Fibroblasts and ascorbate regulate epidermalization in reconstructed human epidermis.
Skin equivalent model provides a new investigating system to study the role of extracellular matrix and dermal factors such as collagen, basement membrane components and fibroblasts (Fb) which contribute to cell-cell and cell-matrix interactions. Although basement membrane factors is known to play an important role in epidermal differentiation and epidermal-matrix adhesion, comparative effects of these extracellular matrix and dermal factors on the reconstruction of epidermis are little known. In this study, we investigated effects of type I collagen (Coll I), type IV collagen plus laminin (LAM) coated Coll I (Coll IV+LAM), and human Fb enriched Coll I (Coll I+Fb) on epidermal reconstruction. When human keratinocytes were cultured on three different gels containing Coll I, Coll IV+LAM and Coll I+Fb, basal keratinocytes were cuboidal and perpendicular to the dermo-epidermal junction only in the gel containing Coll I+Fb. Proliferation marker expression was prominent and differentiation marker expression was similar with those of normal skin in the gel containing Coll I+Fb than in the other gel models. Since ascorbate is suspected to exert an effect as a modulator of proliferation and differentiation in keratinocytes, we tested the effects of ascorbate on human epidermis reconstruction. When 25 microg/ml ascorbate was added, disordered arrangement of epidermis was disappeared and differentiation marker expression was similar with its expression in normal skin. These data indicate that human Fb and a modulator of proliferation and differentiation such as ascorbate are essential for epidermalization in reconstructed epidermis.
Wha Kim
Suk
S
Department of Plastic and Reconstructive Surgery, Seoul National University, Seoul, South Korea.
Lee
Il-Whan
IW
Cho
Hyun-Joo
HJ
Cho
Kwang-Hyun
KH
Han Kim
Kyu
K
Chung
Jin-Ho
JH
Song
Peter I
PI
Chan Park
Kyoung
K
D001205:Ascorbic Acid / Q000494:pharmacology*; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D024042:Collagen Type I / Q000494:pharmacology; D024141:Collagen Type IV / Q000494:pharmacology; D004338:Drug Combinations; D000078404:Epidermal Cells; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007797:Laminin / Q000494:pharmacology; D013194:Staining and Labeling
D015415:Biomarkers; D024042:Collagen Type I; D024141:Collagen Type IV; D004338:Drug Combinations; D007797:Laminin; D001205:Ascorbic Acid