Methylcellulose cell culture as a new cytotoxicity test system for biomaterials
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Abstract
The cytotoxicity of biomaterials can be testedin vitro using various culture systems. Liquid culture systems may detect cytotoxicity of a material either by culture of cells with extracts or with the material itself. In the latter instance, renewing the medium will remove possible released cytotoxic products. The agar-overlay test is a short term semi-solid culture system in which the possible cytotoxicity of biomaterials is identified only by the presence of cell free zones. The aim of this study was to develop a more sensitive cytotoxicity test system for biomaterials, using methylcellulose as a culture gel, mixed with human fibroblasts. The main advantage of the test system is the possibility of evaluating cytotoxicity for a period of up to seven days without renewal of the culture gel. Furthermore it is possible to both quantitatively evaluate by counting absolute cell numbers and to qualitatively evaluate by studying cell morphology with light- and/or electron microscopy. Processed dermal sheep collagen was selected as test material, since contradictory results concerning the cytotoxicity of its extracts have been reported by others [2, 15, 18, 19]. Using our test system, both primary and secondary cytotoxic effects were found. Primary cytotoxicity is due to direct leakage of products from the material, detected by testing, extracts of the collagen or the collagen itself. Secondary cytotoxicity is due to release of cytotoxic products resulting from cell-biomaterial interactions. We conclude that our test system is extremely useful to test materials which are suspected of primary and/or secondary cytotoxicity, either with slow release of cytotoxic products or release of products with late cytotoxic effects.
Keywords
Culture System Cytotoxic Effect Liquid Culture Test Material Human FibroblastPreview
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- 1.H. J. JOHNSON, S. J. NORTHUP, P. A. SEAGRAVES, M. ATALLAH, L. LIN and T. D. DARBY,J. Biomed. Mater. Res. 19 (1985) 489.Google Scholar
- 2.J. B. ULREICH, M. CHVAPIL, ibid.15 (1981) 913.Google Scholar
- 3.A. COOKE, R. F. OLIVIER, M. EDWARD,Brit. J. Exp. Path. 64 (1983) 172.Google Scholar
- 4.P. B. WACHEM VAN, A. H. HOGT, T. BEUGELING, J. FEIJEN, A. BANTJES, J. P. DETMERS and W. G. AKEN VAN,Biomater. 8 (1987) 323.Google Scholar
- 5.P. B. VAN WACHEM, T. BEUGELING, J. FEIJEN, A. BANTJES, J. P. DETMERS and W. G. VAN AKEN, ibid.6 (1985) 403.Google Scholar
- 6.J. A. TAYLOR, R. A. ABODEELY and R. L. FUSON, in “Trans. Amer. Soc. Artif. Int. Organs”, Vol XIX (1973) pp. 175–8.Google Scholar
- 7.A. M. ADAMS, J. V. SOAMES and R. F. SEARLE,Cytotechnol. (1988) 179.Google Scholar
- 8.S. SRIVASTAVA, S. D. GORHAM, J. M. COURTNEY,Biomater. 11 (1990) 133.Google Scholar
- 9.W. L. GUESS, S. A. ROSENBLUTH, B. SCHMIDT and J. AUTIAN,J. Pharmaceutical Sci. 45 (1965) 1545.Google Scholar
- 10.A. HENSTEN-PETTERSEN and K. HELGELAND,Scand. J. Dent. Res. 85 (1977) 291.Google Scholar
- 11.J. AUTIAN, in “Advanced Modern Toxicology, New Concept in Safety Eval-I” (1979) pp. 119–49.Google Scholar
- 12.N. N. ISCOVE and H. SCHREIER, in “Clonal Growth of Cells in Semisolid or Viscous Medium, Immunological Methods” (Academic Press, 1979).Google Scholar
- 13.K. KUBUTA and H. D. PREISTER,Exp. Hematol. 10 (1982) 292.Google Scholar
- 14.K. OZAWA, Y. HASHIMOTO, A. URABE, T. SUD, K. MOTOYOSHI, F. TAKAKU and Y. MIURA,Internat. Soc. Exper. Hematol. 10 (1982) 145.Google Scholar
- 15.C. METSELAAR, PhD Thesis, Amsterdam University, Amsterdam, The Netherlands (1990).Google Scholar
- 16.H. TRÄUBEL, Gerbungen mit Isocyanaten, Teil 1, Das Leder, 150–4, 1977.Google Scholar
- 17.Idem., Gerbungen mit Isocyanatderivaten, Teil 2, Das Leder, 181–5, 1977.Google Scholar
- 18.M. CHVAPIL,J. Biomed. Mater. Res. 16 (1982) 245.Google Scholar
- 19.M. CHVAPIL, D. P. SPEER, W. MORA and C. D. ESKELSON,J. Surg. Res. 35 (1983) 402.Google Scholar
- 20.V. M. SHUBIK, M. A. NEVSTRUYEVA, S. A. KALNITSKIL, G. N. LIVSHITS, G. N. MERKUSHEV, E. M. PILSHCHIK and R. E. PONOMAREVA,J. Hygiene, Epidemiol. Microbiol. and Immunol. 22 (1978) 408.Google Scholar
- 21.G. L. KENNEDY, L. RAYANNE, L. FERENZ and A. BURGESS,J. Appl. Toxicol. 6 (1986) 145.Google Scholar
- 22.W. E. BROWN, A. H. GREEN, M. H. KAROL and Y. C. E. ALARIE,Texicol. and Appl. Pharmacol. 63 (1982) 45.Google Scholar
- 23.K. C. CHANG and M. H. KAROL,Clin. Allergy,14 (1984) 329.Google Scholar
- 24.M. BERODE, P. LEUENBERGER and H. SAVOLAINEN,Biochem. Internat. 17 (1988) 1.Google Scholar
- 25.E. YANO, M. YOSHIOKA and A. KOIZUMI,Jpn J. Ind. Health 23 (1981) 537.Google Scholar
- 26.F. R. JOHANNSEN and G. J. LEVINSKAS,J. Appl. Toxicol. 7 (1987) 259.Google Scholar
- 27.T. M. VAN GULIK,Neth. J. Surg. 39 (1987) 90.Google Scholar
- 28.V. EVERTS, PhD Thesis, University of Amsterdam, Amsterdam, The Netherlands (1987).Google Scholar
- 29.T. YAJIMA,Cell and Tissue Res. 245 (1986) 253.Google Scholar
- 30.D. E. WOOLLEY, in “Extracellular Matrix Biochemistry”, edited by K. A. Piez and A. H. Reddi (Elsevier, New York, 1984) pp. 119–57.Google Scholar
- 31.G. MURPHY, J. J. REYNOLDS,Bio Essays 2 (1985) 55.Google Scholar
- 32.L. WEISS, in “Cell and Tissue Biology and Textbook of Histology”, Ch. 5, 6th Edn (Urban and Schwarzenberg, Baltimore, 1988) pp. 157–88.Google Scholar
- 33.Idem., ibid. pp. 191–209.Google Scholar
- 34.M. E. NIMNI, D. T. CHEUNG, B. STATES, M. KODAMA and K. SHEIKH, in “Collagen”, Ch. 1, Vol. 3, 741–71, edited by M. E. Nimni (CRC Press, Inc., Boca Raton, Florida, 1988).Google Scholar
- 35.E. GENDLER, S. GENDLER and M. E. NIMNI,J. Biomed. Mater. Res. 18 (1984) 727.Google Scholar
- 36.P. K. BAJPAI and P. K. STULL,Int. Res. Commun. Syst. Med. Sci. 8 (1980) 642.Google Scholar
- 37.S. L. WEINBERG, G. B. CIPOLLETTI and R. J. TURNER, in “Biologic and Synthetic Vascular Prostheses”, Ch. 24, edited by J. C. Stanley (Grune and Stratton Inc., New York, 1982) 433–45.Google Scholar
- 38.D. P. SPEER, M. CHVAPIL, C. D. ESKELSON and J. ULREICH,J. Biomed. Mater. Res. 14 (1980) 753.Google Scholar
- 39.M. CHVAPIL, D. SPEER, W. MORA and C. ESKELSON,J. Surg. Res. 35 (1983) 402.Google Scholar
- 40.E. EYBLE, A. GRIESMACHER, M. GRIMM and E. WOLNER,J. Biomed. Mater. Res. 23 (1989) 1355.Google Scholar