Mtm 20

SEM images for fish scale gelatin nanofibers without essential oil emulsions were analyzed using an FEI Inspect S50 Scanning Electron Microscope equipped with an EDX unit for elemental analysis. All samples were covered with a thin gold layer of about 20 nm to avoid the charging effect by using a sputtering Cressington 108 auto sputter coater device, equipped with a Cressington mtm 20 thickness controller. The secondary electron images were obtained at a 10 mm distance, using 10 kV acceleration voltage and magnification from 50× up to 10,000×. The nanofibers’ average thickness was reported as the mean diameter of a minimum of 50 nanofibers, using Origin’s built-in Gaussian fitting curve software, and the thickness measurement was processed using ImageJ software, version no. 1. 54d.

The analysis of fabrics was conducted employing an ultra-high-resolution field emission gun scanning electron microscope (FEG-SEM) system from NOVA 200 Nano SEM, FEI. A 10 kV acceleration voltage was selected in order to acquire secondary electron images. The surfaces of the samples were coated with a 25 nm film of Au-Pd (80–20 weight percent) using a sputter coater of high-resolution (208HR Cressington Company, which was fixed to an MTM-20 Cressington high-resolution thickness controller). An analysis was conducted on the morphology of the HAp layer that was applied to the PVA-braided architectures using the identical SEM technique. Subsequently, the elemental (atomic) compositions of the composite braids were analyzed, which included the Ca/P ratio of HAp. In order to accomplish this, the SEM apparatus utilized energy-dispersive X-ray spectroscopy (EDS) analysis with an EDAX Si (Li) detector and a 5 kV acceleration voltage.

All microscopy images were examined using a FEI Inspect model S50 apparatus-Scanning Electron Microscope equipped with an EDX unit for elemental analysis. Secondary electrons (SE) images and Energy-dispersive X-ray spectra (EDX) were obtained. The SEM images were obtained at a 10 mm working distance, at 5 kV acceleration voltage and for magnifications from 1000× up to 10,000×. The nanofibres thickness represents the mean diameter of minimum 50 nanofibres measurements and the results were processed using ImageJ software. The average fibre diameters and their standard errors were calculated using Origin 2016 built-in Gaussian fitting curve software. The EDX spectra are obtained for image magnification of 10,000×. Prior to any investigations, all the samples were coated with a thin Au layer (~5 nm), in order to avoid charging effects. The Au layer that covers sample surfaces is obtained using a sputtering Cressington 108 auto sputter coater device, equipped with a Cressington mtm 20 thickness controller.

An ultrahigh
resolution field-emission gun scanning electron microscope (NOVA 200
Nano SEM, FEI Company) was used to conduct morphological analyses
of fabrics. Acceleration voltages of 5 and 15 kV were used to obtain
secondary electron images and backscattering electron images, respectively.
Samples were covered with a film of Au–Pd (80–20 weight
%) in a high-resolution sputter coater, 208HR Cressington Company,
coupled to an MTM-20 Cressington high-resolution thickness controller.

AgNPs suspensions were dropped on single-side-polished silicon support for scanning electron microscopy (SEM) analysis, or copper grids coated with a carbon film for transmission electron microscopy (STEM) analysis and dried at room temperature. AgNPs for SEM analysis were coated with a thin film (5 nm) of Au-Pd (80–20 weight %) in a high-resolution sputter coater, 208HR (USA), coupled to an MTM-20 Cressington High-Resolution Thickness Controller. Morphological analysis of dried suspension was conducted by collecting secondary electron (SEM) or transmitted electrons (STEM) images by using an Ultrahigh-resolution Field Emission Gun Scanning Electron Microscope (FEG-SEM), NOVA 200 Nano SEM, FEI Co. (USA). The obtained micrographs were elaborated using ImageJ software and the particles size distribution was measured.
AFM analysis was carried out using L018W46 (Dimension Icon, Bruker, France), depositing single drops of the suspension on a microscope glass slide and allowing water evaporation at room temperature (24 °C). Scans were acquired in SCANASYST-AIR mode, with non-contacting silicon tips on nitride lever from Nanosensor (Switzerland). Images obtained were elaborated using Nanoscope software and the particles size distribution was calculated. At least 150 nanoparticles were selected from different acquired images.

For the Compound I implant, the sample was dissolved in 1 ml of DCM, and DCM was evaporated under a steady stream of nitrogen. For the EFdA implant, the sample was dissolved in 1 ml of chloroform, and 1 ml water was added to it. The mixture was vortexed and let it sit to extract the drug into the water phase. Centrifugation was carried out to separate layer of chloroform and water. The upper water layer containing the sample was removed and lyophilized to dry out the water. The dried pellets were dissolved in ACN for HPLC analysis. The LODs have been detailed above. Reverse‐phase HPLC was used to measure DL (%), defined as the measured mass of Compound I per mass of PLGA NP/implants, and EE (%), which is defined as the ratio of the compounds loaded to the total drugs used for fabricating the NPs/implants as described earlier.¹⁸Compound I and EFdA implants were also evaluated under an ultra‐high‐resolution Hitachi scanning electron microscopy (SU7000). Implants were flash frozen in liquid nitrogen. Implants were then broken in half using tweezers and cross‐sectioned using a razor blade to about 1 mm thickness. Samples were placed on a stub using carbon tape, with razor blade edge facing down. Samples were coated with platinum to a thickness of 5 nm using a high resolution sputter coater (Cressington, 208HR) with rotary planetary tilt stage and thickness controller MTM‐20.