脱细胞猪小肠黏膜下层的组织相容性

Histocompatibility of the decellularized porcine small intestinal submucosa

目的:制备脱细胞猪小肠黏膜下层(SIS),研究其用作组织工程支架材料的组织相容性。方法:0.1%十二烷基磺酸钠(SDS)处理新鲜猪SIS 30min制备脱细胞SIS,H-E、4,6-二脒基-2-苯基吲哚(DAPI)及天狼星红染色观察脱细胞前后SIS的生物学特性,将其植入SD大鼠背部皮下,检测其组织相容性,对照组SD大鼠植入未脱细胞的正常SIS作为阳性对照。结果:脱细胞SIS呈白色、半透明的膜状物,具有良好的弹性及柔韧性;H-E、DAPI及天狼星红染色光学显微镜观察结果显示脱细胞SIS无细胞及DNA残留,组织结构保留完整,胶原纤维间孔隙率增加;皮下埋植术后全部动物存活,各时间段脱细胞SIS植入组大鼠创口周围皮肤愈合良好,无红肿、渗出等炎症反应。组织学观察脱细胞SIS植入早期结构完整;随着植入时间的延长,脱细胞SIS逐渐降解,结构松散。脱细胞SIS植片周围炎症细胞数明显少于正常SIS植入组。结论:脱细胞猪SIS植入SD大鼠体内具有良好的组织相容性,未引起明显的炎症反应,是一种理想的组织工程支架材料。

Decellularized porcine small intestinal submucosa(SIS)was prepared and its histocompatibility was investigated as a scaffold in tissue engineering.Methods:Fresh porcine SIS was decellularized with the concentration of 0.1%SDS for 30 min to produce an acellular porcine SIS scaffold.After decellularization,the scaffolds were examined by H-E,DAPI and picrosirius red staining to observe the biological characteristics of SIS.Then SIS was implanted in the back of SD rats to investigate its histocompatibility.In addition,the implanted normal SIS was used as positive control.Results:Acellular SIS was white and semitransparent membrane with good flexibility.H-E,DAPI and picrosirius red staining with a light microscopy revealed that the acellular SIS was decellularized completely,the overall histoarchitecture was preserved,and the porosity of collagen fibers was increased.The SD rats in the subcutaneous implant test all survived,without inflammatory responses such as red swelling of the skin and effusion.The acellular SIS kept the integral structure under optical microscope with H-E staining at 7 days after implantation.Degradations were observed at 14 and 28 days after implantation.The average density of inflammatory cells around acellular SIS was significantly lower than the normal group.Conclusion:Acellular SIS has good in vivo histocompatibility and no severe inflammatory reaction.It is an ideal scaffold for tissue engineering.