ESTABLISHMENT OF A PROTOCOL FOR COLLECTION AND PREPARATION OF WISTAR RATS CALCANEAL TENDON SLIDES – A HISTOLOGICAL EVALUATION
DOI:
https://doi.org/10.25110/arqsaude.v30i2.2026-12315Keywords:
Histology, Histopathology, Protocol, Tendon.Abstract
The calcaneal tendon is the largest and strongest tendon in the body and is commonly affected by tendinopathies. Studies involving this tissue are mainly focused on injuries and therapeutic approaches; therefore, careful tendon harvesting and histological processing are essential to avoid sample loss. However, the literature lacks detailed protocols describing these procedures. Thus, the aim of this study was to establish the most appropriate protocol for collection and histological slide preparation of the calcaneal tendon in Wistar rats. Eighty Wistar rats were euthanized, followed by immediate tendon collection. The animals were allocated into 16 groups, varying according to the association of the tendon with skin, muscle group, and the tibiotarsal joint, as well as tissue positioning within the paraffin block (top or bottom). Samples were fixed in 10% formalin, processed using conventional histological techniques, and stained with hematoxylin and eosin. Histological evaluation was performed using ordinal scoring systems for fibroblast presence, collagen fiber arrangement, stromal disposition, and cellular organization. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s post hoc test (p<0.05). The results demonstrated descriptive variation in histological quality among the evaluated protocols, particularly regarding cellular organization and collagen arrangement. Positioning the tendon with skin at the bottom of the paraffin block resulted in better structural preservation in a descriptive manner. No statistically significant differences were observed between the 2 µm and 5 µm section thicknesses. It is concluded that standardization of tissue positioning within the block and cutting orientation is the main determinant of quality and reproducibility in histopathological evaluation of the calcaneal tendon.
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