Title: Classical Autoflourescence Reduction Approaches are Ineffective for Human Enamel Tufts
Monil Mehta (Presenter)
University of Pittsburgh
Xu Yang, University of Pittsburgh
Elia Beniash, University of Pittsburgh
Objectives: Non-specific fluorescence from demineralized enamel matrix can significantly compromise the immunofluorescence studies and lead to false positives. The goal of this study was to assess the effectiveness of autoflourescence reduction methods in mature human enamel matrix and forming dental tissues under different conditions.
Methods: We compared two methods of background fluorescence elimination, Sodium borohydride (SBH) and Sudan Black (SBB) treatments on the sections of forming mouse incisor and human enamel tufts. Human enamel tufts were collected from demineralized healthy teeth, extracted for clinical reasons and embedded in paraffin. Mandibles were collected from 10 days old mice, demineralized and embedded in paraffin. Ten micron thick sections of both samples were treated with SBH and SBB according to standard protocols. Non-treated samples were used as controls. The samples were imaged in the epifluorescence mode and the fluorescence intensities were measured.
Results: We demonstrated that SBB is far superior to SBH in reducing the background fluorescence in mouse forming dental tissues, including the forming enamel matrix, but that neither SBB nor SBH treatments were effective in reducing background fluorescence in mature human enamel tufts.
Conclusions: The differences observed in the effects of the autoflourescence reduction methods might reflect the differences in the composition and amount of crosslinks between forming and mature enamel matrix. Further studies will be needed to optimize and standardize immunochemical procedures for the mature enamel matrix.