Jähn Katharina, Braunstein Volker, Furlong Pamela I, Simpson Angharad E, Richards R Geoff, Stoddart Martin J
AO Research Institute, Clavadelerstrasse 8, 7270 Davos, Switzerland.
J Orthop Surg Res. 2010 May 10;5:32. doi: 10.1186/1749-799X-5-32.
Bone graft studies lack standardized controls. We aim to present a quick and reliable method for the intra-operative generation of acellular bone explants.
Therefore, ovine cancellous bone explants from the iliac crest were prepared and used to test several methods for the induction of cell death. Over night heat inactivation was used as positive treatment control, methods to be investigated included UV light, or X- ray exposure, incubation in a hypotonic solution (salt-free water) and a short cycle of repeated freezing and thawing.
Viability of treated and 2 days cultured bone explants was investigated by lactate dehydrogenase assay. Non-treated cultured control explants maintained around 50% osteocyte viability, while osteocyte survival after the positive treatment control was abolished. The most dramatic loss in cell viability, together with a low standard deviation, was a repeated cycle of freezing and thawing.
To summarize, we present a freeze-thaw method for the creation of acellular bone explants, which is easy to perform, not time-consuming and provides consistent results.
骨移植研究缺乏标准化对照。我们旨在提出一种在术中快速可靠地生成脱细胞骨外植体的方法。
因此，制备了来自羊髂嵴的松质骨外植体，并用于测试几种诱导细胞死亡的方法。过夜热灭活用作阳性处理对照，待研究的方法包括紫外线或X射线照射、在低渗溶液（无盐水）中孵育以及短周期反复冻融。
通过乳酸脱氢酶测定法研究处理后及培养2天的骨外植体的活力。未处理的培养对照外植体维持约50%的骨细胞活力，而阳性处理对照后骨细胞存活被消除。细胞活力丧失最显著且标准差低的是反复冻融循环。
总之，我们提出了一种用于创建脱细胞骨外植体的冻融方法，该方法易于实施，不耗时且结果一致。
