Histomorphology of bone bridge formation after epiphyseal injury observed by three staining methods

doi:10.3969/j.issn.2095-4344.1308

Abstract

Abstract:

BACKGROUND: Existing evidence has shown that combined usage of various staining methods contributes to exploring the morphological performance of bone and cartilage diseases. Histomorphology of bone bridge formation after epiphyseal injury using various staining methods is rarely reported.
OBJECTIVE: To observe the histomorphology of bone bridge formation after epiphyseal injury by hematoxylin-eosin staining, safranin O-fast green staining and Masson staining.
METHODS: Forty Sprague-Dawley rats were provided by the Laboratory Animal Center of Fujian University of Traditional Chinese Medicine, and the study was approved by the Laboratory Ethics Committee of Fujian University of Traditional Chinese Medicine. The rats were randomly divided into model (proximal tibia epiphyseal injury model) and control groups (n=20/group). X-ray was conducted at 1, 7 and 28 days after modeling, and MRI was scanned at 7 and 28 days after modeling. The tibia damage tissues were removed under anesthesia at 1, 7, 14, and 28 days after modeling. Hematoxylin-eosin staining, safranin O-fast green staining and Masson staining were performed to observe the histomorphology of the growth plate damage area at postoperative different time points.
RESULTS AND CONCLUSION: (1) X-ray films: at 1 day after modeling, the tibial metaphysis injury and widening growth plate space surrounding the perichondrium were observed. On day 7, periosteal reaction in the injured metaphysis was obvious. On day 28, periosteal reaction disappeared, and the space between metaphysis and growth plate returned to be normal. MRI T2WI at 28 days after modeling showed that epiphyseal plate became obscure and a blank line low signal was observed, suggesting bone bridge formation. (2) At 1 day after modeling, all staining methods revealed blood vessels, especially Masson staining. On day 7, the morphological changes of chondrocytes in the injury region were more obvious in hematoxylin-eosin staining compared with the safranin O-fast green staining and Masson staining. On day 14, under safranin O-fast green staining, the growth plate was clearly contrasted with the bone bridge, while unossified fibrous vascular tissue in the bone bridge can be observed by Masson staining. On day 28, hematoxylin-eosin staining showed obvious periodic morphological changes of chondrocytes. Safranin O-fast green staining showed clear morphology of bone bridge and growth plate, and the compressive change of the growth plate in the damaged area clearly. (3) These results indicate that the combined application of the three staining methods can comprehensively and objectively explore the histomorphological manifestations of the bone bridge formation after epiphyseal injury.

Key words: staining methods, epiphyseal injury, bone bridge, periosteum, histology, hematoxylin-eosin staining, safranin O-fast green staining, Masson staining

CLC Number:

R446
R318

Pan Yuancheng, Zhang Xinzhao, Chen Shunyou. Histomorphology of bone bridge formation after epiphyseal injury observed by three staining methods[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(23): 3649-3653.

Figures/Tables 2

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