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[利用组织培养方法对软骨移植物进行重要保存]

[Vital preservation of cartilage transplants with tissue culture methods].

作者信息

Bujía J, Osete J M, Sprekelsen C, Wilmes E

机构信息

HNO-Klinik und Poliklinik, Ludwig-Maximilians-Universität München.

出版信息

Laryngorhinootologie. 1994 May;73(5):249-52. doi: 10.1055/s-2007-997123.

Abstract

Cartilage grafting is one of the most common procedures in plastic surgery. Since storage of both autologous and allogenic cartilage is necessary, different preservation methods have been used with more or less success. The use of chemical preservation procedures like formaldehyde, Merthiolate or Cialit lead to a loss of the vitality of the graft. This work presents a study of the cell vitality and the matrix of cartilage grafts stored in different solutions (formaldehyde, saline, RPMI 1640, Ham F-12 and DMEM 4500) during 150 days. The cell viability was assessed using tissue sections (neutral red supravital staining and trypan blue dye exclusion test) and isolated cells (trypan blue dye exclusion test and cell adhesion in monolayer culture). The state of the cartilage matrix was analysed by means of the azan, alcian and toluidine blue staining). Cartilage immersed in formaldehyde solution lost 100% of the vitality after a storage period of 10 days, the one immersed in saline solution after 30 days. Cartilage stored in tissue culture media retained its vitality (> 85%) during the whole storage time. Histological staining methods showed a decrease of the staining intensity after 10 days storage in formaldehyde and after 30 days storage in saline solution. No differences in vitality and the matrix staining were found among all three culture media. Our results suggest that viable cartilage tissue can be successfully stored for a long time using tissue culture methods.

摘要

软骨移植是整形手术中最常见的手术之一。由于自体和异体软骨都需要储存,因此人们使用了不同的保存方法,或多或少都取得了一些成功。使用甲醛、硫柳汞或西力生等化学保存方法会导致移植物活力丧失。这项工作展示了一项关于在150天内储存在不同溶液(甲醛、生理盐水、RPMI 1640、Ham F-12和DMEM 4500)中的软骨移植物的细胞活力和基质的研究。使用组织切片(中性红活体染色和台盼蓝拒染试验)和分离细胞(台盼蓝拒染试验和单层培养中的细胞黏附)评估细胞活力。通过偶氮、阿尔辛蓝和甲苯胺蓝染色分析软骨基质的状态。浸泡在甲醛溶液中的软骨在储存10天后失去了100%的活力,浸泡在生理盐水溶液中的软骨在30天后失去活力。储存在组织培养基中的软骨在整个储存期间保持其活力(>85%)。组织学染色方法显示,在甲醛中储存10天后和在生理盐水溶液中储存30天后,染色强度降低。在所有三种培养基中,未发现活力和基质染色有差异。我们的结果表明,使用组织培养方法可以成功地长期储存有活力的软骨组织。

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