Bh2 uma

The microstructure of oil droplets in the emulsions were determined by an optical microscope (BH2-UMA, Olympus Corporation, Tokyo, Japan) with 50-fold eyepiece, and the images were taken at room temperature.

The accumulated calcium deposition was analyzed using Alizarin Red S staining, following a method developed for a previous study [22 (link)]. In brief, the specimens were fixed with 4% paraformadedyde (Sigma-Aldrich) for 15 min and then incubated in 0.5% Alizarin Red S (Sigma-Aldrich) at pH of 4.0 for 15 min at room temperature. Then, the photographs were observed using an optical microscope (BH2-UMA; Olympus, Tokyo, Japan) equipped with a digital camera (Nikon, Tokyo, Japan) at 200 magnifications. After this, the scaffolds were washed with PBS and quantified using a solution of 20% methanol and 10% acetic acid in water. After 15 min, the liquid was transferred to a 96-well plate, and the quantity of Alizarin Red was determined using a spectrophotometer at 450 nm.

Isothermal crystallization process of FAPbI₃ solutions was recorded by using optical microscope (BH2‐UMA, OLYMPUS) with an objective lens, IC20 (objective magnification, 20×). The solutions were dropped onto the substrates and then were spin coated at 6000 rpm for 15 s before transferring to a hot plate of 100 °C. Considering the crystal growth to be a 2‐demensional growth, fractional crystallinity curves were obtained by carefully observing the growth of the crystal over a specific area of (2000 um × 1500 um). Crystallization rates were calculated by the Avrami plot of fractional crystallization. With the Avrami model, fraction of transformed material (Y) after a certain time (t) at a constant temperature, Avrami equation can be calculated by the following form.
$$Y=1-\exp \left(-Z_{\mathrm{t}}{t}^n\right)$$ The equation can be transformed by taking logarithm on both sides.
$$\ln \left[-\ln \left(1-X\left(t\right)\right)\right]=n\ln t+\ln Z_{\mathrm{t}}$$
Z_t and n represent the kinetic rate constant and Avrami exponent, respectively. With ln [ − ln (1 − X(t))] plotted against ln t, the intercept of the graph is Z_t and the slope is n.

The joint samples, heart, liver, spleen, lung, kidney and skin tissue were collected from the rats and routine HE staining was performed. Knee joint samples were fixed in 4% (v/v) neutral buffered formalin for 24 h and decalcified for 1 month at room temperature in neutral 10% EDTA solution. Subsequently, the sample was dehydrated in an ethanol gradient, clarified, and embedded in a paraffin block. Tissue sections (8 μm) were prepared. Six representative sections of the joints from various depths were mounted on glass slides, stained with Safranin-O, and photographed under a microscope (BH2 UMA, Olympus). After overnight incubation with MMP13 (ABCAM, ab39012), COL2A1 (ABCAM, ab34712), ADAMTS-5 (ABCAM, ab41037), and AGGRECAN (ABCAM, ab3778) primary antibodies at 4°C, sections were incubated with secondary antibodies for 2 h at room temperature. Color development was performed using a DAB substrate system. Hematoxylin was used to stain the nucleus of cells.

The morphological structure of the pristine and plasma-treated PVA were examined using field-emission scanning electron microscopy (S-4800, Hitachi, Tokyo, Japan) and OM (BH2-UMA, Olympus, Tokyo, Japan). The molecular structures of the PVA matrices were characterized using an FT-IR instrument (Nicolet iS10, Thermo Fisher Scientific, Waltham, MA, USA). The surface oxidation states of the PVA matrices were investigated using XPS (K-alpha, Thermo Fisher Scientific, Waltham, MA, USA). The hydrophilicity of the plasma-treated PVA matrices was investigated by measuring the angle of the water droplet using a CA analyzer (Phoenix 300, SEO, Suwon, Republic of Korea. The swelling ratios were determined by immersing the pristine and plasma-treated PVAs in aqueous solution, EtOH, and commercial EC/EMC 3:7 (1.0 M LiPF₆) electrolyte.