Abbas Fazel Anvari Yazdi, Kobra Tahermanesh, Maryam Ejlali, Amin Babaei-Ghazvini, Bishnu Acharya, Ildiko Badea, Daniel J MacPhee, Xiongbiao Chen
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引用次数: 0
摘要
导言：脱细胞子宫细胞外基质已成为生物材料领域的一个关键焦点，为子宫组织再生、疾病诊断和治疗研究以及最终的子宫移植提供了一个前景广阔的来源。在这项研究中，我们考察了猪子宫组织脱细胞的各种方案，旨在揭示 DNA 去除、生物活性分子保存和微结构改变的复杂动态：用十二烷基硫酸钠（SDS）、Triton® X-100、过乙酸+乙醇和 DNase I 对猪子宫组织进行了 6 种不同的处理，这些处理方案都是经过严格筛选和设计的。脱细胞后，我们检测了 DNA 定量、组织学染色（H&E 和 DAPI）、糖胺聚糖（GAG）检测、扫描电子显微镜（SEM）、傅立叶变换红外光谱（FT-IR）、X 射线光电子能谱（XPS）和热重分析（TGA）：对所有 6 种方案进行了比较分析，结果表明所有方案都实现了脱细胞；而 0.1% SDS + 1% Triton® X-100 加搅拌的 DNA 去除效率最高。此外，研究还发现，DNase I 在提高脱细胞过程的效率方面发挥了关键作用，它在消化细胞内容物和清除细胞碎片方面的作用显著，达到 99.79% （19.63 ± 3.92 纳克/毫克干重）：我们的研究结果加深了对脱细胞子宫细胞外基质中 DNA 清除、GAG 保留、微结构改变和蛋白质分解的细微理解，同时强调了为预期应用而设计的脱细胞方案的重要性。这项研究和我们的发现标志着子宫移植和相关组织工程/再生医学领域取得了重大进展。
Comparative analysis of porcine-uterine decellularization for bioactive-molecule preservation and DNA removal.
Introduction: Decellularized uterine extracellular matrix has emerged as a pivotal focus in the realm of biomaterials, offering a promising source in uterine tissue regeneration, research on disease diagnosis and treatments, and ultimately uterine transplantation. In this study, we examined various protocols for decellularizing porcine uterine tissues, aimed to unravel the intricate dynamics of DNA removal, bioactive molecules preservation, and microstructural alterations.
Methods: Porcine uterine tissues were treated with 6 different, yet rigorously selected and designed, protocols with sodium dodecyl sulfate (SDS), Triton® X-100, peracetic acid + ethanol, and DNase I. After decellularization, we examined DNA quantification, histological staining (H&E and DAPI), glycosaminoglycans (GAG) assay, scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and Thermogravimetric Analysis (TGA).
Results: A comparative analysis among all 6 protocols was conducted with the results demonstrating that all protocols achieved decellularization; while 0.1% SDS + 1% Triton® X-100, coupled with agitation, demonstrated the highest efficiency in DNA removal. Also, it was found that DNase I played a key role in enhancing the efficiency of the decellularization process by underscoring its significance in digesting cellular contents and eliminating cell debris by 99.79% (19.63 ± 3.92 ng/mg dry weight).
Conclusions: Our findings enhance the nuanced understanding of DNA removal, GAG preservation, microstructural alteration, and protein decomposition in decellularized uterine extracellular matrix, while highlighting the importance of decellularization protocols designed for intended applications. This study along with our findings represents meaningful progress for advancing the field of uterine transplantation and related tissue engineering/regenerative medicine.
期刊介绍：
The translation of new discoveries in medicine to clinical routine has never been easy. During the second half of the last century, thanks to the progress in chemistry, biochemistry and pharmacology, we have seen the development and the application of a large number of drugs and devices aimed at the treatment of symptoms, blocking unwanted pathways and, in the case of infectious diseases, fighting the micro-organisms responsible. However, we are facing, today, a dramatic change in the therapeutic approach to pathologies and diseases. Indeed, the challenge of the present and the next decade is to fully restore the physiological status of the diseased organism and to completely regenerate tissue and organs when they are so seriously affected that treatments cannot be limited to the repression of symptoms or to the repair of damage. This is being made possible thanks to the major developments made in basic cell and molecular biology, including stem cell science, growth factor delivery, gene isolation and transfection, the advances in bioengineering and nanotechnology, including development of new biomaterials, biofabrication technologies and use of bioreactors, and the big improvements in diagnostic tools and imaging of cells, tissues and organs.
In today`s world, an enhancement of communication between multidisciplinary experts, together with the promotion of joint projects and close collaborations among scientists, engineers, industry people, regulatory agencies and physicians are absolute requirements for the success of any attempt to develop and clinically apply a new biological therapy or an innovative device involving the collective use of biomaterials, cells and/or bioactive molecules. “Frontiers in Bioengineering and Biotechnology” aspires to be a forum for all people involved in the process by bridging the gap too often existing between a discovery in the basic sciences and its clinical application.
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