Ab217608

ELISA kits were used to measure the concentrations of IL-1β (cat. no. ab217608; Abcam), IL-6 (cat. no. ab46027; Abcam) and TNF-α (cat. no. ab181421; Abcam) as per the manufacturer's instructions. Briefly, 50 µl samples or standards were added to appropriate wells. This was followed by the addition of antibody cocktails from the ELISA kits into each well and incubation at room temperature for 1 h. Subsequently, 3,3′,5,5′-tetramethylbenzidine substrate from the kits was added to each well and incubated for 10 min in the dark with shaking at 400 rpm. Following incubation with stop solution, the optical density values of each well were measured at a wavelength of 450 nm using a microplate reader.

Arterial blood samples were drawn from each patient at three time points: T1 (before the surgery), T2 (the end of the surgery), and T3 (24 hours after the surgery). The blood samples were stored in tubes containing ethylene diamine tetraacetate acid and were further processed within 12 hours. Each sample was centrifuged for 15 min at 3000rpm to obtain the plasma. Then, the levels of BMP4 (ab231930, Abcam, Cambridge, UK), Noggin (JYM2457Hu, Wuhan, CHN), IL-1β (ab217608, Abcam, Cambridge, UK), TNF-α (ab181421, Abcam, Cambridge, UK), and IL-10 (ab185986, Abcam, Cambridge, UK) were determined using the enzyme-linked immunosorbent assays method (SimpleStep ELISA^® kits, Abcam, Cambridge, UK) by a researcher who was blind to the group allocation.