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Gene Expression of Growth Differentiation Factors in the Developing Periodontium of Rat Molars

¹ Periodontology and
² Biostructural Science, Department of Hard Tissue Engineering, Graduate School of Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyou-ku, Tokyo 113-8549, Japan;

View larger version (94K): [in this window] [in a new window]   Figure 1. Buccal-lingual sections of a two-week-old rat mandible through the first molar tooth germ (M1) stained with Azan (A-C) or treated for detecting GDF-5 (D,G,J), -6 (E,H,K), and -7 (F,I,L) transcripts. (A) Whole view of M1 before eruption. Small rectangles of B and C indicate the areas enlarged in B and C, respectively. (B) Obliquely oriented thin collagen fibers (stained blue) show initial signs of attachment to the root surface as matrix fibers of acellular extrinsic fiber cementum. (C) Collagen fibers are poorly oriented in the apical region. (D-F) Cells expressing the GDF genes (stained brown) are aligned on the surface of the alveolar bone (ab) and scattered in the periodontal ligament (arrowheads). (J-L) In the apical region, few GDF-positive cells are detectable only on the alveolar bone surface. (G-I) Control with sense probes showed no positive reactions. ab, alveolar bone; d, dentin; pul, pulp. Bar = 500 µm (A), 50 µm (B-L).

View larger version (94K): [in this window] [in a new window]   Figure 2. Buccal-lingual sections of the M1 in three-week-old rat stained with Azan (A-C) or treated for detecting GDF-5 (D,G), -6 (E,H), and -7 (F,I) transcripts. Small rectangles of B and C (A) indicate the areas enlarged in B and C, respectively. (B) Collagenous matrix fibers of acellular extrinsic fiber cementum are shown to be continuous with thick periodontal ligament fiber bundles extending obliquely from acellular cementum toward the alveolar bone (ab). (C) Cellular cementum is deposited on the root dentin surface near the apical end (arrows). Only scattered thin collagen fibers are seen in this region. (D-I) GDFs mRNA expression in regions B and C indicated in Fig. 2A. GDF-positive cells are widely distributed in the periodontal ligament, on the surfaces of the alveolar bone and the cementum (arrowheads). ab, alveolar bone; d, dentin; pdl, periodontal ligament; pul, pulp. Bar = 500 µm (A), 50 µm (B-I).

View larger version (93K): [in this window] [in a new window]   Figure 3. Buccal-lingual sections of the M1 in an eight-week-old rat stained with Azan (A,C,D) or treated for detecting GDF-5 (E,H), -6 (F,I), and -7 (G,J) transcripts. Small rectangles of C and D (A) indicate the areas enlarged in C and D, respectively. (C) Periodontal ligament fiber bundles appear thicker and denser than in the previous stage. (D) Thick cellular cementum (cc) on the root dentin surface. (B) Fluorescent green lines of labeled calcein follow the course of mineralizing fronts of dentin (d), alveolar bone (ab), and cellular cementum (cc) in the area outlined by the red rectangle in Fig. 3A. (E-J) Cells showing GDFs gene expression are observed in the cells on the surfaces of the cementum and the alveolar wall and also on those scattered in the periodontal ligament (pdl) (arrowheads). ab, alveolar bone; cc, cellular cementum; d, dentin; pdl, periodontal ligament; pul, pulp. Bar = 500 µm (A and B), 50 µm (C-J).

View larger version (79K): [in this window] [in a new window]   Figure 4. Buccal-lingual sections of the M1 in a 36-week-old rat stained with Azan (A,C,D) or treated for detecting GDF-5 (E,H), -6 (F,I), and -7 (G,J) transcripts. (A) Apically, one-fourth of the root is fully covered by thick cellular cementum. Small rectangles of C and D indicate the areas enlarged in C and D, respectively. (B) The fluorescent green line of labeled calcein is almost undetectable in the alveolar bone (ab), dentin (d), and cementum (cc) in the area outlined by the red rectangle in Fig. 4A. (E,F) GDF-positive cells are not visible in the periodontal tissues in sections hybridized with the GDFs mRNA probe. ab, alveolar bone; cc, cellular cementum; d, dentin; pdl, periodontal ligament; pul, pulp. Bar = 500 µm (A and B), 50 µm (C-J).

Journal of Dental Research, Vol. 82, No. 3, 166-171 (2003)
DOI: 10.1177/154405910308200304


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