Multifunctional enzyme marker

UHPLC system (Vanquish, Thermo Fisher Scientific, United States); Chromatographic column (ACQUITY UPLC BEH C18 (1.7 μm 2.1*100 mm), Waters, United States); High resolution mass spectrum (Orbitrap Exploris 120, Thermo Fisher Scientific, United States); Analytical Balance (Mettler Toledo, Swiss); High-throughput tissue grinder (Shanghai, China); Pure water filter (Merck Millipore, United States); Ultrasonic cleaner (Fangao Co., Ltd., Shenzhen, China); CO₂ incubator (ThermoFisher, United States); Multifunctional enzyme marker (PerkinElmer, United States); Ultra low temperature refrigerator (Haier, China).

The CCK-8 kit was obtained from the Tong Ren Chemical Research Institute, Japan. The human prostate hyperplasia cell line (BPH-1) and RPMI-1640 medium were obtained from Wuhan Service Biotechnology Co. The capsular tab, urethral end piece, and sutures were sterilised and added to RPMI-1640 complete medium at a ratio of 0.2 g/ml and incubated at 37 °C for 48 h to prepare the implant extracts.
A suspension of 2 × 10³ BPH-1 cells, in a volume of 100 μL, was seeded into 96-well plates and incubated for 12 h in a 5% CO₂ environment to allow for adhesion. The medium was then replaced with the implant extract. The control group was maintained in RPMI-1640 complete medium, whereas the experimental group was exposed to the capsular tab, urethral end piece, and suture extracts, each with three replicates per group. According to previous studies^{12 (link),13 (link)}, following incubation for 1, 3, 5, and 7 days, 10 μL of CCK-8 reaction solution was added to each well. The samples were then incubated at 37 °C for 1 h and Absorbance, A (Absorbance, A) was measured at 450 nm using a multifunctional enzyme marker (Perkin Elmer, USA). The relative cell viability was determined as [(A sample group)/(A control group) × 100].