Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
J Tissue Eng Regen Med. 2021 Feb;15(2):116-128. doi: 10.1002/term.3150. Epub 2020 Nov 23.
The aim of this study was evaluating the effects of vacuum on microstructure and biocompatibility of bovine decellularized pericardium. So the bovine pericardia were decellularized and then the vacuum was applied for two periods of time; 90 and 180 min. DNA, glucose amino glycan, collagen and elastin content assay, scanning electron microscopy (SEM) examination, hematoxylin and eosin (H&E) and Masson's trichrome stainings performed to evaluate microstructure of tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, subcutaneous implantation, and tensile test were used to assay biocompatibility and mechanical properties of decellularized tissues. The results showed that applying vacuum reduced residual DNA significantly. Vacuum after 180 min reduced more residual DNA. There were no significant differences in the content of glucose amino glycan (GAG), collagen, and elastin between the vacuumed and control groups. SEM examination was revealed that vacuum for 180 min increased pore size and porosity more than 90 min and control groups. H&E and Masson's trichrome stainings revealed extracellular matrix preservation after decellularization in all groups. Cell viability was increased in vacuumed samples significantly after 72 h in vaccumed samples. H&E staining and tensile test after implantation of tissues were showed less inflammation in the vacuum applied tissues and increased durability. The vacuum increased DNA removal, pore size, porosity, and biocompatibility in vitro and in vivo and durability of bovine decellularized pericardium in vivo. Considering the important role of time, more studies should be performed to optimize time, intensity, and method of application of vacuum in decellularization of different tissues as well as bovine pericardium.
本研究旨在评估真空对牛去细胞心包组织的微观结构和生物相容性的影响。为此，将牛心包组织进行去细胞化处理，然后分别进行 90 分钟和 180 分钟的真空处理。通过 DNA、葡萄糖氨基聚糖、胶原蛋白和弹性蛋白含量测定、扫描电子显微镜（SEM）检查、苏木精和伊红（H&E）及 Masson 三色染色评估组织的微观结构。通过 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）试验、皮下植入和拉伸试验评估去细胞组织的生物相容性和力学性能。结果表明，应用真空可显著减少残留 DNA。180 分钟的真空处理可减少更多的残留 DNA。与对照组相比，实验组葡萄糖氨基聚糖（GAG）、胶原蛋白和弹性蛋白的含量无显著差异。SEM 检查显示，180 分钟的真空处理比 90 分钟和对照组增加了更多的孔径和孔隙率。H&E 和 Masson 三色染色显示所有组的细胞外基质在去细胞化后均得到保留。细胞活力在真空处理样本中 72 小时后显著增加。组织植入后的 H&E 染色和拉伸试验显示，应用真空的组织炎症反应较少，耐久性增加。真空处理在体外和体内增加了牛去细胞心包组织的 DNA 去除率、孔径、孔隙率、生物相容性和耐久性。考虑到时间的重要作用，应该进行更多的研究，以优化不同组织和牛心包去细胞化过程中真空的时间、强度和应用方法。
