Optimization of decellularization methods using human small intestinal submucosa for scaffold generation in regenerative medicine

IF 1.8 4区 医学 Q3 PATHOLOGY International Journal of Experimental Pathology Pub Date : 2023-08-25 DOI:10.1111/iep.12492
Shumei Mineta, Shunji Endo, Tomio Ueno
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Abstract

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.

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人小肠粘膜下层脱细胞制备再生医学支架方法的优化。
猪小肠粘膜下层虽然在再生医学中被成功地用作支架,但它具有先天的生物力学异质性。在这项研究中,我们假设人类小肠粘膜下层可能是一种可行的替代生物支架。我们首次描述了人小肠粘膜下提取的特征,并研究了适当的脱细胞方法。总共获得了16份人小肠粘膜下样本,并使用三种已报道的猪脱细胞方法:Abraham、Badylak和Luo进行了脱细胞。对于每种方法,四个标本被脱细胞。其余4个标本被指定为非脱细胞。我们测量了去细胞人肠道样本中残留DNA和生长因子的数量。另外,将去细胞化的人小肠黏膜下层与小鼠骨髓间充质干细胞共培养,观察间充质干细胞的存活和增殖情况。脱细胞组织中残余DNA量的参考值为细胞外基质干重50 ng/mg或更少。亚伯拉罕的方法最成功地满足了这个标准。残余生长因子的测量显示,在使用亚伯拉罕和巴迪拉克方法去细胞的样品中观察到低水平。每个小肠粘膜下样本与小鼠骨髓来源的间充质干细胞共同培养,证实了使用Abraham和Badylak的方案获得的样本中细胞存活和增殖。亚伯拉罕的方法最成功地满足了高效组织脱细胞和细胞活力和增殖的标准。因此,我们认为这种方法最适合于人小肠粘膜下层的脱细胞。
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来源期刊
CiteScore
4.50
自引率
3.30%
发文量
35
审稿时长
>12 weeks
期刊介绍: Experimental Pathology encompasses the use of multidisciplinary scientific techniques to investigate the pathogenesis and progression of pathologic processes. The International Journal of Experimental Pathology - IJEP - publishes papers which afford new and imaginative insights into the basic mechanisms underlying human disease, including in vitro work, animal models, and clinical research. Aiming to report on work that addresses the common theme of mechanism at a cellular and molecular level, IJEP publishes both original experimental investigations and review articles. Recent themes for review series have covered topics as diverse as "Viruses and Cancer", "Granulomatous Diseases", "Stem cells" and "Cardiovascular Pathology".
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