Keratinocytes Influence the Maturation and Organization of the Elastin Network in a Skin Equivalent1 Florence Duplan-Perrat, Odile Damour, Caroline Montrocher,

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Keratinocytes Influence the Maturation and Organization of the Elastin Network in a Skin Equivalent1 Florence Duplan-Perrat, Odile Damour, Caroline Montrocher, Simone Peyrol, Guillaume Grenier, Marie-Paule Jacob, Fabienne Braye Journal of Investigative Dermatology Volume 114, Issue 2, Pages 365-370 (February 2000) DOI: 10.1046/j.1523-1747.2000.00885.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Histologic analysis of a DE and SE. (a, c) DE, (b, d) SE. DE were harvested after 30 d (a) and 60 d (c) of fibroblast culture and SE after 30 d (b) and 60 d (d) after seeding (Hematoxylin Phloxin Safron staining). Ep, epidermis; D, dermis; DEJ, dermal–epidermal junction; ECM, extracellular matrix; Fb, fibroblast. Scale bar: 70 μm. Journal of Investigative Dermatology 2000 114, 365-370DOI: (10.1046/j.1523-1747.2000.00885.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Detection of fibrillin-1 and elastin in human skin, DE, and SE. (A, B) Human skin, (C–F) DE, (G–J) SE. Ep, epidermis; D, dermis; DEJ, dermal–epidermal junction. Fibrillin labeling in DE at both 30 d (C) and 75 d (E) of fibroblast culture and in SE at both 30 d (G) and 45 d (I) of fibroblast culture corresponding to 15 and 30 d of keratinocytes. Elastin labeling in DE at 30 d (D) and 75 d (F) and in SE at 30 d (H) and 45 d (J). Scale bar: 24 μm. Journal of Investigative Dermatology 2000 114, 365-370DOI: (10.1046/j.1523-1747.2000.00885.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 TEM observations of DE and SE. (A) DE, (B–G) SE. In DE on day 60 (A), rare hollow microfibrils (arrow) are only detected close to fibroblasts (F) into collagen-GAG-chitosan (*). Scale bar: 200 nm. SE on day 60 (B), deeper in the dermis, microfibrillar material (arrow) is abundant among collagen fibers, around active fibroblasts (F). Scale bar: 500 nm. In SE on day 60 (C), hollow microfibrils (open arrow) are closely associated with collagen fibers (black arrow). Scale bar: 200 nm. In 60 d SE (D), presence of a segment of basal lamina (arrow head) disposed along the basal keratinocytes (K). Groups of microfibrils form oxytalan fibers (arrow), which already appeared perpendicularly oriented to DEJ. Scale bar: 500 nm. In 30 d SE (E), microfibrils show hollow cross-sections (arrow). Scale bar: 200 nm. In 45 d SE (F), closely parallel microfibrils form bundles, probably because of new elastin embedding deposit. In SE on day 60 (G), amorphous elastin (arrow) has deposited on microfibrils. Scale bar: 200 nm. Journal of Investigative Dermatology 2000 114, 365-370DOI: (10.1046/j.1523-1747.2000.00885.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions