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2 protocols using mtt 3 4 5 dimethylthiazol 2 yl diphenyltetrazolium bromide

1

Chitosan-Hydroxyapatite Composite for BMP-2 Delivery

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Chitosan (deacetylation degree: ≥92.6%) and acetic acid were obtained from Heppe MEDICAL CHITOSAN GmbH (USA) and Riedel de Haen (Germany), respectively. Ethanol was purchased from Sigma (Germany) and hydroxyapatite (HAp) in bead form (55–110 μm) was obtained from Science Application Industries (France). Phosphate buffer saline (PBS) tablets and sodium azide were bought from Sigma-Aldrich (Germany). Recombinant BMP-2 and BMP-2 ELISA kit were obtained from Thermo Fisher Scientific (USA) and MEL was obtained from Sigma-Aldrich (Germany). For cell culture studies, RAW 264.7 cell line (ATCC® TIB-71™) used in was purchased from American Type Culture Collection (ATCC). RANKL, MCSF, MTT (3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide) and isopropanol were all bought from Sigma-Aldrich (Germany). Dulbecco’s Modified Eagle Medium (DMEM-high glucose), fetal bovine serum (FBS), penicillin-streptomycin, l-glutamine and Dulbecco’s phosphate buffer saline were obtained from Capricorn Scientific (Germany). Glutaraldehyde, hexamethyldisilazane (HMDS), p-nitrophenyl phosphate (pNPP) tablets, iron chloride hexahydrate, naphthol AS-MX phosphate and fast red violet LB salt were all purchased from Sigma (Germany). Disodium tartrate dehydrate and N, N-dimethylformamide were bought from Merck (Germany). Sodium acetate anhydrous was obtained from Riedel de Haen (Germany).
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2

Cytotoxicity Assay of Pharmaceutical Compounds

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Lansoprazole, omeprazole, pantoprazole, and raloxifene were purchased from Sigma-Aldrich (USA) as powder and dissolved in DMSO. They were further diluted with Dulbecco's Modified Eagle Medium to reach the desired final concentrations immediately before use. Doxorubicin hydrochloride was purchased from Ebewe Untrerach (Austria) as a solution at a concentration of 2 mg mL -1 . MTT (3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide) was purchased from Sigma-Aldrich (USA).
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