibers of 200 μm diameter. The resulting skin responses were evaluated visually and investigated with a reflectance confocal microscope and imaging. The erythema response to the microscopic challenge was always diffuse with no clear boundaries extending to several times the exposed site diameter at doses greater than 2 MED. The skin returned to normal appearance from the microscopic challenge after two weeks of exposure while change in appearance for the larger areas persisted for several weeks to months. This new modality of testing provides the possibility to study skin at the microscopic level with a rapid recovery following challenge.