engleski

Histological structure of mineralised red deer antler: application of three staining methods

Red deer (Cervus elaphus) antlers are unique structures and only mammalian tissue capable of complete regeneration. The fact that their growth is completed within 4 months, makes them one of the fastest growing tissues in mammals. Starting from the near second year of life, antlers are casted and regrown each year in red deer, in a so-called antler growth cycle. It consists of 4 phases: growth, mineralisation, velvet shedding and casting. Mineralised antlers are made of a central cancellous (spongy) bone surrounded by a compact (cortical) bone extending longitudinally through the main beam and tines. We have collected antlers of red deer spikers 10 days after velvet shedding. Tissue samples were fixed with 10% buffered formalin and 70% alcohol. Demineralisation was made using Osteosoft® (mild decalcifier solution based on EDTA). Differences in the histological structure were shown by routine staining method (H&E) and special staining methods: modified method for ossification process, Alizarin Red S - Alcian Blue and Villanueva’s tetrachrome bone stain. Following the modified staining method for ossification process bone was presented as orange, while cartilage was blue. Alizarin Red S - Alcian Blue staining showed bone as red and cartilage as blue. Villanueva’s tetrachrome bone stain showed osteoid as green, bone orange to red, osteocytes and canaliculi red, while nuclei of osteoblasts were dark purple. The results of our staining showed that mineralisation process is not completed with velvet shedding. Parts of the compact and cancellous bone are still containing remnants of non-mineralised cartilage. External thin layer of antlers still resembles to non-calcified tissue, which may eventually be lost during the maturation of antlers.

antler, red deer, staining methods, histology

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