Dulbecco s modified eagle s medium nutrient mixture f 12 dmem f 12

Agarose (Aga, agarose with low electroendosmosis), calcein/PI live/dead cell assay kit, Hoechst 33342 staining solution, cell counting kit-8 (CCK-8), DiO cell plasma membrane staining kit, and hematoxylin/eosin staining kit were purchased from Beyotime Biotechnology Co. (Shanghai, China). Polyethylenimine (PEI, MW = 25,000) and fluorescein diacetate (FDA) were obtained from Sigma-Aldrich (St Louis, MO, United States). Doxorubicin hydrochloride (Dox) and advanced antifade mounting medium with DAPI were procured from Meilun Biotechnology Co., Ltd. (Dalian, China). Fetal bovine serum (FBS), 0.05% trypsin-ethylenediaminetetraacetic acid solution (Trypsin–EDTA), penicillin–streptomycin solution, Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F-12; 1:1 ratio), phosphate-buffered saline (PBS), and other cell culture-related reagents were purchased from Invitrogen (Waltham, MA, United States). Human lens epithelial cell line HLE B3 (CRL-11421™, LEC) was originated from American Type Culture Collection. Foldable hydrophobic IOL were obtained from 66 Vision Tech Co., Ltd. (Suzhou, China). The clinical postoperative administration drugs were supplied by the Eye Hospital of Wenzhou Medical University (Wenzhou, China). All other chemicals were of analytical grade and used without further purification.

The MSC extraction and use were approved by the Ethics Research Committee of Instituto Israelita de Ensino e Pesquisa Albert Einstein (Sao Paulo, Brazil) under CAAE number: 64288917.0.0000.0071. The MSC isolation, culture and immunophenotypic characterization were performed in previous study [17 (link)]. In brief, mononuclear cells were obtained from bone marrow of healthy donors and separated by density gradient using Ficoll-Paque™ Premium (1.084 g/mL) (GE Healthcare, Uppsala, Sweden). After isolation, mononuclear cells were cultivated in Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 (DMEM/F-12) (GIBCO^® Invitrogen Corporation, Carlsbad, CA, USA) supplemented with fetal bovine serum (FBS) (GIBCO^® Invitrogen Corporation, CA, USA), and 1% antibiotic-antimycotic solution (GIBCO^® Invitrogen Corporation, CA, USA), incubated at 37 °C with 5% de CO₂. When required, the cells were trypsinized using TrypLE™ Express Enzyme (GIBCO^® Invitrogen Corporation, CA, USA). The immunophenotypic characterization was performed after third cell passage using flow cytometry (FACSAria™ III, BD Biosciences, San Jose, CA, USA), and data were analyzed through FlowJo software (Tree Star, Ashland, OR, USA).

Epigallocatechin gallate with a purity of 99.87% was purchased from MedChemExpress Co., Ltd. (#: HY-13653, Shanghai, China). Primary antibodies against phosphorylated (P)-AKT, total (T)-AKT, P-mTOR, T-mTOR, and GAPDH were provided by Abcam (Cambridge, UK), while secondary antibody was obtained from LI-COR Biosciences (Lincoln, USA). Trypsin-EDTA (0.25%) phenol red and Dulbecco's modified Eagle's medium nutrient mixture F-12 (DMEM/F-12), fetal bovine serum (FBS), and trypsin-ethylenediaminetetraacetic acid (0.25%) phenol were provided by Invitrogen-Gibco (Grand Island, NY, USA).

MCC950 sodium and dihydroethidium (DHE) were purchased from MedChemExpress (MCE R; China). (3,4-dihydroxyphenethyl) methacrylamide (DPMA) was purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). Poly (ethylene glycol) methacrylate (PEGMA) was purchased from Aladdin Reagent Co., Ltd. and passed through an Al₂O₃ column to remove the inhibitor before use. 3-acrylamidopropyl trimethylammonium chloride (APTAC, 75wt% in H₂O) and lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP, 98%) were obtained from Aladdin Reagent Co., Ltd and used without further purification. Fluorescein sodium (FS) was purchased from Abmole Co., Ltd. (United States). Scopolamine (Scop) was purchased from Sigma–Aldrich Co., Ltd. Human corneal epithelial cells (HCECs) were obtained from ATCC (Manassas, VA, United States). Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM/F-12; 1: 1 ratio), FBS, and insulin were purchased from Invitrogen, Gibco Co., Ltd. Triton X-100 was purchased from Sigma Co., Ltd. (Darmstadt, Germany). CM-H2DCFDA (General Oxidative Stress Indicator) was obtained from Invitrogen Molecular Probe Co., Ltd. Cell Counting Kit-8 (CCK-8) was purchased from Beyotime Biotechnology Co. (Shanghai, China). The In situ Cell Death Detection kit (TUNEL) was purchased from Roche Co., Ltd. (Germany).

Oxypaeoniflorin (Oxy) (purity > 98.06%) was purchased from MedChemExpress (#HY-N0748, Shanghai, China). Fetal bovine serum (FBS), trypsin-EDTA (0.25%) phenol red, and Dulbecco's modified Eagle's medium, Nutrient Mixture F-12 (DMEM/F-12), were purchased from Invitrogen-Gibco (Grand Island, NY). Anti-GAPDH (ab8245), anti-Sirt1 (ab189494), anti-Bcl-2 (ab32124), anti-Bax (ab32503), and anti-4-HNE (ab48506) antibodies were obtained from Abcam (Cambridge, MA, USA). The secondary antibody was obtained from LI-COR Biosciences (Lincoln, United States). The QuantiPro ™ Bicinchoninic Acid Protein Assay Kit from Sigma-Aldrich LLC (St. Louis, MO, USA) was employed to analyze protein concentrations. Lipopolysaccharide (LPS) was obtained from Sigma-Aldrich (#L6386-25MG, Beijing Merck Limited, China). All chemical reagents used were of analytical grade in the present study.

Farrerol (Farr), purity>98%, was obtained from Chengdu Pufei De Biotech Co., Ltd. (Chengdu, China). H₂O₂, 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), and Cell Counting Kit-8 (CCK-8) were purchased from Sigma Chemical Company (St. Louis, MO, USA). Antibodies against P-Akt, Akt, P-ERK, ERK, P-P38, P38, JNK, P-JNK, and β-actin were purchased from Cell Signaling Technology (Boston, MA, USA). In addition, antibodies against Nrf2, HO-1, NQO1, GCLC, GCLM, PARP, Bax, Bcl-2, caspase-3, caspase-8, and caspase-9 were obtained from Proteintech Group and Abcam (Cambridge, MA, USA). Dulbecco's modified Eagle's medium : Nutrient Mixture F-12 (DMEM/F-12), the antibiotic-antimycotic solution, and trypsin-EDTA were obtained from Invitrogen-Gibco, MBI, and Biofil, respectively. In addition, MDA, GSH, and SOD test kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Unless otherwise stated, all chemicals were purchased from Sigma Chemical Co. (St. Louis. MO, USA). MES23.5 cells were provided by Professor Wei-Dong Le (Baylor College of Medicine, TX, USA). Nesfatin-1 was obtained from Phoenix Pharmaceuticals Inc. (Burlingame, CA, USA). Dulbecco’s modified Eagle’s medium nutrient mixture-F12 (DMEM/F12) was purchased from Gibco (Grand Island, NY, USA). The PE-conjugated monoclonal active-caspase-3 antibody apoptosis kit was from BD Bioscience (San Diego, CA, USA). PD98059 and the BCA kit was from Beyotime (Shanghai, CN). KT5720 and anti-rabbit secondary antibody conjugated to horseradish peroxidase were from Santa Cruz (Santa Cruz, CA). The cell fractionation kit was from Clontech (Mountain View, CA, USA). Anti-Cox IV monoclonal antibody was from Clontech (CA, USA). The primary antibody against TH was from Millipore (Darmstadt, Germany). Primary antibodies against ERK1/2 and phospho-ERK1/2 were from cell signaling technology (MA, USA). Alexa Fluor 555 donkey anti-rabbit secondary antibody was from Eugene (OR, USA). GW5074 was from Selleck (TX, USA).