Cytotoxicity measurements of tissue engineered scaffold

Abstract

Cytotoxicity measurements on materials are extremely important in tissue engineering field. The objective of this study is to measure and compare the cytotoxicity of scaffold for tissue engineered using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT), 2-(methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2-4disulfophenyl)-H-tetrazolium(WST-8) and Alamar Blue cytotoxicity assay on cytotoxixity of scaffold for tissue engineering. In order to evaluate the sensitivity of rat bone marrow stromal cell(rBMSC) for cytotoxicity assays, a series of two-fold dilution of rBMSC were plated in 96-wells. rBMSC suspension was seeded on poly(lactide-co-glycolide) (PLGA) scaffold and then cytotoxicity was measured in vitro by MTT, WST-8 and alamar blue assay. The MTT assay was more sensitive than WST-8, alamarblue assay for rBMSC. rBMSC suspension seeded on various content of demineralized bon particle(DBP) impregnated PLGA scaffold was measured to confirm on affect to natural material by MTT assay. In the MTT assay results, optical density was influenced by content of DBP Finally, cytotoxicity of DBP/PLGA scaffold was assayed using modified method of ASTM F813-07. We confirmed that cell viability was significantly high in PLGA scaffold including DBP of 20 and 40% compared to other scaffolds. This report may be useful as guideline in tissue engineered scaffolds.