使用聚乙二醇和海藻糖的组合冷冻干燥牛心包的性能影响  

Effects of freeze-drying bovine pericardium using a combination of polyethylene glycol and trehalose

作　　者：黄伟 李维杰[1,2,3] 刘宝林 HUANG Wei;LI Weijie;LIU Baolin(Institute of Biothermal Science and Technology,University of Shanghai for Science and Technology,Shanghai 200093,P.R.China;Shanghai Co-innovation Center for Energy Therapy of Tumors,Shanghai 200093,P.R.China;Shanghai Technical Service Platform for Cryopreservation of Biological Resources,Shanghai 200093,P.R.China)

机构地区：[1]上海理工大学生物系统热科学研究所,上海200093 [2]上海市肿瘤能量治疗技术与器械协同创新中心,上海200093 [3]上海市生物资源低温保存技术服务平台,上海200093

出　　处：《生物医学工程学杂志》2024年第2期368-375,共8页Journal of Biomedical Engineering

基　　金：上海市肿瘤能量治疗技术与器械协同创新中心。

摘　　要：冷冻干燥是一种使生物样本在干燥状态下保存的技术,有利于储存、运输并节省成本。本文使用聚乙二醇(PEG)和海藻糖(Tre)组成的冻干保护剂处理牛心包,进行冷冻干燥。结果表明:使用PEG+10%w/v Tre处理牛心包进行冷冻干燥后,其力学性能优于使用戊二醛(GA)固定的牛心包。卡尔费休法测定,牛心包复水后的湿态含水量为(74.81±1.44)%,与GA固定的牛心包无明显差异。干态含水量为(8.64±1.52)%,说明在冷冻干燥过程中能有效脱水。差示扫描量热仪(DSC)的测试结果显示,牛心包的热皱缩温度为(84.96±0.49)℃比GA固定的牛心包(83.14±0.11)℃高,说明具有更高的热稳定性。傅里叶变换红外光谱(FTIR)结果显示,在冷冻干燥过程中,蛋白质结构并未受到破坏。苏木精-伊红(HE)染色表明,冷冻干燥过程可以减少孔隙的产生,防止冰晶的生长,使组织纤维结构排列更紧密。细胞存活率与溶血率检测结果显示,细胞增殖率为(77.87±0.49)%,毒性分级为1级,溶血率为(0.17±0.02)%,低于5%的标准,即经过冷冻干燥处理的牛心包在细胞毒性和溶血方面表现良好,符合相关标准。综上,使用PEG+10%w/v Tre处理牛心包进行冷冻干燥后的性能符合要求。The freeze-drying is a technology that preserves biological samples in a dry state,which is beneficial for storage,transportation,and cost saving.In this study,the bovine pericardium was treated with a freeze-drying protectant composed of polyethylene glycol(PEG)and trehalose(Tre),and then freeze-dried.The results demonstrated that the mechanical properties of the pericardium treated with PEG+10%w/v Tre were superior to those of the pericardium fixed with glutaraldehyde(GA).The wet state water content of the rehydrated pericardium,determined using the Karl Fischer method,was(74.81±1.44)%,which was comparable to that of the GA-fixed pericardium.The dry state water content was significantly reduced to(8.64±1.52)%,indicating effective dehydration during the freeze-drying process.Differential scanning calorimetry(DSC)testing revealed that the thermal shrinkage temperature of the pericardium was(84.96±0.49)℃,higher than that of the GA-fixed pericardium(83.14±0.11)℃,indicating greater thermal stability.Fourier transform infrared spectroscopy(FTIR)results showed no damage to the protein structure during freeze-drying.Hematoxylin and eosin(HE)staining demonstrated that the freeze-drying process reduced pore formation,prevented ice crystal growth,and resulted in a tighter arrangement of tissue fibers.The frozen-dried bovine pericardium was subjected to tests for cell viability and hemolysis rate.The results revealed a cell proliferation rate of(77.87±0.49)%,corresponding to a toxicity grade of 1.Additionally,the hemolysis rate was(0.17±0.02)%,which is below the standard of 5%.These findings indicated that the frozen-dried bovine pericardium exhibited satisfactory performance in terms of cytotoxicity and hemolysis,thus meeting the relevant standards.In summary,the performance of the bovine pericardium treated with PEG+10%w/v Tre and subjected to freeze-drying could meet the required standards.

关 键 词：冷冻干燥 力学性能 热皱缩温度 聚乙二醇 海藻糖 

分 类 号：R318.08[医药卫生—生物医学工程]