For patients with extensive full-thickness burns who do not have sufficient autologous split-thickness skin for skin grafts, the application of biological skin substitutes may be considered. The aim of this study was to find an optimal new type method for the production of a biovital skin substitute based on acellular dermal matrix (ADM) and preclinical evaluations. In this work, 25 methods of ADM production were assessed. The proposed methods are based on the use of the following enzymes: papain, Carica papaya lipase (CPL), and purification using a polymer/salt aqueous two-phase system. The obtained ADM samples were characterized via scanning electron microscopy (SEM), porosity measurement and water vapor transmission test. Results showed that the collagen bundles of ADM microparticles were intact and orderly. Through differential scanning calorimetry (DSC), thermo gravimetric analysis (TGA) and biocompatibility tests, the results indicated that the proportion of papain and CPL was the same and 5 h processing time are the optimum conditions for ADM preparation and the material showed good biocompatibility. Our results suggested that the potential of developing this kind of decellularization process to manufacture ADM scaffolds for clinical application.

中文翻译：

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基于植物酶和水相两相法制备的脱细胞细胞外基质胶原微粒的生物学评价

对于大面积全层烧伤患者，没有足够的自体二层皮进行植皮，可考虑应用生物皮肤替代物。本研究的目的是找到一种基于脱细胞真皮基质 (ADM) 和临床前评估的生物活性皮肤替代品生产的最佳新型方法。在这项工作中，评估了 25 种 ADM 生产方法。建议的方法基于使用以下酶：木瓜蛋白酶、番木瓜脂肪酶 (CPL)，以及使用聚合物/盐水两相系统进行纯化。所获得的 ADM 样品通过扫描电子显微镜 (SEM)、孔隙率测量和水蒸气透过率测试进行了表征。结果表明，ADM微粒的胶原束完整有序。通过差示扫描量热法（DSC）、热重分析（TGA）和生物相容性测试，结果表明木瓜蛋白酶和CPL的比例相同，5 h处理时间是制备ADM的最佳条件，材料表现出良好的生物相容性。我们的研究结果表明，开发这种去细胞化工艺来制造用于临床应用的 ADM 支架具有潜力。