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Figure 4. Cyclin and tenascin expression was up-regulated in upper lips cultured in the absence of serum and supplemented only with exogenous TGF-β3. Cleft upper lips were isolated and explanted into serumless cultures in the absence (a,d,g,j) or presence of 10 (b,e,h,k) or 100 ng/mL (c,f,i,l) of TGF-β3. Explants cultured for 60 hrs in the absence of TGF-β3 exhibited a significant cleft (between arrowheads) remaining between the 2 halves of the cleft lip (a,d). In the presence of 10 ng/mL TGF-β3, 1 in 10 of the cleft lips achieved partial fusion (b,e) in which an epithelial seam (asterisks) demarcated the 2 halves of the cleft lip. In the presence of 100 ng/mL TGF-β3, 4 in 10 of the cleft lips fused (c,f), with small remnants of the epithelium remaining (asterisk) and continuity of the mesenchyme throughout the fusion area. In the presence of TGF-β3 (h,i,k,l), the number of cells positive for cyclin (g-i) or extracellular matrix positive for tenascin (j-l) was apparently increased when compared with those in controls (g,j). The total number of mesenchymal cells at the fusion area in lips treated with 100 ng/mL TGF-β3 was increased slightly but significantly. However, the percentage of cyclin-positive mesenchymal cells was substantially increased: 4.5% (33.75 + 8.75 cyclin-positive cells compared with 753.68 + 98.95 total cells, N = 5), 6.8% (55.42 + 7.92 cyclin-positive cells compared with 818.95 + 143.16 total cells, N = 5), and 16.5% (157.5 + 14.58 cyclin-positive cells compared with 953.68 + 77.89 total cells, N = 5) for 0, 10, and 100 ng/mL TGF-β3, respectively (m). TGF-β3 inceased cyclin expression dose-dependently (0.47 + 0.28 for control, 2.63 + 0.45 at 10 ng/mL and 4.32 + 0.58 at 100 ng/mL TGF-β3, N = 5, for all groups) (n). Similarly, TGF-β3 promoted tenascin-C expression (5.06 + 1.03 for control, 6.92 + 1.24 at 10 ng/mL, and 9.51 + 1.32 at 100 ng/mL TGF-β3, N = 5, for all groups) (n). Scale bar in (a) for (a-c) is 1 mm; those in (d) for (d-f) and in (g) for (g-l) are 100 µm.  p < 0.01 compared with 0 ng/mL TGF-β3 group and  p < 0.01 compared with 10 ng/mL TGF-β3 group.
J DENT RES, Vol. 81, No. 10,
688-694 (2002)
DOI: 10.1177/154405910208101007
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