Objective: The aim of the present study was to investigate the responses of heterogeneous populations of macrophages in order to determine its effect on dentin bridge formation using calcium hydroxide during the healing process after a pulpotomy. Method: The first maxillary molars from the Wistar rats (n=30), 56 days old, were obtained. The rats were sacrificed at 1, 3, 7, 14, and 28 days, postoperatively. The residual pulp of the first maxillary molars was evaluated by both immunohistochemistry and immunoelectron microscopy to anti-class Φέ molecules (OX6 antibody), monocyto/macrophage lineage cells (ED1, ED2 and ED3 antibody). Result: By 1-3 days postoperatively, ED1- and OX6-immunopositive cells were widely distributed under the demarcation zone and a small number of ED2- and ED3-positive cells were also found. And many ED1-positive cells appeared phagocytic vesicles at 3 days postoperatively. By 7 days postoperatively, some ED1- and OX6-positive cells accumulated along the differentiating odontoblast layer to be found. By 14-28 days postoperatively, the pulp progressively lost the ED1-, ED2-, ED3- and OX6-positive cells and a few of them were also found in the odontoblast layer of the dentin bridge.Conclusion: This study may provide a basis for investigating the participation and changes and variety functions of macrophages in wound healing and represent strong evidence that the macrophages may be related to the proliferation and cytodifferentiation processes of odontoblasts for dentin bridge formation.

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