An efficient protocol for decellularization of the human endometrial fragments for clinical usage.

IF 4.4 3区 医学 Q2 ENGINEERING, BIOMEDICAL Progress in Biomaterials Pub Date : 2021-06-01 Epub Date: 2021-05-21 DOI:10.1007/s40204-021-00156-5
Zinat Sargazi, Saeed Zavareh, Mina Jafarabadi, Mojdeh Salehnia
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引用次数: 6

Abstract

The present study was aimed to compare different decellularization protocols for human endometrial fragments. The freeze-thaw cycles in combination with treatment by Triton X-100 and four concentrations of sodium dodecyl sulfate (SDS; 0.1, 0.5, 1, and 1.5%) with two exposure times (24 and 72 h) were applied for tissues decellularization. After analysis the morphology and DNA content of tissues the group with better morphology and lower DNA content was selected for further assessments. The nucleus by Acridine orange and extracellular matrix (ECM) using Masson's trichrome, Alcian blue, and periodic acid-Schiff staining were studied. The amount of tissues collagen types I and IV, fibronectin, glycosaminoglycans (GAGs), and elastin was analyzed by Raman spectroscopy. The ultrastructure and porosity of decellularized scaffold were studied by scanning electron microscopy (SEM). The MTT assay was applied for assessments of cytotoxicity of scaffold. The treated group with 1% SDS for 72 h showed the morphology similar to native control in having the minimum level of DNA and well preserved ECM. Raman spectroscopy results demonstrated, the amount of collagen types I and IV, GAG, and fibronectin was not significantly different in decellularized scaffold compared with native group but the elastin protein level was significantly decreased (P < 0.001). SEM micrographs also showed a porous and fiber rich ECM in decellularized sample similar to the native control. This combined protocol for decellularization of human endometrial tissue is effective and it could be suitable for recellularization and clinical applications in the future.

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一种用于临床的人子宫内膜碎片脱细胞的有效方案。
本研究旨在比较不同的脱细胞方案的人子宫内膜碎片。冻融循环联合Triton X-100和四种浓度的十二烷基硫酸钠(SDS;0.1, 0.5, 1和1.5%),两次暴露时间(24和72小时)用于组织脱细胞。分析组织形态和DNA含量后,选择形态较好、DNA含量较低的组进行进一步评价。细胞核采用吖啶橙染色,细胞外基质(ECM)采用马松三色、阿利新蓝和周期性酸-希夫染色。用拉曼光谱分析组织中I型和IV型胶原蛋白、纤维连接蛋白、糖胺聚糖(GAGs)和弹性蛋白的含量。采用扫描电镜对脱细胞支架的超微结构和孔隙率进行了研究。采用MTT法评价支架的细胞毒性。1% SDS处理72 h后,细胞形态与自然对照相似,DNA水平最低,ECM保存良好。拉曼光谱结果显示,去细胞支架中I型和IV型胶原、GAG和纤维连接蛋白的数量与天然支架组相比无显著差异,但弹性蛋白水平明显降低(P
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来源期刊
Progress in Biomaterials
Progress in Biomaterials MATERIALS SCIENCE, BIOMATERIALS-
CiteScore
9.60
自引率
4.10%
发文量
35
期刊介绍: Progress in Biomaterials is a multidisciplinary, English-language publication of original contributions and reviews concerning studies of the preparation, performance and evaluation of biomaterials; the chemical, physical, biological and mechanical behavior of materials both in vitro and in vivo in areas such as tissue engineering and regenerative medicine, drug delivery and implants where biomaterials play a significant role. Including all areas of: design; preparation; performance and evaluation of nano- and biomaterials in tissue engineering; drug delivery systems; regenerative medicine; implantable medical devices; interaction of cells/stem cells on biomaterials and related applications.
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