University of São Paulo, São Paulo, Brazil.
J Tissue Eng Regen Med. 2018 Apr;12(4):e1880-e1888. doi: 10.1002/term.2618. Epub 2017 Dec 13.
Technically produced scaffolds are common to establish transplantable tissues for regenerative medicine, but also biological ones that are closer to the natural condition become of interest. Placentas are promising, because they represented available, complete organs with rich extracellular matrix (ECM) and well-developed vasculature that easily could build anastomoses to a host's organ. Only placentas from larger animal models such as the bovine meet the dimensions large enough for most organs but are not adequately described yet. We here studied the nature of the ECM in 27 natural and decellularized bovine cotyledons, that is, the fetal part of the placentomes, by means of histology, immunohistochemistry, and electron microscopy. Successful decellularization was done by perfusion with 0.01%, 0.1%, and 0.5% sodium dodecyl sulfate each and subsequent immersion in 1% Triton X-100, resulting in a removal of cells and DNA, whereas the structure of the allantochorionic surface and villi was preserved. Although some fibres disappeared, also the arrangement of the main ECM proteins was largely similar before and after decellularization: Along the larger vessels, a densely packed network of thick fibres occurred, organized in layers without cells or spaces in between. Collagen IV, fibronectin, and laminin contributed to those areas. In contrast, collagen I and III characterized the meshwork of medium-sized and thin fibres in the mesenchyme, respectively. In conclusion, decellularized bovine cotyledons indeed had characteristics of a biological scaffold and provide an interesting alternative to develop large-scale scaffolds with complex vascular architecture for tissue engineering purposes.
从技术角度来看,用于再生医学的移植组织通常采用人工合成支架,而更接近自然条件的生物支架也逐渐受到关注。胎盘是很有前途的,因为它们是现成的、完整的器官,具有丰富的细胞外基质(ECM)和发育良好的脉管系统,很容易与宿主器官建立吻合。只有像牛这样的大型动物模型的胎盘才能达到大多数器官所需的足够大的尺寸,但目前对其还没有充分的描述。我们通过组织学、免疫组织化学和电子显微镜研究了 27 个天然和去细胞化的牛胎膜小叶(即胎盘的胎儿部分)的 ECM 性质。通过分别用 0.01%、0.1%和 0.5%十二烷基硫酸钠灌注,然后用 1%Triton X-100 浸泡,成功地实现了去细胞化,去除了细胞和 DNA,而胎盘绒毛膜表面和绒毛的结构得以保留。尽管一些纤维消失了,但去细胞化前后主要 ECM 蛋白的排列方式也基本相似:在较大的血管周围,出现了一层紧密堆积的厚纤维网络,无细胞或无间隙地排列成层。胶原 IV、纤维连接蛋白和层粘连蛋白参与了这些区域。相比之下,I 型和 III 型胶原分别构成了间质中中、小纤维的网格结构。总之,去细胞化的牛胎膜小叶确实具有生物支架的特性,为开发具有复杂血管结构的大规模支架以用于组织工程目的提供了一种有趣的替代方案。