USE OF CRYSTAL VIOLET STAIN FOR THE ASSESSMENT OF MITOTIC FIGURES IN ORAL EPITHELIAL DYSPLASIA AND ORAL SQUAMOUS CELL CARCINOMA

Abstract

INTRODUCTION: Histopathological grading depends on several features, of which appearance of mitotic figures and apoptotic bodies in the tissue sections plays an important role. The mitotic figures and apoptotic bodies help assess the cellular proliferation and the turnover of the tumor. As mitotic figures (MF) and apoptotic bodies are difficult to assess under routine H and E sections, other methods such as immunohistochemistry (IHC) and flow cytometry have been implemented but, these methods are expensive and more time-consuming and are highly technique sensitive. Hence, stains, which should be cheap, and can be used routinely, and can be tried in identifying the mitotic figures and apoptotic bodies, which has come out with a better result. Routine histological procedures cannot differentiate between pyknotic nuclei, apoptotic cells and MFs which makes the diagnosis difficult.
 MATERIAL AND METHODS: Study samples were divided in to 3 groups, group 1- case group in which 40 samples of paraffin blocks diagnosed as oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) and group 2- control group in which 40 samples from the healthy individuals. 5 micron thickness sections were made from each tissue specimen. One section was stained with H & E stain nd another section was stained with 1% crystal violet stain.
 RESULTS: In case group there were 24(60%) male and 16(40%) female while in control group there were 22 (55%) male and 18(45%) female. Mean age in case group was 36.2±15.4 and in control group was25.7±12.7. There is a significant increase in mitotic figures of H & E (P< 0.01) and crystal violet (P < 0.01). In case, there was a significant (P<0.05) increase in mitotic figures when stained with crystal violet. As compared to H&E stain there was a significant (P < 0.01) increase in metaphase observed in crystal violet stain. Mean mitotic figure by H&E in case group was 4.15±1.07 while in crystal violet it was 5.97±1.87.
 CONCLUSION: % crystal violet stain is the cheaper stain and can be performed in small laboratories and resource constraint settings. Crystal violet can be used for the localization of mitotic figures and assessing proliferation.