十二烷基醚硫酸钠和十二烷基硫酸钠对大鼠肠道脱细胞作用的组织学和共聚焦拉曼显微镜比较

Seyed Hossein Asadi‐Yousefabad, Sajad Daneshi, M. Pourentezari, N. Tanideh, Mohammad Zamani Rarani, Elias Kargar-Abarghouei, Hengameh Dortaj, M. Salari, Zeinolabedin Sharifian Dastjerdi
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摘要

背景:十二烷基硫酸钠(SDS)洗涤剂广泛应用于组织脱细胞制备组织工程支架。尽管它具有很强的脱细胞作用,但这种物质具有相对高的毒性,并引起组织成分的改变。十二烷基醚硫酸钠(SLES)是一种新型聚阴离子洗涤剂,其毒性比SDS小,但比SDS弱。本研究旨在利用SDS和SLES溶液使肠组织脱细胞,形成细胞支架,并检测从该组织中获得的支架。方法:雄性Sprague-Dawley大鼠18只分为3组。所有大鼠麻醉后均取肠。第一组(对照组)将大鼠肠道置于10%福尔马林溶液中。在第二组中,使用SLES溶液对肠道进行脱细胞。第三组动物的肠道用SDS溶液去细胞化。为了评估脱细胞作用,用苏木精-伊红染色和阿利新蓝染色检测糖胺聚糖(GAGs),用马松三色染色检测胶原纤维。用共聚焦拉曼显微镜比较胶原蛋白、脂质、GAG和基因含量。结果:苏木精-伊红染色显示,SDS或SLES均能去除去细胞支架的细胞核和DNA。与SDS组相比,SLES组上皮组织的变化较少,两种支架的肌肉层都得到了很好的保存。共聚焦拉曼显微镜结果显示,两种洗涤剂均能分解色氨酸、脂质、糖原和蛋白质;然而,两种物质的糖原残留量相同,但SLES脱细胞肠中蛋白质、羟脯氨酸和脂质的二硫键大部分被保留。结论:两种物质均适用于肠道脱细胞,去除肠道组织的整体结构,但SLES保留的胶原蛋白和GAG含量优于SDS。
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Comparison of Sodium Lauryl Ether Sulfate and Sodium Dodecyl Sulfate in the Intestinal Decellularization of Rats Using a Histological Method and Confocal Raman Microscopy
Background: Sodium dodecyl sulfate (SDS) detergent is widely used in tissue decellularization to produce scaffolds for tissue engineering. Despite its strong decellularization, this substance has relatively high toxicity and causes changes in tissue composition. Sodium lauryl ether sulfate (SLES) is a new poly anionic detergent that is less toxic than SDS but weaker than it. The present study aimed to decellularize the intestinal tissue using SDS and SLES solutions, forming a cell scaffold, and examining scaffolds obtained from this tissue. Methods: Eighteen male Sprague-Dawley rats were divided into three groups. The intestines of all rats were removed after anesthesia. In the first group (controls), rats’ intestines were placed in a 10% formalin solution. In the second group, intestines were decellularized using an SLES solution. In the third group animals’ intestines were decellularized using an SDS solution. To evaluate decellularization, samples were stained with hematoxylin-eosin staining and Alcian blue staining for glycosaminoglycans (GAGs), and Masson’s trichrome for collagen fibers. A confocal Raman microscope was used to compare collagen, lipid, GAG, and genetic content. Results: Hematoxylin-eosin staining showed that the nucleus and DNA were removed in the decellularized scaffolds by SDS or SLES. The SLES group, compared to the SDS group, showed fewer changes in the epithelial tissue, and muscle layers in both scaffolds were well preserved. The results of confocal Raman microscopy showed that tryptophan, lipid, glycogen, and protein were broken down by both detergents; however, the residual amount of glycogen was the same in both substances, but disulfide bonds of proteins, hydroxyproline, and lipids in the decellularized intestine with SLES were mostly preserved. Conclusion: Both substances were suitable for intestinal decellularization and removed the overall structure of intestinal tissue, but SLES retained collagen and GAG content better than SDS.
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