Hansbrough J F, Morgan J, Greenleaf G, Parikh M, Nolte C, Wilkins L
Department of Surgery, University of California, San Diego Medical Center 92103.
J Burn Care Rehabil. 1994 Jul-Aug;15(4):346-53. doi: 10.1097/00004630-199407000-00010.
We used a living bilayered cultured skin replacement to close full-thickness wound defects on the dorsum of athymic mice. The skin replacement is composed of human fibroblasts that condense a bovine collagen lattice; the lattice is then seeded with cultured human keratinocytes. The collagen lattice with fibroblasts serves as a dermal template, and the overlying human keratinocytes form the epidermal component of this composite skin replacement. Twenty-four animals were grafted, and groups of six were killed and biopsied at 6, 15, 30, and 60 days after graft replacement. Twenty-four mice in the control group receiving grafts of fresh, split-thickness, human cadaver skin were biopsied at the same time points. "Take" of all grafts was excellent, with only one graft loss in the 48 mice (one Graftskin graft, at 15 days). Light microscopy revealed that vascular ingrowth into Graftskin occurred rapidly, and discrete dermal and epidermal layers were seen at all time points. Evidence of basement membrane formation was seen at 6 days after grafting by immunohistochemical staining for laminin and by electron microscopic visualization of lamina lucida and lamina densa zones at the dermal-epidermal junction. The results demonstrated that Graftskin formed a structurally complete skin replacement within 1 week of placement on full-thickness wounds on athymic mice, and effective skin coverage was provided for the 60-day observation period after grafting.
我们使用一种活的双层培养皮肤替代物来闭合无胸腺小鼠背部的全层伤口缺损。该皮肤替代物由浓缩了牛胶原蛋白晶格的人成纤维细胞组成；然后在晶格上接种培养的人角质形成细胞。带有成纤维细胞的胶原蛋白晶格作为真皮模板，覆盖其上的人角质形成细胞形成这种复合皮肤替代物的表皮成分。对24只动物进行了移植，在移植替代后第6、15、30和60天，将6只一组的动物处死并进行活检。在相同时间点对接受新鲜的、断层的人尸体皮肤移植的24只对照组小鼠进行活检。所有移植的“成活率”都很高，48只小鼠中只有1只移植失败（15天时1只Graftskin移植失败）。光学显微镜检查显示，Graftskin中血管长入迅速，在所有时间点都可见离散的真皮和表皮层。通过层粘连蛋白免疫组织化学染色以及真皮 - 表皮交界处透明板和致密板区域的电子显微镜观察，在移植后6天可见基底膜形成的证据。结果表明，Graftskin在放置于无胸腺小鼠全层伤口上1周内形成了结构完整的皮肤替代物，并在移植后60天的观察期内提供了有效的皮肤覆盖。
