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The bioactivity of calcium silicate mineral trioxide aggregate (MTA) cements has been attributed to their ability to produce apatite in presence of phosphate-containing fluids. This study evaluated surface morphology and chemical transformations of an experimental accelerated calcium-silicate cement as a function of soaking time in different phosphate-containing solutions. Cement discs were immersed in Dulbecco's phosphate-buffered saline (DPBS) or Hank's balanced salt solution (HBSS) for different times (1-180 days) and analysed by scanning electron microscopy connected with an energy dispersive X-ray analysis (SEM-EDX) and micro-Raman spectroscopy. SEM-EDX revealed Ca and P peaks after 14 days in DPBS. A thin Ca- and P-rich crystalline coating layer was detected after 60 days. A thicker multilayered coating was observed after 180 days. Micro-Raman disclosed the 965-cm(-1) phosphate band at 7 days only on samples stored in DPBS and later the 590- and 435-cm(-1) phosphate bands. After 60-180 days, a layer approximately 200-900 μm thick formed displaying the bands of carbonated apatite (at 1,077, 965, 590, 435 cm(-1)) and calcite (at 1,088, 713, 280 cm(-1)). On HBSS-soaked, only calcite bands were observed until 90 days, and just after 180 days, a thin apatite-calcite layer appeared. Micro-Raman and SEM-EDX demonstrated the mineralization induction capacity of calcium-silicate cements (MTAs and Portland cements) with the formation of apatite after 7 days in DPBS. Longer time is necessary to observe bioactivity when cements are immersed in HBSS.

Background: Cell-substratum interactions play a peculiar role in cell proliferation, differentiation and migration. They are regulated by various glycoproteins, among which fibronectin, and by receptors connecting cells to the extracellular matrix, i.e. integrins. Therefore, the aim of this study was to correlate the proliferation rates of the human fibroblast line WI-38 cultured in presence of titanium dental implants to cell adhesion capability to substrata., Methods: WI-38 fibroblasts were cultured in presence of four dental implants in titanium (one hydroxyapatite coated) for 48, 72 and 96 hours. Cell proliferation was evaluated by detecting 5-bromodeoxyuridine incorporation. Fibronectin organization and alpha5beta1 integrin expression were evidenced by indirect immunofluorescence., Results: A correlation between fibronectin organization and cell proliferation rates was demonstrated: cultures showing fibronectin mainly organized in fibrils presented the highest cell proliferation degrees. After 96 hours, the observed decrease of the number of proliferating cells corresponded to a different fibronectin organization. In presence of the hydroxyapatite coated implant, colocalization of fibronectin and alpha5beta1 integrin was represented in focal contacts in cultures exhibiting the highest proliferation rate, while cells with the lowest proliferation one expressed alpha5beta1 integrin in point contacts., Conclusions: Evidences obtained in this work showed that both the organization of fibronectin and the expression of alpha5beta1 integrin are strictly correlated to cell proliferation rates. Therefore, these parameters could be useful for evaluating the biocompatibility of dental materials in vitro.

The nature of the bone-implant interface has been much focused in investigating dental implant materials, whereas the relationship between implant and fibroblasts has received much less attention. To evaluate the biocompatibility degree of an implant material, both cell adhesion and cell growth must be tested in the presence of the implant. Four dental implant (A,B,C,D) made in titanium alloy and one of them (C) hydroxyapatite (HA)-coated in fibroblast cultures (48 and 72 h) were tested, by performing immunocytochemical techniques and then by observing fibronectin (FN) arrangement for cell adhesion and counting 5-bromodeoxyuridine (5-BrdU) to evaluate cell proliferation. Different FN arrangements were observed-i.e. organized in fibrils, or in focal adhesion plaques, as well as dispersed in the intercellular space-which varied for the different implants employed at the various culture stages. Equally, the per cent ratio of 5-BrdU positive cells was different, with a more significant increase (p < 0.001) between 48 and 72 h for implant C and the controls. It was observed that the higher percentages of 5-BrdU positive cells were in cultures where FN was organized mainly in focal adhesions, as well as 5-BrdU positive cells increased after 72 h in cultures, which after 48 h presented much FN dispersed in the intercellular space. It may be assumed that a correlation exists between FN arrangement and the percentage of 5-BrdU positive cells and that these two parameters vary in the presence of the different implants. Moreover, the HA-coated implant seems to be the most biocompatible in fibroblast cultures.