去细胞组织水凝胶,尤其是模仿天然组织,具有很高的组织工程潜力,三维(3D)细胞培养,生物打印,和治疗剂封装由于其优异的生物相容性和促进细胞生长的能力。重要的是要注意,脱细胞过程显著影响细胞外基质的结构完整性和性质,这又在大分子水平上塑造了所得水凝胶的特性。因此,我们的研究旨在确定一种有效的绵羊肺组织化学脱细胞方法,使用混合/搅拌技术与一系列洗涤剂,通常包括[十二烷基硫酸钠(SDS),TritonX-100和3-((3-胆酰氨基丙基)二甲基铵)-1-丙磺酸盐](CHAPS),和很少使用(胆酸钠水合物,NP-40和3-[N,N-二甲基(3-肉豆蔻酰基氨基丙基)铵]丙磺酸酯(ASB-14)。通过组织学和生化方法确定所用洗涤剂对去细胞化的有效性后,肺源性脱细胞细胞外基质转化为水凝胶。我们使用增殖试验研究了肺细胞和脱细胞细胞外基质之间的相互作用,扫描电子显微镜,和免疫荧光显微镜方法对气-液界面BEAS-2B细胞的研究。值得注意的是,这项研究强调了ASB-14在去细胞化过程中的有效性,展示了其在去除细胞成分,同时保留重要的细胞外基质生物大分子的关键作用,包括糖胺聚糖,胶原蛋白,和弹性蛋白。所得的水凝胶表现出良好的机械性能,并且与细胞-细胞和细胞-细胞外基质相互作用相容。
Decellularized tissue hydrogels, especially that mimic the native tissue, have a high potential for tissue engineering, three-dimensional (3D) cell culture, bioprinting, and therapeutic agent encapsulation due to their excellent biocompatibility and ability to facilitate the growth of cells. It is important to note that the
decellularization process significantly affects the structural integrity and properties of the extracellular matrix, which in turn shapes the characteristics of the resulting hydrogels at the macromolecular level. Therefore, our study aims to identify an effective chemical
decellularization method for sheep lung tissue, using a mixing/agitation technique with a range of detergents, including commonly [Sodium dodecyl sulfate (SDS), Triton X-100, and 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate] (CHAPS), and rarely used (sodium cholate hydrate, NP-40, and 3-[N,N-Dimethyl(3-myristoylaminopropyl)ammonio]propanesulfonate) (ASB-14). After the effectiveness of the used detergents on
decellularization was determined by histological and biochemical methods, lung derived decellularized extracellular matrix was converted into hydrogel. We investigated the interactions between lung cells and decellularized extracellular matrix using proliferation assay, scanning electron microscopy, and immunofluorescence microscopy methods on BEAS-2B cells in air-liquid interface. Notably, this study emphasizes the effectiveness of ASB-14 in the
decellularization process, showcasing its crucial role in removing cellular components while preserving vital extracellular matrix biological macromolecules, including glycosaminoglycans, collagen, and elastin. The resulting hydrogels demonstrated favorable mechanical properties and are compatible with both cell-cell and cell-extracellular matrix interactions.