脱细胞技术在组织工程气管构建过程中的应用与特点

doi:10.3969/j.issn.2095-4344.2773

中国组织工程研究 ›› 2020, Vol. 24 ›› Issue (35): 5695-5700.doi: 10.3969/j.issn.2095-4344.2773

• 组织构建综述 tissue construction review • 上一篇下一篇

脱细胞技术在组织工程气管构建过程中的应用与特点

王志豪，武聪，吴思华，史宏灿

扬州大学临床医学院，扬州大学转化医学中心，江苏省中西医结合老年病防治重点实验室，江苏省扬州市 225001

收稿日期:2019-12-30 修回日期:2020-01-07 接受日期:2020-02-26 出版日期:2020-12-18 发布日期:2020-10-19
通讯作者: 史宏灿，主任医师，教授，博士生导师，扬州大学医学院，江苏省扬州市 225001
作者简介:王志豪，男，1994年生，河南省焦作市人，汉族，扬州大学医学院在读硕士，主要从事组织工程气管脱细胞基质研究。
基金资助:
国家自然科学基金(81770018)；江苏省研究生科研创新计划(XKYCX19_153)

Application and characteristics of decellularization technology in tissue engineered trachea

Wang Zhihao, Wu Cong, Wu Sihua, Shi Hongcan

Yangzhou University School of Clinical Medicine, Yangzhou University Transformation Medical Center, Jiangsu Provincial Key Laboratory of Geriatric Disease Prevention and Treatment with Integrated Traditional Chinese and Western Medicine, Yangzhou 225001, Jiangsu Province, China

Received:2019-12-30 Revised:2020-01-07 Accepted:2020-02-26 Online:2020-12-18 Published:2020-10-19
Contact: Shi Hongcan, Chief physician, Professor, Doctoral supervisor, Yangzhou University School of Clinical Medicine, Yangzhou University Transformation Medical Center, Jiangsu Provincial Key Laboratory of Geriatric Disease Prevention and Treatment with Integrated Traditional Chinese and Western Medicine, Yangzhou 225001, Jiangsu Province, China
About author:Wang Zhihao, Master candidate, Yangzhou University School of Clinical Medicine, Yangzhou University Transformation Medical Center, Jiangsu Provincial Key Laboratory of Geriatric Disease Prevention and Treatment with Integrated Traditional Chinese and Western Medicine, Yangzhou 225001, Jiangsu Province, China
Supported by:
the National Natural Science Foundation of China, No. 81770018; the Postgraduate Research Innovation Program of Jiangsu Province, No. XKYCX19_153

摘要/Abstract

摘要：

文题释义：

脱细胞技术：是物理、化学和生物酶法去除组织/器官中的细胞成分，消除抗原性，保留器官的三维结构和含有生物活性的细胞外基质。物理法通过超声、高压、冻融等物理手段破坏细胞膜达到脱细胞效果；化学法利用去污剂改变细胞膜的通透性，使细胞溶胀破裂完成脱细胞过程；生物酶法主要使核酸酶、胰蛋白酶对组织及器官进行脱细胞处理。

去污剂联合酶法：是以脱氧胆酸钠和脱氧核糖核酸酶为基础的脱细胞方法，通过脱氧胆酸钠和脱氧核糖核酸酶破坏细胞膜、消除细胞DNA，消除组织和器官的免疫原性，并保留细胞外基质，为细胞生长和增殖提供物质基础。通过化学法和生物酶法结合，可有效提高脱细胞效率。

背景：气管长段损伤的修复一直是气管治疗的重大挑战，组织工程气管脱细胞技术可为气管长段损伤修复提供有效治疗途径。

目的：综述组织工程气管中不同脱细胞技术的优缺点及其在气管替代治疗中的应用。

方法：以“tissue engineered trachea, decellularization, tracheal reconstruction，tracheal replacement therapy；组织工程气管，脱细胞，气管重建，气管替代治疗”为关键词，检索CNKI、万方、PubMed数据库2006年1月至2019年11月的相关文献进行分析总结。

结果与结论：①气管长段损伤常因肿瘤、创伤、狭窄、感染等病变引起，需气管替代物进行气道重建治疗，而组织工程研究中脱细胞技术制备的脱细胞气管基质是其治疗的有效途径；②脱细胞技术通过物理、化学和生物酶等工艺去除组织或器官的免疫原性，并有效保留组织器官的细胞外基质，为重塑功能性气管提供支持环境，术后患者无需依赖免疫抑制剂；③目前已有脱细胞技术用于临床气管移植的案例，但患者术后出现较多并发症，其功能尚需进一步完善；④随着脱细胞技术研究的不断深入，脱细胞方法变得更加成熟、精细、多样，为未来临床气管治疗提供了方向和途径。

ORCID: 0000-0002-4808-9644(王志豪)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 组织工程气管, 脱细胞, 气管替代治疗, 气管重建, 细胞外基质, 去污剂联合酶法, 真空脱细胞法, 十二烷基硫酸钠

Abstract:

BACKGROUND: Repair of long tracheal injury has always been a major challenge in tracheal therapy. Tissue engineering tracheal decellularization technology can provide an effective treatment approach for long tracheal injury repair.

OBJECTIVE: To review the advantages and disadvantages of different acellular techniques in tissue engineered trachea and their application in tracheal replacement therapy.

METHODS: Using the terms “tissue engineered trachea, decellularization, tracheal reconstruction, tracheal replacement therapy” as key words, CNKI, Wanfang and PubMed database from January 2006 to November 2019 were analyzed and summarized.

RESULTS AND CONCLUSION: Long tracheal injury is mainly caused by tumor, trauma, stenosis, and infection. It requires tracheal replacement for airway reconstruction therapy, and the decellularized tracheal matrix prepared by acellular technology in tissue engineering research is an effective way for its treatment. Decellularization technology removes the immunogenicity of tissues or organs through physical, chemical and biological enzymes, and effectively retains the extracellular matrix of tissues and organs, providing a supportive environment for remodeling functional trachea. After surgery, patients do not need to rely on immunosuppressants. At present, there are cases of decellularization technology used in clinical tracheal transplantation, but patients have more complications after surgery, and its functions need to be further improved. With the development of decellularization technology, acellular methods have become more mature, sophisticated and diversified, providing directions and approaches for future clinical tracheal treatment.

Key words: tissue engineered trachea, decellularization, tracheal replacement therapy, tracheal reconstruction, extracellular matrix, detergent combined with enzyme method, vacuum-assisted decellularization, sodium dodecyl sulfate

中图分类号:

王志豪, 武聪, 吴思华, 史宏灿.

脱细胞技术在组织工程气管构建过程中的应用与特点 [J]. 中国组织工程研究, 2020, 24(35): 5695-5700.

Wang Zhihao, Wu Cong, Wu Sihua, Shi Hongcan. Application and characteristics of decellularization technology in tissue engineered trachea[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(35): 5695-5700.

图/表（结果） 2

参考文献

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引言

材料方法

讨论

组织工程气管重建为气管长段损伤患者带来希望，有望帮助患者重建正常的呼吸功能，其中细胞外基质的保留是气管功能重建的关键因素，其与细胞生物功能密切相关，是细胞微环境的主要成分，可更好地模拟体内细胞微环境。气管经脱细胞技术处理后获得的脱细胞基质不仅可以有效地保留细胞外基质，去除抗原性，还具有良好的生物相容性和机械性能^[45-46]。其内含有多种活性成分与生长因子，可主动诱导并支持细胞的黏附、增殖、分化，迁入的细胞对材料进一步改造、降解，促进组织修复，重塑血管化、功能化的组织与器官，实现组织的形成和结构重塑^[47]。气管组织可通过物理、化学或生物等多种脱细胞方法联合应用，可以尽可能地保持细胞外基质生物活性的完整，促进气管组织功能的修复和重建。

目前去污剂联合酶的脱细胞方法已应用于气管疾病的临床研究，2008年MACCHIARINI等^[34]完成临床上首例成功组织工程气管置换的病例，改善患者因肺结核浸润继发晚期左主支气管软化症所致的严重呼吸困难症状。随后，国外在2010年进行了第1例儿童脱细胞气管同种异体替换术以修复7 cm的受损气管，并进行4年的随访记录，患儿为10岁先天性长段气管狭窄的儿童。术后42个月可见移植段完整的黏膜层和长段通畅的气道，患儿目前临床表现良好并继续接受教育，摆脱长期医疗干预。证实了脱细胞气管治疗儿童气管疾病的可行性，脱细胞气管基质具有一定的生长潜力^[48]。但患者在此阶段因需清除分泌物、肉芽组织，术后多次手术治疗，治疗费用昂贵，未能在临床气管替代治疗中普及。2017年国内TAN等^[49]在体外通过将猪脱细胞真皮基质包裹镍钛合金支架构建复合气管支架，用于1例晚期肺癌侵及左主支气管患者的替代治疗术中，并将受者自身作为“体内生物反应器”，通过体外泵系统向气管复合支架内反复灌注血管内皮生长因子、促血管生成素等生长因子和种子细胞进行修饰，促进气管支架血管化、上皮化，术后4个月支气管镜随访及活检证实血运重建和上皮再生，术后6个月肺癌复发，患者接受放化疗后，患者的生存时间延长和呼吸功能改善，术后13个月死于肺癌复发，但未出现与气管替代物植入有关的并发症。

理想的气管脱细胞技术是可以制备出满足患者正常的气道和肺功能、高质量的生活以及消除反复手术干预的脱细胞气管基质。虽然目前脱细胞方法可避免气管基质引起的宿主免疫排斥反应，但脱细胞技术在组织工程气管的临床应用还需进一步研究，气管脱细胞过程中许多问题仍需解决，如：化学试剂和酶对组织的渗透性都较差，需较长时间作用；组织内试剂残留可引起严重的炎症反应；长时间洗涤浸泡可导致基质中胶原纤维间距增大，组织尺寸发生改变、机械性能下降；去污剂和酶对细胞外基质的损伤等。虽然脱细胞技术还存在许多缺点，但脱细胞气管基质本身依然有着许多的优点及优势，其可以保留的气管三维结构及细胞外基质成分是最好的天然支架；其还具有良好的生物相容性，通过将患者的自体细胞的植入基质内实现再细胞化。相信随着学者们对组织工程中脱细胞技术的不断开发以及与其他气管支架相互结合，可以完美的解决脱细胞技术现有的缺点，并进一步优化气管支架性能，获得更高效、安全的气管支架，扩展脱细胞技术在组织工程气管中的临床应用，攻克气管长段损伤修复这一巨大挑战，提高患者的生活质量。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

脱细胞技术在组织工程气管构建过程中的应用与特点

Application and characteristics of decellularization technology in tissue engineered trachea

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