壳多糖基质网架复层组织工程皮肤的动物移植实验(英文)

Animal grafting experiment of reconstructive tissue engineered artificial skin made by chitosan as stromal scaffold

背景:治疗由烧伤或疾病等引起的大面积全层皮肤缺失最为常见的和有效的方法就是皮肤移植修复,但目前面临的最大障碍是皮肤供体不足。以自体皮肤细胞作为种子细胞构建的组织工程皮肤是解决这一矛盾的最理想途径。目的:探讨组织工程皮肤动物移植实验的效果。设计:随机对照实验。单位:一所大学再生医学科学研究所及皮肤科。材料:实验于1998-09/2001-07在吉林大学细胞工程研究室完成。选用新生Wistar大鼠20只;8周龄雄性裸鼠24只。方法:应用以壳多糖为基质网架构建的组织工程化复层人工皮肤,对裸鼠大面积(直径20mm)全层皮肤缺损模型进行移植修复。24只8周龄裸鼠分为组织工程皮肤移植(artificialskin,AS)组、壳多糖膜覆盖物(chitosan,CH)组及对照组(controlgroup,CG,术后进行大体观察,并在3,)7,14和21d应用组织学、红外热像扫描分析等手段对修复组织进行动态监测。主要观察指标:①实验动物大体观察情况。②修复区血供情况的红外热像观察。③组织学观察。结果:AS组在移植第3天,移植的组织工程皮肤与自体皮肤能够很好地融合,有少量毛细血管长入移植物,皮肤移植物颜色与自体皮肤颜色接近;随着时间的延长,移植物中毛细血管的数量逐渐增多,表皮层清晰可见基底层、棘细胞层、颗粒层和角质层,角化现象加强。

BACKGROUND:Skin transplantation is the most effective conventional method to c ure large area full thickness skin damage caused by empyrosis or some diseases, but present deficiency of skin donator is the largest barrier in front. The most ideal way to solve this problem is to use tissue engineering skin reconstructe d by self skin cells as seed cells. OBJECTIVE:To investigate the effects of tissue engineered artificial animal sk in in animal grafting experiment. DESIGN:A randomized controlled trial SETTING:Institute of frontier medical sciences and department of dermatology i n a university. MATERIALS:Study was performed in the Cell Engineering Institute of Jilin Univ ersity from September 1998 to July 2001.Totally 20 newborn Wistar rats and 24 8 week old male nude mice were selected into our study. METHODS:Full thickness wounds(diameter:20 mm)were made on the backs of twen ty four nude mice to establish full thickness skin defect animal model for gra fting by the tissue engineered reconstructive artificial skin made by chitosan( CH) as stromal scaffold. Twenty four 8 week old nude mice were divided into ar tificial skin (AS) group,chitosan membrane(CH) group and control group (CG). All wounds were covered with AS,CH or petrolatum gauze correspondingly.The wounds o f each groups were observed daily,techniques like histology and infrared ray sc an were used for a dynamical surveillance on the 3rd,7th,14th and 21st days. MAIN OUTCOME MEASURES:①general observation;②blood supply in recipient area u nder infrared ray observation;③histological observation. RESULTS:Transplanted AS had a favorable fusion between tissue engineered skin and self skin on the 3rd day after grafting with a few of ingrowths of capilla ries.The color of the AS was closed to self skin. The capillaries gradually inc reased in the grafts over time. The new epidermis was clearly consisted of strat um basale,stratum spinosum,stratum granulosum,and stratum corneum.Keratinization enhanced with exfoliation.Cells in dermis increased and the scaffold gradually degraded.The secreted extracellular matrix increased as well.On the 14th day aft er grafting,the wounds almost recovered. The color of the grafted artificial ski n was more similar to the nature skin with very little scaring,which indicated t hat a second grafting was unnecessary.The scabs did not completely fall off in C H group until the 14th day,and the wound was not healed.The color was darker tha n that of AS group.The scabs fell off in CG,and the wounds were big and deep wit h dark red color. CONCLUSION:The new reconstructive tissue engineered artificial skin with CH a s stromal scaffold has good histocompatibility,which can be applied in grafting for full thickness wounds.