Department of Digestive Surgery, Kawasaki Medical School, Kurashiki, Japan.
Int J Exp Pathol. 2023 Dec;104(6):313-320. doi: 10.1111/iep.12492. Epub 2023 Aug 25.
Porcine small intestinal submucosa, despite its successful use as a scaffold in regenerative medicine, has innate biomechanical heterogeneity. In this study, we hypothesized that human small intestinal submucosa could be a viable alternative bio-scaffold. For the first time, we characterize submucosal extraction from human small intestine and examine appropriate decellularization methods. In total, 16 human small intestinal submucosal samples were obtained and decellularized using three reported methods of porcine decellularization: Abraham, Badylak, and Luo. For each method, four specimens were decellularized. The remaining four specimens were designated as non-decellularized. We measured the amount of residual DNA and growth factors in decellularized human intestinal samples. Additionally, decellularized human small intestinal submucosa was co-cultured with mouse bone marrow-derived mesenchymal stem cells to examine mesenchymal stem cell survival and proliferation. The reference value for the amount of residual DNA deemed appropriate in decellularized tissue was established as 50 ng/mg of extracellular matrix dry weight or less. Abraham's method most successfully met this criterion. Measurement of residual growth factors revealed low levels observed in samples decellularized using the Abraham and Badylak methods. Co-culture of each small intestinal submucosal sample with mouse bone marrow-derived mesenchymal stem cells confirmed viable cell survival and proliferation in samples derived using protocols by Abraham and Badylak. Abraham's method most successfully met the criteria for efficient tissue decellularization and cell viability and proliferation. Thus, we consider this method most suitable for decellularization of human small intestinal submucosa.
猪小肠黏膜下层尽管已成功用作再生医学中的支架，但具有固有生物力学异质性。在这项研究中，我们假设人小肠黏膜下层可以作为一种可行的替代生物支架。我们首次对人小肠黏膜下层的提取进行了特征描述，并研究了适当的脱细胞方法。总共获得了 16 个人小肠黏膜下层样本，并使用三种已报道的猪脱细胞方法：Abraham、Badylak 和 Luo 对其进行脱细胞处理。对于每种方法，都有四个样本进行脱细胞处理。其余四个样本被指定为非脱细胞处理。我们测量了脱细胞的人肠样本中残留 DNA 和生长因子的含量。此外，将脱细胞的人小肠黏膜下层与人骨髓间充质干细胞共培养，以研究间充质干细胞的存活和增殖情况。在脱细胞组织中，认为适当的残留 DNA 量的参考值为 50ng/mg 细胞外基质干重或更少。Abraham 方法最符合该标准。对残留生长因子的测量显示，使用 Abraham 和 Badylak 方法脱细胞的样本中观察到低水平的生长因子。将每个小肠黏膜下层样本与人骨髓间充质干细胞共培养，证实了源自 Abraham 和 Badylak 方案的样本中细胞的存活和增殖。Abraham 方法最符合高效组织脱细胞和细胞存活及增殖的标准。因此，我们认为该方法最适合人小肠黏膜下层的脱细胞处理。
