人新型脱细胞真皮制备及性状分析  

Preparation and properties of a new human acellular dermal matrix

作　　者：姜涛[1] 张爱君[1] 李雪阳[1] 马志兵[1] 沈才齐[1] 金培生[1] 

机构地区：[1]徐州医学院附属医院整形外科,江苏省徐州市221002

出　　处：《中国组织工程研究》2016年第7期1006-1012,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:早期快速血管化是创面修复的关键。高孔隙率、大孔径的真皮支架可以促进种子细胞更快更好的黏附、生长、迁移并促进创面修复中血管化的快速形成。目的:通过改良人脱细胞真皮基质制备方法,以期得到孔隙率高、细胞渗透性更好、组织相容性良好的同种异体皮。方法:健康成人皮肤,分别用传统方法及改良方法脱去细胞成分。改良方法采用无菌条件下剪除皮下脂肪组织,1 mol/L NaC l溶液37℃24 h,去表皮,2%NaO H 45℃摇床处理4 h,PBS冲洗至溶液成中性,冷冻干燥,4℃保存备用。对两种方法制备脱细胞真皮基质孔隙率、体外降解时间的差异及材料浸润液对脂肪干细胞毒性进行检测;苏木精-伊红染色、扫描电镜检测其脱细胞效率、胶原完整性、脂肪干细胞细胞相容性及支架孔隙等。结果与结论:两种处理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;改良型脱细胞真皮基质孔隙率达(93.22±0.99)%明显高于传统(74.28±2.06)%(P<0.001);体外降解时间无明显差异(P>0.05);改良方法制备脱细胞真皮材料浸润液对干细胞无明显细胞毒性;改良脱细胞真皮基质基底面接种细胞3-7 d后细胞可突破基底层并迁移、游走至真皮深面,传统脱细胞真皮基质中细胞在基底面单-复层生长,无明显真皮浸润。结果提示新型脱细胞真皮基质拥有较传统脱细胞真皮基质更大的孔径、孔隙率,更适合细胞的浸润生长。BACKGROUND： Early vascularization is crucial for wound healing. A high-porosity, macrovoid allogeneic skin leads to the rapid vascularization and cellular infiltration. OBJECTIVE： To obtain a new allogeneic skin product with high porosity, good cell permeability and good histocompatibility using an improved preparation method of human acellular dermal matrix. METHODS： Cell components of healthy human skins were removed by the improved method and the traditional method, respectively. The improved method was to remove the subcutaneous fat, eliminate the epidermis（1 mol/L Na Cl solution at 37 ℃ for 24 hours） followed by shaking processing（2% Na OH at 45 ℃ for 4 hours）, and then, the solution was neutralized with PBS rinsing, dried and stored at 4 ℃ for standby. We detected the porosity and degradation time in vitro of the acellular dermal matrices prepared by two methods and the cytotoxicity of the material infiltration liquid on the adipose-derived stem cells. Hematoxylin-eosin staining was used for the detection of the cell residual, the integrity of collagen and cell biocompatibility. Scanning electron microscopy was used to detect the pore size. RESULTS AND CONCLUSION： Both the two methods could completely remove the cell components, and maintain the integrity of the collagen scaffold. The porosity of acellular dermal matrix with the improved method was（93.22±0.99）%, which was significantly higher than that with the traditional method [（74.28±2.06）%; P 0.001]. However, there was no significant difference in in vitro degradation time between the two kinds of acellular dermal matrices（P 0.05）. No obvious cytotoxicity of the acellular dermal matrix prepared with the improved method was detected. At 3-7 days of co-culture, the adipose-derived stem cells cultured on the acellular dermal matrix prepared with the improved method could penetrate the basement membrane to the deep dermis, while there was no obvious cell invasion and growth in the deep dermis prepared by the traditional

关 键 词：真皮 干细胞 脂肪组织 皮肤 组织构建 组织工程 脱细胞真皮 种子细胞 脂肪干细胞 生物支架 组织相容性 创面修复 组织工程皮肤 

分 类 号：R318[医药卫生—生物医学工程]