Orthodontic force: Immunohistochemistry versus hematoxylin-eosin

J. Pharm. Pharmacogn. Res., vol. 10, no. 4, pp. 695-700, July-August 2022.

Short Communication

Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin

[Análisis histológicos de la fuerza ortodóncica en Cavia porcellus: Comparación entre inmunohistoquímica y hematoxilina-eosina]

Erliera Sufarnap1,3*, Syafruddin Ilyas2, Nazruddin Nazruddin3, Deddi P. Putra4, Aditya Rachmawati3

1Doctorate Program, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

2Department of Biology, Faculty of Mathematics and Natural Science, Universitas Sumatera Utara, Medan, Indonesia.

3Department of Orthodontic, Faculty of Dentistry, Universitas Sumatera Utara, Medan, Indonesia.

4Faculty of Pharmacy, Universitas Andalas, Padang, Indonesia.

*E-mail: erliera@usu.ac.id

Abstract

Context: Histological quantification of osteoclasts and osteoblasts can evaluate biological responses to orthodontic tooth movement. Histological analysis of bone samples can be technically challenging.

Aims: To evaluate the differences between hematoxylin and eosin (HE) staining and immunohistochemistry (IHC) in quantifying osteoblast and osteoclast cells following the application of static orthodontic force.

Methods: Orthodontic force was applied using a rubber separator around the maxilla incisor of Cavia porcellus. Tooth samples were taken at 0, 4, 8, 14, 21, and 28 days after applying orthodontic force. HE and IHC staining quantify osteoblast and osteoclast cells in the alveolar bone. IHC staining, i.e., Tartrate-resistant acid phosphatase (TRAP) staining, was used to identify osteoclasts, and osteocalcin (OCN) staining was used to identify osteoblasts.

Results: Significantly higher numbers of osteoclasts and osteoblasts were observed with IHC compared to HE staining (p<0.05). Significant positive linear correlations in the numbers of osteoclasts (r = 0.757) and osteoblasts (r = 0.622) identified were observed between IHC and HE staining.

Conclusions: The results of this study indicate HE staining may represent an acceptable alternative method of quantifying osteoclasts and osteoblasts in the preliminary research of orthodontic tooth movement (OTM).

Keywords: hematoxylin; immunohistochemistry; orthodontic; osteocalcin; tartrate-resistant acid phosphatase.

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Resumen

Contexto: La cuantificación histológica de osteoclastos y osteoblastos puede evaluar las respuestas biológicas al movimiento dental ortodóncico. El análisis histológico de muestras de hueso puede ser técnicamente desafiante.

Objetivos: Evaluar las diferencias entre la tinción con hematoxilina y eosina (HE) y la inmunohistoquímica (IHC) en la cuantificación de células de osteoblastos y osteoclastos después de la aplicación de fuerza ortodóncica estática.

Métodos: Se aplicó fuerza de ortodoncia utilizando un separador de goma alrededor del incisivo maxilar de Cavia porcellus. Se tomaron muestras de dientes a los 0, 4, 8, 14, 21 y 28 días después de aplicar la fuerza de ortodoncia. La tinción con HE e IHC cuantifica las células de osteoblastos y osteoclastos en el hueso alveolar. Se usó tinción IHC, es decir, tinción con fosfatasa ácida resistente a tartrato (TRAP), para identificar osteoclastos, y tinción con osteocalcina (OCN) para identificar osteoblastos.

Resultados: Se observaron números significativamente más altos de osteoclastos y osteoblastos con IHC en comparación con la tinción con HE (valor de p<0,05). Se observaron correlaciones lineales positivas significativas en el número de osteoclastos (r = 0,757) y osteoblastos (r = 0,622) identificados entre la tinción IHC y HE.

Conclusiones: Los resultados de este estudio indican que la tinción HE puede representar un método alternativo aceptable para cuantificar osteoclastos y osteoblastos en la investigación preliminar del movimiento dental ortodóncico (OTM).

Palabras Clave: fosfatasa ácida tartrato resistente; hematoxilina; inmunohistoquímica; ortodoncia; osteocalcina.

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Citation Format: Sufarnap E, Ilyas S, Nazruddin N, Putra DP, Rachmawati A (2022) Histological analyses of orthodontic force in Cavia porcellus: Comparison between immunohistochemistry and hematoxylin-eosin. J Pharm Pharmacogn Res 10(4): 695–700.
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