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. 2023 Aug 27;15(8):e44228.
doi: 10.7759/cureus.44228. eCollection 2023 Aug.

A Comparative Study of Morphometric Analysis of Nucleolar Organizer Regions in Oral Leukoplakia and Oral Squamous Cell Carcinoma and Significance of AgNOR as a Diagnostic Tool

Arun Singh  1 , Shalini Singh  2 , Virendra Soni  2 , Dhirendra K Srivastava  2
Affiliations

A Comparative Study of Morphometric Analysis of Nucleolar Organizer Regions in Oral Leukoplakia and Oral Squamous Cell Carcinoma and Significance of AgNOR as a Diagnostic Tool

Arun Singh et al. Cureus. .

Abstract

Background Silver stainable nucleolar organizer regions (AgNORs) have proven to exhibit utmost importance due to their higher occurrence in the nucleus especially in malignant cells than in normal. Thus, they assist in the examination of nucleolar structures and variations in nucleolar activity. Aim Quantitative and qualitative analysis in relation to the number and area of AgNOR in tissue sections of the normal oral mucosa (NOM), oral leukoplakia (OL), and oral squamous cell carcinoma (OSCC) was the main aim of the study. Materials & method A total of 50 cases comprising 20 OL with dysplasia, 20 OSCC cases, and 10 samples of normal oral mucosa were taken. Silver nitrate (Sol A) & gelatin (Sol B) solutions were freshly prepared for staining the lesional slides. Results The mean value of nuclear profile area (A Nuc) was comparatively higher in oral leukoplakia i.e. 41.97 and in oral squamous cell carcinoma i.e. 62.36 in comparison to the control group where it was 36.19. The mean value of a single AgNOR profile area per nucleus (A NOR) was found to be comparatively lower in both study groups i.e. oral leukoplakia (2.76) and oral squamous cell carcinoma (1.61) in comparison to the control group (3.45) . The mean value of total AgNOR profile area per nucleus (TA NOR) and the number of profiles of AgNORs per nucleus (n NOR) were found higher in both study groups (oral leukoplakia and oral squamous cell carcinoma) as compared to normal oral mucosa of the control group. However, the findings of all four parameters of morphometric analysis were found to be significantly associated with disorder of oral mucosa i.e. cases of oral leukoplakia and oral squamous cell carcinoma (P value <0.01). Conclusion It can thus be suggested that the mean AgNOR count displayed a higher value in OSCC. Hence, the number of AgNORs in nuclei increases as epithelial cells undergo malignant transformation which is designated that mean AgNOR count may contribute to establishing the prognosis of a lesion.

Keywords: dysplasia; microscopic fields; morphometric analysis; nuclear profile area; nucleolar organizer region; oral leukoplakia; oral squamous cell carcinoma; profile area per nucleus; silver colloid; silver nitrate.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Intraoral photograph of oral leukoplakia and oral squamous cell carcinoma
A: Intraoral photograph showing oral leukoplakia (OL) on the right buccal mucosa; B: Intraola photograph showing oral squamous cell carcinoma (OSCC) on the left retromolar area
Figure 2
Figure 2. AgNOR stained section of normal oral mucosa and oral leukoplakia
A: AgNOR stained normal oral mucosa (NOM) (measurement of nuclear area) (x100); B: AgNOR stained section of oral leukoplakia (OL) (measurement of single AgNOR area, total AgNOR area and number of AgNOR) (x100)
Figure 3
Figure 3. AgNOR stained section of moderately differentiated OSCC and poorly differentiated OSSC
A: AgNOR stained section of moderately differentiated oral squamous cell carcinoma (OSCC) (measurement of single AgNOR area, total AgNOR area, and number of Agnor) (x100); B: AgNOR stained section of  poorly differentiated oral squamous cell carcinoma (OSSC) (measurement of single AgNOR area, total AgNOR area, and number of Agnor) (x100)
Figure 4
Figure 4. AgNOR stained section of oral leukoplakia and well differentiated OSSC
A: AgNOR stained section of oral leukoplakia (OL) (measurement of nuclear area) (x100); B: AgNOR stained section of well-differentiated oral squamous cell carcinoma (OSSC) (measurement of nuclear area) (x100)
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