Kuranobu T
Department of Orthopedic Surgery, Faculty of Medicine, Tottori University, Japan.
Nihon Seikeigeka Gakkai Zasshi. 1993 Dec;67(12):1130-9.
This study investigated optimal conditions for preservation and intramuscular allografting of vertebral body and disc units. First, thirty-one vertebral body and disc units were removed from rabbits and divided into three groups: a slow freezing group, a quick freezing group, and a fresh non-freezing group. In the slow freezing group, disc units were slowly cryopreserved by decreasing temperatures from 4 to -80 degrees C using a program freezer. In the quick freezing group, they were rapidly cryopreserved by freezing to -80 degrees C without a program freezer. Second, to determine favorable duration of immersion in cryoprotectant (10% dimethyl sulfoxide), another 20 vertebral body and disc units were divided into four groups: immersed at 4 degrees C for 30, 90, 240 and 1440 minutes. After immersion, they were cryopreserved slowly using a program freezer. These disc units were thawed and examined histologically and biochemically. In the slow freezing group, both cell number in the anulus fibrosus and the value of 35SO4 incorporation in the disc were higher than those in the quick freezing group, and the disc viability of the 90-minute group was the highest. On the basis of these experiments, vertebral body and disc units were grafted into the back muscles of rabbits. To investigate the viability of the disc after allografting, another 38 specimens were grafted into the back muscles and examined after 4 or 12 weeks. The values for 35SO4 incorporation of each group at 4 weeks were about 80% of the pre-grafting level and those at 12 weeks were about 60%, and the viability of discs maintained. There were no significant differences between the slow freezing group, the quick freezing group, and the fresh non-freezing group after grafting. Further study is needed to determine a method in maintaining viability of the disc after grafting.
本研究调查了椎体与椎间盘单元保存及肌内同种异体移植的最佳条件。首先，从兔体内取出31个椎体与椎间盘单元并分为三组：慢速冷冻组、快速冷冻组和新鲜未冷冻组。在慢速冷冻组中，椎间盘单元使用程序降温冰箱从4℃缓慢降至-80℃进行冷冻保存。在快速冷冻组中，它们在没有程序降温冰箱的情况下快速冷冻至-80℃。其次，为确定在冷冻保护剂（10%二甲亚砜）中浸泡的适宜时长，另外20个椎体与椎间盘单元被分为四组：在4℃下浸泡30、90、240和1440分钟。浸泡后，使用程序降温冰箱对它们进行慢速冷冻保存。这些椎间盘单元解冻后进行组织学和生物化学检查。在慢速冷冻组中，纤维环中的细胞数量和椎间盘内35SO4掺入值均高于快速冷冻组，且90分钟组的椎间盘活力最高。基于这些实验，将椎体与椎间盘单元移植到兔的背部肌肉中。为研究同种异体移植后椎间盘的活力，另外38个标本被移植到背部肌肉中，并在4周或12周后进行检查。每组在4周时的35SO4掺入值约为移植前水平的80%，在12周时约为60%，且椎间盘活力得以维持。移植后，慢速冷冻组、快速冷冻组和新鲜未冷冻组之间无显著差异。需要进一步研究以确定移植后维持椎间盘活力的方法。
