Generation and Transplantation of an Autologous Vascularized Bioartificial Human Tissue
Generation and Transplantation of an Autologous Vascularized Bioartificial Human Tissue
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DOI：
10.1097/tp.0b013e3181ac15e1
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发表时间：
2009-07-27
期刊：
影响因子：
6.2
通讯作者：
Walles, Thorsten
中科院分区：
文献类型：
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作者：
Mertsching, Heike;Schanz, Johanna;Walles, Thorsten
背景移植血管化的缺乏阻碍了生物人工组织的产生和临床应用。我们开发了一种技术来生成具有所有先天血管化的生物人造人体组织。临床上植入该组织作为概念验证，以评估移植后血管网络血栓形成和组织活力。在两步程序中对猪小碗段进行脱细胞，保留其血管结构。细胞外基质的特征在于定量的DNA残基和蛋白质组成。在动态组织培养中，将血管内皮细胞与人内皮细胞重新融合。通过（1）组织学、（2）免疫组织学、（3）活-死测定和（4）代谢活性对工程化组织进行表征。为评价其组织性能，将其植入临床，1周后恢复正常。用脱氧胆酸钠一水合物溶液制备组织导致不完全脱细胞化。通过用DNA酶进行额外的组织孵育来去除细胞残留物。人内皮细胞在主要的猪细胞外基质内形成了一个有活力的内皮，表达CD 31、Flk-1和血管内皮钙粘蛋白。生物人工组织的代谢活性在体外随时间持续增加。临床组织移植证实血管通畅和组织存活1周。结论。已经在体外和临床环境中显示了生物工程化具有所有先天血管化的人类组织的可行性。这些结果可能为各种复杂的生物人工组织替代物和器官替代物的临床应用打开大门。
Background. The lack of transplant vascularization forecloses the generation and clinical implementation of bioartificial tissues. We developed techniques to generate a bioartificial human tissue with all innate vascularization. The tissue was implanted clinically as proof of concept to evaluate vascular network thrombogenicity and tissue viability after transplantation.Methods. A porcine small bowl segment was decellularized in a two-step procedure, preserving its vascular structures. The extracellular matrix was characterized quantitatively for DNA residues and protein composition. The vascular remainings were reseeded with human endothelial cells in a dynamic tissue culture. The engineered tissue was characterized by (1) histology, (2) immune-histology, (3) life-dead assay, and (4) metabolic activity. To evaluate the tissue capabilities, it was implanted clinically and recovered after 1 week.Results. Tissue preparation with sodium desoxycholate monohydrate solution resulted in an incomplete decellularization. Cell residues were removed by additional tissue incubation with DNAse. The human endothelial cells formed a viable endothelium inside the primarily porcine extracellular matrix, expressing CD31, Flk-1, and vascular endothelium-cadherin. The metabolic activity of the bioartificial tissue increased continuously over time ill vitro. Clinical tissue transplantation confirmed vessel patency and tissue viability for 1 week.Conclusions. The feasibility to bioengineer a human tissue with all innate vascularization has been shown in vitro and the clinical setting. These results may open the door for the clinical application of various sophisticated bioartificial tissue substitutes and organ replacements.