Wong S Y, Dunstan C R, Evans R A, Hills E
Pathology. 1982 Oct;14(4):439-42. doi: 10.3109/00313028209092124.
We have developed a technique for assessing bone viability by the histochemical demonstration of lactate dehydrogenase (LDH) activity in osteocytes. Fresh sawn and ground (75 microns) sections were prepared from femoral heads removed at operation from patients with osteoarthritis of the hip. The sections were decalcified overnight in cold 10% EDTA, pH 7.0. LDH activity was shown by the tetrazolium-formazan reaction with nitroblue tetrazolium as indicator and lithium lactate as substrate. Osteocytes were regarded as viable if their cytoplasm stained dark blue, indicating LDH activity; lacunae containing non-viable osteocytes could be identified by interference contrast illumination. Nearly all osteocytes were viable in the samples studied. Small trabecular fragments, such as could be obtained by needle biopsy, were also suitable for staining after grinding to approximately 50 microns. The method should have application both in research and in diagnosis of ischemic bone disease.
我们已经开发出一种通过组织化学方法显示骨细胞中乳酸脱氢酶（LDH）活性来评估骨活力的技术。从髋关节骨关节炎患者手术切除的股骨头制备新鲜锯切和研磨（75微米）切片。切片在冷的10% EDTA（pH 7.0）中脱钙过夜。以硝基蓝四氮唑为指示剂、乳酸锂为底物，通过四氮唑-甲臜反应显示LDH活性。如果骨细胞的细胞质染成深蓝色，表明有LDH活性，则认为骨细胞是存活的；通过干涉对比照明可以识别含有非存活骨细胞的腔隙。在所研究的样本中，几乎所有骨细胞都是存活的。小的小梁碎片，如通过针吸活检获得的，在研磨至约50微米后也适合染色。该方法在缺血性骨病的研究和诊断中均有应用价值。
