ABSTRACT
BACKGROUND: Oral leukoplakia is the most common precancerous lesion. Various grading systems based on histological findings have been proposed for dysplasia. Recently, computer-assisted morphometric analysis has been established to reduce the interobserver and intraobserver variability during the histological grading of epithelial dysplasia. This study was undertaken to establish the morphometric changes in the suprabasal cell layer of different grades of oral epithelial dysplasia. MATERIALS AND METHODS: Forty paraffin-embedded tissue sections (10 normal mucosa, 10 mild dysplasia, 10 moderate dysplasia, and 10 severe dysplasia cases) were stained using hematoxylin and eosin stain, and analyzed for cellular and nuclear morphometry using binocular microscope. RESULTS: Our results showed that values of nuclear area, nuclear perimeter, nuclear volume density, and nuclear/cytoplasmic (N/C) ratio were increased gradually in dysplasia compared to control groups and the values were statistically significant (P = 0.001). Nuclear diameter and cellular area were increased in dysplasia when compared to control group (P = 0.001). Mild and moderate dysplasia showed decreased value of nuclear form factor compared to control group, whereas severe dysplasia showed highest value. A fair correlation was found when comparing histological grading and grouping based on nuclear area, nuclear perimeter, N/C ratio, and nuclear volume density. CONCLUSION: Nuclear features reflect cell behavior, and its morphometric analysis can be considered as a reliable tool for differentiating various grades of epithelial dysplasia.
ABSTRACT
BACKGROUND: Xylene is an aromatic hydrocarbon that is routinely used in histopathological laboratories. It is biohazardous and produces many toxic effects like carcinogenesis. A safer substitute for xylene is necessary to minimize its usage. AIM: The aim of this study was to evaluate the effectiveness of 1.7% dishwashing solution, 95% lemon water, and 100% coconut oil when compared to xylene as a deparaffinizing agent during hematoxylin and eosin (H&E) staining. MATERIALS AND METHODS: Fifteen paraffin-embedded tissue blocks were selected. Four sections were made from each block. One section was stained with conventional H&E method using xylene (group A) as deparaffinizing agent and other three sections were stained with xylene-free H&E method using 1.7% dishwashing solution (group B), 95% lemon water (group C), and 100% coconut oil (group D), respectively. Slides were scored blindly by a single pathologist considering the parameters such as nuclear and cytoplasmic staining; uniformity, clarity, and crispness of staining; and presence or absence of wax retention. RESULTS: Adequate nuclear staining was noted in 100% of sections of groups A, B, C, and D (P < 0.001), whereas adequate cytoplasmic staining was noted in 93.33% each in groups A, C, and D when compared with 100% in group B (P > 0.05). Uniform staining was present in 80% of groups A and B and in 73.33% of groups C and D (P > 0.05). Clarity of staining was present in 86.66% of groups A and B and in 80% of groups C and D (P > 0.05), whereas crispness of staining was seen in 73.33% of groups A and D, 86.66% of group B, and 80% of group C (P > 0.05). Wax retention was noted in 20% of groups A and B, and 26.66% of groups C and D (P > 0.05). Adequate staining for diagnosis was noted in 100% of group A sections followed by 93.33% in group B, 86.66% in group C, and 80% in group D as compared with 90% in group B (P > 0.05). CONCLUSION: Dishwashing solution, lemon water, and coconut oil can be used as safer and cost-effective substitutes to xylene for deparaffinization in H&E staining procedure.
