Bone regeneration can be possible through grafts or engineered bone replacement when bone defects are larger than the critical size. Decellularized bone extracellular matrix (ECM) is an alternative that is able to accelerate tissue regeneration, while decellularization protocols influence engineered bone quality. The objective of this study was to compare the quality of decellularized bone produced through different methods. Four decellularization methods were employed using (a) sodium lauryl ether sulfate (SLES), (b) sodium dodecyl sulfate (SDS) 0.5%, (c) SDS 1% and (d) trypsin/EDTA. All samples were then washed in triton X‐100. DNA quantification, hematoxylin and eosin, and Hoechst staining showed that although DNA was depleted in all scaffolds, treatment with SLES led to a significantly lower DNA content. Glycosaminoglycan quantification, Raman confocal microscopy, alcian blue and PAS staining exhibited higher carbohydrate retention in the scaffolds treated with SLES and SDS 0.5%. Raman spectra, scanning electron microscopy and trichrom Masson staining showed more collagen content in SLES and SDS‐treated scaffolds compared to trypsin/EDTA‐treated scaffolds. Therefore, although trypsin/EDTA could efficiently decellularize the scaffolds, it washed out the ECM contents. Also, both MTT and attachment tests showed a significantly higher cell viability in SLES‐treated scaffolds. Raman spectra revealed that while the first washing procedure did not remove SLES traces in the scaffolds, excessive washing reduced ECM contents. In conclusion, SLES and, to a lesser degree, SDS 0.5% protocols could efficiently preserve ultrastructure and ECM constituents of decellularized bone tissue and can thus be suggested as nontoxic and safe protocols for bone regeneration.

中文翻译：

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用于骨组织工程的各种脱细胞剂效率的比较评估。

当骨缺损大于临界尺寸时，可以通过移植物或工程骨替代物进行骨再生。脱细胞骨细胞外基质 (ECM) 是一种能够加速组织再生的替代方案，而脱细胞方案会影响工程骨质量。本研究的目的是比较通过不同方法生产的脱细胞骨的质量。四种脱细胞方法使用 (a) 十二烷基醚硫酸钠 (SLES)、(b) 十二烷基硫酸钠 (SDS) 0.5%、(c) SDS 1% 和 (d) 胰蛋白酶/EDTA。然后用triton X-100洗涤所有样品。DNA 定量、苏木精和伊红以及 Hoechst 染色表明，尽管所有支架中的 DNA 都已耗尽，但 SLES 处理导致 DNA 含量显着降低。糖胺聚糖定量，拉曼共聚焦显微镜、阿尔新蓝和 PAS 染色在用 SLES 和 SDS 0.5% 处理的支架中表现出更高的碳水化合物保留。拉曼光谱、扫描电子显微镜和三色马松染色显示，与胰蛋白酶/EDTA 处理的支架相比，SLES 和 SDS 处理的支架中胶原含量更高。因此，虽然胰蛋白酶/EDTA 可以有效地使支架脱细胞，但它会洗掉 ECM 的内容。此外，MTT 和附着测试均显示 SLES 处理的支架具有显着更高的细胞活力。拉曼光谱显示，虽然第一次洗涤程序没有去除支架中的 SLES 痕迹，但过度洗涤会降低 ECM 含量。总之，SLES 和在较小程度上的 SDS 0。