Journal articles: 'In vitro toxicity tests'

1

Lambré, Claude R., Michaela Aufderheide, Robert E. Bolton, Bice Fubini, Henk P. Haagsman, Paul M. Hext, Mark Jorissen, et al. "In Vitro Tests for Respiratory Toxicity." Alternatives to Laboratory Animals 24, no. 5 (September 1996): 671–81. http://dx.doi.org/10.1177/026119299602400506.

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Cinelli, S., A. Falezza, C. Meli, P. Ciliutti, and J. A. Vericat. "Alternative methods in toxicology tests: In vitro toxicity." Cytotechnology 5, S1 (1991): 51–54. http://dx.doi.org/10.1007/bf00736807.

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Martin, Angela, and Martin Clynes. "Acid phosphatase: Endpoint for in vitro toxicity tests." In Vitro Cellular & Developmental Biology - Animal 27, no. 3 (March 1991): 183–84. http://dx.doi.org/10.1007/bf02630912.

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Flint, Oliver P. "In Vitro Toxicity Testing: Purpose, Validation and Strategy." Alternatives to Laboratory Animals 18, no. 1_part_1 (November 1990): 11–18. http://dx.doi.org/10.1177/026119299001800103.1.

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The fullest potential for in vitro evaluation of toxicity will be realised in the context of the process of assessing the risk of human toxicity. This article is an attempt to clarify what contributions can be made by in vitro tests and what types of in vitro test can best be used. In vitro tests are clarified according to the type of biological endpoint evaluated, first into tests for general (‘basal’) cytotoxicity and, secondly, into tests for differentiated cell function. The role of each type of test is analysed and it is suggested that tests for general cytotoxicity, as opposed to differentiated function, are difficult to interpret in terms of in vivo toxicity. A general approach to evaluating in vitro tests is described, and a strategy for using these tests is proposed.

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Clemedson, Cecilia, Elisabeth McFarlane-Abdulla, Marianne Andersson, Frank A. Barile, Mabel C. Calleja, Christophe Chesné, Richard Clothier, et al. "MEIC Evaluation of Acute Systemic Toxicity." Alternatives to Laboratory Animals 24, no. 1_part_1 (June 1996): 251–72. http://dx.doi.org/10.1177/026119299602400102.1.

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The multicentre evaluation of in vitro cytotoxicity (MEIC) study is a programme designed to evaluate the relevance of in vitro toxicity tests for predicting human toxicity, and is organised by the Scandinavian Society for Cell Toxicology. The project started in 1989 and is scheduled to be finished by June 1996. MEIC is a voluntary effort by international laboratories to test the same 50 reference chemicals in their own in vitro toxicity systems. At present, 31 laboratories have submitted results for the first 30 reference chemicals from a total of 68 in vitro cytotoxicity tests. In the definitive evaluation of the MEIC programme, these in vitro results will be compared with human lethal blood concentrations and other relevant acute systemic toxicity data, and the results will be published as a series of articles. This paper, which is the first article in this series, describes and analyses the methodologies used in the 68 tests. The origins and purities of the test chemicals, the biological systems and the toxicity endpoints are also discussed. Since MEIC is not centrally directed, the selection of tests was entirely dependent on the preferences of the individual laboratories. Thus, the collection of tests is not representative of the full range of existing in vitro toxicity tests. In our study, basal cytotoxicity tests and ecotoxicological tests are prevalent, while tests for toxicity to primary cultures of differentiated cells, measured by organotypic toxicity endpoints, are clearly under-represented.

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Isomaa, Boris, Henrik Lilius, and Christina Råbergh. "Aquatic Toxicology in Vitro: A Brief Review." Alternatives to Laboratory Animals 22, no. 4 (July 1994): 243–53. http://dx.doi.org/10.1177/026119299402200405.

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There is an urgent need for effective in vitro tests in aquatic toxicology, because only a very small proportion of the chemicals in common use have been adequately tested for their toxicity to aquatic organisms and aquatic ecosystems. Toxicity tests with higher animals, besides being time-consuming and expensive, are ethically questionable, which further increases the importance of developing efficient in vitro toxicity tests. In developing in vitro tests for toxicity assessments, aquatic toxicology lags behind mammalian toxicology. Aqueous environmental chemistry is complex, and the sensitivity of the organisms living in a particular aquatic environment may vary considerably. The predictive value of single-species or cell culture tests is therefore generally considered to be low. Nevertheless, single-species tests, utilising bacteria, algae, protozoans and invertebrates, have frequently been used in in vitro toxicity studies of aquatic pollutants (mainly as screening tests). Attempts at large-scale validations are few. Such attempts seem to be hampered by the complexity of the aquatic ecosystem. Although cells from aquatic organisms have been isolated and cultured for many years, the use of isolated or cultured cells in aquatic toxicology has been limited. However, during the last few years, interest in the use of fish cells in toxicity testing has grown rapidly. For aquatic in vitro toxicology to develop further, a more comparative and mechanistic approach needs to be adopted.

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Northup, Sharon J. "Perspectives on In Vitro Toxicity for Medical Devices." Journal of the American College of Toxicology 7, no. 4 (July 1988): 481–89. http://dx.doi.org/10.3109/10915818809019521.

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In vitro toxicity testing has found widespread application in its use for screening materials for medical devices. Cytotoxicity tests, which have been in use for nearly 20 years, have been validated for intralaboratory repeatability, interlaboratory reproducibility, and correlation with acute animal toxicity assays. The three primary cytotoxicity assays, i.e., direct contact, agar diffusion, and elution tests, allow a selection between assay and material characteristics. Mutagenicity assays have had limited application to materials testing because of the insoluble nature of the materials and the low level of extractable chemicals, which are generally below the sensitivity limit of these assays. In vitro blood compatibility tests for hemolysis and complement activation are used primarily for blood contacting materials in applications where there is a large surface area of material for ex vivo applications.

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Edwards, James, Gerlinde Pappa, Michael Török, Ann Fowler, and Jochen Bausch. "α-Mangostin: Developmental toxicity in vitro and in screening tests." Toxicology Letters 189 (September 2009): S144. http://dx.doi.org/10.1016/j.toxlet.2009.06.809.

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Goldberg, Alan M., John M. Frazier, David Brusick, Michael S. Dickens, Oliver Flint, Stephen D. Gettings, Richard N. Hill, et al. "Framework for validation and implementation of in vitro toxicity tests." In Vitro Cellular & Developmental Biology - Animal 29, no. 9 (September 1993): 688–92. http://dx.doi.org/10.1007/bf02631424.

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Clemedson, Cecilia, Marianne Andersson, Yasunobu Aoki, Frank A. Barile, Anna Maria Bassi, Marbel C. Calleja, Argelia Castano, et al. "MEIC Evaluation of Acute Systemic Toxicity." Alternatives to Laboratory Animals 26, no. 1_suppl (March 1998): 131–83. http://dx.doi.org/10.1177/026119299802601s03.

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Results from tests on the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) reference chemicals 31–50 in 67 different in vitro toxicity assays are presented in this paper as a prerequisite to in vitro/in vivo comparisons for all MEIC in vitro toxicity data in forthcoming papers, i.e. the final MEIC evaluation of the relevance of the tests. With the aim of increasing knowledge about the relative significance of some in vitro methodological factors, the strategies and methods of the preceding parts in the MEIC series (Parts II and III) were again employed to enable comparative cytotoxicity analysis of the new in vitro results presented in this paper. A principal components analysis (PCA) of the results from tests of the 20 chemicals in 67 assays demonstrated a dominating first component describing as much as 74% of the variance in the toxicity data, indicating a similar ranking of the cytotoxicities of the chemicals in most of the tests. The influence on the general variability of the results of a few, key methodological factors was also evaluated by using linear regression comparisons of the results of all pairs of methods available in the study, i.e. methods which were similar in all respects except for the factor being analysed. Results from this “random probe” analysis were: a) the cytotoxicities of 11 of the 20 chemicals increased considerably with exposure time (> 10 times over 4–168 hours); b) in general, human cell line toxicity was well predicted by cytotoxicity in animal cells; c) prediction of human cell line toxicity by most ecotoxicological tests was only fairly good; d) 14 comparisons of similar assays with different cell lines showed similar toxicities (mean R2 = 0.83); e) nine comparisons of similar assays employing different primary cultures and cell lines shared similar toxicities (mean R2 = 0.71); and f) 16 comparisons of similar assays with different growth/viability endpoints showed similar toxicities (mean R2 = 0.71). Results b, d, e and f must contribute to the PCA-documented high general similarity of the in vitro toxicity data. Results a and c, together with factors which were not analysed, such as different protocols and inter-laboratory variability of tests, could explain the 26% dissimilarity. To provide background information to the planned final MEIC evaluation of the relevance of the 61 methods in which all 50 chemicals have been tested, an additional PCA was made of the 50 chemical-61 assay in vitro database (from Parts II and III and the present paper). This supplementary PCA demonstrated an 80% similarity of results. Compared with the previous analysis of the tests of the first 30 MEIC reference chemicals (MEIC Part III), the present analysis of the tests of the last 20 MEIC chemicals indicates a somewhat higher variation in the results. Correspondingly, some deviating endpoint measurements and cell line responses were demonstrated by the pairwise comparisons in the present study. As a result, the analysis revealed a high correlation (R2 = 0.73) between the average human cell line toxicity and the results from a new protein denaturation test. These preliminary results suggest that intracellular protein denaturation may be a frequently occurring mechanism in basal cytotoxicity.

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Kolarova, J., J. Velisek, and Z. Svobodova. "Comparison of in vitro (fish cell line) and in vivo (fish and crustacean) acute toxicity tests in aquatic toxicology." Veterinární Medicína 66, No. 8 (July 5, 2021): 350–55. http://dx.doi.org/10.17221/161/2020-vetmed.

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The use of in vitro (fish cell lines) is a cost-effective, very rapid, and informative tool for toxicological assessments. Using the neutral red (NR) assay, we compared the in vitro acute toxicity (20hEC50) of twenty-six chemical substances on a rainbow trout gonad cell line (RTG-2) with their in vivo acute toxicity to Barbados Millions Poecilia reticulata (48hLC50, OECD 203) and crustacean Daphnia magna (48hEC50, OECD 202). The 20hEC50 values obtained by the NR assay were higher in nearly all the cases when compared to the 48hLC50 in P. reticulata and the 48hEC50 in D. magna, indicating that the sensitivity of the RTG-2 cell line was lower compared to P. reticulata and D. magna. A high (r = 0.89) and significant (P < 0.001) correlation was recorded between the 20hEC50 values of the RTG-2 and the 48hEC50 values of D. magna. The correlation between the 20hEC50 values of the RTG-2 and the 48hLC50 values of P. reticulata was lower (r = 0.65; P < 0.001), but also significant. The authors recommend use of the NR assay on the RTG-2 cell lines as a screening protocol to evaluate the toxicity of xenobiotics in aquatic environments to narrow the spectrum of the concentrations for the fish toxicity test.

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12

Ekwall, Björn, Inger Bondesson, Sven Hellberg, Johan Högberg, Lennart Romert, Kjell Stenberg, and Erik Walum. "Validation of In Vitro Cytotoxicity Tests — Past and Present Strategies." Alternatives to Laboratory Animals 19, no. 2 (April 1991): 226–33. http://dx.doi.org/10.1177/026119299101900215.

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In recent years, conventional toxicity testing in animals has been reinforced by in vitro methods. As a result, toxicity testing in some sectors has become more effective and at the same time more ethical. This trend is probably only at its beginning, as many of the newly-developed methods have not yet won general acceptance as a basis for the large-scale replacement and reduction of animal experimentation. What limits the wider use of these methods is validation, i.e. the evaluation of their reliability and relevance. The present paper is a short review of the validation efforts made hitherto, including projects being planned and under discussion. Our own MEIC approach is compared with other strategies. Finally, our opinion on the effectiveness of one large consensus project relative to several different smaller validation programmes is expressed — we advocate the latter strategy, because it will save time and reduce costs.

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Aydιn, Ahmet, Göknur Aktay, and Erdem Yesilada. "A Guidance Manual for the Toxicity Assessment of Traditional Herbal Medicines." Natural Product Communications 11, no. 11 (November 2016): 1934578X1601101. http://dx.doi.org/10.1177/1934578x1601101131.

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Herbal remedies have been used for thousands of years in worldwide traditional medicines for their potential health benefits. Although they are generally presumed safe unless a significant risk has been identified in humans, increasing number of case reports notify acute or chronic intoxications resulting from their use. This study aims to produce a scientific guide for the evaluation of traditional herbal medicines (THMs) in terms of their toxicity risks based on the published regulatory documents. For this purpose recommended in vitro and in vivo toxicity tests on medicinal products for human use issued by the international regulatory bodies are overviewed and they are then adopted to be used for the toxicity assessment of THMs. Accordingly, based on compilation of these issued regulations, the following tests are recommended for the toxicity assessment of THMs; in vitro cytotoxicity, genotoxicity, acute and repeated dose toxicity, carcinogenicity, reproductive and developmental toxicity, local tolerance tests, toxicokinetic studies, and additional toxicity tests including safety pharmacology, immunotoxicity and antigenicity, endocrine system toxicity, gastro-intestinal toxicity, renal and hepatotoxicity, and drug interaction studies. This study describes and discusses the applicability of these tests for the risk assessment in THMs.

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Klaric, Martina. "Current status of human pluripotent stem cell based in vitro toxicity tests." Frontiers in Bioscience S5, no. 1 (2013): 118–33. http://dx.doi.org/10.2741/s361.

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Liu, Weiying, Jing Xi, Yiyi Cao, Xinyue You, Ruixue Chen, Xinyu Zhang, Li Han, Guoyu Pan, and Yang Luan. "An Adaption of Human-Induced Hepatocytes to In Vitro Genetic Toxicity Tests." Mutagenesis 34, no. 2 (December 24, 2018): 165–71. http://dx.doi.org/10.1093/mutage/gey041.

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Parchment, R. E., M. Gordon, C. K. Grieshaber, C. Sessa, D. Volpe, and M. Ghielmini. "Predicting hematological toxicity (myelosuppression) of cytotoxic drug therapy from in vitro tests." Annals of Oncology 9, no. 4 (April 1998): 357–64. http://dx.doi.org/10.1023/a:1008245906772.

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Garle, M. J., J. H. Fentem, and J. R. Fry. "In vitro cytotoxicity tests for the prediction of acute toxicity in vivo." Toxicology in Vitro 8, no. 6 (December 1994): 1303–12. http://dx.doi.org/10.1016/0887-2333(94)90123-6.

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Shaliutina, Olena, Anna Materiienko, Anna Shaliutina-Kolešová, and Ievgeniia Gazo. "Using fish spermatozoa in in vitro toxicity tests: A potential toxicology tool." Aquaculture 539 (June 2021): 736647. http://dx.doi.org/10.1016/j.aquaculture.2021.736647.

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19

Clemedson, Cecilia, Marika Nordin-Andersson, Henning F. Bjerregaard, Jørgen Clausen, Anna Forsby, Helena Gustafsson, Ulrika Hansson, et al. "Development of an In Vitro Test Battery for the Estimation of Acute Human Systemic Toxicity: An Outline of the EDIT Project." Alternatives to Laboratory Animals 30, no. 3 (May 2002): 313–21. http://dx.doi.org/10.1177/026119290203000309.

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The aim of the Evaluation-guided Development of New In Vitro Test Batteries (EDIT) multicentre programme is to establish and validate in vitro tests relevant to toxicokinetics and for organ-specific toxicity, to be incorporated into optimal test batteries for the estimation of human acute systemic toxicity. The scientific basis of EDIT is the good prediction of human acute toxicity obtained with three human cell line tests (R2 = 0.77), in the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) programme. However, the results from the MEIC study indicated that at least two other types of in vitro test ought to be added to the existing test battery to improve the prediction of human acute systemic toxicity — to determine key kinetic events (such as biotransformation and passage through biological barriers), and to predict crucial organ-specific mechanisms not covered by the tests in the MEIC battery. The EDIT programme will be a case-by-case project, but the establishment and validation of new tests will be carried through by a common, step-wise procedure. The Scientific Committee of the EDIT programme defines the need for a specific set of toxicity or toxicokinetic data. Laboratories are then invited to perform the defined tests in order to provide the “missing” data for the EDIT reference chemicals. The results obtained will be evaluated against the MEMO (the MEIC Monograph programme) database, i.e. against human acute systemic lethal and toxicity data. The aim of the round-table discussions at the 19th Scandinavian Society for Cell Toxicology (SSCT) workshop, held in Ringsted, Denmark on 6–9 September 2001, was to identify which tests are the most important for inclusion in the MEIC battery, i.e. which types of tests the EDIT programme should focus on. It was proposed that it is important to include in vitro methods for various kinetic events, such as biotransformation, absorption in the gut, passage across the blood–brain barrier, distribution volumes, protein binding, and renal clearance/accumulation. Models for target organ toxicity were also discussed. Because several of the outlier chemicals (paracetamol, digoxin, malathion, nicotine, paraquat, atropine and potassium cyanide) in the MEIC in vivo–in vitro evaluation have a neurotoxic potential, it was proposed that the development within the EDIT target organ programme should initially be focused on the nervous system.

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Clothier, Richard H., Karen A. Atkinson, Michael J. Garle, Rachel K. Ward, and Angela Willshaw. "The Development and Evaluation of In Vitro Tests by the FRAME Alternatives Laboratory." Alternatives to Laboratory Animals 23, no. 1 (January 1995): 75–90. http://dx.doi.org/10.1177/026119299502300111.

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This review outlines the work which has been conducted in the FRAME Alternatives Laboratory during the first ten years of the FRAME Research Programme. A number of in vitro tests, including the kenacid blue, neutral red release and fluorescein leakage assay methods, have been evaluated and have subsequently been included in validation schemes organised by the US Soap and Detergent Association, the US Cosmetic, Toiletry and Fragrance Association, the European Commission and the European Cosmetic, Toiletry and Perfumery Association, as well as in the Scandinavian multicentre evaluation of in vitro cytotoxicity testing scheme. More recently, research has been undertaken in the areas of phototoxicity, immunotoxicity, dermal toxicity and intercellular communication, in addition to investigations into fundamental mechanisms of toxicity.

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Dodd, Darol E., Gary M. Hoffman, and Colin J. Hardy. "Perfluoro-n-Butyl Iodide: Acute Toxicity, Subchronic Toxicity and Genotoxicity Evaluations." International Journal of Toxicology 23, no. 4 (July 2004): 249–58. http://dx.doi.org/10.1080/10915810490502050.

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Perfluoro-n-butyl iodide (PFBI) is a promising alternative to chlorofluorocarbon solvents used in aircraft ground maintenance operations and other military and commercial operations, because it cleans well, has zero ozone depletion potential, and has extremely low global warming properties. Toxicity tests were performed with PFBI to determine and evaluate its health hazard. Using standard testing guidelines (e.g., Organization for Economic Cooperation and Development [OECD]), tests included acute (4-h) and 4-week (6 h/day, 5 days/week) inhalation (nose-only) toxicity studies in rats, acute (10-min) inhalation cardiac sensitization study in dogs, in vitro chromosomal aberrations experiments in human lymphocytes, and in vitro mutagenic experiments in Salmonella typhimurium and Escherichia coli. There were no mortalities in rats ( n = 10) exposed for 4 h to 10,000 ppm PFBI, but all rats ( n = 10) died within 2 h when exposed to 20,000 ppm PFBI. The 4-h LC50 (95% confidence limits) was 14,000 ppm (13,000 ppm to 16,000 ppm). Signs (nasal discharge and labored breathing) observed in the rats exposed to 10,000 ppm returned to normal within 48 h. PFBI has the potential to cause cardiac sensitization in epinephrine-challenged dogs at 6200 ppm. A concentration of 3900 ppm was a no-observed-adverse-effect level (NOAEL) in the cardiac sensitization study. In the 4-week inhalation study (5 rats/sex/group), respiratory mucosal hypertrophy/hyperplasia was observed in rats of the 10,000-ppm group. A NOAEL of 1000 ppm was selected for the 4-week study on the basis that the mild increase in T4 observed at 1000 ppm was considered adaptive, not adverse, because of the absence of frank effects in the thyroid. In the in vitro studies, PFBI showed no evidence of either mutagenic or clastogenic activity. The toxicity profile of PFBI was compared to trifluoroiodomethane. In conclusion, the results of these studies indicate a low order of general toxicity and an absence of genotoxicity following PFBI exposure.

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Vippola, M., GCM Falck, HK Lindberg, S. Suhonen, E. Vanhala, H. Norppa, K. Savolainen, A. Tossavainen, and T. Tuomi. "Preparation of nanoparticle dispersions for in-vitro toxicity testing." Human & Experimental Toxicology 28, no. 6-7 (June 2009): 377–85. http://dx.doi.org/10.1177/0960327109105158.

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Studies on potential toxicity of engineered nanoparticle (ENP) in biological systems require a proper and accurate particle characterization to ensure the reproducibility of the results and to understand biological effects of ENP. A full characterization of ENP should include various measurements such as particle size and size distribution, shape and morphology, crystallinity, composition, surface chemistry, and surface area of ENP. It is also important to characterize the state of ENP dispersions. In this study, four different ENPs, rutile and anatase titanium dioxides and short single- and multi-walled carbon nanotubes, were characterized in two dispersion media: bronchial epithelial growth medium, used for bronchial epithelial BEAS cells, and RPMI-1640 culture media with 10% of fetal calf serum (FCS) for human mesothelial (MeT-5A) cells. The purpose of this study was to determine the characteristics of ENPs and their dispersions as well as to compare dispersion additives suitable for toxicity tests and thus establish an appropriate way to prepare dispersions that performs well with the selected ENP. Dispersion additives studied in the media were bovine serum albumin (BSA) as a protein resource, dipalmitoyl phosphatidylcholine (DPPC) as a model lung surfactant, and combination of BSA and DPPC. Dispersions were characterized using optical microscopy and transmission electron microscopy. Our results showed that protein addition, BSA or FCS, in cell culture media generated small agglomerates of primary particles with narrow size variations and improved the stability of the dispersions and thus also the relevance of the in-vitro genotoxicity tests to be done.

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Borges de Menezes, José Vitor Nogara, Esther Rieko Takamori, Maria Francisca Thereza Borro Bijella, and José Mauro Granjeiro. "In vitro toxicity of MTA Compared with other Primary Teeth Pulpotomy Agents." Journal of Clinical Pediatric Dentistry 33, no. 3 (April 1, 2009): 217–22. http://dx.doi.org/10.17796/jcpd.33.3.cq7677j4l532r1rg.

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Objective: The main goal of this work is to compare the In vitro toxicity of MTA with other primary teeth pulpotomy agents. Study Design: The In vitro toxicity of MTA, calcium hydroxide, ferric sulphate solution,diluted formocresol and Buckley's formocresol were tested using MTT and Neutral Red Uptake cell viability assays. The results for MTA were compared to those obtained for the other substances using ANOVA and Tukey statistical tests (p<0,05). Results: MTA had the lower in vitro toxicity and Buckley's formocresol, the higher, with statiscally significant difference. Conclusion: Among the primary teeth pulpotomy agents tested, MTA showed the lower In vitro toxicity, standing as the most promising substitute to formocresol.

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Stoian, A., S. Druon–Bocquet, H. Groux, and J. Sanchez. "New Membrane Device for in Vitro VOC Toxicity Tests: Experimental and Modelling Study." Procedia Engineering 44 (2012): 377–78. http://dx.doi.org/10.1016/j.proeng.2012.08.421.

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Shelby, Michael D., James W. Allen, William J. Caspary, Steven Haworth, James Ivett, Andrew Kligerman, Carol A. Luke, et al. "Results of in vitro and in vivo genetic toxicity tests on methyl isocyanate." Environmental Health Perspectives 72 (June 1987): 183–87. http://dx.doi.org/10.1289/ehp.8772183.

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Maier, P. "Development of in vitro toxicity tests with cultures of freshly isolated rat hepatocytes." Experientia 44, no. 10 (October 1988): 807–17. http://dx.doi.org/10.1007/bf01941176.

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Balls, M., and R. H. Clothier. "Comments on the scientific validation and regulatory acceptance of in vitro toxicity tests." Toxicology in Vitro 5, no. 5-6 (January 1991): 535–38. http://dx.doi.org/10.1016/0887-2333(91)90088-u.

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Combes, Robert, Christina Grindon, Mark T. D. Cronin, David W. Roberts, and John F. Garrod. "Integrated Decision-tree Testing Strategies for Acute Systemic Toxicity and Toxicokinetics with Respect to the Requirements of the EU REACH Legislation." Alternatives to Laboratory Animals 36, no. 1 (February 2008): 45–63. http://dx.doi.org/10.1177/026119290803600107.

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Liverpool John Moores University and FRAME conducted a joint research project, sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for acute systemic toxicity and toxicokinetic testing. The paper reviews in vitro tests based on basal cytotoxicity and target organ toxicity, along with QSAR models and expert systems available for this endpoint. The use of PBPK modelling for the prediction of ADME properties is also discussed. These tests are then incorporated into a decision-tree style, integrated testing strategy, which also includes the use of refined in vivo acute toxicity tests, as a last resort. The implementation of the strategy is intended to minimise the use of animals in the testing of acute systemic toxicity and toxicokinetics, whilst satisfying the scientific and logistical demands of the EU REACH legislation.

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Combes, Robert, Christina Grindon, Mark T. D. Cronin, David W. Roberts, and John F. Garrod. "Integrated Decision-tree Testing Strategies for Acute Systemic Toxicity and Toxicokinetics with Respect to the Requirements of the EU REACH Legislation." Alternatives to Laboratory Animals 36, no. 1_suppl (October 2008): 91–109. http://dx.doi.org/10.1177/026119290803601s08.

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Liverpool John Moores University and FRAME conducted a joint research project, sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for acute systemic toxicity and toxicokinetic testing. The paper reviews in vitro tests based on basal cytotoxicity and target organ toxicity, along with QSAR models and expert systems available for this endpoint. The use of PBPK modelling for the prediction of ADME properties is also discussed. These tests are then incorporated into a decision-tree style, integrated testing strategy, which also includes the use of refined in vivo acute toxicity tests, as a last resort. The implementation of the strategy is intended to minimise the use of animals in the testing of acute systemic toxicity and toxicokinetics, whilst satisfying the scientific and logistical demands of the EU REACH legislation.

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Laue, Heike, Lu Hostettler, Gordon Sanders, Georg Kreutzer, and Andreas Natsch. "PeBiToSens™: A Platform for PBT Screening of Fragrance Ingredients Without Animal Testing." CHIMIA International Journal for Chemistry 74, no. 3 (March 25, 2020): 168–75. http://dx.doi.org/10.2533/chimia.2020.168.

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The determination of persistence (P), bioaccumulation (B) and toxicity (T) plays a central role in the environmental assessment of chemicals. Persistence is typically evaluated via standard microbial biodegradation tests. Bioaccumulation refers to the accumulation of chemicals in organisms and is usually assessed in fish exposed to the test chemical. Toxicity is determined at three trophic levels, with fish toxicity as the highest trophic level assessed. Thus, animal tests are classically needed for both B and T assessment. In vitro systems based on fish liver cells or liver S9 fractions ('RT-S9 assay') have been recently adopted by OECD to measure the biotransformation rates for the chemicals for B assessment. Biotransformation drives clearance from the body and reduces bioaccumulation. For T assessment, an assay based on in vitro toxicity on fish gill cells has been established ('RTgill-W1 assay'). Here we summarize our findings indicating that these tests are highly predictive for fragrance ingredients, and show with two case studies of our latest new registered substances how we apply these tests in particular during development and also for chemical registration. This platform of tests (PeBiToSens™) could fully replace animal tests in ecotoxicological assessment and is key in the Givaudan Safe by Design™ approach to develop safer and environmentally compatible novel fragrance ingredients.

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Ekwall, Björn, Frank A. Barile, Argelia Castano, Cecilia Clemedson, Richard H. Clothier, Paul Dierickx, Barbro Ekwall, et al. "MEIC Evaluation of Acute Systemic Toxicity." Alternatives to Laboratory Animals 26, no. 2_suppl (August 1998): 617–58. http://dx.doi.org/10.1177/026119299802602s03.

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The Multicenter Evaluation of In Vitro Cytotoxicity (MEIC) programme was set up to evaluate the relevance for human acute toxicity of in vitro cytotoxicity tests. At the end of the project in 1996, 29 laboratories had tested all 50 reference chemicals in 61 cytotoxicity assays. Five previous articles have presented the in vitro data and the human database to be used in the evaluation. This article presents three important parts of the final evaluation: a) a comparison of rat and mouse oral LD50 with human acute lethal doses for all 50 chemicals; b) a display of the correlations between IC50 (concentration causing 50% inhibition) values from all 61 assays and three independent sets of human acute lethal blood concentrations, i.e. clinical lethal concentrations, forensic lethal concentrations, and peak concentrations; and c) a series of comparisons between average IC50 values from ten human cell line 24-hour assays and human lethal blood concentrations. In the latter comparisons, results from correlations were linked with known human toxicity data for the chemicals, to provide an understanding of correlative results. This correlative/mechanistic approach had the double purpose of assessing the relevance of the in vitro cytotoxicities, and of testing a series of hypotheses connected with the basal cytotoxicity concept. The results of the studies were as follows. Rat LD50 predictions of human lethal dosage were only relatively good (R2 = 0.61), while mouse LD50s gave a somewhat better prediction (R2 = 0.65). Comparisons performed between IC50 values from the 61 assays and the human lethal peak concentrations demonstrated that human ceil line tests gave the best average results (R2 = 0.64), while mammalian and fish cell tests correlated less well (R2 = 0.52–0.58), followed by non-fish ecotoxicological tests (R2 = 0.36). Most of the 61 assays underpredicted human toxicity for digoxin, malathion, carbon tetrachloride and atropine sulphate. In the correlative/mechanistic study, the 50 chemicals were first separated into three groups: A = fast-acting chemicals with a restricted passage across the blood–brain barrier; B = slow-acting chemicals with a restricted passage across the blood–brain barrier; and C = chemicals which cross the blood–brain barrier freely, while inducing a non-specific excitation/depression of the central nervous system (CNS). The IC50 values for chemicals in group C were divided by a factor of ten to compensate for a hypothetical extra vulnerability of the CNS to cytotoxicity. Finally, the average human cell line IC50 values (24-hour IC50 for groups A and C, and after 48-hour for group B) were compared with relevant human lethal blood concentrations (peak concentrations for groups A and C, and 48-hour concentrations for group B). As a result, in vitro toxicity and in vivo toxicity correlated very well for all groups (R2 = 0.98, 0.82 and 0.85, respectively). No clear overprediction of human toxicity was made by the human cell tests. The human cell line tests underpredicted human toxicity for only four of the 50 chemicals. These outlier chemicals were digoxin, malathion, nicotine and atropine sulphate, all of which have a lethal action in man through interaction with specific target sites not usually found in cell lines. Potassium cyanide has a cellular human lethal action which cannot be measured by standard anaerobic cell lines. The good prediction of the human lethal whole-blood concentration of this chemical was not conclusive, i.e. was probably a “false good correlation”. Another two chemicals in group C resulted in “false good correlations”, i.e. paracetamol and paraquat. The comparisons thus indicated that human cell line cytotoxicities are relevant for the human acute lethal action for 43 of the 50 chemicals. The results strongly support the basal cytotoxicity concept, and further point to the non-specific CNS depression being the obligatory reaction of humans to cytotoxic concentrations of chemicals, provided that the chemicals are able to pass the blood–brain barrier.

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Babı́n, M. M., P. Garcı́a, C. Fernández, C. Alonso, G. Carbonell, and J. V. Tarazona. "Toxicological characterisation of sludge from sewage treatment plants using toxicity identification evaluation protocols based on in vitro toxicity tests." Toxicology in Vitro 15, no. 4-5 (August 2001): 519–24. http://dx.doi.org/10.1016/s0887-2333(01)00057-1.

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Cheli, F., C. Giromini, and A. Baldi. "Mycotoxin mechanisms of action and health impact: ‘in vitro’ or ‘in vivo’ tests, that is the question." World Mycotoxin Journal 8, no. 5 (September 30, 2015): 573–89. http://dx.doi.org/10.3920/wmj2014.1864.

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The aim of this paper is to present examples of in vitro and in vivo tests for mycotoxin mechanisms of action and evaluation of health effects, with a focus on the gut environment and toxicity testing. In vivo investigations may provide information on the net effects of mycotoxins in whole animals, whereas in vitro models represent effective tools to perform simplified experiments under uniform and well-controlled conditions and a suitable alternative to in vivo animal testing providing insights not achievable with animal studies. The main limits of in vitro models are the lack of interactions with other cells and extracellular factors, lack of hormonal or immunological influences, and lack or different levels of in vitro expression of genes involved in the overall response to mycotoxins. The translation of in vitro data into meaningful in vivo effects remains an unsolved problem. The main issues to be considered are the mycotoxin concentration range in accordance with levels encountered in realistic situations, the identification of reliable biomarkers of mycotoxin toxicity, the measurement of the chronic toxicity, the evaluation of single- or multi-toxin challenge. The gastrointestinal wall is the first barrier preventing the entry of undesirable substances. The intestinal epithelium can be exposed to high concentrations of mycotoxins upon ingestion of contaminated food and the amount of mycotoxin consumed via food does not always reflect the amount available to exert toxic actions in a target organ. In vitro digestion models in combination with intestinal epithelial cells are powerful tools to screen and predict the in vivo bioavailability and digestibility of mycotoxins in contaminated food and correctly estimate health effects. In conclusion, in vitro and in vivo tests are complementary approaches for providing a more accurate picture of the health impact of mycotoxins and improved understanding and evaluation of relevant dietary exposure and risk scenarios.

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Cajaraville, M. P. "Assessment of toxic effects of metal nanoparticles using biomarkers and in vitro toxicity tests." Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 157 (September 2010): S22. http://dx.doi.org/10.1016/j.cbpa.2010.06.061.

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Clemedson, Cecilia, Elisabeth McFarlane-Abdulla, Marianne Andersson, Frank A. Barile, Mabel C. Calleja, Christophe Chesné, Richard Clothier, et al. "MEIC Evaluation of Acute Systemic Toxicity." Alternatives to Laboratory Animals 24, no. 1_part_1 (June 1996): 273–311. http://dx.doi.org/10.1177/026119299602400103.1.

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Results from tests of the first 30 MEIC reference chemicals in 68 different toxicity assays are presented as a prerequisite to subsequent in vitro/in vivo comparisons of acute toxicity data. A comparative cytotoxicity study was also carried out. Firstly, the variability of all of the results was analysed by using principal components analysis (PCA), analyses of variance (ANOVAs) and pairwise comparisons of means according to Tukey's method. The first PCA component described 80% of the variance of all of the cytotoxicity data. Tukey's ANOVA indicated a similar sensitivity for the assays, of approximately 80%. Secondly, the influence of five major methodological components on the general variability of the results was evaluated by linear regression and ANOVA linear contrast analyses. The findings were that: a) the toxicity of many chemicals increased with exposure time; b) in general, human cytotoxicity was predicted well by animal cytotoxicity tests; c) this prediction was poor for two chemicals; d) the prediction of human cytotoxicity by the ecotoxicological tests was only fairly good; e) one organotypic endpoint used, i.e. contractility of muscle cells, gave different results to those obtained according to viability/growth toxicity criteria; f) twelve comparisons of similar test systems involving different cell types (including highly differentiated cells) showed similar toxicities regardless of cell type; and g) nine out often comparisons of test systems with identical cell types and exposure times revealed similar toxicities, regardless of the viability or growth endpoint measurement used. Factors b, f and g must be the main causes of the remarkable similarity between the total results, while factors a, c, d and e, together with other minor factors that were not analysed, contributed to the 20% dissimilarity. The findings strongly support the basal cytotoxicity concept, and will facilitate future in vitro toxicity testing.

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Frazier, John M. "Validation of In Vitro Models." Journal of the American College of Toxicology 9, no. 3 (May 1990): 355–59. http://dx.doi.org/10.3109/10915819009078744.

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The 1980s have seen major research efforts focused on the development of alternative toxicity testing methodologies for the safety evaluation of chemicals. A difficult problem which has slowed the implementation of these methodologies has been validation. Validation means different things to different toxicologists; however, successful validation can take place only within the context of how the methodology is to be used: as a screen, an adjunct, or a replacement. The strategy for performing a validation study as well as the criteria for evaluation of the test methodologies will vary for each of these use categories. Correlative tests are adequate for screening purposes but mechanistic tests are required for adjuncts and replacements. The time has come to define the scientific basis of validation so that new methodologies can be efficiently processed through validation programs and incorporated into the safety evaluation process.

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Simeonova, Rumyana, Dimitrina Zheleva, Iva Valkova, Georgi Stavrakov, Irena Philipova, Mariyana Atanasova, and Irini Doytchinova. "A Novel Galantamine-Curcumin Hybrid as a Potential Multi-Target Agent against Neurodegenerative Disorders." Molecules 26, no. 7 (March 25, 2021): 1865. http://dx.doi.org/10.3390/molecules26071865.

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The acetylcholinesterase (AChE) inhibitors are the main drugs for symptomatic treatment of neurodegenerative disorders like Alzheimer’s disease. A recently designed, synthesized and tested hybrid compound between the AChE inhibitor galantamine (GAL) and the antioxidant polyphenol curcumin (CU) showed high AChE inhibition in vitro. Here, we describe tests for acute and short-term toxicity in mice as well as antioxidant tests on brain homogenates measured the levels of malondialdehide (MDA) and glutathione (GSH) and in vitro DPPH, ABTS, FRAP and LPO inhibition assays. Hematological and serum biochemical analyses were also performed. In the acute toxicity tests, the novel AChE inhibitor given orally in mice showed LD50 of 49 mg/kg. The short-term administration of 2.5 and 5 mg/kg did not show toxicity. In the ex vivo tests, the GAL-CU hybrid performed better than GAL and CU themselves; in a dose of 5 mg/kg, it demonstrates 25% reduction in AChE activity, as well as a 28% and 73% increase in the levels of MDA and GSH, respectively. No significant changes in blood biochemical data were observed. The antioxidant activity of 4b measured ex vivo was proven in the in vitro tests. In the ABTS assay, 4b showed radical scavenging activity 10 times higher than the positive control butylhydroxy toluol (BHT). The GAL-CU hybrid is a novel non-toxic AChE inhibitor with high antioxidant activity which makes it a prospective multitarget drug candidate for treatment of neurodegenerative disorders.

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Kolossa, Marike, and Hasso Seibert. "Toxicity Testing by Means of Cryopreserved Bovine Spermatozoa." Alternatives to Laboratory Animals 19, no. 2 (April 1991): 204–8. http://dx.doi.org/10.1177/026119299101900210.

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The aim of the present study was to investigate the suitability of bovine spermatozoa cryopreserved in a “defined” medium as an in vitro model for the assessment of the cytotoxic potential of chemicals. The endpoints used for this purpose were motion activity and cellular ATP content. The evaluation of properties of cryopreserved sperm shortly after thawing and at the end of a one-hour incubation period, shows that the cryoprotective medium developed is able to provide suitable cellular material for cytotoxicity tests. Results from experiments employing substances with known modes of action are presented, and suggest that cryopreserved sperm can be used as successfully as native sperm in cytotoxicity tests.

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Blaauboer, Bas J. "Biokinetic and Toxicodynamic Modelling and its Role in Toxicological Research and Risk Assessment." Alternatives to Laboratory Animals 31, no. 3 (May 2003): 277–81. http://dx.doi.org/10.1177/026119290303100310.

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Toxicological risk assessment for chemicals is still mainly based on highly standardised protocols for animal experimentation and exposure assessment. However, developments in our knowledge of general physiology, in chemicobiological interactions and in (computer-supported) modelling, have resulted in a tremendous change in our understanding of the molecular mechanisms underlying the toxicity of chemicals. This permits the development of biologically based models, in which the biokinetics as well as the toxicodynamics of compounds can be described. In this paper, the possibilities are discussed of developing systems in which the systemic (acute and chronic) toxicities of chemicals can be quantified without the heavy reliance on animal experiments. By integrating data derived from different sources, predictions of toxicity can be made. Key elements in this integrated approach are the evaluation of chemical functionalities representing structural alerts for toxic actions, the construction of biokinetic models on the basis of non-animal data (for example, tissue–blood partition coefficients, in vitro biotransformation parameters), tests or batteries of tests for determining basal cytotoxicity, and more-specific tests for evaluating tissue or organ toxicity. It is concluded that this approach is a useful tool for various steps in toxicological hazard and risk assessment, especially for those forms of toxicity for which validated in vitro and other non-animal tests have already been developed.

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Walum, Erik, Anna Forsby, Cecilia Clemedson, and Björn Ekwall. "Dynamic Qualities of Validation and the Evolution of New In Vitro Toxicological Tests." Alternatives to Laboratory Animals 24, no. 3 (June 1996): 333–38. http://dx.doi.org/10.1177/026119299602400306.

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This review summarises some aspects of the dynamics of the evolution of toxicological test methods based on cell biology. Within the multicentre evaluation of in vitro cytotoxicity (MEIC) programme, some general principles for validation have been proposed: a) a human database should be used as the source of reference data in the validation of new methods aimed at predicting human toxicity; b) the relevance of a test should be determined before its reliability is assessed; c) a parallel validation of methods is preferable to a serial validation; and d) toxicokinetic data should be included in the validation process to improve the predictivity of cytotoxicity test results. These toxicokinetic data can be used to extrapolate the cytotoxic in vitro concentrations to provide human toxic exposure levels. As part of test development, the cytotoxic concentration can also be compared directly with the critical toxic human blood or tissue concentration. This approach is being explored in the ERGATT/CFN integrated toxicity testing scheme (ECITTS) prevalidation project. The critical toxic concentrations are determined by using a set of neurospecific cellular tests, chosen and combined on the basis of knowledge of common neurotoxicity mechanisms. Another approach to selecting tests for prevalidation is through the development of tests that are found to be necessary and complementary to existing tests. Such a programme has been initiated on the basis of the results of the MEIC study. The progress made so far in this “missing tests programme” is presented in this paper.

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Araújo, Gabrielle Luck de, Maria Augusta Amaral Campos, Maria Anete Santana Valente, Sarah Cristina Teixeira Silva, Flávia Dayrell França, Miriam Martins Chaves, and Carlos Alberto Tagliati. "Alternative methods in toxicity testing: the current approach." Brazilian Journal of Pharmaceutical Sciences 50, no. 1 (March 2014): 55–62. http://dx.doi.org/10.1590/s1984-82502011000100005.

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Alternative methods are being developed to reduce, refine, and replace (3Rs) animals used in experiments, aimed at protecting animal welfare. The present study reports alternative tests which are based on the principles of the 3Rs and the efforts made to validate these tests. In Europe, several methodologies have already been implemented, such as tests of irritability, cell viability, and phototoxicity as well as in vitro mathematical models together with the use of in silico tools. This is a complex process that spans from development to regulatory approval and subsequent adoption by various official entities. Within this regulatory framework is REACH, the European Community Regulation for chemicals and their safe use. In Brazil, the BraCVAM (Brazilian Center for the Validation of Alternative Methods) was recently established to validate alternative methods and stimulate incorporation of new methodologies. A new vision of toxicology is emerging for the 21st century (Tox-21), and the subsequent changes are shaping a new paradigm.

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Hellberg, Sven, Lennart Eriksson, Jörgen Jonsson, Fredrik Lindgren, Michael Sjöström, Svante Wold, Björn Ekwall, et al. "Analogy Models for Prediction of Human Toxicity." Alternatives to Laboratory Animals 18, no. 1_part_1 (November 1990): 103–16. http://dx.doi.org/10.1177/026119299001800114.1.

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Estimating the toxicity to humans of chemicals by testing on human subjects is not considered to be ethically acceptable, and toxicity testing on laboratory animals is also questionable. Therefore, there is a need for alternative methods that will give estimates of various aspects of human toxicity. Batteries of in vitro tests, together with physicochemical and toxicokinetic data, analysed by efficient data analytical methods, may enable analogy models to be constructed that can predict human toxicity. It may be possible to model non-specific toxicity relating to lipophilicity, or basal cytotoxicity, for a series of diverse compounds with large variation in chemical structure and physicochemical properties. However, local models for a series of similar compounds are generally expected to be more accurate, as well as being capable of modelling more-specific interactions. Analogy models for the prediction of human toxicity are discussed and exemplified with physicochemical and cytotoxicity data from the first ten chemicals in the multicenter evaluation of in vitro cytotoxicity (MEIC) project.

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Calleja, Mabel C., and Guido Persoone. "Cyst-Based Toxicity Tests. IV. The Potential of Ecotoxicological Tests for the Prediction of Acute Toxicity in Man as Evaluated on the First Ten Chemicals of the MEIC Programme." Alternatives to Laboratory Animals 20, no. 3 (July 1992): 396–405. http://dx.doi.org/10.1177/026119299202000306.

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In the framework of the multicentre evaluation of in vitro cytotoxicity (MEIC) programme, the first ten chemicals of the prescribed list were tested for acute toxicity in four standardised cyst-based aquatic invertebrate tests, consisting of two rotifer species (the estuarine Brachionus plicatilis and the freshwater Brachionus calyciflorus) and two crustacean species (the halophilic anostracan Artemia salina and the freshwater anostracan Streptocephalus proboscideus). Mortality was the test criterion and toxic effects, expressed as 24-hour LC50 values, were correlated with rodent and human acute oral toxicity data. Generally, a good correlation was obtained between any of the invertebrate tests and the rodent data. Likewise, the predictive screening potential of the aquatic invertebrate tests for acute oral toxicity in man was slightly better than the rat test for eight (excluding diazepam and digoxin) and nine (including diazepam, excluding digoxin) of the ten substances. The aquatic test systems, however, appear to be more suitable for compounds soluble in water.

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Phalen, Robert F. "Commentary on ‘‘Toxicity Testing in the 21st Century: A vision and a Strategy’’." Human & Experimental Toxicology 29, no. 1 (January 2010): 11–14. http://dx.doi.org/10.1177/0960327109354660.

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Toxicity Testing in the 21st Century: A Vision and a Strategy (NRC, 2007) presents a bold plan for chemical toxicity testing that replaces whole-animal tests with cell-culture, genetic, other in-vitro techniques, computational methods, and human monitoring. Although the proposed vision is eloquently described, and recent advances in in-vitro and in-silico methods are impressive, it is difficult believe that replacing in-vitro testing is either practical or wise. It is not clear that the toxicity-related events that occur in whole animals can be adequately replicated using the proposed methods. Protecting public health is a serious endeavor that should not be limited by denying animal testing. Toxicologists and regulators are encouraged to read the report, carefully consider its implications, and share their thoughts. The vision is for too important to ignore.

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Sbarbati-DelGuerra, Rosella, Simona Maltinti, Piero Cerrai, Mario Tricoli, and Giulio D. Guerra. "In Vitro Cytotoxicity Testing of a Candidate Drug Delivery Material." Alternatives to Laboratory Animals 24, no. 4 (August 1996): 573–80. http://dx.doi.org/10.1177/026119299602400418.

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Samples of a newly synthesised class of macromolecular compound, poly(N-allyl acrylamide)s, obtained by thermal condensation at 200°C of a polyelectrolyte complex, polyfallylammonium acrylate), were tested for both cytotoxicity and water absorption. The cytotoxicity tests (Level I), carried out on both an unmodified complex and a fully condensed polyamide, indicate that these materials exert no acute toxicity on 3T3 fibroblast cells. The water absorption tests, carried out on both partially condensed and fully condensed polyamides, at three different pH values, indicate that these materials are suitable to be used for the controlled delivery of drugs containing ionic or polar groups.

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Dreanca, Alexandra, Codruta Sarosi, Alina Elena Parvu, Mihai Blidaru, George Enacrachi, Robert Purdoiu, Andras Nagy, et al. "Systemic and Local Biocompatibility Assessment of Graphene Composite Dental Materials in Experimental Mandibular Bone Defect." Materials 13, no. 11 (May 31, 2020): 2511. http://dx.doi.org/10.3390/ma13112511.

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The main objective of this research is to demonstrate the biocompatibility of two experimental graphene dental materials by in vitro and in vivo tests for applications in dentistry. The novel graphene dental materials, including one restorative composite and one dental cement, were subjected to cytotoxicity and implantation tests by using a rat model of a non-critical mandibular defect. In vitro cytotoxicity induced by materials on human dental follicle stem cells (restorative composite) and dysplastic oral keratinocytes (dental cement) was investigated at 37 °C for 24 h. After in vivo implantation, at 7 weeks, bone samples were harvested and subjected to histological investigations. The plasma biochemistry, oxidative stress, and sub-chronic organ toxicity analysis were also performed. The resulting cytotoxicity tests confirm that the materials had no toxic effects against dental cells after 24 h. Following graphene dental materials implantation, the animals did not present any symptoms of acute toxicity or local inflammation. No alterations were detected in relative organ weights and in correlation with hepatic and renal histological findings. The materials’ lack of systemic organ toxicity was confirmed. The outcomes of our study provided further evidence on the graphene dental materials’ ability for bone regeneration and biocompatibility.

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Danzeisen, Ruth, David Lee Williams, Vanessa Viegas, Michael Dourson, Steven Verberckmoes, and Arne Burzlaff. "Bioelution, Bioavailability, and Toxicity of Cobalt Compounds Correlate." Toxicological Sciences 174, no. 2 (February 14, 2020): 311–25. http://dx.doi.org/10.1093/toxsci/kfz249.

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Abstract Based on the wide use of cobalt substances in a range of important technologies, it has become important to predict the toxicological properties of new or lesser-studied substances as accurately as possible. We studied a group of 6 cobalt substances with inorganic ligands, which were tested for their bioaccessibility (surrogate measure of bioavailability) through in vitro bioelution in simulated gastric and intestinal fluids. Representatives of the group also underwent in vivo blood kinetics and mass balance tests, and both oral acute and repeated dose toxicity (RDT) testing. We were able to show a good correlation between high in vitro bioaccessibility with high in vivo bioavailability and subsequent high in vivo toxicity; consequently, low in vitro bioaccessibility correlated well with low in vivo bioavailability and low in vivo toxicity. In vitro bioelution in simulated gastric fluid was the most precise predictor of the difference in the oral RDT lowest observed adverse effect levels of 2 compounds representing the highly and poorly bioaccessible subset of substances. The 2 compounds cobalt dichloride hexahydrate and tricobalt tetraoxide differed by a factor of 440 in their in vitro bioaccessibility and by a factor of 310 in their RDT lowest observed adverse effect level. In summary, this set of studies shows that solubility, specifically in vitro bioelution in simulated gastric fluid, is a good, yet conservative, predictor of in vivo bioavailability and oral systemic toxicity of inorganic cobalt substances. Bioelution data are therefore an invaluable tool for grouping and read across of cobalt substances for hazard and risk assessment.

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Walum, E. "A Comparison of Different Endpoints in Neuroblastoma Cell Toxicity Tests of Three Heavy Metal Compounds." Alternatives to Laboratory Animals 14, no. 3 (March 1987): 164–66. http://dx.doi.org/10.1177/026119298701400312.

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The cytotoxicities of HgCl2 (MC), CH3Hg Cl (MMC) and (C2H5)3SnCl (TET) were investigated in a mouse neuroblastoma cell line (C1300, clone NB41A3). General cytotoxicity was determined as TD25 and TD50 values in a cell detachment test and a monolayer growth test, respectively. Membrane toxicity was studied as increase in the efflux of tritiated 2-deoxy- D-glucose-6-phosphate from exposed cells, and metabolic effects as inhibition of oxygen consumption. It was found that the general endpoints (detachment and growth) correlated better to both animal and human in vitro toxicity than the more specific endpoints.

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Walum, Erik, Michael Balls, Vera Bianchi, Bas Blaauboer, George Bolcsfoldi, Andre Guillouzo, Gregory A. Moore, Lena Odland, Christoph Reinhardt, and Horst Spielmann. "ECITTS: An Integrated Approach to the Application of In Vitro Test Systems to the Hazard Assessment of Chemicals,." Alternatives to Laboratory Animals 20, no. 3 (July 1992): 406–28. http://dx.doi.org/10.1177/026119299202000307.

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As a result of a workshop held at Täljöviken, Åkersberga, Sweden, on 27–29 May 1991, a multicentre collaborative research project was established, with the purpose of developing the concept of integrated in vitro toxicity testing. The first priority was the selection of tests within eight appropriate areas: basal cytotoxicity, irritancy, developmental toxicity, hepatotoxicity, nephrotoxicity, immunotoxicity, neurotoxicity and biokinetics. An ideal battery of tests for each area was identified. Since it was realised that it would not be feasible to include the full ideal list of tests in the project, a minimum test list was also agreed. For each area, ten calibration chemicals were selected. From these 80 compounds, 30 were selected for inclusion, together with 20 of the OECD test programme chemicals, in a first test set of chemicals. The toxicity of these 50 test set chemicals will be investigated in the minimum integrated test scheme. The aim of the project is to employ non-animal methods to assess the toxicological properties of chemicals, and to improve this assessment through the use of knowledge about mechanisms of toxic action. The information produced will contribute to the establishment of a more-scientific and more-efficient toxicological procedure for hazard assessment. Questions concerning which parameters need to be investigated and combined to make hazard assessments, and which parameters relevant to in vivo toxicity can be determined in non-whole animal test systems, will also be addressed.

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Ekwall, Björn, Inger Bondesson, José V. Castell, Maria José Gómez-Lechón, Sven Hellberg, Johan Högberg, Ramiro Jover, et al. "Cytotoxicity Evaluation of the First Ten MEIC Chemicals: Acute Lethal Toxicity in Man Predicted by Cytotoxicity in Five Cellular Assays and by Oral LD50 Tests in Rodents." Alternatives to Laboratory Animals 17, no. 2 (December 1989): 83–100. http://dx.doi.org/10.1177/026119298901700205.

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The MEIC (multicentre evaluation of in vitro cytotoxicity) programme is a five-year programme to validate in vitro tests for general toxicity, and is organised by the Scandinavian Society for Cell Toxicology. Interested laboratories are invited, on an international basis, to test 50 published reference chemicals in their respective assays. Submitted results will then be evaluated yearly by the MEIC Committee for their relevance to various types of human toxicity, including an evaluation for the same chemicals of the prediction by animal tests of human toxicity. To establish the validation methods, a preliminary validation cycle is being performed in 1989/90 with submitted results for the first ten MEIC chemicals. The present paper is the very first step of this preliminary validation process. The prediction of human toxicity by five cytotoxicity assays (altogether 14 different cell systems/endpoints) has been evaluated, and also compared with the predictive value of rodent LD50 tests. Mouse LD50 prediction of human lethal dosage for these substances was good, while rat LD50 prediction was less satisfactory. The collective predictions by all 14 cell systems/endpoints of human toxicity in the form of a multivariate PLS (partial least squares) model of human acute lethal blood concentrations, as well as the corresponding prediction by a HeLa cell assay, were comparable to the efficiency of mouse LD50 prediction of human lethal dosage. When combined with simple toxicokinetic data (absorption of chemicals in the intestine and distribution volumes), the PLS model and the HeLa assay were able to predict human lethal dosage of the ten chemicals as accurately as the mouse LD50 value. The small number of chemicals studied to date means that general conclusions cannot be drawn from these results. Further validation of more chemicals with the in vitro methods is essential and promises to be worthwhile.

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Journal articles on the topic 'In vitro toxicity tests'

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