After the cavity filling, the roots of each tooth were removed 3 mm below the enamel–cementum junction with a diamond disc (Disc DS022, Clinique, manufacturer information is not provided), under continuous water cooling.
Then, the pulp tissue was removed using small excavators, and a 17% EDTA solution (Endo-Solution, Cerkamed, Stalowa Wola, Poland) was applied to the pulp chamber for 1 min. Then the pulp chamber was thoroughly rinsed with distilled water.
Each tooth fragment was fixed using a cyanoacrylate adhesive (Loctite, Westlake, OH, USA) to the inner face of the metal cap of a glass container. Through the cap, a 3 mm high syringe needle was inserted into the pulp chamber. The needle was connected to a perfusor coupled to a 100 mL saline solution vial. The vial was fixed at a height of 20 cm to produce a hydrostatic pressure of 20 cm H2O in the pulp chamber [29 ,30 (link)].
Then, the pulp tissue was removed using small excavators, and a 17% EDTA solution (Endo-Solution, Cerkamed, Stalowa Wola, Poland) was applied to the pulp chamber for 1 min. Then the pulp chamber was thoroughly rinsed with distilled water.
Each tooth fragment was fixed using a cyanoacrylate adhesive (Loctite, Westlake, OH, USA) to the inner face of the metal cap of a glass container. Through the cap, a 3 mm high syringe needle was inserted into the pulp chamber. The needle was connected to a perfusor coupled to a 100 mL saline solution vial. The vial was fixed at a height of 20 cm to produce a hydrostatic pressure of 20 cm H2O in the pulp chamber [29 ,30 (link)].
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