Kapila Supriya Nikita, Natarajan Srikant, Boaz Karen, Pandya Jay Ashokkumar, Yinti Shanmukha Raviteja
Postgraduate Student, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India .
Associate Professor, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India .
J Clin Diagn Res. 2015 Sep;9(9):ZC93-7. doi: 10.7860/JCDR/2015/14641.6569. Epub 2015 Sep 1.
Quicker decalcification is essential for faster diagnosis of hard tissue pathology. Heat and mechanical agitation are known to hasten decalcification.
To compare the rate of decalcification, cellular and staining characteristics of decalcified specimens of bone and teeth by using the conventional method (10% formal formic acid), heating to 45(o)C and by physical agitation with magnetic stirrer.
Weight-matched samples of caprine-origin bone (n=15) and teeth (n=15) were decalcified using three methods namely: a) Gooding and Stewart's fluid; b) Gooding and Stewart's fluid heated to 45(o)C for 6 hours daily; and c) Gooding and Stewart's fluid agitated using a magnetic stirrer for 6 hours daily. Non-lesional skin tissue samples were placed along with each specimen. End point of decalcification (chemical test) was noted; 4 micron sections were taken and stained with H&E.
Differences in rate of decalcification and staining characteristics were assessed by Kruskal Wallis test and chi-square test respectively.
Hard tissues decalcified faster with stirring and heating methods. The amount of osteocyte retraction noted in bone was significantly reduced in the stirring method. In tooth specimens, modified techniques resulted in poorer nuclear-cytoplasmic contrast of pulp cells. Heating affected the odontoblast layer. Soft tissues exhibited higher eosinophilia in stirring and conventional methods, whereas nuclear-cytoplasmic contrast and chromatin staining was poorest in heating and conventional methods.
Physical agitation of decalcifying fluid may be recommended while maintaining satisfactory quality of tissue morphology and staining.
更快的脱钙对于硬组织病理学的快速诊断至关重要。已知加热和机械搅拌可加速脱钙。
通过使用传统方法（10% 甲醛 - 甲酸）、加热至45℃以及用磁力搅拌器进行物理搅拌，比较骨和牙齿脱钙标本的脱钙速率、细胞及染色特征。
使用三种方法对来自山羊的重量匹配的骨样本（n = 15）和牙齿样本（n = 15）进行脱钙，即：a）古丁和斯图尔特氏液；b）将古丁和斯图尔特氏液加热至45℃，每天加热6小时；c）使用磁力搅拌器搅拌古丁和斯图尔特氏液，每天搅拌6小时。每个标本均放置非病变皮肤组织样本。记录脱钙终点（化学测试）；制作4微米切片并用苏木精和伊红染色。
分别通过克鲁斯卡尔 - 沃利斯检验和卡方检验评估脱钙速率和染色特征的差异。
采用搅拌和加热方法时硬组织脱钙更快。在搅拌法中，骨中观察到的骨细胞收缩量显著减少。在牙齿标本中，改良技术导致牙髓细胞核质对比度变差。加热影响成牙本质细胞层。在搅拌法和传统方法中，软组织表现出更高的嗜酸性，而在加热法和传统方法中，核质对比度和染色质染色最差。
在保持组织形态和染色质量令人满意的同时，可推荐对脱钙液进行物理搅拌。
