Crossbeam 340

To examine the micromorphology of the hydrogel array, the gels were dried in a vacuum-drying machine at −80°C for 24 hours. The samples were then treated with a metal spraying machine and observed under a scanning electron microscope (Zeiss Crossbeam 340, Carl Zeiss AG, Oberkochen) at an operating voltage of 2 kV. ImageJ analyzed the pore size.

Cells were imaged using a confocal laser scanning microscope (Leica, SP5X, 20 x dip-in water-immersion objective; numerical aperture (NA) 1.0) with laser power set to 1.2 mW to avoid photobleaching. F-Actin Alexa 488 was detected with an Argon laser at 488 nm; F-Actin Red 555 (Tetramethylrhodamine (TRITC)) with an Argon laser at 540 nm, and DRAQ5 with a He-Ne laser at 633 nm. Hoechst 33342 was detected with a UV laser at 350 nm. Images of cells with scaffolds represent an overlay of 35–40 z-stack sections. Confocal images of cells on aligned microfibers represent an overlay ranging from 18–21 z-stack sections (each step 0.99 μm equaling 17.82–20.79 μm with a line average of 3).
For scanning electron microscopy, a Zeiss Crossbeam 340 scanning electron microscope (Carl Zeiss Microscopy GmbH, Oberkochen, Germany) was used. Aligned microfibers were washed with PBS and incubated in 6% glutaraldehyde (Sigma-Aldrich, Schnelldorf, Germany) for 15 min. After a washing step in ice-cold PBS, the samples were dehydrated by increasing concentrations of ethanol from 50 to 100%. The samples were incubated in hexamethyldisilazane (Sigma-Aldrich, Schnelldorf, Germany) for 15 min and dried overnight. All samples were sputter-coated with a 4 nm layer of platinum with a Leica EM ACE600 (Leica Microsystems, Wetzlar, Germany) before imaging.

Following μCT imaging, the cortical bone samples were embedded in polymethyl methacrylate, grinded to a coplanar state, and polished to provide a smooth surface of the bone tissue. Quantitative back‐scattered electron imaging (qBEI) was performed using a scanning electron microscope (Zeiss crossbeam 340, Carl Zeiss AG, Germany) with a back‐scattered electron detector (Fig. 1). Prior to scanning, the embedded bone samples were sputtered with carbon. Gray scale images were obtained through a constant beam current determined using a Faraday cup and calibration standard with aluminum and carbon (MAC Consultants Ltd., England), in combination with a constant working distance of 20 mm and a voltage of 20 kV. In total, five images per sample around the crack were acquired and analyzed using a custom‐made MATLAB script routine (MATLAB, R2019b, MathWorks Inc., USA). Based on the BMD distribution the following parameters were determined: average calcium concentration (CaMean, wt%), most frequent calcium concentration (CaPeak, wt%), and mineralization heterogeneity, which is based on the standard deviation of the BMDdistribution curve (CaWidth, wt%).

After washing thoroughly with PBS five times, cells were fixed overnight at 4°C in 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.2). Samples were then washed five times in 0.1 M cacodylate buffer (pH 7.2), dehydrated through a graded series of 30%, 50%, 70%, 90% acetone solution in 10 min incubations, and incubated six times in 100% acetone for 10 min, followed by critical point drying with CO₂. Dried specimens were sputter coated with gold/palladium. Samples were analyzed with a JEOL JSM-7500F (JEOL GmbH) or a Zeiss Crossbeam 340 (Zeiss) field emission scanning electron microscope operating at 5Kv. Representative images were selected from 3 independent experiments.

One-piece whiteSKY zirconia implants (Bredent GmbH & Co. KG, Senden, Germany), 4 mm in diameter and 8 mm in length, were used in this study (Figure 1). The implant design is considered to be particularly stable because there is a screw connection to fix an abutment on the implant. The manufacturer specifies a flexural strength of 1250 MPa +/− 120 MPa, a modulus of elasticity of 200 GPa and a fracture toughness of 6–8 MPa/m for these implants. In total, 54 implants were used in this study, of which 18 implants used non-treated as a control group. The surface structure was investigated using a scanning electron microscope (Zeiss Crossbeam 340, Carl Zeiss AG, Oberkochen, Germany).

Cu₅₀Zr₅₀ MG samples with in-plane dimensions of 2 × 2 mm² and thickness of approximately 50–100 μm were prepared for laser shock experiments using the single-roller melt spinning method. X-ray diffraction (Empyrean XRD, Malvern Panalytical Ltd) was performed to verify the amorphous state of the samples (Supplementary Fig. 6a and 9a). After laser shock tests, the MG samples were retrieved, and microscopy (Hitachi FE-SEM S4800) was used to characterize the plane where spalling occurred. Then, the void distribution underneath the spall plane was characterized using a focused ion beam (FIB) to mill a rectangular well in the sample (ZEISS Crossbeam 340). HRTEM and selected-area electron diffraction (SAED) characterizations were performed using Tecnai G2 F20 S-TWIN (FEI, US) at an accelerating voltage of 200 kV.

The morphologies of the pristine TPU and TPU/NS membranes were observed using scanning electron microscopy (SEM, Crossbeam 340, Zeiss, Germany), and the compositional analysis was conducted by energy-dispersive X-ray spectrometry (EDS). Based on the SEM images, the average pore size, porosity and diameters of the nano-silver particles were measured using Image J software and was read by two independent investigators. X-ray diffraction (XRD) spectra of the TPU and TPU/NS membranes were obtained with a 2θ range between 10° and 80° using an X-ray diffractometer (X’Pert Power, Panalytical B.V., Netherlands). The attenuated total reflectance Fourier transform-infrared (ATR-FTIR) spectra of the samples were obtained using an ATR-FTIR spectrometer (Nicolet 460, USA). The water contact angles of the samples were measured using a contact angle analyser (Theta Lite 101, Biolin Scientific, Sweden).