脱细胞猪小肠黏膜下层粉末用于大鼠脾部分切除创面止血的研究

Study on acellular porcine small intestinal submucosa powders for hemostasis of wound in rats after partial splenectomy

目的探讨脱细胞猪小肠黏膜下层(SIS)粉末在大鼠脾部分切除创面的止血效果。方法经机械刮除、脱脂脱细胞、胃蛋白酶酶解、冻干等步骤获得SIS粉末。HE染色检测脱细胞SIS细胞残留情况,检测SIS粉末吸水性并在扫描电镜下观察其超微结构。将40只SD大鼠随机分为A、B、C、D组,切除大鼠部分脾组织,并将SIS粉末、福和泰止血粉、瞬时止血粉、无菌纱布压迫分别用于A、B、C、D组大鼠创面止血。扫描电镜下观察止血粉在创面上的情况,记录出血时间和出血量。于第20天、30天每组各处死5只大鼠,肉眼以及HE染色镜下观察创面止血粉末吸收情况。结果经过脱脂脱细胞处理后,SIS无细胞成分残留,镜下见SIS粉末呈单层的长薄片状。SIS粉末吸水率在120 s时达到饱和,是瞬时止血粉的1.87倍,差异具有统计学意义(P<0.01)。脾部分切除使用止血粉后,A、B、C组创面均很快停止出血。A、B、C组的出血时间和出血量均显著短/少于D组(P<0.05);A组的出血时间和出血量均明显短/少于C组(P<0.05),A组的出血时间明显短于B组(P<0.05)。肉眼及镜下可见在第20天A、B、C组均可见未吸收完的止血粉末,第30天止血粉末较第20天明显吸收。结论脱细胞SIS不含细胞成分,在体内可降解,具有极强的吸水性,对大鼠脾部分切除创面具有令人满意的止血效果。

Objective To explore the hemostatic effect of acellular porcine small intestinal submucosa(SIS)powders on the wound of rats with partial spleen excision.Methods SIS powders were obtained by mechanical scraping,degreasing and decellularization,pepsin thawing,and freeze-drying.The residual cells of decellularized SIS was detected by HE staining,and the water absorption of SIS powders was examined and its ultrastructure was observed under a scanning electron microscope.A total of 40 SD rats were randomly divided into group A,B,C,D,with partial splenic tissue removed.SIS powders,Fuhetai hemostatic powders,instantaneous hemostatic powders and sterile gauze compression were used for wound hemostasis in group A,B,C and D,respectively.The ultrastructure was observed under the scanning electron microscope,and the bleeding time and amount of the wound were recorded.Five rats were killed at the 20th day and 30th day in each group.The absorption of hemostatic powders on the wound were observed by naked eyes and HE staining under the microscope.Results After the degreasing and decellularization,the SIS had no cellular components remaining,and the SIS powders were seen in the shape of a single layer of long flakes under the microscope.The water absorption rate of SIS powders reached saturation at 120 seconds,which was 1.87 times on that of instantaneous hemostatic powders,and the difference was statistically significant(P<0.01).After spleenectomy and using hemostatic powders,the wounds of group A,B,C stopped bleeding quickly.The bleeding time and volume of group A,B,and C were significantly shorter/lower than those of group D(P<0.05).The bleeding time and volume of group A were significantly shorter/lower than those of group C,the bleeding time of group A was significantly shorter than that of group B(P<0.05).The unabsorbed hemostatic powders were observed in group A,B and C on the 20th day,and the powders were significantly absorbed on the 30th day.Conclusion The decellularized SIS contains no cellular components,is degradable in the body,has extremely strong water absorption,and has a satisfactory hemostatic effect on wound of rats with partial splenectomy.