Enhancing Histological Visualization of Tooth and Bone Ground Sections Using Terminalia chebula Stain: A Comparative Analysis

BACKGROUND: Visualization of mineralized tissues such as tooth and bone in ground sections is widely used for histological studies. However, unstained ground sections often provide limited contrast and obscure microscopic details. Natural plant-based stains have been explored as cost-effective and safer alternatives to synthetic dyes. Terminalia chebula, rich in tannins and polyphenolic compounds, has shown potential staining affinity for hard tissues. OBJECTIVES: The aim of this study is to evaluate the staining efficacy of T. chebula on ground sections of tooth and bone by comparing stained and unstained sections, and to assess contrast, structural detail, and observer acceptance under stereomicroscope and compound microscope. METHODS: Forty extracted normal teeth and 40 bone samples from human skulls were prepared into thin ground sections (~50 μm). Each section was initially observed unstained (Group 2) and then stained with T. chebula extract for 4 h (Group 1). Photomicrographs were taken under both microscopes and evaluated by blinded oral pathologists and students for contrast, structural detail, and acceptance using standardized grading and scoring systems. RESULTS: T. chebula staining significantly improved contrast in tooth sections under both stereomicroscope and compound microscope (p < 0.001). Enhanced visualization of enamel rods, dentinal tubules, and cementum was observed. Bone sections demonstrated improvement in contrast under stereomicroscopy; however, under compound microscopy, unstained bone sections showed relatively higher Grade 1 contrast. Improvements in structural detail were more pronounced in tooth sections than bone sections. Observer acceptance was significantly higher for stained tooth sections (p < 0.001), whereas moderate but significant improvement was noted for bone sections. CONCLUSION: T. chebula stain enhances contrast and improves visualization of microstructural features in tooth ground sections, supporting its potential as an effective and economical adjunct for teaching and research in dental histology. Further studies are required to optimize staining protocols and to evaluate its broader applicability, particularly in bone specimens.