Model for quantitative immunohistochemical assessment of pulpal response to biomaterials

The purpose of this study was to propose a new method for quantitative evaluation of the pulpal inflammation to restorative materials using immunohistochemistry and image analysis. Class V cavities were prepared and filled with different restorative materials in 20 healthy premolars to be extracted for orthodontic reasons. Teeth were extracted at different time intervals, fixed, demineralized, and embedded in paraffin. Six-micrometer-thick sections were cut serially and mounted on slides and every fifth section was stained using hematoxylin and eosin or Masson's trichrome, and served to localize the inflammatory reaction. The slides corresponding to the bulk of the inflammatory reaction were then used for immunohistochemical detection of the inflammatory cells using monoclonal antibodies: CD15 (granulocytes and histiocytes), CD45RO (T lymphocytes and monocytes), Pan-B cell (B lymphocytes, macrophages, and a subpopulation of T lymphocytes), CD45RA (B lymphocytes and monocytes), and EMA (plasma cells). The slides were observed and submitted to computerized image analysis using a SAMBA 2000 system for counting of the cells. The CD15, CD45RO, Pan-B-cell, and CD45RA antibodies positively stained the target cells, which could be counted with the computer. The EMA antibody did not permit staining of cells. These results indicate that polymorphonuclears, T lymphocytes, and B lymphocytes are present in inflamed human dental pulp. The immunologic detection of inflammatory cells followed by computerized image analysis allows an accurate characterization of pulpal pathology, and could be useful for the study of pulp reactions to restorative biomaterials.