[Technique for undecalcified bone tissue. Immunohistochemistry and in situ hybridization].

Abstract

Objective: To investigate the feasibility of applying immunohistochemistry (IH) and in situ hybridization (ISH) technique to the undecalcified bone sections.

Methods: The proximal tibiae of SD rats were embedded with the modified methylmethacrylate low-temperature embedding method and then made into undecalcified bone sections. The sections were stained with Goldner's Masson Trichrome, and the expression of IGF-1 mRNA and protein in the bone tissue cells were detected by IH and ISH.

Results: Goldner's staining demonstrated the structure of trabeculae bone was intact and osteoid at the edge of trabeculae stained red. Osteoblasts and osteoclasts in bone sections could be seen clearly. The positive signals of IGF-1 mRNA and protein were found in the cytoplasm of osteoblasts, chondrocytes in epiphyseal plate and some mononuclear cells in the bone marrow. The matrix of trabeculae also showed positive expression of IGF-1 protein.

Conclusion: The undecalcified bone sections prepared by the method can meet the needs of IH and ISH. The establishment of this method will provide technological platform for the study of molecular pathology on metabolic bone diseases such as osteoporosis.