RESUMO
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 microm day(-1), mean +/- SEM) of that of authentic skin (521 +/- 17 microm 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 mm2 of the dermal substrate while bioengineered skin produced a thinner, less organized epithelium covering 20.4 +/- 3.0 mm2. At this later stage, a higher expression of beta-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.
