Human plasma as a dermal scaffold for the generation of a completely autologous bioengineered skin
Human plasma as a dermal scaffold for the generation of a completely autologous bioengineered skin
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DOI：
10.1097/01.tp.0000112381.80964.85
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发表时间：
2004-02-15
期刊：
影响因子：
6.2
通讯作者：
Meana, A
中科院分区：
文献类型：
--
作者：
Llames, SG;Del Rio, M;Meana, A
背景角质形成细胞培养物已用于治疗严重烧伤患者。在这里，我们描述了一种新的培养的生物工程皮肤的基础上（1）角质形成细胞和成纤维细胞从一个单一的皮肤活检和（2）真皮基质的基础上，人血浆。报道了在这种基于血浆的基质上生长的角质形成细胞所实现的高扩增能力。此外，成功的临床前和临床试验的结果。通过双酶消化（分别为胰蛋白酶和胶原酶）获得角质形成细胞和成纤维细胞。在这种情况下，将人成纤维细胞包埋在凝固的基于血浆的基质中，该基质用作三维支架。将人角质形成细胞接种在基于血浆的支架上以形成皮肤构建体的表皮组分。作为临床前方法，在免疫缺陷小鼠身上测试了基于血浆的生物工程皮肤的再生性能。最后，将此皮肤替代物移植到两名严重烧伤患者身上。接种在基于血浆的支架上的角质形成细胞生长至汇合，在培养24至26天后允许1,000倍的培养面积扩增。在工程化血浆支架上扩增的人角质形成细胞的实验移植产生了最佳的表皮结构和表型，包括结构性细胞内蛋白和基底膜组分的表达。此外，我们在此报告了两名严重烧伤患者在1年和2年随访时的成功植入和稳定的皮肤再生。我们的数据表明，这种新的真皮等效物允许（1）产生大的生物工程皮肤表面，（2）恢复表皮和真皮皮肤区室，和（3）功能性表皮干细胞保存。
Background. Keratinocyte cultures have been used for the treatment of severe burn patients. Here, we describe a new cultured bioengineered skin based on (1) keratinocytes and fibroblasts obtained from a single skin biopsy and (2) a dermal matrix based on human plasma. A high expansion capacity achieved by keratinocytes grown on this plasma-based matrix is reported. In addition, the results of successful preclinical and clinical tests are presented.Methods. Keratinocytes and fibroblasts were obtained by a double enzymatic digestion (trypsin and collagenase, respectively). In this setting, human fibroblasts are embedded in a clotted plasma-based matrix that serves as a three-dimensional scaffold. Human keratinocytes are seeded on the plasma-based scaffold to form the epidermal component of the skin construct. Regeneration performance of the plasma-based bioengineered skin was tested on immunodeficient mice as a preclinical approach. Finally, this skin equivalent was grafted on two severely burned patients.Results. Keratinocytes seeded on the plasma-based scaffold grew to confluence, allowing a 1,000-fold cultured-area expansion after 24 to 26 days of culture. Experimental transplantation of human keratinocytes expanded on the engineered plasma scaffold yielded optimum epidermal architecture and phenotype, including the expression of structural intracellular proteins and basement-membrane components. In addition, we report here the successful engraftment and stable skin regeneration in two severely burned patients at 1 and 2 years follow-up.Conclusions. Our data demonstrate that this new dermal equivalent allows for (1) generation of large bioengineered skin surfaces, (2) restoration of both the epidermal and dermal skin compartments, and (3) functional epidermal stem-cell preservation.