Ldh kit

The interaction between GCSF/CT and red blood cell (RBC) or platelet adhesion was investigated according to a previous study [37 (link)]. Before the test, RBCs and platelet-rich plasma (PRP) were separated by centrifugation (1,500 rpm, 10 min). For RBC adhesion, 100 μl of RBCs was added to the surface of various aerogels (10 × 10 × 10 mm) and incubated at 37 °C for 1 h. Subsequently, aerogels were rinsed with PBS solution and placed into 4 ml of deionized water to lyse adhered RBCs. RBC adhesion in different groups was evaluated based on the OD₅₄₀ value of the supernatant.
For the platelet adhesion experiment, various aerogels (10 × 10 × 10 mm) containing 100 μl of PRP were incubated at 37 °C for 1 h. Next, 4 ml of Triton X-100 solution was used to lyse adhered platelets, releasing the lactate dehydrogenase (LDH) enzyme. Subsequently, an LDH kit (Biyuntian, China) was used to measure the LDH concentration. The percentage of platelet adhesion was calculated according to the manufacturer’s instructions.
After treatment with RBCs/PRPs, various aerogels were fixed with 2.5% of glutaraldehyde and dehydrated by gradient alcohol (50%, 60%, 70%, 80%, 90%, and 100%) to observe the interaction between various aerogels and RBCs/platelets. The cross-sectional and longitudinal sections of the various aerogels were observed using SEM.

Samples were sterilized by Co-60 irradiation (15 kGy, Changzhou Atomic Hi-Tech Radiation Co., Ltd., Changzhou, China). Platelet-rich plasma (PRP) was obtained by centrifugation of the CWB (400 g, 10 min). CS, YST, Celox™, and CLP were each weighed by 0.1 g for the test. A suspension of 100 μL of PRP was added to the sample. Samples were incubated for 1 h at 37 °C. Non-adherent platelets were removed by washing with PBS (pH = 7.4). Then were soaked in 1% Triton X-100 solution to lyse platelets to release lactate dehydrogenase (LDH). LDH was determined using the LDH kit (Shanghai Biyuntian Biotechnology Co., Ltd., Shanghai, China), according to instructions. Finally, the OD490 nm value measured the supernatant absorbance value. The OD490 nm value of the solution consisting of 100 μL of PRP not exposed to the hemostatic agent was measured and used as a reference value. The percentage of adherent platelets was calculated by the following equation [23 (link)]: