Mouse Subcutaneous Tissue Reaction to Calcium Hydroxide-based Root Canal Filling Material

Mouse subcutaneous tissue reaction to an embedded calcium hydroxide-based root canal filling material was analyzed histopathologically. After the material was placed within the mouse dorsal subcutaneous tissues, we performed examinations using histopathological, histochemical and immunohistochemical techniques. Two weeks after embedment, the proliferation of granulation tissue had already begun to surround the calcification. Most of the cells observed were macrophages. Likewise, multinucleated giant cells increased significantly. The multinucleated giant cells were observed as two types. In one, the centers of the giant cells were vacuoles, while in the others there were deeply stained calcifications with hematoxylin. Twelve weeks after embedment of the materials, further growth of multinucleated giants cells were sighted. Histochemically, von Kossa-stain-positive granules were observed within the macrophages and multinucleated giant cells as black fine granules. According to the TRAP stained specimens, the multinucleated giant cells especially reacted strongly at 4 weeks. However, the reaction became very weak at 12 weeks. CD68 immunohistochemical staining showed positive reactions in the cytoplasm of the proliferating macrophages and multinucleated giant cells. These results suggest that multinucleated giant cells are present in the surrounding tissues due to implantation of the calcium hydroxide-based root canal filling material, and that the presence of ACP in the cells is due to ingested calcium during active phagocytosis, which would disappear later on.