Abstract
Bone is a very complex tissue that is constantly changing throughout the lifespan. The precise mechanism of bone regeneration remains poorly understood. Large bone defects can be caused by gunshot injury, trauma, accidents, congenital anomalies and tissue resection due to cancer. Therefore, understanding bone homeostasis and regeneration has considerable clinical and scientific importance in the development of bone therapy. Macrophages are well known innate immune cells secreting different combinations of cytokines and their role in bone regeneration during bone healing is essential. Here, we present a method to identify mRNA transcripts in cryosections of non-decalcified rat bone using in situ hybridization and hybridization chain reaction to explore gene expression in situ for better understanding the gene expression of the bone tissues.
Highlights
The particular mechanism of bone regeneration is under active examination
The success of the in situ hybridization (ISH) technique realization extremely depends on the quick preparation of good-quality bone tissue sections
The aim of our work was to develop a new approach for obtaining high-quality undecalcified bone sections applicable to various ISH analyses
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. For successful bone healing, coordinated cross talk is needed between inflammatory and bone-forming cells [1–5]. The exact mechanisms of bone regeneration remains to be elucidated [6–9]. Bones and bone marrow contain different types of macrophages: (i) erythroid island macrophages; (ii) hematopoietic stem cell macrophages; and (iii) osteoclasts [10]. Macrophages play an important role both in osteoblast-mediated bone formation [9] and in osteoclast development [10]. The newly-discovered osteal macrophages, so called “osteomacs”, have a fundamental role during bone regeneration [6,9,11,12]. The exact role of these cells is still under study. The cytokines and other soluble factors secreted by macrophages can induce the bone formation in vitro [2,5,9,13]
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