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Matrix Metalloproteinases have a Role in Palatogenesis

Division of Child Dental Health, University of Bristol Dental School, Lower Maudlin Street, Bristol, BS1 2LY, UK;

View larger version (116K): [in this window] [in a new window]   Figure 1. Hematoxylin and eosin (H&E)-stained sections showing palate development in vivo alongside palates cultured in vitro. Scale bar represents 0.1 mm. The tongue (T), palatal shelves (P), nasal septum (NS), Meckel's cartilage (M), and medial edge epithelium (MEE) are labeled. (A) Vertical orientation of palatal shelves. (B) Horizontal orientation of palatal shelves, no contact. (C) Palatal shelves in contact, medial edge epithelium present. (D) Palatal shelves fused, mesenchyme continuous.

View larger version (89K): [in this window] [in a new window]   Figure 2. Gelatin gel zymograms for MMP-2 assessment. Each lane represents a single palate. A 5-µg quantity of protein was loaded per sample. Pro and active (72 and 62 kDa) MMP-2 bands were present on all gels. MMP-9 was not detected by gelatin gel zymography at any of these stages of palate development. (A) MMP-2 levels during in vivo murine palate development. Each of the key stages of palatogenesis is represented in duplicate by embryonic day 13, 14, and 15 palates. Pro and active levels of MMP-2 increase from embryonic day 13. (B) MMP-2 levels during in vitro murine palate development. Elevated active MMP-2 levels with increasing time in culture, suggesting that the in vitro model of palate development was representative of in vivo palatogenesis with respect to MMP-2. (C) MMP-2 levels during in vitro murine palate development with BB3103, a synthetic MMP inhibitor. Reduction of active MMP-2 (62 kDa) to levels undetectable with gelatin gel zymography at 20 µM BB3103.

View larger version (78K): [in this window] [in a new window]   Figure 3. MMP-3 expression during in vitro palatogenesis, with immunohistochemistry. Scale bar represents 0.1 mm. (A) MMP-3 was expressed within the medial edge epithelium during its breakdown. MMP-3 staining was minimal preceding and following this event, similar to the pattern of staining seen in vivo (data not shown). (B) MMP-3 expression in the palate cultured with 20 µM BB3103 that fused. High expression of MMP-3 leads to medial edge epithelium breakdown and palatal shelf fusion in the presence of the inhibitor. (C) MMP-3 expression in a palate cultured with 20 µM BB3103 that attained contact without fusion. Medial edge epithelium breakdown was inhibited. MMP-3 was less prominent in palates that failed to fuse.

Journal of Dental Research, Vol. 81, No. 12, 826-830 (2002)
DOI: 10.1177/154405910208101206


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