冻干辐照对猪腹膜体外稳定性的影响

Freeze-drying and irradiation effects on stability of the porcine peritoneum in vitro

背景：有实验表明60Coγ射线灭菌处理的胶原膜会促使胶原分子产生交联,影响其稳定性。目的：通过测定冻干辐照处理后猪腹膜胶原酶酶解时间,了解冻干辐照猪腹膜在体外的稳定性。设计、时间及单位：动物实验观察,于2007-07/2008-06在南昌大学烧伤研究所、南昌大学-新三思公司联合力学实验室、江西中医学院中药固体制剂国家工程研究中心与江西天兆科技发展公司完成。材料：选用健康活体猪6只,无菌条件下取新鲜猪腹膜。60Coγ射线辐照装置为江西天兆科技发展公司生产。方法：新鲜猪腹膜按照处理方式分成4组,新鲜组：将新鲜猪腹膜放入含有庆大霉素1×105U/L的生理盐水中清洗修剪平整后,再放入保护液中浸泡约15min,最后放入0-4℃冰箱中保存、备用。冻干组：将新鲜猪腹膜放入EMDE＋10%甘油中的猪腹膜浸泡15min后,取出放入-78℃的冰箱中,使其形成冰晶,再放入真空冷冻干燥机内冻干6h,取出室温保存、备用。辐照组：在冻干组基础上将猪腹膜放入剂量为25kGy的60Coγ辐照装置内,动态辐照72h后取出,放入0-4℃冰箱中保存、备用。冻干＋辐照组：在辐照组基础上将猪腹膜放入剂量为25kGy的60Coγ辐照装置内,动态辐照72h后取出室温保存、备用。主要观察指标：同时取直径为5mm各组猪腹膜圆片用胶原酶Ⅰ型酶解,计算各片材料完全酶解时间。结果：4组猪腹膜胶原酶酶解时间：（9.6±1.2）,（6.1±1.6）,（29.2±3.0）,（23.5±2.6）mins。辐照组和冻干＋辐照组猪腹膜酶解时间比新鲜组和冻干组明显延长（P〈0.05）,但新鲜组和冻干组之间比较及辐照组和冻干＋辐照组之间比较,猪硬脑膜酶解时间无明显差别（P〉0.05）。结论：经60Coγ射线照射后的猪腹膜抗酶解能力增强;辐照均能增加猪腹膜的稳定性,冻干对猪腹膜的稳定性无明显影响。

BACKGROUND： Collagen membrane following ^60Co γ ray germicidal treatment can promote the crosslinking of collagen molecule and affect its stability. OBJECTIVE： To determine the in vitro stability of freeze-dried and irradiated porcine peritoneum by assessing enzymolysis time of collagenase in porcine peritoneum following irradiated and freeze-dried treatment. DESIGN, TIME AND SETTING： This animal experiment was performed at the Burn Institute, Nanchang University, Association Mechanics Laboratory, Nanchang University-New Sans Company, Chinese Crude Drug Solid Preparation Nation Project Research Center of Jiangxi Traditional Chinese Medical College and Jiangxi Tianzhao Technology Development Company from July 2007 to June 2006. MATERIALS： Six healthy living pigs were used to sterilely obtain fresh porcine peritoneum. ^60Co γ ray irradiation equipment was purchased from Jiangxi Tianzhao Technology Development Company. METHODS： Fresh porcine peritoneum was assigned into 4 groups. In the fresh group, fresh porcine peritoneum was washed in saline containing 1×10^5 U/L gentamicin, trimmed, and then immersed in protective solution for 15 minutes. Finally, samples were placed in 0 4 ℃ for use. In the freeze-dried group, fresh porcine peritoneum was placed in EMDE＋10% glycerol for 15 minutes, placed in -78 ℃ to form ice crystal, froze and dried in a vacuum freeze-drying machine for 6 hours, and then took out at room temperature for use. In the irradiation group, on the basis of freeze-dried group, the porcine peritoneum was irradiated with 25 kGy ^60Co γ ray in an irradiation equipment for 72 hours, and then maintained in 0-4℃ for use. In the freeze-dried ＋ irradiation group, on the basis of irradiation group, the porcine peritoneum was irradiated with 25 kGy ^60Co γ ray in an irradiation equipment for 72 hours, and then maintained at room temperature for use. MAIN OUTCOME MEASURES： 5-mm wafer of each group porcine peritoneum were dealt with type I collagenase, and digested time was recorded. RESULTS： Collagenase enzymolysis time of the porcine peritoneum was respectively （9.6±1.2）, （6.1±1.6）, （29.2±3.）, （23.5±2.6） minutes in four groups. Enzymolysis time was significantly longer in the irradiation group and the freeze-dried ＋ irradiation group compared with the fresh group and the freeze-dried group （P 〈 0.05）. However, no significant differences in enzymolysis time of the porcine peritoneum were detected between the fresh group and the freeze-fried group, between the irradiation group and the freeze-dried ＋ irradiation group （P 〉 0.05）. CONCLUSION： Following ^60Co γ-ray irradiation, antienzymolysis ability became strong. Irradiation can increase the stability of the porcine peritoneum. Freeze-frying does not have significant effects on porcine peritoneum stability.