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[复合组织的冷冻保存:原理、文献综述及我们自身实验的初步结果]

[The cryopreservation of composite tissues: principle, literature review and preliminary results of our own experiments].

作者信息

Bakhach J, Casoli V, Guimberteau J-C

机构信息

Institut Aquitain de Chirurgie Plastique, Microchirurgie et Chirurgie de la main, 56, allée des tulipes, 33600 Pessac, Bordeaux, France.

出版信息

Ann Chir Plast Esthet. 2007 Oct;52(5):531-47. doi: 10.1016/j.anplas.2007.06.006. Epub 2007 Jul 23.

Abstract

The cryopreservation of cells and human tissues has generated a great interest from the scientific community since 1949 when the cryoprotective activity of glycerol was discovered. For a homogeneous cellular group or a one-layer cellular tissue it is easy to define the optimal technique conditions of its cryopreservation (cryoprotective agents, speed and steps of freezing, speed of warming). It is considered successful when a high recovery of the cellular structures and tissue components after warming is achieved. The cryopreservation of a whole composite tissue is less easy to obtain. Each tissue presents its own parameters and its own reactivity during the cryopreservation process. The challenge consists in, on the one hand, the selection of the ideal cryoprotective agents'combination which can fit the needs of the different tissues and on the other hand, the definition of adequate technical parameters. The aim of this work is to demonstrate the feasability to cryopreserve a composite tissue in order to carry out surgical reconstructive procedures of particular anatomical and functionnal units (metacarpo-phalangeal and proximal interphalangeal joints, flexor system apparatus, extensor system, median nerve, etc.) with complete revitalization of the allograft using vascular microsurgical procedures. To do so, our present work is divided into three different parts. The first chapter deals with the fundamental principles of the cryobiology of biological structures with special interest in the liquid transfer process between the extracellular and intracellular compartments and ice initiation and agregation during the freezing process. The different physical and chemical reactions and their consequences on the biological tissues are described according to the different cryoprotective agents used, should they belong to the extracellular or intracellular cryoprotective groups. The second chapter makes a review of the litterature concerning the results of all experiments made on the cryopreservation of the different tissue structures as skin, vessels, bones, cartilage, periosteum, nerves, cornea, on the one hand, and the different organs as kidneys, liver, heart, trachea, lung, parathyroid glands and ovaries, on the other hand. We are reporting the results of these experiments focusing on the immunomodulation effect of cryopreservation on the antigenic response of biological structures. These experiments were made either on organs or on the cells involved in the immunogenic process. In the third chapter, we are reporting the results of our experiments carried out in the Aquitaine Hand Institute in the field of the cryopreservation of the xenografts of digital segments on the rabbit. These digital segments were cryopreserved, then warmed and revitalized through vascular microsurgical techniques. The preliminary results are very encouraging and pave the way to the allotransplantation of cryopreserved composite organs in our common surgical activity.

摘要

自1949年甘油的冷冻保护活性被发现以来,细胞和人体组织的冷冻保存引起了科学界的极大兴趣。对于均匀的细胞群或单层细胞组织,很容易确定其冷冻保存的最佳技术条件(冷冻保护剂、冷冻速度和步骤、复温速度)。当复温后细胞结构和组织成分的回收率很高时,就认为是成功的。整个复合组织的冷冻保存则不太容易实现。每种组织在冷冻保存过程中都有其自身的参数和反应性。挑战在于,一方面要选择能够满足不同组织需求的理想冷冻保护剂组合,另一方面要确定适当的技术参数。这项工作的目的是证明冷冻保存复合组织以进行特定解剖和功能单位(掌指关节和近端指间关节、屈肌系统装置、伸肌系统、正中神经等)的外科重建手术的可行性,并通过血管显微外科手术使同种异体移植完全恢复活力。为此,我们目前的工作分为三个不同的部分。第一章论述生物结构低温生物学的基本原理,特别关注细胞外和细胞内隔室之间的液体转移过程以及冷冻过程中的冰晶形成和聚集。根据所使用的不同冷冻保护剂,描述了不同的物理和化学反应及其对生物组织的影响,这些冷冻保护剂属于细胞外或细胞内冷冻保护组。第二章综述了有关不同组织结构(如皮肤、血管骨骼、软骨、骨膜、神经、角膜)以及不同器官(如肾脏、肝脏、心脏、气管、肺、甲状旁腺和卵巢)冷冻保存的所有实验结果。我们报告这些实验的结果,重点是冷冻保存对生物结构抗原反应的免疫调节作用。这些实验是在器官或参与免疫原过程的细胞上进行的。在第三章中,我们报告了在阿基坦手部研究所进行的关于兔指节异种移植冷冻保存的实验结果。这些指节经过冷冻保存,然后通过血管显微外科技术复温和恢复活力。初步结果非常令人鼓舞,为在我们的普通外科手术中冷冻保存复合器官的同种异体移植铺平了道路。

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