To date, the decellularized scaffold has been widely explored as a source of biological scaffolds for regenerative medicine. However, the acellular matrix derived from natural tissues and organs has a lot of defects, including the limited amount of autogenous tissue and surgical complication such as risk of blood loss, wound infection, pain, shock, and functional damage in the donor part of the body. In this study, we prepared acellular matrix using adipose-derived stem cell (ADSC) sheets and evaluate the cellular compatibility and immunoreactivity. The ADSC sheets were fabricated and subsequently decellularized using repeated freeze-thaw, Triton X-100 and SDS decellularization. Oral mucosal epithelial cells were seeded onto the decellularized ADSC sheets to evaluate the cell replantation ability, and silk fibroin was used as the control. Then, acellular matrix was transplanted onto subcutaneous tissue for 1 week or 3 weeks; H&E staining and immunohistochemical analysis of CD68 expression and quantitative real-time PCR (qPCR) were performed to evaluate the immunogenicity and biocompatibility. The ADSC sheet-derived ECM scaffolds preserved the three-dimensional architecture of ECM and retained the cytokines by Triton X-100 decellularization protocols. Compared with silk fibroin in vitro, the oral mucosal epithelial cells survived better on the decellularized ADSC sheets with an intact and consecutive epidermal cellular layer. Compared with porcine small intestinal submucosa (SIS) in vivo, the homogeneous decellularized ADSC sheets had less monocyte-macrophage infiltrating in vivo implantation. During 3 weeks after transplantation, the mRNA expression of cytokines, such as IL-4/IL-10, was obviously higher in decellularized ADSC sheets than that of porcine SIS. A Triton X-100 method can achieve effective cell removal, retain major ECM components, and preserve the ultrastructure of ADSC sheets. The decellularized ADSC sheets possess good recellularization capacity and excellent biocompatibility. This study demonstrated the potential suitability of utilizing acellular matrix from ADSC sheets for soft tissue regeneration and repair.

中文翻译：

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干细胞片技术生产的潜在生物材料的制备和评估，以用于未来的再生医学。

迄今为止，脱细胞支架已被广泛地探索为再生医学的生物支架的来源。然而，源自天然组织和器官的脱细胞基质存在许多缺陷，包括自体组织数量有限和外科手术并发症，例如失血，伤口感染，疼痛，休克和供体部分功能受损的风险。身体。在这项研究中，我们使用脂肪干细胞（ADSC）片材制备了脱细胞基质，并评估了细胞相容性和免疫反应性。制备ADSC片材，随后使用重复的冻融，Triton X-100和SDS脱细胞作用脱细胞。将口腔粘膜上皮细胞接种到脱细胞的ADSC片上以评估细胞的再植能力，并以丝心蛋白为对照。然后，将无细胞基质移植至皮下组织1周或3周；H＆E染色和CD68表达的免疫组织化学分析和定量实时PCR（qPCR）进行了评估免疫原性和生物相容性。ADSC片材衍生的ECM支架通过Triton X-100脱细胞方案保留了ECM的三维结构，并保留了细胞因子。与体外丝素蛋白相比，口腔粘膜上皮细胞在具有完整且连续的表皮细胞层的脱细胞ADSC片上存活得更好。与猪体内小肠粘膜下层（SIS）相比，均质的脱细胞ADSC片体内植入的单核细胞-巨噬细胞浸润较少。移植后3周内，IL-4 / IL-10等细胞因子的mRNA表达，在脱细胞的ADSC床单中明显高于猪SIS。Triton X-100方法可以有效去除细胞，保留主要的ECM成分，并保留ADSC板的超微结构。脱细胞的ADSC片材具有良好的再细胞化能力和优异的生物相容性。这项研究证明了利用ADSC片中的脱细胞基质进行软组织再生和修复的潜在适用性。