Fig. 4
Characterization of regenerated tissues after transplantation of reconstituted teeth. Twenty-eight days after ectopic transplantation of MDPSCs (a, e, j, n) in the native autoclaved tooth, in the autoclaved teeth reconstituted (b, f, k, o) with the EDTA extracts, (c, g, l, p) with the CM, and (d, h, m, q) with the EDTA extracts and the CM. a–d In-situ hybridization analysis of expression of thyrotropin-releasing hormone degrading enzyme (TRH-DE) as a pulp marker. e–h In-situ hybridization analysis of enamelysin. Odontoblastic processes (arrows) extending into the tubular dentin. i Comparison of the numbers of enamelysin-positive cells along the dentinal wall. j–m Immunostaining with periodontal ligament-associated protein 1 (PLAP-1, green) merged with Hoechst 33342 (blue). A typical PLAP-1-positive cell (arrow). n–q Immunostaining with periostin (green) merged with Hoechst 33342 (blue). r Ratio of the PLAP-1-positive cell number to the total cell number. Data are expressed as means ± SD of four determinations. *P < 0.05. # P < 0.001 to nonextracted tooth, reconstituted tooth with the MDPSC CM alone, and the MDPSC CM with the EDTA extracts. CM conditioned medium (Color figure online)