Cell Biology and Toxicology

, Volume 24, Issue 4, pp 315–319 | Cite as

Correlation of visual in vitro cytotoxicity ratings of biomaterials with quantitative in vitro cell viability measurements

Article

Abstract

Medical devices and implanted biomaterials are often assessed for biological reactivity using visual scores of cell–material interactions. In such testing, biomaterials are assigned cytotoxicity ratings based on visual evidence of morphological cellular changes, including cell lysis, rounding, spreading, and proliferation. For example, ISO 10993 cytotoxicity testing of medical devices allows the use of a visual grading scale. The present study compared visual in vitro cytotoxicity ratings to quantitative in vitro cytotoxicity measurements for biomaterials to determine the level of correlation between visual scoring and a quantitative cell viability assay. Biomaterials representing a spectrum of biological reactivity levels were evaluated, including organo-tin polyvinylchloride (PVC; a known cytotoxic material), ultra-high molecular weight polyethylene (a known non-cytotoxic material), and implantable tissue adhesives. Each material was incubated in direct contact with mouse 3T3 fibroblast cell cultures for 24 h. Visual scores were assigned to the materials using a 5-point rating scale; the scorer was blinded to the material identities. Quantitative measurements of cell viability were performed using a 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay; again, the assay operator was blinded to material identities. The investigation revealed a high degree of correlation between visual cytotoxicity ratings and quantitative cell viability measurements; a Pearson’s correlation gave a correlation coefficient of 0.90 between the visual cytotoxicity score and the percent viable cells. An equation relating the visual cytotoxicity score and the percent viable cells was derived. The results of this study are significant for the design and interpretation of in vitro cytotoxicity studies of novel biomaterials.

Keywords

3T3 cell line Adhesives Biomaterials Cytotoxicity Fibroblasts 

Abbreviations

MTT

3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide

Notes

Acknowledgements

This study was performed as part of a DuPont Six Sigma project. We are grateful to Deana DiCosimo, Harvey Gold, and Lourdes Puig from the DuPont Six Sigma organization for their guidance and suggestions.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  1. 1.Biochemical Sciences and Engineering, Central Research and DevelopmentDuPont Experimental StationWilmingtonUSA
  2. 2.DuPont Experimental StationWilmingtonUSA

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