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



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.


3T3 cell line Adhesives Biomaterials Cytotoxicity Fibroblasts 



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



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.


  1. AAMI (Association for the Advancement of Medical Instrumentation). Biological evaluation of medical devices–Part 5: Tests for in vitro cytotoxicity. Arlington, VA; 2003.Google Scholar
  2. Food and Drug Administration, US Department of Health and Human Services. Use of International Standard ISO 10993, biological evaluation of medical devices–Part 1: Evaluation and testing; G95-1. Rockville, MD; 1995.Google Scholar
  3. Furst W, Banerjee A. Release of glutaraldehyde from an albumin-glutaraldehyde tissue adhesive causes significant in vitro and in vivo toxicity. Ann Thorac Surg. 2005;79:1522–8.PubMedCrossRefGoogle Scholar
  4. International Organization for Standardization. Biological evaluation of medical devices – ISO 10993, Part 5: Tests for in vitro cytotoxicity. Geneva, Switzerland; 1999.Google Scholar
  5. Kim JC, Bassage SD, Kempski MH, del Cerro M, Park SB, Aquavella JV. Evaluation of tissue adhesives in closure of scleral tunnel incisions. J Cataract Refract Surg. 1995;21:320–5.PubMedGoogle Scholar
  6. Kodokian GK, Arthur SD. Polysaccharide-based polymer tissue adhesive for medical use. U.S. Patent Application 11/244,756;2006.Google Scholar
  7. Leggat PA, Smith DR, Kedjarune U. Surgical applications of cyanoacrylate adhesives: a review of toxicity. ANZ J Surg. 2007;77:209–13.PubMedCrossRefGoogle Scholar
  8. Ministry of Health and Welfare, Pharmaceutical Affairs Bureau. Japanese guidelines for basic biological tests for medical devices and materials, notification no. 99. Tokyo, Japan; 1995.Google Scholar
  9. Ratner BD, Hoffman AS, Schoen FJ, Lemons JE (eds) Biomaterials science, 2nd ed. New York: Academic; 1996.Google Scholar
  10. Sgouras D, Duncan R. Methods for the evaluation of biocompatibility of soluble synthetic polymers which have potential for biomedical use: 1 – Use of the tetrazolium-based colorimetric assay (MTT) as a preliminary screen for evaluation of in vitro cytotoxicity. J Mater Sci Mater Med. 1990;1:61–8.CrossRefGoogle Scholar
  11. Toriumi DM, Raslan WF, Friedman M, Tardy ME. Variable histotoxicity of histoacryl when used in a subcutaneous site: an experimental study. Laryngoscope. 1991;101:339–43.PubMedGoogle Scholar

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

Personalised recommendations