The pulpal survival after injury is mediated amongst others by changing of pulp fibroblasts to odontoblast like cells for building and mineralizing new dentin matrix. The aim of this study was to detect a relationship between root development and mineralizing capacity of human pulp fibroblasts in vitro.
Methods:
Single cultured human pulp fibroblasts (D-MEM, 10% FCS, 50µg/ml Gentamycin) were divided in group A – incomplete and group B – complete root development. Stimulation of mineralization in vitro was done by adding calcium hydroxide (1.7g/ml), beta-glycerophosphate (10mM) and ascorbic acid (50µg/ml) individually or combined together. Determination of viability was done at day 1 by using an MTT-test. At day 4, amount of collagen typ I was measured with METRA-CIP EIA Kit and alkaline phosphatase activity was determined by the 4-NPP method at days 1, 4, 8, 11 and 15. Deposition of calcium phosphate by pulp fibroblasts was detected with an X-ray microanalysis.
Results:
Compared to control cells, a significant increase in pulp fibroblasts viability was obtained by adding beta-glycerophosphate and ascorbic acid (p = 0.05). The amount of collagen typ I as well as the deposition of calcium phosphate were significantly influenced with all stimulating agents (p = 0.05), while no significant effect of the stimulating agents was detectable in alkaline phosphatase activity. Root development did not influence any measured parameter.
Conclusion:
Compared to the stimulating agents used, root development had no significant influence on the mineralizing capacity of human pulp fibroblasts in this in-vitro study as shown by viability, alkaline phosphatase activity, collagen typ I amount and deposition of calcium phosphate.