Calcein am pi double staining kit

BMSCs were seeded on the scaffolds (5 × 10⁵ cells/scaffold) in 96-well plates. After 3 days of incubation at 37 °C and 5% CO₂, the scaffolds were stained with a Calcein-AM/PI double staining kit (Solarbio). According to the instructions, the scaffolds were rinsed with PBS, stained with dye solution for 20 min in the dark. At the wavelength of 490 ± 10 nm, Calcein-AM detected yellow-green living cells, and propidium iodide (PI) detected red dead cells. The number and proportion of living and dead cells on the scaffold were analyzed by a confocal laser scanning microscope (LSM 780, AxioObserver, Zeiss).

Phosphate-buffered saline (PBS), Trypsin/EDTA solution, Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and penicillin/streptomycin (P/S) double antibody were purchased from Gibco. Accutase® solution and NaAsO₂ were purchased from Sigma-Aldrich. Endothelial cell medium (ECM) was purchased from ScienCell Research Laboratories. Primary antibodies (anti-vimentin and anti-E-cadherin) were purchased from Cell Signaling Technologies. Additional primary antibodies (anti-CD34, anti-N-cadherin) were purchased from Abcam. A Cell Counting Kit-8 (CCK-8) was purchased from DOJINDO. Zn(NO₃)₂·6H₂O, 2-methylimidazole, polyethylene glycol (2000), indocyanine green (ICG) and fluorescein isothiocyanate (FITC) were purchased from Aladdin. Methanol was purchased from Guangzhou Chemical Reagent Factory. An Annexin V-FITC/PI apoptosis detection kit was purchased from BD Biosciences. A calcein-AM/PI double staining kit was purchased from Solarbio, Beijing.

CuCl₂·2H₂O, sodium citrate, Na₂S·9H₂O, tetraethyl orthosilicate (TEOS), triethanolamine (TEA), hydrochloric acid (HCl, 37%), ethanol, and ammonia solution (25–28%) were purchased from Sinopharm Chemical Reagent Co. (Shanghai, China). Disulfiram (DSF), cetyltrimethylammonium chloride (CTAC, 25 wt%), 3-mercaptopropyltrimethoxysilane (MPTMS), and 2, 2′-dipyridyl disulfide (DPDS) were obtained from Sigma-Aldrich. Bis(3-triethoxysilylproyl)disulfide (BTES) was bought from Lark Chemical Technology Co., Ltd. HS-PEG-SH was purchased from Shanghai Yarebio Technology Co., Ltd. Calcein-AM/PI double staining kit was purchased from Beijing Solarbio Science & technology Co., Ltd. Cell counting kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies. Annexin V-FITC/PI apoptosis detection kit was purchased from Vazyme (Nanjing, China). Cell culture medium 1640, trypsin–EDTA and fetal bovine serum (FBS) were provided by Gibco-BRL (Burlington, Canada). H&E and Ki-67 cell proliferation kit were purchased from Beyotime Biotechnology Co. (Haimen, China). The primary antibody rabbit anti-NPL4 antibody was bought from Abcam. The second antibody anti-rabbit IgG (horseradish peroxidase (HRP)-linked antibody) was bought from Cell Signaling Technology, Inc. All chemicals were used as received without further purification, and their aqueous solutions were prepared using deionized water.