Bovine serum albumin 0.1

Whatman chromatography paper No.1 was purchased from Zhengcheng Scientific Experimental Equipment Mall. NiFe₂O₄ nanoparticles (Chemical Abstracts Service (CAS: 12168-54-6), human serum albumin (HSA, CAS: 70024-90-7) and anti-HSA antibody were purchased from Shanghai Yugong Biotechnology Co., Ltd. Cysteamine (CYS, CAS: 60-23-1), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, CAS: 25952-53-8), N-hydroxysulfosuccinimide (NHS, CAS: 6066-82-6), phosphate buffered saline (PBS, 0.01 M, pH = 7.4) and bovine serum albumin (BSA, 0.1 %, CAS: 9048-46-8) were purchased from Sigma-Aldrich Corporation (Saint Louis, MO, USA). Carcinoembryonic antigen (CEA) was purchased from Beijing Key-Bio Biotech Co., Ltd. Hemoglobin (HGB), uric acid (UA, CAS: 69-93-2), and creatinine (CRE, CAS: 60-27-5) were purchased from Shanghai Fanke Biological Technology Co., Ltd. The water used in the experiments was obtained from an ultra-pure water manufacturing system (URT-11-10T).

Normal human keratinocytes were grown until 60% confluence in KGM medium. 24 h treatment with human recombinant neurotrophin nerve growth factor (NGF), brain-derived neurotrophic Factor (BDNF), and beurotrophin 3 (NT3) (all purchased from Sigma-Aldrich, St. Louis, MO, USA, and used at 100 ng/mL) was performed in KBM (keratinocyte basal Mmdium) plus bovine serum albumin (BSA) 0.1% (Sigma-Aldrich, St. Louis, MO, USA) and penicillin/streptomycin/amphotericin B (Lonza, Walkersville, MD, USA).

5 × 10³ keratinocytes/well, of either stem cell-like or proliferative progeny cells, were seeded in 96-well culture plates in KGM. At 48 h from plating, cells were treated with human recombinant neurotrophin nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin 3 (NT3) (all purchased from Sigma-Aldrich, St. Louis, MO, USA, and used at 100 ng/mL) in KBM (keratinocyte basal medium) plus bovine serum albumin (BSA) 0.1% (Sigma-Aldrich, St. Louis, MO, USA) and Penicillin/Streptomycin/Amphotericin B (Lonza, Walkersville, MD, USA). After 48 h, cells were incubated with 0.5% MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) for 4h at 37°C and then dissolved with 100 μL isopropanol. The plate was read at 560 nm. Results are expressed as viability percentage, as compared to control.