title: A short-term screening protocol, using fibrillin-1 as a reporter molecule, for photoaging repair agents.
abstract: Photoaged skin is characterized by coarse and fine wrinkles. The mechanisms of wrinkle formation are undetermined, but appear to be due to changes within the matrix of the dermis and at the dermal-epidermal junction. Previous studies have identified marked reductions in procollagens I and III, collagen VII, and the fibrillin-rich microfibrillar apparatus in this area. Topically applied all-trans retinoic acid can repair photoaged dermal matrix, but this takes at least 6 mo of treatment. In this study, we have examined the abundance and distribution of fibrillin-1 prior to, and following, 192 wk of all-trans retinoic acid treatment. We have further developed a short-term protocol to determine the utility of potential repair agents, using fibrillin-1 as the marker for outcome. Individuals with clinically assessed severe photoaging were recruited to the study (n = 8). 0.025% all-trans retinoic acid, 5% sodium lauryl sulfate (irritant control), or vehicle were applied under occlusion to photoaged extensor forearm. A fourth control area was also occluded. After 96 h, punch biopsies were taken under local anesthesia and processed for either transmission electron microscopy or snap frozen. Frozen sections were prepared for immunohistochemistry and in situ hybridization immunohistochemistry. Electron microscopy revealed aberrant elastic fibers in the papillary dermis of photoaged forearm skin, with sparse microfibrillar apparatus and interstitial collagen. After application of 0.025% all-trans retinoic acid, there was increased deposition of both these dermal matrix components, with the aberrant elastic fibers no longer apparent. Significant increases (p < 0.05) were observed at the protein and mRNA levels for fibrillin-1 following all-trans retinoic acid and sodium lauryl sulfate treatments, with all-trans retinoic acid having a significantly greater effect than irritant control (p < 0.001); however, neither application had significant effect on the abundance of collagen VII or its mRNA. Investigation of collagen I synthesis revealed no difference following treatments. To ascertain the clinical relevance of using fibrillin-1 as a marker for photoaging, facial skin was biopsied at baseline and after long-term (192 wk) topical all-trans retinoic acid treatment (n = 5). Biopsies were wax-embedded and sections prepared for immunohistochemistry for fibrillin-1. Significant increases in the abundance of the microfibrillar apparatus was observed proximal to the dermal- epidermal junction (p < 0.001) following long-term all-trans retinoic acid application. This study indicates that all-trans retinoic acid can significantly affect fibrillin-1 content in photoaged skin. Furthermore, fibrillin-1 can be used as a "reporter" molecule in short-term protocols for testing the utility of topical agents in the repair of photoaged skin.
authors.affiliation: Dermatology Centre, The University of Manchester, Hope Hospital, Salford, Manchester, UK.
authors.forename: R E
authors.initials: RE
authors.lastname: Watson
authors.forename: N M
authors.initials: NM
authors.lastname: Craven
authors.forename: S
authors.initials: S
authors.lastname: Kang
authors.forename: C J
authors.initials: CJ
authors.lastname: Jones
authors.forename: C M
authors.initials: CM
authors.lastname: Kielty
authors.forename: C E
authors.initials: CE
authors.lastname: Griffiths
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers; D004334:Drug Administration Schedule; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005260:Female; D000071838:Fibrillin-1; D000071837:Fibrillins; D006651:Histocytochemistry; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008875:Middle Aged; D012867:Skin / Q000378:metabolism / Q000473:pathology; D015595:Skin Aging / Q000187:drug effects*; D013997:Time Factors; D014212:Tretinoin / Q000627:therapeutic use; D054392:Adipokines
chemical_list: D015415:Biomarkers; D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; D014212:Tretinoin; C000605722:FBN1 protein, human; D054392:Adipokines