Chemicals including calcium nitrate tetrahydrate (Ca(NO3)2·4H2O, 98%), calcium chloride (CaCl2, 95.0%), diammonium hydrogen phosphate ((NH4)2HPO4, 98%), sodium hydroxide (NaOH, 97%), phosphoric acid (H3PO4, 85% aqueous solution), ammonia solution (25% aqueous solution), and acetic acid (99.7%), were purchased from FUJIFILM Wako Pure Chemical Corp., Osaka, Japan. Pyromellitic acid (98.0%) was purchased from Tokyo Chemical Industry Co., Ltd., Tokyo, Japan. Furthermore, calcium carbonate (CaCO3 (calcite), 99.5%) and hydrochloric acid solution (HCl, 1.0 mol dm−3) were purchased from Nacalai Tesque Inc., Kyoto, Japan. All chemicals were used without further purification.
Calcium chloride
Manufactured by Fujifilm
Sourced in Japan
Calcium chloride is a common laboratory chemical that serves as a desiccant, or drying agent. It is used to remove moisture from air or other gases, and can be found in various laboratory applications where controlled drying is required.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
U 3310 spectrophotometer, by Hitachi (2 mentions)
Sodium hydrogen carbonate, by Fujifilm (2 mentions)
Protease from aspergillus oryzae, by Tokyo Chemical Industry (2 mentions)
Sodium chloride (nacl), by Fujifilm (2 mentions)
D glucose, by Fujifilm (2 mentions)
Magnesium chloride, by Fujifilm (2 mentions)
Ammonia solution, by Fujifilm (1 mentions)
Sodium hydroxide (naoh), by Fujifilm (1 mentions)
Calcium nitrate tetrahydrate, by Fujifilm (1 mentions)
Phosphoric acid, by Fujifilm (1 mentions)
Acetic acid, by Fujifilm (1 mentions)
Diammonium hydrogen phosphate, by Fujifilm (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (1 mentions)
β glycerophosphate disodium salt hydrate, by Merck Group (1 mentions)
Rhvegf165, by R&D Systems (1 mentions)
15 protocols using calcium chloride
1
Synthesis and Characterization of Calcium Phosphate Compounds
2
Differentiation Potential of Dental and Bone Stem Cells
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Human BMSCs derived from iliac crests (Lonza, Basel, Switzerland) and DPSCs isolated from adult third molars (Lonza) were cultured in growth medium (GM) comprising Dulbecco's modified Eagle's medium (DMEM; Wako, Osaka, Japan) supplemented with 20% fetal bovine serum (Thermo Fisher Scientific, MA, USA) and 1% penicillin-streptomycin (Sigma-Aldrich, MO, USA). Green fluorescent protein- (GFP-) labelled DPSCs were used to track their localization inside cell constructs. DPSCs at passages 3–5 were transduced with GFP using a lentivirus vector (Sigma-Aldrich) and selected by 1 μg/mL puromycin (Sigma-Aldrich) for at least 1 week.
To induce the endothelial differentiation, each cell type and cell constructs were treated with an endothelial differentiation medium (EM; EGM2-MV (Lonza) supplemented with 50 ng/mL rhVEGF165 (R&D Systems, MN, USA)). For osteogenic differentiation, cell constructs were maintained in osteogenic differentiation medium (OM) comprising GM supplemented with dexamethasone (1 × 10−6 mol/L; Sigma-Aldrich), β-glycerophosphate disodium salt hydrate (1 × 10−2 mol/L; Sigma-Aldrich), ascorbic acid (50 μg/mL; Sigma-Aldrich), and calcium chloride (1 × 10−2 mol/L; Wako). Cells and cell constructs were incubated in a humidified incubator at 37°C with 5% CO2.
To induce the endothelial differentiation, each cell type and cell constructs were treated with an endothelial differentiation medium (EM; EGM2-MV (Lonza) supplemented with 50 ng/mL rhVEGF165 (R&D Systems, MN, USA)). For osteogenic differentiation, cell constructs were maintained in osteogenic differentiation medium (OM) comprising GM supplemented with dexamethasone (1 × 10−6 mol/L; Sigma-Aldrich), β-glycerophosphate disodium salt hydrate (1 × 10−2 mol/L; Sigma-Aldrich), ascorbic acid (50 μg/mL; Sigma-Aldrich), and calcium chloride (1 × 10−2 mol/L; Wako). Cells and cell constructs were incubated in a humidified incubator at 37°C with 5% CO2.
3
Fabrication of Alginate Hydrogel Beads
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Alginate hydrogel beads were obtained as reported in a previous study [8] . Tiny droplets of 1.5% w/v sodium alginate (Sigma–Aldrich, St. Louis, MO, USA) solution were discharged from an inkjet system (Cluster Technologies, Osaka, Japan) into a reservoir flask filled with 5% calcium chloride (Wako, Osaka Japan) solution. The droplets of the alginate solution change into a gel immediately after they have been dropped into a solution containing calcium ions. When a 25-μm diameter nozzle was used, the size of the droplet was approximately 20 μm. Alginate hydrogel beads were washed with phosphate-buffered saline and suspended in fresh culture medium.
4
Comprehensive Proteomics Sample Preparation
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Ammonium bicarbonate, tris(hydroxymethyl)aminomethane hydrochloride, SDC, sodium N-lauroylsarcosinate, Ammonium bicarbonate, tris(2-carboxyethyl)phosphine, 2-chloroacetamide, calcium chloride, ethyl acetate, acetonitrile (ACN), acetic acid, TFA, and other chemicals were purchased from Fujifilm Wako. RapiGest was purchased from Waters Corporation. Modified trypsin was from Promega. TrypN was from Protifi. Styrene divinylbenzene (SDB-XC) Empore disk was purchased from GL Sciences. Water was purified by a Millipore Milli-Q system.
5
Peptide Nanogel Biodegradability Evaluation
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The peptide nanogel biodegradability in enzyme-containing buffer solution was investigated. The peptide nanogel solution (10 mL) was added to 10 mL 0.02 mol L−1 sodium hydrogen carbonate (guaranteed reagent grade, Fujifilm Wako Pure Chemical), 1 mmol L−1 calcium chloride (90% purity, Fuji-film Wako Pure Chemical), and 0.01 wt.% protease (from Aspergillus oryzae, Tokyo Chemical Industry, Tokyo, Japan). The mixed solutions were incubated at 37 °C for 6 days. Then, the absorbance of the treated solutions was measured using a spectrophotometer (U-3310 spectrophotometer, Hitachi, Tokyo, Japan). The degradation rate was determined from the absorbance at 273 nm using Equation (1):
where Abs1 and Abs0 are the absorbance values of the filtrate and residue, respectively.
where Abs1 and Abs0 are the absorbance values of the filtrate and residue, respectively.
6
Gelatin-based Biomaterial Synthesis
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BeMatrix™, an alkaline-treated gelatin (AlGltn) derived from porcine skin, was kindly donated by Nitta Gelatin Inc. (Osaka, Japan). Ethanol (EtOH), 1,1,1,3,3,3-hexafluoroisopropanol (HFIP), dimethylsulfoxide (DMSO), triethylamine (TEA), 2,4,6-trinitrobenzoylsulfonic acid (TNBS), hydrochloric acid (HCl), sodium dodecyl sulfide (SDS), calcium chloride, 10% formalin neutral buffer solution, 4′,6-diamidino-2-phenylindole (DAPI), tris(hydroxymethyl)aminomethane, tert-butylalcohol, and glycine were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Hexanoyl (Hx: C6) chloride, decanoyl (Dec: C10) chloride, and stearyl (Ste: C18) chloride were purchased from Sigma Chemical Co. (St. Louis, MO, USA). A porcine aorta was purchased from Funakoshi Corporation (Tokyo, Japan). L929 cells were purchased from (RIKEN Bio Resource Center Cell Bank, RBRC-RCB2619, Ibaraki, Japan). All chemicals were used without further purification.
7
Enzymatic Sensing Platform Chemicals
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glycerol kinase (GK) (Grade III, 20 U/mg-solid) and L-α-glycerophosphate oxidase (LGOx) (Grade III, 15 U/mg-solid) were purchased from Toyobo CO. Ltd. (Osaka, Japan). Horseradish peroxidase (HRP) (>250 units/mg-solid), ferrocenyl methanol (FcMeOH) (95%), poly-L-lysine (PLL) (Mw: 70,000~150,000), poly (sodium 4-styrene sulfonate) (PSS) (Mw: ~70,000), acetylcholine chloride (Ach) (~99%), gamma-aminobutyric acid (GABA) (>99%), and D-serine (>98%) were purchased from Sigma-Aldrich Inc. Sodium (St. Louis, MO, USA) 4-(2-hydroxyethyl) piperazine-1-ethanesulfonate (HEPES) (>99%) was purchased from Dojindo Laboratories. Adenosine-5′-triphosphate (98%), adenosine diphosphate (ADP) (97%), adenosine monophosphate (AMP) (99.3%), adenosine (Ado) (98%), L-glutamic acid sodium (99%), sodium chloride (99.5%), potassium chloride (99.5%), calcium chloride (95%), magnesium chloride, D-glucose (98%), potassium hexacyanoferrate (III) [K4Fe(CN)6] (99%), potassium hexacyanoferrate (II) [K3Fe(CN)6] (99.5%), glycerol (>99%), ethanol (95%), and sodium hydroxide solution were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). The FcMeOH solution was prepared using 95% ethanol, and all other reagents were prepared using ultrapure water (18.2 MΩ cm at 25 °C) produced with a Milli-Q water system (Tokyo, Japan).
8
Enzymatic Biodegradation of Peptide Nanoparticles
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The biodegradability of the peptide nanoparticles in a buffer solution containing enzymes was next elucidated. The peptide nanoparticle solution (0.01 dm3) was added to 0.01 dm3 of an aqueous solution of 0.02 mol dm−3 sodium hydrogen carbonate (guaranteed reagent grade, Fujifilm Wako Pure Chemical), 0.001 mol dm−3 calcium chloride (90%, Fujifilm Wako Pure Chemical), and 0.01 wt.% protease (from Aspergillus oryzae, Tokyo Chemical Industry, Tokyo, Japan). The mixed solutions were incubated at 37 °C for six days, after which the absorbances of the treated solutions were measured with a spectrophotometer (U-3310 spectrophotometer, Hitachi). The degradation rate was determined from the absorbances at 273 nm using Equation (1).
where Abs1 and Abs0 are the absorbances of the filtrate and filtrate, respectively.
where Abs1 and Abs0 are the absorbances of the filtrate and filtrate, respectively.
9
Fabrication and Characterization of Alginate-Based Bone Tissue Scaffolds
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Human MG-63 cell line was provided by the Iranian National Cell Bank (Pasture Institute; Iran). Pen/Strep, FBS, PBS, DMEM/LG, and 0.25% Trypsin–EDTA solution were obtained from Gibco (UK). Sodium alginate (I-1G, high content of guluronic acid, and MW: 70 kDa) was purchased from Kimica (Tokyo, Japan). BCIP/NBT alkaline phosphates color development kit, lysozyme, nano-hydroxyapatite, p-NPP, barium chloride, strontium chloride, DMSO, and MTT powder were purchased from Sigma-Aldrich. Alizarin Red S, calcium chloride, magnesium chloride, Tris-Based, sodium chloride, paraformaldehyde, and trisodium citrate dehydrate were obtained from Wako Pure Chemical Corporation (Osaka, Japan). Collagen type I was obtained from SBPE Company (Tabriz, Iran). SOD, TAC, and Superoxide dismutase assay kits were purchased from Randox (Crumlin, United Kingdom). Sodium sulfate was obtained from Merck (Germany). RIPA lysis buffer kit, HRP-conjugated anti-IgG, B-actin, COL1A1, COL2A1, OCN, and SOX-9 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).
10
Synthesis of PEG-based Polymer Compounds
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2-1. Material 2,2′-Azobisisobutyronitrile (AIBN) (98%), calcium chloride (95%), and 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) (99%) were purchased from FUJIFILM Wako Pure Chemical Corporation. Osmium tetroxide (4% in water) and polyethylene glycol monomethyl ether (Mw: 400, 2000, and 4000) were purchased from Tokyo Chemical Industry. Sodium chloride (99%), magnesium chloride hexahydrate (99%), potassium chloride (99.5%), and dry N,N-dimethylformamide (DMF) (99.5%) were purchased from Kanto Chemical. Dimethyl sulfoxide (DMSO, 98%) was purchased from Kishida.
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