An increasing number of in vitro tests are being proposed as alternatives for animals in ocular safety testing. These in vitro systems use a variety of approaches and measure a broad range of endpoints, including cellular cytotoxicity and metabolism. We will briefly review the development and validation of these model systems. Several tests appear promising, and may prove useful as adjuncts in our current ocular safety assessment schemes. We will also discuss how these tests can most rapidly be brought to use in ocular safety assessment, and where efforts should be directed to develop more-highly predictive in vitro model systems for the future.
ScaifeM.C. (1982). An investigation of detergent action on cells in vitro and possibe correlations with in vivo data.International Journal of Cosmetic Science4, 179–193.
2.
ScaifeM.C. (1985). An in vitro cytotoxicity test to predict the ocular irritation potential of detergents and detergent products.Food and Chemical Toxicology23, 253–258.
3.
ReinhardtC.A., AeschbacherM., BracherM., and SpenglerJ. (1987). Validation of three cell toxicity tests and the Hen's egg test with guinea-pig eye and human skin irritation data. In In vitro toxicology. Alternative Methods in Toxicology, Vol. 5, Approaches to Validation (ed. GoldbergA.M.) pp. 463–469. New York: Mary Ann Liebert.
4.
AeschbbacherM., ReinhardtC.A., and ZbindenG. (1986). A rapid cell membrane permeability test using fluorescent dyes and flow cytometry.Cell Biology and Toxicology2, 247–255.
5.
ShadduckJ.A., EverittJ., and BayP.H.S. (1985). Use of in vitro cytotoxicity to rank ocular irritation of six surfactants. In A Progress Report from the Johns Hopkins Center for Alternatives to Animal Testing. Alternative Methods in Toxicology, Vol. 3, (ed. GoldbergA.M.), pp. 641–649. New York: Mary Ann Liebert.
6.
ShadduckJ.A., RenderJ., EverittJ., MeccoliR.A., & Essex-SorlieD. (1987). An approach to validation: comparison of six materials in three tests. In Alternative Methods in Toxicology, Vol. 5, Approaches to Validation (ed. GoldbergA.M.), pp. 75–78. New York: Mary Ann Liebert.
7.
BorenfreundE., and ShopsisC. (1985). Toxicity monitored with a correlated set of cell-culture assays.Xenobiotica15, 705–711.
8.
ShopsisC., BorenfreundE., WalbergJ., and StarkD.M. (1985). A battery of potential alternatives to the Draize test: uridine uptake inhibition, morphological cytotoxicity, macrophage chemotaxis and exfoliative cytology.Food and Chemical Toxicology23, 259–266.
9.
StarkD.M., BorenfreundE., WalbergJ., and ShopsisC. (1985). Comparison of several alternative assays for measuring potential toxicants. In A Progress Report From The Johns Hopkins Center for Alternatives to Animal Testing. Alternative Methods in Toxicology, Vol. 3, (ed. GoldbergA.M.), pp. 373–390. New York: Mary Ann Liebert.
10.
ShopsisC. (1984). Inhibition of uridine uptake in cultured cells: a rapid, sublethal cytotoxicity test.Journal of Tissue Culture Methods9, 19–22.
11.
ShopsisC., and SatheS. (1984). Uridine uptake inhibition as a cytotoxicity test: correlations with the Draize test.Toxicology29, 195–206.
12.
BorenfreundE., ShopsisC., BorreroO., and SatheS. (1983). In vitro alternative irritancy assays: comparison of cytotoxic and membrane transport effects of alcohols.Annals of the New York Academy of Sciences407, 416–419.
13.
BorenfreundE., and BorreroO. (1984). In vitro assays: potential alternatives to the Draize ocular irritancy test.Cell Biology and Toxicology1, 55–65.
14.
BorenfreundE., and PuernerJ.A. (1984). A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NR-90).Journal of Tissue Culture Methods9, 7–9.
15.
BorenfreundE., and PuernerJ.A. (1985). Toxicity determined in vitro by morphological alterations and neutral red absorption.Toxicology Letters24, 119–124.
16.
BorenfreundE., and PuernerJ.A. (1986). Cytotoxicity of metals, metal-metal and metal-chelator combinations assayed in vitro.Toxicology39, 121–134.
17.
BabichH., ShopsisC., and BorenfreundE. (1986). In vitro cytotoxicity testing of aquatic pollutants (cadmium, copper, zinc, nickel) using established fish cell lines.Ecotoxicology and Environmental Safety11, 91–99.
18.
ReaderS.J., BlackwellV., O'HaraR., ClothierR.H., GriffinG., and BallsM. (1989). A vital dye release method for assessing the short-term cytotoxic effects of chemicals and formulations.ATLA17, 28–37.
19.
BulichA.A. (1986). Bioluminescence assays. In Toxicity Testing Using Micro Organisms, Vol. I (ed. BittonG., DutkaB.J.), pp. 57–74. Florida: CRC Press.
20.
BrunerL.H., and ParkerR.D. (1990). Evaluation of six in vitro alternatives for ocular irritancy testing.Toxicologist10, 259–259.
21.
ReinhardtC.A., PelliD.A., and ZbindenG. (1985). Interpretation of cell toxicity data for the estimation of potential irritation.Food and Chemical Toxicology23, 247–252.
22.
North-RootH., YackovichF., DemetruliasJ., GaculaM. Jr., and HeinzeJ.E. (1982). Evaluation of an in vitro cell toxicity test using rabbit corneal cells to predict the eye irritation potential of surfactants.Toxicology Letters14, 207–212.
23.
North-RootH., YackovichF., DemetruliasJ., GaculaM. Jr., and HeinzeJ.E. (1985). Prediction of the eye irritation potential of shampoos using the in vitro SIRC cell toxicity test.Food and Chemical Toxicology23, 271–273.
24.
ShopsisC., and EngB. (1985). Rapid cytotoxicity testing using a semi-automated protein determination on cultured cells.Toxicology Letters26, 1–8.
25.
BallsM., and HornerS.A. (1985). The FRAME interlaboratory programme on in vitro cytotoxicology.Food and Chemical Toxicology23, 209–213.
26.
KnoxP., UphillP.F., FryJ.R., BenfordJ., and BallsM. (1986). The FRAME multicentre project on in vitro cytotoxicology.Food and Chemical Toxicology24, 457–463.
27.
HornerS.A., FryJ.R., ClothierR.H., and BallsM. (1985). A comparison of two cytotoxicity assays for the detection of metabolism-mediated toxicity in vitro: a study with cyclophosphamide.Xenobiotica15, 681–686.
28.
SilvermanJ. (1983). Preliminary findings on the use of protozoa (Tetrahymena thermophila) as models for ocular irritation testing in rabbits.Laboratory Animal Science33, 56–59.
29.
SilvermanJ., and PennisiS. (1987). Evaluation of Tetrahymena thermophila as an in vitro alternative to ocular irritation studies in rabbits.Journal of Toxicology — Cutaneous and Ocular Toxicology6, 33–42.
30.
TchaoR. (1988). Trans-epithelial permeability of fluorescein in vitro as an assay to determine eye irritants. In Progress in In Vitro Toxicology. Alternative Methods in Toxicology, Vol. 6, (ed. GoldbergA.M.), pp. 271–283. New York: Mary Ann Liebert.
31.
LeightonJ., NassauerJ., and TchaoR. (1985). The chick embryo in toxicology: an alternative to the rabbit eye.Food and Chemical Toxicology23, 293–298.
32.
LeightonJ., NassauerJ., TchaoR., and VerdoneJ. (1983). Development of a procedure using the chick egg as an alternative to the Draize rabbit test. In Product Safety Evaluation. Alternative Methods in Toxicology, Vol. 1, (ed. GoldbergA.M.) pp. 165–177. New York: Mary Ann Liebert.
33.
BoomanK.A., De ProspoJ., DemetruliasJ., DriedgerA., GriffithJ.F., GrochoskiG., KongB.M., McCormickW.C., North-RootH., RozenM.G., and SedlakR.I. (1989). The SDA alternatives program: comparison of in vitro data with Draize test data.Journal of Toxicology — Cutaneous and Ocular Toxicology8, 35–49.
34.
LuepkeN.P., and WallatS. (1987). HET-CAM-reproducibility studies. In In vitro toxicology: Alternative Methods in Toxicology Vol. 5, Approaches to validation (ed. GoldbergA.M.), pp. 353–363. New York: Mary Ann Liebert.
35.
LuepkeN.P. (1986). The hen's egg test (HET)—an alternative toxicity test.British Journal of Dermatology Suppl.115, 133–135.
36.
LeupkeN.P., and KemperF.H. (1986). The HET-CAM test: an alternative to the Draize eye test.Food and Chemical Toxicology24, 495–496.
37.
LuepkeN.P. (1985). Hen's egg chorio-allantoic membrane test for irritation potential.Food and Chemical Toxicology23, 287–291.
38.
LuepkeN.P. (1985). HET-Chorionallantois-Test: an alternative to the Draize rabbit eye test. In In Vitro toxicology. Alternative Methods in Toxicology Vol. 3, A progress report for The Johns Hopkins Center for Alternatives to Animal Testing (ed. GoldbergA.M.), pp. 593–605. New York: Mary Ann Lierbert.
39.
BurtonA.B., YorkM., and LawrenceR.S. (1981). The in vitro assessment of severe eye irritants.Food and Cosmetic Toxicology19, 471–480.
40.
PriceJ.B., and AndrewsI.J. (1985). The in vitro assessment of eye irritancy using isolated eyes.Food and Chemical Toxicology23, 313–315.
41.
WeteringsP.J., & Van ErpY.H.M. (1987). Validation of the BE/CAM assay — an eye irritancy screening test. In In vitro toxicology, Alternative Methods in Toxicology, Vol. 5, Approaches to Validation (ed. GoldbergA.M.), pp. 515–521. New York: Mary Ann Liebert.
42.
MuirC.K., FlowerC., & Van AbbeN.J. (1986). The effect of shampoos on rabbit ileum in vitro compared to eye irritancy in vivoJournal of the American College of Toxicology.5, 113–116.
43.
MuirC.K., FlowerC., & Van AbbeN.J. (1983). A novel approach to the search for in vitro alternatives to in vivo eye irritancy testing.Toxicology Letters18, 1–5.
44.
MuirC.K. (1983). The toxic effect of some industrial chemicals on rabbit ileum in vitro compared with eye irritancy in vivo.Toxicology Letters19, 309–312.
45.
OliverG.J., and PembertonM.A. (1985). An in vitro epidermal slice technique for identifying chemicals with potential for severe cutaneous effects.Food and Chemical Toxcicology23, 229–232.
46.
MuirC.K. (1984). A simple method to assess surfactant-induced bovine corneal opacity in vitro: preliminary findings.Toxicology Letters22, 199–203.
47.
MuirC.K. (1985). Opacity of bovine cornea in vitro induced by surfactants and industrial chemicals compared with ocular irritancy in vivo.Toxicology Letters24, 157–162.
48.
BrooksD., and MauriceD. (1987). A simple fluorometer for use with a permeability screen. In In vitro toxicology, approaches to validation. Alternative Methods in Toxicology, Volume 5, (ed. GoldbergA.M.) pp. 173–177. Mary Ann Liebert, Inc., New York.
49.
VerstraetenT.V., WilcoxD.K., FribergT.R., and ReelC. (1990). Effects of silicon oil and hyaluronic acid on cultured human retinal pigment epitheliumInvestigative Ophthalmology and Visual Science31, 102–107.
50.
HafemanD.G., ParceW.J., and McConnellH.M. (1988). Light-addressable potentiometric sensor for biochemical systems.Science240, 1182–1185.
51.
ParceJ.W., OwickiJ.C., KercsoK.M., SigalG.B., WadaH.G., MuirV.C., BousseL.J., RossK.L., SikicB.I., and McConnellH.M. (1989). Detection of cell-affecting agents with a silicon biosensor.Science246, 243–247.
52.
ChanK.Y. (1985). An in vitro alternative to the Draize test. In In vitro toxicology. Alternative Methods in Toxicology Vol. 3, A Progress Report from The Johns Hopkins Center for Alternatives to Animal Testing (ed. GoldbergA.M.), pp. 407–422. New York: Mary Ann Liebert.
53.
ChanK.Y. (1986). Chemical injury to an in vitro ocular system: differential release of plasminogen activator.Current Eye Research5, 357–362.
54.
JumblattM.M., and NeufeldA.H. (1986). A tissue culture assay of corneal epithelial wound closure.Investigative Ophthalmology and Visual Science. Vis. Sci.27, 8–13.
55.
SimmonsS.J., JumblattM.M., and NeufeldA.H. (1987). Corneal epithelial wound closure in tissue culture: an in vitro model of ocular irritancy.Toxicology and Applied Pharmacology88, 13–23.
56.
JumblattM.M., and NeufeldA.H. (1985). A tissue culture model of the human corneal epithelium. In In Vitro Toxicology, Alternative Methods in Toxicology Vol. 3, (ed. GoldbergA.M.), pp. 393–404. New York: Mary Ann Liebert.
57.
ChanK.Y., PattonD., and CosgroveY.T. (1989). Time-lapse videomicroscopic study of in vitro wound closure in rabbit corneal cells.Investigative Ophthalmology30, 2488–2498.
58.
FrazierJ.M., GadS.C., GoldbergA.M., and McCulleyJ.P. (1987). In Alternative Methods in Toxicology Vol. 4, A Critical Evaluation of Alternatives to Acute Ocular Irritation Testing (ed. GoldbergA.M.), pp. 1–119New York: Mary Ann Liebert.
59.
KellyC.P. (1989). EYTEX: An in vitro method of predicting ocular safety.Pharmacopeial Forum15, 4815–4824.
60.
SotoR.J., ServiM.J., and GordonU.C. (1989). Evaluation of an alternative method for ocular irritaiton. In Alternative Methods in Toxicology Vol. 7, In Vitro Toxicology: New Directions (ed. GoldbergA.M.), pp. 289–296. New York: Mary Ann Liebert.
61.
GordonV.C., KellyC.P., and BergmanH.C. (1989). Applications of the EYTEX Method for determination of the ocular irritation of surfactant formulations. In Alternative methods in Toxicology Vol. 7, In vitro toxicology: New directions. (ed. GoldbergA.M.), pp. 337–345. New York: Mary Ann Liebert.
62.
WeilC.S., and ScalaR.A. (1971). Study of intra-and interlaboratory variability in the results of rabbit eye and skin irritation tests.Toxicology and Applied Pharmacology19, 276–360.
63.
PatrickE., MaibachH.I., and BurkhalterA. (1985). Mechanisms of chemically induced skin irritation. I. Studies of time-course, dose-response, and components of inflammation in the laboratory mouse.Toxicology and Applied Pharamacology81, 476–490.
64.
PatrickE., BurkhalterA., and MaibachH.I. (1987). Recent investigations of mechanisms of chemically-induced skin irritation in laboratory mice.Journal of Investigative Dermatology88, 24–31.
65.
JumblattM.M., SimmonsS.J., and NeufeldA.H. (1987). Corneal epithelial wound closure: A tissue culture model of ocular irritancy. In In Vitro Toxicology. Alternative Methods in Toxicology Vol. 5, Approaches to Validation (ed. GoldbergA.M.), pp. 139–145. New York: Mary Ann Liebert.
66.
BellE., IvarssonB., and MerrillC. (1979). Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro.Proceedings of the National Academy of Sciences of the United States of America76, 1274–1278.
67.
BellE., EhrlichH.P., ButtleD.H., and NakatsujiT. (1981). Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness.Science211, 1052–1054.
68.
WeinbergC.B., and BellE. (1986). A blood vessel model constructed from collagen and cultured vascular cells.Science231, 397–400.
69.
StephensT.J., SilberP., ReeceB., BryanB., LongD., PakesS., GayR., and HaimesH. (1990). Testskin: an in vitro tool for detecting cytotoxicity and inflammation.Toxicologist10, 79.
