摘要
背景:天然生物材料(如同种异体材料)生产过程中极易受到细菌污染,其中革兰阴性菌释放出的细菌内毒素可以引起人体发热、凝血、休克,严重的会导致死亡。如何去除同种异体材料中的内毒素,同时又能保证材料的天然结构和生物活性,目前尚未有相关报道。目的:探究去除同种异体材料等天然生物材料中内毒素的方法及效果,并对H_(2)O_(2)残留进行控制。方法:采用H_(2)O_(2)溶液协同超声处理方法去除同种异体骨材料的内毒素,研究超声条件及H_(2)O_(2)浓度对内毒素去除效果的影响程度,其中H_(2)O_(2)浓度分别设定为1%,2%,3%,4%,5%,超声功率分别设定为80,120,140,160,200 W,超声时间分别设定为20,30,40,50,60 min,超声水浴温度分别设定为30,35,40,45,50℃,通过内毒素含量检测选择最优的组合条件处理同种异体骨材料,进行后续实验。对去除内毒素后同种异体骨材料的微观结构、H_(2)O_(2)残留及细胞毒性进行检测,同时探讨不同浓度H_(2)O_(2)残留对L929小鼠成纤维细胞存活率的影响。结果与结论:①从内毒素去除效果来看,实验条件为H_(2)O_(2)浓度4%、超声功率200 W、超声时间50 min、超声水浴温度50℃时,为去除同种异体骨材料中内毒素的最佳条件,选择该最优组合去除同种异体骨材料的内毒素;②去除内毒素处理后同种异体骨材料在60℃水浴、换6次液后H_(2)O_(2)残留量小于47.551 mg/kg,对L929小鼠成纤维细胞活性无影响;③光学显微镜及扫描电镜观察显示,采用H_(2)O_(2)溶液协同超声处理对同种异体骨材料的结构无影响;④结果表明,H_(2)O_(2)溶液协同超声处理方式具有高效快速去除内毒素的特点,可保证同种异体骨材料的安全性。
BACKGROUND:Natural biomaterials such as allogeneic materials are highly susceptible to bacterial contamination during the production process.Among them,bacterial endotoxin released by Gram-negative bacteria can cause human fever,coagulation,shock,and even death.There is no relevant research on the method of endotoxin removed from allogeneic materials while maintaining their natural structure and biological activity.OBJECTIVE:To explore the method and effect of endotoxin removed from allogeneic materials and other natural biological materials and to control the residual H_(2)O_(2).METHODS:The endotoxin was removed from allogeneic bone materials by H_(2)O_(2)/ultrasound.The influence of ultrasound and the concentration of H_(2)O_(2)on the endotoxin removal process was explored.The concentration of H_(2)O_(2)was set as 1%,2%,3%,4%,and 5%by mass.The power of ultrasound was set as 80,120,140,160,and 200 W.The duration of ultrasound was 20,30,40,50,and 60 minutes.The temperature was 30,35,40,45,and 50℃.The optimal combination of conditions for treating allogeneic bone materials was selected by endotoxin content detection for subsequent experiments.The microstructure,H_(2)O_(2)residue,and cytotoxicity of allogeneic bone materials after endotoxin removal were detected,and the effects of different concentrations of H_(2)O_(2)residue on the survival rate of L929 mouse fibroblasts were explored.RESULTS AND CONCLUSION:(1)According to the results of the endotoxin removal process,when the concentration of H_(2)O_(2) was 4%,the power of ultrasound was 200 W,with the duration of ultrasound of 50 minutes,and the temperature 50℃,it would effectively remove endotoxin from allogeneic bone materials.The optimal combination was selected to remove endotoxins from allogeneic bone materials.(2)After endotoxin removal,the residual H_(2)O_(2) content of allogeneic bone materials was less than 47.551 mg/kg after being placed in a 60℃water bath and changing the solution 6 times,which had no effect on the viability of L929 mouse fibrobl
作者
侯立存
胡凯
邵怡然
Hou Licun;Hu Kai;Shao Yiran(Shanghai Gencong Biomedical Materials Research Center,Shanghai 201201,China)
出处
《中国组织工程研究》
CAS
北大核心
2025年第28期5985-5993,共9页
Chinese Journal of Tissue Engineering Research
关键词
同种异体材料
内毒素去除
超声
过氧化氢
联合处理
工程化骨材料
allogeneic material
endotoxin removal
ultrasound
hydrogen peroxide
combined treatment
engineered bone material
