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基于单一去污剂的心脏组织去细胞化方案的优化技术

Optimization Techniques of Single-Detergent Based Protocols for Heart Tissue Decellularization.

作者信息

Meșină Mihai, Mîndrilă Ion, Meșină-Botoran Mihaela Iustina, Mîndrilă Laura Andreea, Farhangee Arsalan, Pirici Ionica

机构信息

Doctoral school, University of Medicine and Pharmacy of Craiova, Romania.

Department of Anatomy, Faculty of Medicine, University of Medicine and Pharmacy of Craiova, Romania.

出版信息

Curr Health Sci J. 2023 Apr-Jun;49(2):156-162. doi: 10.12865/CHSJ.49.02.156. Epub 2023 Jun 30.

DOI:10.12865/CHSJ.49.02.156
PMID:37779828
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10541079/
Abstract

The extracellular matrix (ECM) scaffolds are considered a gold standard for the engineering of appropriate grafts used in regenerative medicine for tissue repair, and decellularization of myocardial tissue is one of the most studied processes for obtaining natural ECM to date. Decellularization methods, agents used, or treatment durations can be varied to optimize cardiac tissue decellularization parameters. In this work we performed a morphological and morphometric analysis of cardiac tissue subjected to decellularization protocols based on Sodium Deoxycholate (SD) or Sodium Lauryl Sulfate (SLS) to identify factors that allow optimization of single-detergent based protocols for cardiac ECM manufacturing. For this, Wistar rat hearts (n=10) were subjected to 5 different decellularization protocols (n=2) and then histologically processed to achieve H&E or Azan trichrome stained sections for the morphological and morphometric analysis of the obtained ECM. The results of this study showed that SLS alters the spatial distribution of cardiac ECM collagen fibers, and SD can be successfully used in tailoring single-based detergent decellularization protocols by appropriately adjusting the application times of hypo/hyperosmotic shocks, which increases the lytic action of the detergent, and the washing times for the efficient elimination of cellular residues.

摘要

细胞外基质(ECM)支架被认为是用于组织修复的再生医学中合适移植物工程的金标准,心肌组织去细胞化是迄今为止获取天然ECM研究最多的过程之一。去细胞化方法、使用的试剂或处理持续时间可以变化,以优化心脏组织去细胞化参数。在这项工作中,我们对基于脱氧胆酸钠(SD)或十二烷基硫酸钠(SLS)的去细胞化方案处理的心脏组织进行了形态学和形态计量学分析,以确定能够优化基于单一去污剂的心脏ECM制造方案的因素。为此,将Wistar大鼠心脏(n = 10)进行5种不同的去细胞化方案(n = 2)处理,然后进行组织学处理,以获得苏木精-伊红(H&E)或阿赞三色染色切片,用于对所得ECM进行形态学和形态计量学分析。本研究结果表明,SLS会改变心脏ECM胶原纤维的空间分布,通过适当调整低渗/高渗休克的应用时间以及洗涤时间,SD可成功用于定制基于单一去污剂的去细胞化方案,以提高去污剂的裂解作用并有效去除细胞残留物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/a001680eaff3/CHSJ-49-02-156-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/1eb8b7866063/CHSJ-49-02-156-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/6c0dc58763b5/CHSJ-49-02-156-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/853ddf1da948/CHSJ-49-02-156-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/a001680eaff3/CHSJ-49-02-156-fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/1eb8b7866063/CHSJ-49-02-156-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/6c0dc58763b5/CHSJ-49-02-156-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/853ddf1da948/CHSJ-49-02-156-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1ef/10541079/a001680eaff3/CHSJ-49-02-156-fig4.jpg

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