# Assessment and Comparison of Natural Dyes Like Lawsonia inermis, Zingiber officinale, Curcuma longa, Beta vulgaris, Kumkum, and Hibiscus rosa-sinensis With Eosin as a Cytoplasmic Stain in Oral Histopathologies

## TL;DR

This study compares natural dyes to synthetic eosin for histopathology staining and finds that Lawsonia inermis performs similarly to the standard.

## Contribution

The study identifies Lawsonia inermis as a promising natural alternative to eosin for histopathological staining.

## Key findings

- Lawsonia inermis showed staining intensity comparable to eosin (15.05 vs. 15.85).
- Hibiscus rosa-sinensis and Beta vulgaris had the lowest staining intensity (7.9 and 8.4).
- Lawsonia inermis provided detailed cellular and nuclear staining similar to eosin.

## Abstract

Background

Hematoxylin and eosin (H&E) staining is a commonly used technique in histology and histopathology. However, the synthetic dye eosin poses risks to both human and animal health. The growing ecological concern has led to efforts to find natural, less toxic, biodegradable, and cost-effective alternatives to eosin.

Aim

This study aimed to compare and evaluate the staining characteristics of natural dyes as cytoplasmic stain to eosin as a standard in oral histopathologies.

Materials and methods

A total of 140 histological sections from four tissue types (normal mucosa, oral squamous cell carcinoma, ameloblastoma, and odontogenic cysts) were stained with natural dyes. The groups were divided based on the type of stain: Group A (Lawsonia inermis), Group B (Zingiber officinale), Group C (Curcuma longa), Group D (kumkum), Group E (Hibiscus rosa-sinensis), Group F (Beta vulgaris), and Group G (eosin as control). The staining intensity and nuclear and cytoplasmic detail were evaluated and compared using one-way ANOVA. A significance level of 0.01 was set for statistical analysis.

Results

One-way ANOVA revealed a significant difference in staining intensity across the seven groups (p=0.001). Eosin showed the highest mean staining intensity (15.85), followed by Lawsonia inermis (15.05), while Hibiscus rosa-sinensis and Beta vulgaris displayed the lowest values (7.9 and 8.4, respectively). Post-hoc analysis identified Group A (mean=15.05) shows no significant difference from the control suggesting that the staining performance of Group A is comparable to the control at (p<0.01).

Conclusion

The utilization of Lawsonia inermis as a natural dye in histopathological staining procedures has demonstrated promising results compared to other natural stains. Its ability to provide excellent contrast, detailed cellular and nuclear staining, and effective staining of acellular structures, comparable to eosin, underscores its potential as a viable alternative in diagnostic pathology.