抗生素-过氧乙酸对甲状腺去细胞生物支架消毒效果评价

Evaluation of the antibiotics-peracetic acid sterilization method for decellularized thyroid gland scaffolds

目的:采用抗生素-过氧乙酸两步消毒法对甲状腺去细胞支架进行消毒并对其评价。方法:震荡法制备兔甲状腺去细胞生物支架,分别采用PBS、抗生素及抗生素-过氧乙酸处理去细胞支架,观察处理后支架的三维结构、细胞毒性以及微生物含量。结果:H-E染色和电镜扫描提示3组支架结构无破坏,三维空间结构完整;CCK-8检测提示处理后3组支架无细胞毒性,3组间差异无统计学意义;微生物定量检测分析提示抗生素-过氧乙酸组支架内微生物含量为0 CFU/g,显著低于抗生素组3.45×10^3(2.4×10^3~4.7×10^3)CFU/g和正常对照组2.30×10^8 CFU/g(0.9×10^8~4.1×10^8),3组间微生物含量有显著性差异。结论:抗生素-过氧乙酸两步法成功对甲状腺去细胞生物支架进行消毒、灭菌,方法简便、有效,灭菌后支架无毒性残留及结构破坏。

Objective:To investigate the efficiency of a two-step antibiotics and peracetic acid sterilization method on thyroid gland decellularized biologic scaffolds and to provide experimental data for decellularized thyroid gland scaffolds transplantation.Methods:Thyroid gland decullularized biologic scaffolds were prepared by soaking with rotating and sterilized by PBS,antibiotics,antibiotics-peracetic acid respectively.The three-dimensional structure,cytotoxicity and bacterial bioburden of decellularized biologic scaffolds were observed after disinfection treatment.Results:The H-E staining and scanning electron microscopy showed the three-dimensional structure was preserved well in three groups.The CCK-8 detection showed there was no cytotoxicity in three groups.The absorbance values in three groups had no significant differences.Bacterial bioburden detection showed that antibiotics-peracetic acid decellularized scaffolds were sterile,with a calculated bioburden of zero,antibiotics decellularized scaffolds and PBS decellularized scaffolds were not sterile,with a mean bioburden of 3.45×10^3(2.4×10^3-4.7×10^3)CFU/g and 2.30×10^8 CFU/g(0.9×10^8-4.1×10^8)respectively.There were significant differences in the three groups.Conclusion:Antibiotic-peracetic acid sterilization method successfully disinfects and sterilizes the thyroid decellularized biological scaffold.The method is simple and effective,and the scaffold has no toxicity residue and structural damage after sterilization.