Sem evo ma 10

The cross-section of the prepared HFs was observed using a scanning electron microscope (SEM EVO|MA 10, Zeiss, Milano, Italy). The HFs were fractured in liquid nitrogen and then sputter-coated with gold. Sample images were acquired in high-vacuum mode, working at 20 kV.

Each group was cut with a size of 5 × 5 mm. The surface of the PVA–collagen–HA composite membrane was coated with 10-nm-thick gold using a sputter coater (Quorum Q150R ES, Quorum, East Sussex, UK) and observed using a scanning electron microscope (SEM) (SEM EVO MA 10, ZEISS, Jena, Germany) at an accelerating voltage of 16 kV. The photo taken is the membrane surface with ×1,000 and ×5,000 magnification. The interpretation results of SEM were analyzed using ImageJ software.

Unstained polished sections of the resin-embedded implants were imaged using backscattered scanning electron microscopy (SEM-EVO MA10, Zeiss, White Plains, NY, USA) equipped with Oxford Aztec (Zeiss, USA). To increase the conductivity of the materials and, consequently, image quality, copper SEM tape was attached to each sample and to the stub prior to the imaging and samples were sputter coated with a layer of gold palladium (Polaron e500) (Quorum Technologies, Laughton, UK). SEM parameters chosen for imaging were electron high tension (EHT) 20.00 kV, magnification 26–30× and, I probe 600 pas. Images were analyzed for new bone formation (%) using ImageJ v. 1.8.0 (NIH, Madison, WI, USA). A circular region (ROI), corresponding to the defect area, was initially cropped. Then, both the total area of the defect and the area of the new bone were binarized using an interactive threshold method (default). Thresholding enabled the isolation and quantification of bone formation based on greyscale. The percentage of new bone formation was determined according to the following formula: $$\left(\frac{A bone}{A tot}\right)\times 100$$
where, A bone represents the area (in pixels) of the new bone ingrowth and A tot is the total area of interest.