改良法制备人脱细胞真皮的特性与体外细胞相容性

Characteristics and in vitro cell compatibility of human acellular dermal matrix with improved method

目的探讨改良法制备人脱细胞真皮（ADM）特性及与体外细胞相容性。方法按传统法及改良法分别将健康成人皮肤脱去细胞成分，并分析ADM的孔隙率、体外降解时间及改良法ADM浸润液对脂肪干细胞的细胞毒性；HE染色、扫描电子显微镜（SEM）检测其脱细胞效率、胶原完整性、孔隙大小及脂肪干细胞相容性。结果两种方法均可完整脱去细胞成分，并较好保持胶原支架的完整性；改良法ADM孔隙率93．1％±1．02％明显高于传统法ADM孔隙率74．27％±2．04％（P〈0．05）；改良法ADM浸润液无明显细胞毒性、两种ADM体外降解时间差异无统计学意义（P〉0．05）；两种ADM基底面孔隙直径差异无统计学意义（P〉0．05），而改良法ADM网状面孔隙直径（102．38±15．63）μm明显大于传统法ADM（181．21±66．96）ham（P〈0．05）。结论改良法处理ADM脱细胞效果良好，无明显细胞毒性，具有更高的孔隙率，更大的孔径，脂肪干细胞的细胞渗透性更强。

Objective To detect the characteristics and in vitro cell compatibility of human acellular dermal matrix （ADM） with the improved method. Methods Cell components of healthy human skins were removed by the improved method and the traditional method respectively. The porosity, degradation time in vitro of the ADM prepared by two methods and the cytotoxicity of the material infiltration liquid with the improved method on the adipose derived stem cells were detected. HE staining was used to detect the residual of the cells, the integrity of collagen and cell biocompatibility. Scanning electron microscopy （SEM） was used to detect the pore size. Results Both the two methods could completely remove the cells, and maintain the integrity of the collagen scaffold; The porosity of ADM with the improved method was higher （93.1 ±1.02）; than that of traditional method （74.27±2.04） % （P〈0.05） ; There was no significant difference in the cytotoxicity and in vitro degradation time between the two kinds of ADM; While pore diameter of the improved method was significantly higher [ （ 181.21 ± 66.9） μm] than that [ （ 102.38 ± 15.63） μm] in dermal reticular surface with the traditonal method （P〈0.05） . Conclusions There is no obvious cytotoxicity of the ADM with the improved method, and therefore it is more suitable for cell adhesion growth with higher porosity and larger pore size.