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Nifedipine and Cyclosporin Affect Fibroblast Calcium and Gingiva

¹ Department of Periodontology, Facultad de Odontologia, University of Sevilla, c/Avicena s/n, 41009 Sevilla, Spain;
² Institute of Anatomy and Pathologic Histology, Università Politecnica delle Marche, Ancona, Italy;
³ Institute of Biochemistry, Università Politecnica delle Marche, Ancona, Italy;
⁴ Department of Medical Physiology and Biophysics, University of Sevilla, Spain; and
⁵ Emeritus Professor of Periodontology and Preventive Dentistry, University of London, UK

View larger version (42K): [in this window] [in a new window]   Figure 1. Low-power photomicrograph of the measured periodontal tissues. (a) Hematoxylin-eosin-stained tissue section, 20X. Three measurements of the gingival width were done for each case, considering the distance from the periosteum of the alveolar bone up to the basal membrane underlying the squamous epithelium, as indicated by black arrows. (b) The graph shows an increase in mean width of the nifedipine group (n = 17) and cyclosporin group tissue (n = 7), compared with the control group (n = 8), but only statistical significance for the cyclosporin group vs. the control group (*p < 0.05). The data are expressed as the mean ± SD.

View larger version (103K): [in this window] [in a new window]   Figure 2. Photomicrographs showing gingiva stained with anti-CD34. (a) Few small CD-34-positive vessels in the control group (n = 8) (40X). (b) The nifedipine group (n = 17) shows an increase in small capillaries and some thick-walled vessels (40X). (c) The cyclosporin group (n = 7) shows an increase in small vessels (40X). (d) Histogram showing significantly higher numbers of vessels/mm2 for the cyclosporin group compared with the control and nifedipine groups (*p < 0.05). The data are expressed as the mean ± SD.

View larger version (109K): [in this window] [in a new window]   Figure 3. Photomicrographs showing the different degrees of fibrosis in gingiva stained with Picro-Mallory in the three groups. (a) Control shows few collagen fibers in the subepithelial stroma (n = 8) (10X). (b) The nifedipine group is thickened, with a marked increase in collagen fibers in the subepithelial layer and deeper (n = 17) (20X). (c) The cyclosporin group (n = 7) is also thickened, but with a lesser increase in collagen fibers (20X). (d) Histogram showing differences among groups, with the nifedipine group having the highest increase in collagen (*p < 0.05). The data are expressed as the mean ± SD.

View larger version (41K): [in this window] [in a new window]   Figure 4. 3D representation of changes in fluorescence of FURA 2 AM in (a) the control group (n = 9), (b) the nifedipine group (n = 8), and (c) the cyclosporin group (n = 6). Notice the inhomogeneous distribution of cytoplasmic fluorescence (and therefore of calcium ions) in (a), (b), and (c). Increment in fluorescence is expressed as F/F. Changes in ratiometric values of fluorescence, thought to be proportional to intracellular free calcium, are shown in (d). The nifedipine group showed the lowest [Ca2+]i, but not significantly different from that in the control group. The cyclosporin group showed the highest cytoplasmic calcium concentration under resting conditions (*p < 0.05). The data are expressed as the mean ± SD.