Influence of cryopreservation on the pulpal tissue of immature third molars in vitro

The purpose of this study was to evaluate in vitro the viability of isolated and non-isolated pulpal tissue of immature third molars after cryopreservation. This study was divided in three different experiments. Experiment 1 : Pulpal tissue isolated from 19 third molars was divided in horizontal segments. Each segment was cultured separately in order to evaluate whether differences in growth capacity within the tissue could be found. Experiment 2 : Pulpal tissue isolated from 27 third molars was divided in a mesial and a distal part. One part was cryopreserved before culturing, the other part was cultured immediately. Growth capacity of cryopreserved and non-cryopreserved tissue was evaluated and compared. Experiment 3 : 43 third molars were cryopreserved. After thawing, the dimension of the apical foramen was measured and the pulp was isolated and segmented horizontally. The different parts were cultured and growth capacity was evaluated and compared. Results of experiment 1 and 2 showed no significant difference in growth capacity between fibroblasts originating from different pulpal segments of the same tooth without cryopreservation and between fibroblasts originating from cryopreserved and non-cryopreserved isolated pulpal tissue. In experiment 3 it was demonstrated that the dimension of the apical foramen and pulpal viability after cryopreservation are positively correlated. A minimum dimension of 9.42 mm² enables the cryoprotective agent to penetrate sufficiently and to protect the pulpal tissue from apex to crown. This study proved that cryopreservation of human pulpal tissue is possible if the cryoprotective agent can reach the entire pulp.