RECONSTITUTION OF STRUCTURE AND CELL-FUNCTION IN HUMAN-SKIN GRAFTS DERIVED FROM CRYOPRESERVED ALLOGENEIC DERMIS AND AUTOLOGOUS CULTURED KERATINOCYTES
RECONSTITUTION OF STRUCTURE AND CELL-FUNCTION IN HUMAN-SKIN GRAFTS DERIVED FROM CRYOPRESERVED ALLOGENEIC DERMIS AND AUTOLOGOUS CULTURED KERATINOCYTES
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DOI：
10.1111/1523-1747.ep12476623
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发表时间：
1988-11-01
影响因子：
6.5
通讯作者：
MOELLMANN, GE
中科院分区：
文献类型：
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作者：
LANGDON, RC;CUONO, CB;MOELLMANN, GE
异体真皮加自体表皮细胞培养物移植用于替代大面积烧伤皮肤。将冷冻保存的断层尸体皮肤移植到清创的烧伤创面上，并从未受伤的供体部位培养自体角质形成细胞。几周后，磨损同种异体表皮并用角质形成细胞培养物代替。因此，最终的移植物是自体培养的表皮和异体真皮的复合物。对1例随访28个月的病例进行了组织化学和超微结构观察，研究了真皮-表皮（BMZ.1）和微血管（BMZ.2）基底膜区的重建。移植前，解冻的冷冻保存的皮肤反应正常模式的层粘连蛋白和IV型胶原蛋白的抗体。移植后29天，BMZ.1与两种抗体的反应较弱，BMZ.2不存在抗IV型胶原反应性。然而，BMZ.2的抗层粘连蛋白反应性是中等强度的，与最近的新血管形成一致。在第29天，用自体角质形成细胞培养物替换同种异体移植表皮。25天后（同种异体移植后54天），两种BMZ的染色都很强烈，两种抗体都很强烈。超微结构，在第76天（培养放置后47天）BMZ.1显示只有小的半桥粒，一些初期锚定纤维，和不连续的致密层。然而，BMZ.2完全重组。到124 d，两个BMZ均正常。在76天的真皮中的观察结果包括淋巴细胞、细胞器碎片和过度活跃的胶原纤维生成的存在，所有这些都表明真皮重塑。移植后5、47和95 d，表皮中可见黑素细胞和黑素小体转移。我们的结论是，复合程序重建皮肤具有良好的质地和组织质量。
Grafts of allogeneic dermis plus autologous epidermal cell cultures were used to replace extensively burned skin. Cryopreserved split-thickness cadaveric skin was grafted onto debrided burn wounds, and autologous keratinocytes were cultured from uninjured donor sites. Several weeks later, allograft epidermis was abraded and replaced with the keratinocyte cultures. The final grafts were thus composites of autologous cultured epidermis and allogeneic dermis. In a case with 28 months follow-up, reconstitution of the dermal-epidermal (BMZ.1) and microvascular (BMZ.2) basement membrane zones was studied immunohistochemically and ultrastructurally. Immediately before grafting, thawed cryopreserved skin reacted with antibodies against laminin and type IV collagen in normal patterns. Twenty-nine days after grafting, BMZ.1 reacted weakly with both antibodies, and anticollagen type IV reactivity was absent from BMZ.2. Antilaminin reactivity of BMZ.2, however, was moderately intense, consistent with recent neovascularization. On day 29, the allograft epidermis was replaced with autologous keratinocyte cultures. Twenty-five days later (54 d after allografting), staining of both BMZs was intense with both BMZs was intense with both antibodies. Ultrastructurally, at day 76 (47 d after culture placement) BMZ.1 revealed only small hemidesmosomes, few incipient anchoring fibrils, and a discontinuous lamina densa. BMZ.2, however, was fully reconstituted. By 124 d, both BMZs appeared normal. Observations in the dermis at 76 d included the presence of lymphocytes, organellar debris, and hyperactive collagen fibrillogenesis, all indicative of dermal remodelling. The microvasculature was well differentiated, but no elastic fibers or nerves were found. In the epidermis, melanocytes and evidence of melanosome transfer were seen at 5, 47, and 95 d after grafting of keratinocyte cultures. We conclude that the composite procedure reconstitutes skin with excellent textural and histologic qualities.