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-Smooth-muscle Actin in and Contraction of Porcine Dental Pulp Cells

¹ Harvard School of Dental Medicine, Boston, MA; and
² Department of Orthopaedic Surgery, MRB 106, Brigham and Women's Hospital, 75 Francis Street, Harvard Medical School, Boston, MA 02115;

View larger version (80K): [in this window] [in a new window]   Figure 1. Western blot films and immunohistochemical micrographs demonstrating the presence of -smooth-muscle actin (SMA) in pulp cells. (a) Western blot films showing the SMA content of freshly isolated pulp cells and cells after passages 1 and 3. The control band from the smooth-muscle cells controls is also shown for each run. The amount of protein used for each run is noted under the band. Less protein was used for the higher passage because the SMA content was so high that it blackened the film. (b-d) Immunohistochemistry of pulp cells in monolayer culture after the first (b) and third (d) passages showing the presence of SMA (darkly stained stress fibers) in many of the cells (b, d). The SMA can be seen incorporated into the prominent stress fibers (d). The negative control sample, absent the primary antibody (c), shows no sign of the chromogen. Scale bar, 50 µm.

View larger version (126K): [in this window] [in a new window]   Figure 2. Histology of the pulp cell-seeded matrices after 1, 7, 14, and 28 days of culture, showing the contracture of the constructs with time in culture. Hematoxylin and eosin stain.

View larger version (126K): [in this window] [in a new window]   Figure 3. Micrographs of the 28-day cell-seeded matrices stained with hematoxylin and eosin (a) and with the monoclonal antibody to SMA (b-d). The negative control for the immunohistochemistry is shown in the inset of (b).

View larger version (31K): [in this window] [in a new window]   Figure 4. Data demonstrating the pulp cell-mediated contraction of the collagen-glycosaminoglycan matrices. (a) Graph showing the reduction in diameter of the non-seeded and cell-seeded matrices with time in culture. Mean ± SEM. For the cell-seeded samples: 0 (n = 24), 1 (n = 24), 7 (n = 18), 14 (n = 12), and 28 (n = 6) days. For the non-seeded samples, n = 6. The error bars are smaller than the symbols used for the means. (b) Graph showing the DNA content of the matrices with time in culture (Mean ± SEM; n = 4). (c) Bar chart showing the cell-mediated contraction normalized to DNA content of the matrices. Mean ± SEM. 1 (n = 24), 7 (n = 18), 14 (n = 12), and 28 (n = 6) days.

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