Simulated body fluid

Cocoons of silkworm Bombyx mori (a Chinese strain demoted as 872) were provided by College of Biotechnology, Southwest University, China. Sodium carbonate and sodium biphosphate dihydrate were purchased from KeLong Chemical Reagent Co., Ltd. (Chengdu, China). Sodium dihydrogen phosphate was purchased from Fangzheng reagent Co., Ltd. (Tianjin, China). Calcium chloride was purchased from Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Curcumin (Cur) was purchased from BBI Life Sciences (Shanghai, China). Ethanol was purchased from Chuandong Chemical Co., Ltd. (Chongqing, China). Potassium bromide was purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Simulated body fluid (SBF) was purchased from Beijing Leagene Biotechnology Co. Ltd. (Beijing, China). Dulbecco’s modified Eagle’s medium (DMEM/HG), PBS (1×), and Penicillin Streptomycin 100× solution were purchased from Hyclone (Logan, UT, USA). Fetal bovine serum (FBS) was purchased from Tianhang Biotechnology Co., (Zhejiang, China). All chemicals are analytical grade and used without further purification. Ultrapure water (resistance > 18 MΩ·cm⁻¹) was used in all experiments.

Polyvinyl alcohol, (NH₄)₂HPO₄ (99%), Ca(NO₃)₂•4H₂O (99%), Li3PO4 (99%), CoCl₂ (99%), NH3•H2O (25–28%), and Tris–HCl were purchased from Sinopharm Chemical Reagent Co., Ltd. Simulated body fluid (SBF) was purchased from Leagene (China). Minimum Eagle's medium (MEM), Dulbecco’s modified Eagle’s medium/Ham’s F-12 (DMEM/F-12), and fetal bovine serum (FBS) were purchased from HyClone (USA). Endothelial cell medium was obtained from ScienCell (USA). The NBT/BCIP staining kit and detection kit for ALP activity and calcium content were purchased from Beyotime Company (China). The CCK-8 assay was obtained from Dojindo (Japan). MTT, phalloidin, and 4',6-diamidino-2-phenylindole (DAPI) were purchased from AAT Bioquest (USA). Primary antibodies against ALP, BMP-2, RUNX2, OCN, HIF-1α, VEGF, GAPDH, and HRP-conjugated secondary antibodies were obtained from Affinity (China).