A Short-Term Screening Protocol, Using Fibrillin-1 as a Reporter Molecule, for Photoaging Repair Agents Rachel E.B. Watson, Nicholas M. Craven, Christopher.

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A Short-Term Screening Protocol, Using Fibrillin-1 as a Reporter Molecule, for Photoaging Repair Agents Rachel E.B. Watson, Nicholas M. Craven, Christopher E.M. Griffiths Journal of Investigative Dermatology Volume 116, Issue 5, Pages 672-678 (May 2001) DOI: 10.1046/j.1523-1747.2001.01322.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Occlusion of 0.025% RA and 5% SLS for 4 d produces equivalent, significant increases in epidermal thickness of photoaged skin. Bars represent mean ± SEM; n = 8. Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Long-term treatment of chronically photoaged skin with topical RA increases the abundance of fibrillin-1. Representative photomicrographs of fibrillin-1 staining in baseline (a) and treated (b) skin samples. Scale bar equivalent to 100 μm. (c) Immunohistochemical evaluation of baseline (open bars) chronically photoaged facial skin and after long-term RA use (closed bars). Staining was assessed on a semiquantitative ordinal scale. Bars represent mean ± SEM, n = 5. *p < 0.001. Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Immunohistochemical evaluation of the effect of 4 d occlusion of vehicle, RA, and SLS on procollagen I, collagen VII, and fibrillin-1 in photoaged skin. Immunohistochemical analyses of vehicle-treated (open bars), SLS-treated (hatched bars) and RA-treated (closed bars) photoaged skin. Staining was assessed on a semiquantitative ordinal scale. Bars represent mean ± SEM, n = 8. For fibrillin-1, asterisks denote treatment groups that are significantly different from each other (* p < 0.05; ** p < 0.001). Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Immunohistochemical analysis of the effect of 4 d occlusion of vehicle, RA, and SLS on collagen VII and fibrillin-1 in photoaged skin. Immunohistochemical analysis of collagen VII in (a) baseline photoaged skin, (b) vehicle-treated skin, (c) SLS-treated skin, and (d) RA-treated skin. Little observable difference was seen in any of the treated sites. Immuno histochemical analysis of fibrillin-1 in (e) baseline photoaged skin, (f) vehicle-treated skin, (g) SLS-treated skin, and (h) RA-treated skin. Increases in fibrillin-1 were observed following SLS-treatment, and to a greater extent, after application of RA. Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Transmission electron microscopy reveals the effect of 0.025% RA on the presence of microfibrils at the DEJ using a 4 d occluded patch test protocol. Photomicrographs show the papillary dermis and DEJ as characterized by hemidesmosomes (HD) of the basal keratinocytes and the lamina densa (LD) of the basement membrane. Scale bar: 100 μM. Severely photoaged forearm prior to treatment with 0.025% RA. Dystrophic elastin fibers (E) are observed at high levels within the papillary dermis. Interstitial collagen fibrils (C) are also apparent within this area as are anchoring fibrils (AF; white arrowheads) and some microfibrils (MF; black arrowheads). Following RA treatment, there is an increased presence of microfibrils proximal to the DEJ. Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 0.025% RA significantly increases fibrillin-1 mRNA in photoaged skin using a 4 d occluded patch test protocol. Quantitation of in situ hybridization histochemistry of vehicle-treated (open bars), SLS-treated (hatched bars), and RA-treated (closed bars) photoaged skin. The numbers of positive keratinocytes (a) or dermal fibroblasts (b) were quantitated per high power field (× 40). To account for alterations in epidermal thickness with treatments, epidermal data are expressed as a ratio of positive keratinocytes (KC) per μm of epidermis. Bars represent mean ± SEM, n = 8. Asterisks denote treatment groups that are significantly different from each other (* p < 0.01; ** p < 0.001). Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 0.025% RA significantly increases the number of keratinocytes expressing fibrillin-1 mRNA in a 4 d occluded patch test protocol. Representative photomicrographs of in situ hybridization for fibrillin-1 mRNA in (a) baseline photoaged skin, (b) vehicle-treated skin, (c) SLS-treated skin, and (d) RA-treated skin. Scale bar: 100 μm. Journal of Investigative Dermatology 2001 116, 672-678DOI: (10.1046/j.1523-1747.2001.01322.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions