Perfection v800 photo color scanner

The evaluation of the variation of the marginal bone level over time was performed according to a previous study [14 (link)]. Reproducible intra-oral periapical radiographs were used. When there were no available digital radiographs from the baseline appointment, the analogue periapical radiographs were scanned at 1200 dpi (Epson Perfection V800 Photo Color Scanner; Nagano, Japan).
MBL was measured after calibration based on the inter-thread distance of the implants. Information about the inter-thread distance was obtained from the implant catalogue of each implant manufacturer. Measurements were taken from the implant-abutment junction to the marginal bone level at both mesial and distal sides of each implant (Figure 3), and then the mean value of these two measurements was considered. MBL was calculated by comparing bone-to-implant contact levels to the radiographic baseline examination.
The two types of CIR and implant inclination were calculated according to the aforementioned definitions.
The Image J software (National Institute of Health, Bethesda, MD, USA) was used for all measurements.

The evaluation of the variation of the marginal bone level over time was performed according to a previous study [19 (link)]. Reproducible intra-oral radiographs were used. When there were no available digital radiographies from the baseline appointment, the analogue periapical radiographies were scanned at 1200 dpi (Epson Perfection V800 Photo Color Scanner; Nagano, Japan). Marginal bone level (MBL) was measured after calibration based on the inter-thread distance of the implants. Information about the inter-thread distance was obtained from the implant catalogue of each implant manufacturer. Measurements were taken from the implant-abutment junction to the marginal bone level, at both mesial and distal sides of each implant, and then the mean value of these two measurements was considered. MBL was calculated by comparing bone-to-implant contact levels to the radiographic baseline examination. The Image J software (National Institute of Health, Bethesda, MD, USA) was used for all measurements.
The sets of radiographs for every patient were codified, and the authors who performed the radiological measurements (A.A., A.A.A.) were blinded to the smoking habit for every patient.

For SDS-PAGE gel electrophoresis followed by total protein staining, IP samples were loaded on 4 − 12% Bis-Tris gels (NuPAGE #NP0322BOX) and separated using MOPS buffer (Novex #NP0001). Gels were stained with Imperial Protein Stain (Thermofisher Scientific #24615) per manufacturer’s instructions and images were captured using Epson Perfection V800 Photo Color Scanner. Images were processed by ImageJ software. For western blots, IP samples were loaded (20% of an IP sample/well) and separated on 10% SDS-PAGE gels. The protein was transferred onto a nitrocellulose membrane, and the blot was blocked and incubated with a primary anti-GOA-1 antibody [1:1000 diluted affinity-purified rabbit anti-GOA-1 polyclonal antibody (Patikoglou and Koelle 2002 (link)) at 4°C for overnight], washed, incubated with a secondary antibody (1:3000 HRP-linked Anti-Rabbit antibody Bio-Rad) and protein bands were visualized with SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermofisher Scientific #34580) using a BioRad ChemiDoc MP system. Blots were reprobed for Gβ by stripping and then incubating overnight with 1:200 diluted mouse monoclonal anti-Gβ antibody (Santa Cruz #sc-166123) followed by a secondary incubation with 1:1000 diluted m-IgGk BP-HRP (Santa Cruz #sc-516102), and bands were again visualized by chemiluminescence.