The morphologies of the as-prepared IONP-CM were observed under an AFM (Signal Hill, CA, United States of America) and a field-emission SEM (Zeiss Supra 40 Gemini, Germany). The mechanical characteristics of the prepared IONP-CM were assessed using a universal material testing machine (Shimadzu Co., Japan). The rheological features of the IONP-CM were determined using a modular advanced rheometer system (HAAKE MARS, Thermo Fisher Scientific, United States of America). This involved conducting strain–sweep measurements over a range of 0.01%–10% strain amplitude at a frequency of 6.28 rad/s, as well as frequency–sweep measurements at frequencies ranging from 1 to 30 rad/s with a fixed 1% strain amplitude. For the IONP release test, the IONP-loaded CM scaffolds were precisely shaped into uniform squares (10 mm × 10 mm). After that, they were co-incubated with 1.5 mL of PBS at 37°C. The concentration of Fe ions released from the scaffolds was determined at predetermined time intervals spanning a 7-day period using inductively coupled plasma atomic emission spectrometry (ICP-OES, Agilent 5800, United States).
Universal material testing machine
Manufactured by Shimadzu
Sourced in Japan
The Universal Material Testing Machine is a versatile laboratory equipment designed to perform various mechanical tests on a wide range of materials. Its core function is to apply controlled forces, stresses, or deformations to a sample and measure the resulting response, such as strength, stiffness, or elongation. The machine is capable of conducting tensile, compressive, flexural, and other types of tests to evaluate the mechanical properties of materials.
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3 protocols using universal material testing machine
1
Characterization and Release Kinetics of Iron Oxide Nanoparticle-Loaded Composite Matrices
2
Characterization of Nano-reinforced Scaffolds
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To investigate the structure of these nano-reinforced scaffolds, various characterization methods were utilized. A scanning electron microscope (SEM) was used to observe the surface features. The sample was observed at 100 kV using a JEM-2100 microscope (JEOL Co., Ltd., Tokyo, Japan). Furthermore, five areas of the SEM images were selected to measure the pore size. The X-ray diffraction (XRD) spectra were obtained using a Bruker D8 Discover powder diffractometer (Brook Scientific, Beijing, China) at a diffraction angle of 20°≤ 2θ ≤ 80°. Fourier transform infrared (FTIR) spectra were measured under a Nicolet is50 (Thermo Fisher Scientific, Shanghai, China) to detect the peak shifts of the nanoparticles. A universal material testing machine (Shimadzu, China) was used to test the compressive strength of the scaffold in a wet state, and PBS was used as the soaking liquid. Prepare 10 mm high scaffold samples, load 100 N, load speed of 1 mm/min, load distance of 7 mm and measure five samples in each group. Draw a stress-strain curve, then draw a straight line parallel to the initial linear part from the 1% strain, and the ordinate of the intersection with the stress-strain curve is defined as the compressive strength.
3
Biomechanical Evaluation of Regenerated Tissue
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For biomechanical and biochemical analyses, samples were first trimmed along the rim of the regenerated tissue using a trephine. Then, as per the method described by Hoenig et al. [9 (link)], samples were transferred to a stainless steel dish containing PBS at room temperature for the unconfined compression test (UCC), which was conducted using a universal material testing machine (Shimadzu, Kyoto, Japan). The height of each specimen was determined at a compressive force threshold of 0.05N. Five progressive strain loadings at 4% of the original cartilage height were performed with a test velocity of 0.01 mm/s. After each loading cycle, samples were subjected to a 2000-s relaxation phase to attain equilibrium. The Young's modulus value was then determined using the load and displacement data obtained at the end of each relaxation phase [14 (link)].
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