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The composition of the coating was determined using energy dispersive X-ray analysis.
The phase structure of the MAO Al2O3 coating was determined using X-ray diffraction.
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The surface hardness of substrate and coating were determined using a Vickers hardness tester, and microhardness profile of cross-section was realized by a Vickers microhardness tester.
The rheology of the coating suspensions was determined using a Brookfield Rheometer.
Coat thickness was determined using a 0.05 mm precision pachimeter in three different regions of the animal body.
The coating composition and nanostructure was determined using time-of-flight elastic recoil detection analysis (TOF-ERDA), x-ray diffraction (XRD) and x-ray photoelectron spectroscopy (XPS).
The insulin content in the samples was determined using Coat-a-Count RIA (Millipore Corporation, Bedford, MA, USA).
A standard curve was determined using plates coated with an anti-human IgE (clone LE27) and standard human IgE (World Health Organization) at concentrations ranging from 10 to 0.08 IU/mL.
Testosterone (T) was determined using a COAT-A-COUNT solid-phase radioimmunoassay (RIA Siemens Medical Solutionsns, Malvern, Pennsylvania, USA) and oestradiol by RIA (Pantex, Santa Monica, California, USA).
Serum P4 concentration was determined using a commercial coat-a-count radioimmunoassay according to the manufacturer's instructions (Progesterone Coat-a-Count, TKPG1, Siemens Medical Diagnostics, CA, USA).
Cell invasive ability was determined using transwell chambers coated with a extracellular matrix gel (BD Biosciences).
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