Supriya Nikita Kapila1, Srikant Natarajan2, Karen Boaz3, Jay Ashokkumar Pandya4, Shanmukha Raviteja Yinti1. 1. Postgraduate Student, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India . 2. Associate Professor, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India . 3. Professor and Head, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India . 4. Postgraduate Student, Department of Oral Pathology, Manipal College of Dental Sciences , Mangalore, Manipal University, India.
Abstract
INTRODUCTION: Quicker decalcification is essential for faster diagnosis of hard tissue pathology. Heat and mechanical agitation are known to hasten decalcification. AIM: 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. MATERIALS AND METHODS: 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. STATISTICAL ANALYSIS: Differences in rate of decalcification and staining characteristics were assessed by Kruskal Wallis test and chi-square test respectively. RESULTS: 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. CONCLUSION: Physical agitation of decalcifying fluid may be recommended while maintaining satisfactory quality of tissue morphology and staining.
INTRODUCTION: Quicker decalcification is essential for faster diagnosis of hard tissue pathology. Heat and mechanical agitation are known to hasten decalcification. AIM: 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. MATERIALS AND METHODS: 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. STATISTICAL ANALYSIS: Differences in rate of decalcification and staining characteristics were assessed by Kruskal Wallis test and chi-square test respectively. RESULTS: 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. CONCLUSION: Physical agitation of decalcifying fluid may be recommended while maintaining satisfactory quality of tissue morphology and staining.
Authors: Patricia Goggin; Elaine M L Ho; Helmut Gnaegi; Stuart Searle; Richard O C Oreffo; Philipp Schneider Journal: Bone Date: 2019-10-24 Impact factor: 4.398