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The coronal access of specimens was filled with a temporary filling material (Cavit; 3M ESPE, Seefeld, Germany).
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The root canals of another 70 specimens were filled with Thermafil and AH Plus and the root canal fillings were removed after 24 hours using ProTaper D files and plasma sterilized.
Specimens were filled with a size 60 master gutta-percha cone and accessory gutta-percha cones (Dentsply Maillefer).
Hitherto, the pulp spaces of three specimens were filled with a dye solution and the volume of ultrapure water needed to achieve unstained drain of the pulp space was measured.
The root canals of 70 specimens were filled using a size 25 Thermafil obturator (Dentsply®, Konstanz, Germany) in combination with an epoxy resin-based sealer (AH Plus, Dentsply®, Konstanz, Germany).
The chord member of the test specimen was filled with concrete along its full length.
The pulp space of each specimen was filled with ADF and its nozzle was connected with a pressurized ADF reservoir by a flexible tube (inner diameter 2 mm, no. 1337, Novoplast Schlauchtechnik GmbH, Halberstadt, Germany).
The gap between hydraulic jack and top of the specimen is filled with spacer plates.
The whole process is repeated until the entire specimen is filled with mortar along the height direction.
A 2 mm high cylindrical polyethylene tube with an internal diameter of approximately 8 mm was placed on the surface of each underlying specimen, the tube was filled with composite (either FZ250 or FS), and all samples (except the self-cure samples) were polymerized for 20 s using an LED curing unit, with all procedures carried out at room temperature.
Additionally, two wells were filled with blank alamarBlue solution (without specimens) and one well was filled with centrifuged bacteria from the planktonic inocula in alamarBlue solution to gain values for the maximal metabolic activity.
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CEO of Professional Science Editing for Scientists @ prosciediting.com