3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt

The PLGA copolymer was purchased from the Daigang Biological Co. Ltd. (Jinan, China). Potassium bromide (KBr), Gel, absolute ethanol (99.8% purity), dichloromethane (DCM, 99.8% purity), and glutaraldehyde (99.8% purity) were purchased from the Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). n-HA (70% of purity) was purchased from the Weijing nano Co., Ltd. (Huizhou, China). All other compounds were of analytical purity and were used without any further purification. Cell lysate, an alkaline phosphatase (ALP) assay kit, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the mouse osteocalcin (OC) enzyme-linked immunoassay (ELISA) kit, and the collagen type-I (COL-I) ELISA kit were purchased from Jiancheng Ltd. (Nanjing, China).

Human renal proximal tubular epithelial cells (HK-2 cells) were kindly provided by Shanghai Institute of Materia Medica (China) and cultured in DMEM/F12 culture medium containing 10% fetal bovine serum (FBS) and 1.0% penicillin–streptomycin solution (Gemini Bio Products, West Sacramento, CA, USA).
Aristolochic acid I (AA I) (Purity HPLC ≥ 98%), Dimethyl sulfoxide (DMSO), Hoechst 33,258 were purchased from Solarbio (Solarbio Science & Technology Co., Ltd., Beijing, China); genipin (purity > 98%) was purchased from Sigma. The 3-(4,5-dimethyl thiazol-2-yl-)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) assay kit, lipid peroxidation (MDA) kit, and glutathione peroxidase (GSH-Px) were sourced from Nanjing Jiancheng Bioengineering Institute, Nanjing, China. Caspase-3 assay kit was purchased from Beyotime Biotechnology, Beijing, China. UCP2 polyclonal antibody (Cat # PA5-80203, Lot # AA2D12N), ECL solution and prestained protein marker were obtained from Thermo Fisher Scientific, Shanghai, China. To upregulate the expression of UCP2 in HK-2 cells, UCP2 DNA transfection using lipofectamine 2000 (Thermo Fisher Scientific, Shanghai, China) was performed.