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Insulin-like Growth Factor I Regulates Apoptosis in Condylar Cartilage

¹ Section of Maxillofacial Orthognathics, Department of Maxillofacial Restoration, Division of Maxillofacial/Neck Reconstruction, and
² Section of Pharmacology, Department of Hard Tissue Engineering, Division of Bio-Matrix, Graduate School, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan; and
³ Oral Histology, Graduate School of Dentistry, Health Science University of Hokkaido, Japan

View larger version (15K): [in this window] [in a new window]   Figure 1. Real-time RT-PCR analysis of IGF-1 and IGFBP3 mRNA in the mandibular condyles cultured for 3 days. (A) The mRNA expression of IGF-1 in the AS-ODN group was significantly lower than that in the S-ODN and control groups. (B) The mRNA expression of IGFBP3 in the AS-ODN group was significantly lower than that in the S-ODN and ODN-free groups. Values are means ± SD, derived from triplicate arrays. *p < 0.05 vs. the ODN-free and S-ODN groups.

View larger version (81K): [in this window] [in a new window]   Figure 2. Histological observation. (A) Toluidine blue staining of the mandibular condyle for 1 day (a,b), 3 days (c,d), or 5 days (e,f), treated with IGF-1 AS-ODN (b,d,f) or S-ODN (a,c,e). The condylar cartilage was subdivided into the articular fibrous layer (A), the undifferentiated mesenchymal cell layer (U), and chondrocytes (C). A great many dead cells due to nuclear condensation (arrows) were observed in the undifferentiated mesenchymal cell layer directly beneath the articular fibrous layer (d,f). (a-f) Toluidine blue staining. (B) Toluidine blue staining (a,b,c) and TUNEL staining (d,e,f) of three-day-cultured mandibular condyles treated with ODN-free (a,d), AS-ODN (b,e), and S-ODN (c,f). Many TUNEL-positive cells (large arrows) were observed in the undifferentiated mesenchymal cell layer (b), and nuclear condensation (arrows) was also observed (e). (a-c) Toluidine blue staining. (d-f) TUNEL staining. Scale bars: 10 µm.

View larger version (82K): [in this window] [in a new window]   Figure 3. Electron microscopic images of the cells in the undifferentiated mesenchymal cell layer on day 3 of culture. (A) Electron microscopic image of the S-ODN group. Little heterochromatin (asterisks) was observed. (B) Electron microscopic image of the AS-ODN group. Chromatin condensation (asterisks) was detected in the nucleus and shrunken cytoplasm (arrows). Scale bars: 5 µm.

View larger version (53K): [in this window] [in a new window]   Figure 4. Immunoblotting for cleaved caspase 3. Mandibular condyles were cultured for 3 days in medium containing AS-ODN, S-ODN, or no ODN. Extracted protein was analyzed by immunoblotting analysis with an antibody specific for the cleaved caspase 3. (A) Almost equal amounts of protein were loaded in each sample well. (B) The cleaved products of caspase 3 were increased after treatment with AS-ODN, compared with the ODN-free and S-ODN groups.

Journal of Dental Research, Vol. 87, No. 2, 159-163 (2008)
DOI: 10.1177/154405910808700216


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