Abstract
This study compared the cytotoxicity and the release of nitric oxide induced by collagen membranes in human mononuclear cells. Peripheral blood was collected from each patient and the separation of mononuclear cells was performed by Ficoll. Then, 2x10(5) cells were plated in 48-well culture plates under the membranes in triplicate. The polystyrene surface was used as negative control. Cell viability was assessed by measuring mitochondrial activity (MTT) at 4, 12 and 24 h, with dosage levels of nitrite by the Griess method for the same periods. Data had non-normal distribution and were analyzed by the Kruskal-Wallis test (p<0.05). Statistically significant differences (p<0.05) were observed between the membranes and the control in the experimental period, although there was a significant reduction in viability over time (p<0.01). At 4 and 12 h, the porcine membrane induced a higher release of nitrite compared with the control and bovine membrane, respectively (p<0.01), and this difference was maintained at 24 h (p<0.05). This in vitro study showed that the porcine collagen membrane induces an increased production of proinflammatory mediators by mononuclear cells in the first hours of contact, decreasing with time.
Highlights
Guided tissue regeneration (GTR) is a procedure used to control early periodontal surgical wound healing dynamics in order to promote periodontal regeneration [1,2]
The tests were done in monocyte human primary cell culture plated in bovine and porcine collagen membranes, to evaluate the cytotoxicity and amount of nitrite released along the time
At 4 and 12 h of cell culture, the nitrite levels produced by mononuclear cells cultivated in porcine membrane were significantly higher than in the polystyrene surface (PS) group or in the bovine membrane, respectively (4 h p=0.0070; 12 h p=0.007) (Fig. 3A and 3B, respectively)
Summary
Guided tissue regeneration (GTR) is a procedure used to control early periodontal surgical wound healing dynamics in order to promote periodontal regeneration [1,2]. Several materials have been used as barriers, namely polytetrafluoroethylene (PTFE), collagen, polyglycolic acid and copolymers [3,7], which are classified as non-absorbable or bioabsorbable [3,4]. A number of bioabsorbable barrier materials have been clinically used in GTR procedures [5,6,10]. These barriers are currently produced by collagen reconstitution using cross-linking techniques [4] to reduce the fast resorption and the lack of stability in the surgical area. The techniques used to multiply the links between collagen molecules, increasing the stiffness and slowing down the resorption of the collagen barrier, may increase its cytotoxicity [7]. The properties of collagen barriers may be affected by the origin of the collagen used in their fabrication [7,11]
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