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Epithelial regeneration from bioengineered skin explants in culture
Author(s) -
Mirastschijski U.,
Bugdahl R.,
Rollman O.,
Johansson B.R.,
Ågren M.S.
Publication year - 2006
Publication title -
british journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.304
H-Index - 179
eISSN - 1365-2133
pISSN - 0007-0963
DOI - 10.1111/j.1365-2133.2005.06997.x
Subject(s) - keratinocyte , basement membrane , explant culture , human skin , matrix metalloproteinase , wound healing , regeneration (biology) , pathology , epithelium , artificial skin , chemistry , zymography , in vitro , anatomy , biology , microbiology and biotechnology , medicine , immunology , biochemistry , genetics
Summary Background Artificial skin substitutes are beneficial in the treatment of chronic wounds although their performance relative to authentic human skin is unclear. Objectives We compared the rate of outgrowth and morphology of neoepidermis from a bioengineered skin construct (Apligraf ® ) with normal adult human skin explants on de‐epidermized human dermal growth substrate with or without intact epidermal basement membrane zone. Methods Epithelial outgrowth of air‐exposed cultures in serum‐supplemented keratinocyte medium was quantified by fluorescence imaging, morphology by light microscopy, biomarkers of keratinocyte activation, proliferation and migration by immunohistochemical analysis, and gelatinases by zymography. Results Resurfacing from bioengineereed skin explants started earlier than from normal skin but subsequently, from day 3 to day 9, the rate of epidermalization from bioengineered skin was only 40% (206 ± 23 µm day −1 , mean ± SEM) of that of authentic skin (521 ± 17 µm day −1 , P < 0·001). At culture termination at day 11, normal human skin had formed a multilayered and well‐structured neoepidermis covering 41·0 ± 1·2 mm 2 of the dermal substrate while bioengineered skin produced a thinner, less organized epithelium covering 20·4 ± 3·0 mm 2 . At this later stage, a higher expression of β‐defensin‐2, keratin 16, Ki67 and matrix metalloproteinase (MMP)‐9 was found in neoepidermis formed from authentic skin than from bioengineered skin. Activated MMP‐2 was elevated in bioengineered skin‐derived neoepidermis. Minor epithelial outgrowth was noted with either skin type on the dermal substrate devoid of basement membrane zone. Conclusions Cultured normal skin explants produced a more uniform and expansive in vivo‐ like neoepidermis than bioengineered skin explants.