Dentin adhesives with a low pH (1-2) have been tested non toxic in a dentin barrier test with dentin slices of 500 µm thickness. However, it was advocated to use these materials also in deep cavities. Objective: To evaluate the influence of dentin thickness on the reaction of three-dimensional cultures of SV40 transfected pulp derived cells exposed to dentin bonding agents in an in vitro dentin barrier test system. Methods: Three dimensional cultures were prepared as described (Schmalz et al., J Dent Res 78: 522, 1999) and transferred into an in vitro dentin barrier test system with dentin slices of 100, 200, 300 or 500 µm thickness. After 24 h incubation at 37°C the test materials were applied into the cavity part of the device: (1) Syntac Classic (Vivadent), (2) Prompt L-Pop (3M ESPE) (pH 1), and, as a reference, (3) (Vitrebond, 3M ESPE). A A-silicone impression material was used as a negative control (100% cell viability). The materials were tested with or without perfusion (P, 2 ml MEMa with 20%FCS / h) of the pulpal part of the device. After 24h incubation, cell survival was measured by a routine MTT-tests, and 9 replicate cultures per material were statistically analyzed (Mann-Whitney-Test).

Results:

	Cell Survival (percent of negative control; medians)
Dentin	(1)	(2)	(3)	(1P)	(2P)	(3P)
100 µm	10.6	79.7	19.5	14.0	54.1	24.1
200 µm	10.1	81.7	38.0	13.2	82.3	41.0
300 µm	11.0	72.6	40.6	13.0	78.2	44.4
500 µm	12.6	115.8	53.5	19.7	94.6	80.9

Conclusions: For the low pH dentin bonding agent and the lc glass ionomer dentin thickness significantly influences the toxic effect on target cells. For the glutaraldehyde containing dentin bonding agent no such consistent relation could be established under the given experimental conditions.

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