Effects of two common acellular methods on the physicochemical properties of dermal acellular matrix

目前脱细胞基质是一种治疗皮肤损伤的有效修复替代材料，但是关于其性能的系统性评价研究较少。本研究实验组采用两种脱细胞方法制备基质：一种是过氧乙酸（0.2% PAA/4% 乙醇溶液）灭菌后高渗盐水脱细胞（A 组）；另一种是 γ 射线灭菌后 0.05% Trypsin 酶/EDTA 脱细胞（B 组）；对照组进行 PBS 浸泡（C 组），检测三组组织物理特性和化学成分。不同方法处理后，苏木精伊红（HE）染色表明 B 组脱细胞效果良好；A、B 组组织孔隙率均在 84.9% 以上；A 组压缩弹性模量（9.94 ± 3.81）MPa、B 组压缩弹性模量（12.59 ± 5.50）MPa 分别与 C 组相比差异无统计学意义；A、B 组脱细胞基质中胶原蛋白总含量均显著低于 C 组含量（1.662 ± 0.229）mg/g，但胶原Ⅰ/Ⅲ的比值 B 组与 C 组相比差异无统计学意义；扫描电镜（SEM）、原子力显微镜（AFM）观测组织微观结构无明显差异；定性检测各组纤连蛋白和弹性蛋白与 C 组材料基本一致。因此，B 组脱细胞基质作为支架材料性能更好。实验结果说明，采用 γ 射线灭菌、0.05% Trypsin 酶/EDTA 制备的脱细胞基质可用于组织工程皮肤的构建，还可为异种真皮脱细胞基质的制备及裱衬提供参考，为静电纺丝或三维打印组织工程皮肤支架提供理化参数。.; At present, acellular matrix is an effective replacement material for the treatment of skin damage, but there are few systematic evaluation studies on its performance. The experimental group of this study used two decellularization methods to prepare the matrix: one was the acellular matrix which sterilized with peracetic acid first (0.2% PAA/4% ethanol solution) and then treated with hypertonic saline (group A), the other was 0.05% trypsin/EDTA decellularization after γ irradiation (group B); and the control group was soaked in PBS (Group C). Then physical properties and chemical composition of the three groups were detected. Hematoxylin eosin (HE) staining showed that the acellular effect of group B was good. The porosity of group A and B were both above 84.9%. In group A, the compressive modulus of elasticity was (9.94 ± 3.81) MPa, and the compressive modulus of elasticity was (12.59 ± 5.50) MPa in group B. There was no significant difference between group A or B and group C. The total content of collagen in acellular matrix of group A and B was significantly lower than that of group C (1. 662 ± 0.229) mg/g, but there was no significant difference in the ratio of collagen Ⅰ/Ⅲ between group B and group C. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that there was no significant difference in microstructure. Qualitative detection of fibronectin and elastin in each group was basically consistent with that in group C. Therefore, acellular matrix of group B had better performance as scaffold material. The experimental results show that the acellular matrix prepared by γ-ray sterilization and decellularization of 0.05% Trypsin enzyme/EDTA could be used for the construction of tissue-engineered skin. It could also provide reference for the preparation and mounting of heterogeneous dermal acellular matrix. It was also could be used for electrostatic spinning or three-dimensional printed tissue engineered skin scaffold which could provide physical and chemical parameters for it.