人脐带间充质干细胞注射治疗糖尿病足溃疡的前瞻性、随机和对照研究
1Department of Plastic Surgery,Affiliated Hospital of Xuzhou Medical University, 2Graduate School,Xuzhou Medical University, 3Institute for Tissue Engineering and Regenerative Medicine,The Chinese University of Hong Kong, 4School of Biomedical Sciences,The Chinese University of Hong Kong, 5Center for Neuromusculoskeletal Restorative Medicine, Hong Kong Science Park
Summary
本协议描述了一项前瞻性、随机、对照的临床研究，该研究评估了人脐带间充质干细胞注射液治疗慢性糖尿病足溃疡的效果。
Abstract
随着社会经济的发展，糖尿病足溃疡的发病率不断增加。目前，常规换药清创、高压氧、真空密封引流是临床上主要的保守治疗，大伤口常需植皮或皮瓣移植。但治疗效果并不理想，并发症多。糖尿病足溃疡发病机制复杂、治疗时间长、伴生困难大、致残率高，给患者、社会、医疗造成沉重负担。根据我们之前的研究，人脐带血干细胞的药理作用包括非特异性免疫调节;生长因子、血管活性因子和抗炎因子分泌增加;增强人体抗感染能力;消除炎症;促进血管生成和溃疡愈合。这些效应表明干细胞可能可用于难治性伤口的自体或同种异体治疗。因此，我们正在我们的诊所进行一项临床试验，用人脐带干细胞治疗难治性糖尿病伤口，用于符合纳入标准的糖尿病足溃疡患者。
Introduction
糖尿病是一种影响全球个人的疾病，世界卫生组织（WHO）预测，糖尿病患者人数将从2010年的2.85亿增加到2030年的4.39亿1。糖尿病足溃疡（DFU）是糖尿病最严重的并发症之一，也是全世界非创伤性下肢截肢的主要原因2，3，4，5。
最近，干细胞因其多能性、自我更新和促进再生细胞因子分泌的能力而作为一种疗法蓬勃发展6，7。先前的临床实验表明，脂肪来源的干细胞凝胶对治疗慢性糖尿病足部溃疡有积极作用8。作者验证了使用干细胞治疗59名糖尿病伤口的有效性。在第12周，治疗组和对照组的完全伤口闭合率分别为82%和53%，这表明干细胞对难治性糖尿病伤口的治疗有效。总体而言，干细胞再生、替换、修复和分化的能力给生命科学界带来了无限的希望9。
2008年，Dulchavsky等人10使用含有自体骨髓基质细胞（BMSC）的移植物治疗了20 例不同原因的不愈合伤口。组织学检查显示，18例患者的皮肤伤口完全再上皮化。同样，在明胶支架中使用同种异体非糖尿病间充质干细胞（MSCs）治疗非愈合糖尿病伤口可以促进血管生成，增加再上皮化，并减少溃疡面积11。然而，国内外临床研究中干细胞治疗糖尿病足创面的病例很少;大多数只是病例报告或探索性临床研究，缺乏严格的实验设计，很少有设计好的大样本或随机对照临床试验。由于干细胞不是常见的药物或生物制品，因此不同研究的制备方法和质量控制不同。一项研究的数据可能无法完全反映来自同一物种的所有干细胞的安全性。因此，我们进一步总结了人脐带间充质干细胞（HUCMSC）的相关基础研究和临床前实验，并系统评价了其临床应用的安全性和有效性。在此基础上，开发了从人脐带注射间充质干细胞以修复糖尿病足部伤口的治疗方法。本研究旨在验证干细胞修复糖尿病足创面在临床应用中的有效性和安全性。
综上所述，干细胞治疗应用广泛，潜力巨大，是一种很有前途的新型医学治疗方法。在国家自然科学基金（第81571901、第81501671、第82172224）的持续支持下，我们开展了一系列关于用HUCMSCs治疗糖尿病伤口的研究。在《皮肤病学研究》、《干细胞研究与治疗》、《细胞死亡与疾病》杂志上发表相关文章20余篇，申请国家发明专利3项，积累了较大的研究基础12，13。在这里，我们提供了一种标准方法来评估注射HUCMSCs用于糖尿病足伤口修复。该标准程序已获中药临床试验批准（试验注册号：中药临床试验：MR-32-21-015759，[初始发布：2021年10月20日]）。
Protocol
这项前瞻性、单中心、随机、对照的临床研究获得了徐州医科大学附属医院伦理委员会（XYFY2021-KL124-02）的批准。该研究于 2021 年 7 月开始，将持续到 2023 年 7 月;本实验招募了60名患者。所有患者都签署了知情同意书，允许研究人员使用他们的临床材料和生物数据。 1. 招募患者 纳入标准：确保患者的年龄在 18 岁至 80 岁（含）之间，不分性别。 …
Representative Results
目前，我们的研究仍处于患者招募阶段，目前已完成HUCMSCs治疗组3例患者和对照组3例银离子敷料，共6例慢性糖尿病足部伤口患者。HUCMSCs治疗组患者溃疡区域的平均大小为3.5 cm 2，在用HUCMSC治疗后第8天，第15天和第22天分别减小到2.6 cm 2，1.8 cm 2和1.25 cm 2（图4A-D）。用银离子敷料治疗前对照组患者溃疡的平均大小为1.25c…
Discussion
DFU是一个重大的全球公共卫生问题，也是下肢截肢和健康相关生活质量差的主要原因19，20。目前临床管理仍以常规清创、高压氧治疗、真空密封引流（VSD）和保守治疗为主。较大的伤口通常需要移植皮肤和皮瓣。许多患者患有长期和反复的疾病，造成严重的身心痛苦，并严重限制了他们的生活质量21。这意味着迫切需要对DFU更有效的?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
作者感谢徐州医科大学附属医院的合作，包括糖尿病足创面患者的招募和随访。作者还感谢在设计本研究期间参与患者需求调查的患者。
作者宣布收到以下资金支持，用于本文的研究，作者和/或出版：中国国家自然科学基金82172224，江苏省研究生研究与实践创新计划（SJCX22-1271）和创新与技术委员会（Health@InnoHK）。
Materials
| Silver Iin Wound Dressing | Shandong Cheerain Medical Co.,Ltd. | 20152640521 | Sterile silver ion dressing for medical use (Type F) 10 cm x 10 cm |
| Human Umbilical Cord Mesenchymal Stem Cells Injection | Shandong Qilu Cell Therapy Engineering Technology Co., Ltd. | 32183185-X | Main components: human umbilical cord mesenchymal stem cells. Pharmacological effect: non-specific immunomodulator can enhance the secretion of growth factor, vasoactive factor and anti-inflammatory factor, improve the anti infection ability of human body, eliminate inflammation, promote angiogenesis and ulcer healing. |
| Sterile mesh film transparent dressing | Smith & Nephew | 20162644490 | Sterile mesh film transparent dressing (used for wound area measurement) 6 cm x 7 cm |
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Citazione di questo articolo
Zhang, J., Zhao, B., Wei, W., Wang, D., Wang, H., Zhang, A., Tao, C., Li, X., Li, Q., Jin, P. Prospective, Randomized, and Controlled Study of a Human Umbilical Cord Mesenchymal Stem Cell Injection for Treating Diabetic Foot Ulcers. J. Vis. Exp. (193), e65045, doi:10.3791/65045 (2023).