Elsevier

Journal of Endodontics

Volume 25, Issue 1, January 1999, Pages 24-29
Journal of Endodontics

An in vitro pulp chamber with three-dimensional cell cultures

https://doi.org/10.1016/S0099-2399(99)80394-XGet rights and content

To better simulate the in vivo situation, a three-dimensional fibroblast cell culture was introduced into an in vitro pulp chamber model. The system was evaluated by testing a series of dental filling materials. After a 24-h exposure with (0.3 or 5 ml/h) and without perfusion of the pulp chamber, the tissues were subjected to a routine MTT assay. Zinc phosphate cement, conventional glass ionomer cements, a silicone impression material, and zinc oxide-eugenol did not influence cell viability, compared with untreated controls; but, a light-curing glass ionomer cement significantly reduced cell survival. Perfusion of the chambers did not significantly influence the results, but perfusion conditions of 5 ml/h lead to a general decrease of cell vitality. The three-dimensional cell culture system in an in vitro pulp chamber seems to be a substantial improvement, because zinc oxide-eugenol does not evoke a cellular reaction (as is the case in vivo), and the test system is sensitive enough to detect other toxicants.

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    This refers to the pulp analogue within the 24 well plate and the dentin analogue suspended over this pulp analogue within the hanging culture insert, composed of differentiated layer of odontoblast like cells and the treated-dentin disk. Numerous publications have dealt with incorporating the dentin barrier over an artificial pulp chamber in order to simulate the clinical scenario, with several variations, such as utilization of dentin slices of various thicknesses (100, 200, 300, 500, 700 μm) [21,63], cell seeding in 3D polyamide meshes instead of the initial 2D cell cultures on coverslips [21,64–66], using cells of human origin [67,68], and seeding cells in natural hydrogels, such as fibrin and collagen [68]. In general, these devices became more complex and sophisticated through the years, studying the perfusion of eluates from the dentin barrier [20,69], introducing dynamic conditions in the system [23,64,70] and even reproducing the dentin barrier itself [71].

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    It should also be pointed out that a large portion of the in vitro cytotoxicity/biocompatibility studies have been developed with the use of monolayer cell culture models [15,16]. However, the 3D culture models, in which cells are cultivated in specific types of collagen matrix (scaffolds), appear to provide more favorable conditions for the morphological and phenotypical expression of the cells, and this experimental model may also be used for the direct and indirect evaluation of the biologic effects of new dental materials and techniques [24,32]. Another factor that contributes to the efficacy of the in vitro cytotoxicity/biocompatibility tests for dental materials is the selection of the tests to be performed [11], which may vary according to the type of product to be tested and the following levels of response one wishes to obtain: cytotoxicity, the induction of an inflammatory responses, biostimulation or cell differentiation capacity, among other important cellular functions for the homeostasis and repair of the dentin–pulp complex.

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Drs. Schmalz, Schuster, Nuetzel, and Schweikl are affiliated with the Department of Operative Dentistry and Periodontology, University of Regensburg, Franz-Josef-Strauss-Allee 11, D-93053 Regensburg, Germany.

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