The animals were sacrificed by cervical dislocation, and the left tibia was dissected. Cortical bone sections were prepared and stained using a protocol for undecalcified bone [54 (link)]. Briefly, immediately after dissection, the bones were fixed in Karnovsky’s fixative and cut with a diamond blade saw (Buehler, Co., Dusseldorf, Germany) in the transverse plane at 50% of the bone biomechanical length. The cortical sections were ground to a final thickness of ∼ 50 μm using Carbimet paper discs (800 and 1200 grit; Buehler, Co., Dusseldorf, Germany), dehydrated in ascending graded ethanol and stained with fluorescein isothiocyanate (FITC; Sigma-Aldrich, Co., St. Louis, MO, USA).
Carbimet paper discs
Manufactured by Buehler
Sourced in United States
Carbimet Paper Discs are abrasive discs used for sample preparation in metallographic analysis. They are designed to provide a consistent, uniform surface finish on metal samples during the polishing process.
Automatically generated - may contain errors
6 protocols using carbimet paper discs
1
Undecalcified Bone Histological Analysis
2
Dentin Exposure and Laser Treatment
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After the approval of the Ethics Committee of Research on Human Beings (CAAE 310887/4.1.0000.5417), sixty sound human third molars were obtained and randomized into six groups according to the sizes of the exposed dentin areas (n = 10). To expose dentin, the occlusal face was sectioned perpendicular to the long axis of the tooth using a sectioning machine (Isomet™ Low Speed Saw®, Buehler; Lake Bluff, IL, USA) with a water-cooled diamond disc (Extec Corporation; Enfield, CA, USA). Enamel remaining was removed using #320 grit paper (Carbimet Paper Discs, Buehler; Lake Bluff, IL, USA), and a smear layer was simulated with #600 grit paper for 30 s using a polishing machine (Arotec; Cotia, SP, Brazil).
A condensation-cured silicon mold (Zetalabor, Zhermack; São Paulo, SP, Brazil) was positioned centrally on the dentin surface to obtain a standardized area of 36 mm2 to ensure effective laser treatment. A white nail polish (Revlon, Revlon International Corp; New York, NY, USA) was applied around the mold to limit the testing area.
A condensation-cured silicon mold (Zetalabor, Zhermack; São Paulo, SP, Brazil) was positioned centrally on the dentin surface to obtain a standardized area of 36 mm2 to ensure effective laser treatment. A white nail polish (Revlon, Revlon International Corp; New York, NY, USA) was applied around the mold to limit the testing area.
3
Microstructural Characterization of NiTi Alloy
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The cross-sections of five NiTi cylindrical specimens (one specimens from each group) were polished with a standard sequence of abrasives (Carbimet Paper Discs, Buehler: 240 and 600 grit) and alumina paste (Alpha Micropolish, Buehler: 6 μm, 1 μm and 0.5 μm particle sizes). The polished surfaces were etched using an etching solution of composition HF + HNO3 + H2O with a volume ratio of 1:4:5 by swabbing technique.[16 ] The microstructures of the specimens were examined with an optical microscope (Leica DM4000M LED; Leica Microsystems CMS GmbH, Germany) and an scanning electron microscope (SEM) (CX-200; Coxem Ltd., Daejeon, South Korea). Magnifications ranging from ×50 to ×2000 were used with the optical microscope and SEM. The grain size was calculated using the line intercept method on the optical microscope.[17 ] Forty-five lines per mm were used for the calculation in both vertical and horizontal direction. The elemental composition of different zones namely, the matrix, grain boundaries, martensitic variants and precipitates was determined for the SM alloy using scanning electron microscope (CX-200; Coxem Ltd., Daejeon, South Korea) coupled to energy dispersive X-ray spectrometer (SEM-EDS). The quantitative analysis was performed in nonstandard analysis mode employing ZAF correction methods using the software.
4
Dentin Surface Preparation for DBS
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A total of 80 sound human third molars were extracted for surgical reasons after consent and approval by the local Ethical Committee (protocol number 49812415.1.0000.5417), and were randomized using Excel software (Microsoft Office ® , Redmond, WA, USA) according to the dimensions of exposed dentin area into eight groups (n=10). Teeth were stored in 0.1% tymol solution. After the teeth were completely cleaned, the crowns were transversely sectioned on the occlusal third to expose the dentin using a low-speed diamond saw (Isomet™ Low Speed Saw ® , Buehler, Lake Bluff, IL, USA) with a water-cooled diamond disc (Extec, Enfield, CA, USA). The remaining enamel was removed using #320 grit silicon carbide paper (Carbimet Paper Discs, Buehler) on a polishing machine (Arotec, Cotia, SP, Brazil). The samples were polished for 30 s with a #600 grit silicon carbide paper to simulate smear layer formation and stored in deionized water at 37 °C until the moment of DBS application.
5
Enamel-Dentin Slice Preparation for Analysis
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One 2 mm thick exactly plane-parallel enamel-dentin slice was cut horizontally from the mid-coronal region of 6 third molars (Innenlochsäge Leitz 1600, Leica Mikrosysteme Vertrieb GmbH, Wetzlar, Germany; blade thickness 300 µm) using copious water cooling. A central predetermined breaking line was notched centrally into the pulpal surface of the 6 enamel-dentin slices in bucco-lingual direction using a water-cooled diamond disc (Superdiaflex H 365F 190, Horico Dental, Berlin, Germany). A standard smear layer was produced as follows: The coronal surfaces of each of these 6 specimens were ground wet by one operator (A.B.) for 60 s using a circular motion and finger pressure on the sandpaper lying on a laboratory bench (600 grit ANSI/CAMI; Carbimet Paper Discs, Buehler, Lake Bluff, IL, USA), rinsed with demineralized water (1.82 × 107 µSv; TKA GenPure, TKA xCAD, TKA Wasseraufbereitungssysteme GmbH, Niederelbert, Germany), and gently dried with compressed air up to the point that all visible water was removed. Subsequently, the specimens were fractured by hand along the predetermined breaking line to expose a perpendicular view onto the undisturbed dentin covered with a smear layer and smear plugs.
6
Vickers Hardness Measurement of Cylindrical Specimens
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All cylindrical specimens (3/group) were mounted and the cut surface was ground and polished with sandpaper (Carbimet Paper Discs, Buehler: 240 and 600 grit) in a polishing machine (Bainpol, Chennai Metco Pvt Ltd, Chennai, India). Final polishing was done with alumina paste (Alpha Micropolish, Buehler: 6 μm, 1 μm and 0.5 μm particle sizes). Hardness measurements were performed in each specimen with Vicker's hardness tester (VM-50, Fuel Instruments and Engineers Pvt. Ltd., Kohlapur, India) at room temperature with 5 kg load and 15 s dwell time. Five indentations were made in each specimen with a square-based pyramidal diamond indenter at the center and at equidistant adjacent locations in each specimen (n = 15). The two diagonals of the indentations were measured to the nearest 0.1 μm with a filar micrometer and averaged. Hardness was taken as the maximum force divided by the area of contact.
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