Ocular Toxicity Tests
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I.N.1 I.N.2 I.N.3 I.N.4 I.N.5
I.N.6 I.N.7 I.N.8
General Considerations . . . . . . . . Dendritic Cell Culture . . . . . . . . . . Corneal Epithelial Organ Culture . . . . . . . . . . . . . . . . . . . . . . . . . Surface Biotinylation-Tight Junction Permeability Assay . . . Fluorescein Isothiocyanate-Dextran Retention. . . . . . . . . . . . . . . . . . . . . . . Electrophoretic Mobility Shift Assay. . . . . . . . . . . . . . . . . . . . . . . . . . . Murine Local Lymph Node Assay. . . . . . . . . . . . . . . . . . . . . . . . . . . The Draize Test . . . . . . . . . . . . . . . .
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tively good agreement between certain pre-clinical and clinical observations (Dart 2003). A comprehensive review of various in vitro and in vivo procedures has been published recently by Wilhelmus in 2001.
322 REFERENCES
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Dart J (2003) Corneal toxicity; the epithelium and stroma in iatrogenic and factitious disease. Eye 17:886–892 Wilhelmus KD (2001) The Draize Test. Surv Ophthalmol 45:493–515
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I.N.2 Dendritic Cell Culture PURPOSE AND RATIONALE
I.N.1 General Considerations Ocular drugs, cosmetics or all chemicals, which can come in contact with the eye carry the potential of harming various ocular structures. Thus, their possible toxic effects on ocular structures must be evaluated in tests before use in humans. At present, it has been suggested that this be done in 3 steps. First, a thorough review of the literature of similar compounds should be conducted to obtain theoretical knowledge about ocular toxicity. Second, tests of this substance are performed in various in vitro procedures to obtain possible serious toxic effects, which might stop further testing. Third, if no serious toxic effects have been observed tests are conducted in the eyes of one animal followed by subsequent tests in more animals. Fourth, the substance if an ocular drug will then be subjected to clinical trials. An important feature of these pre-clinical tests is that they correctly predict or are in agreement with findings in humans. A recent study concluded that there is indeed a relaIntroduced by Wolfgang H. Vogel, Jefferson Medical College, Thomas Jefferson University, Department of Pharmacology, Philadelphia, USA
This test uses an in vitro human dendritic cell culture to obtain information of the potential for various chemicals to induce allergic contact dermatitis. This test is used as an alternative to the Local Lymph node assay (LLNA) to minimize or replace the use of live animal testing for predicting skin sensitization (Kimber et al. 2002, see below). The test allows for evaluation of skin sensitization by examining the presence of cell surface markers on Periperal Blood Mononuclear Cell (PBMC)-derived dendritic cells (DC) that are known to be involved in the development of allergic contact dermatitis. PROCEDURE
The method contains three stages including first establishing a cell line followed by test chemical exposure and finally evaluated for expression of cell surface markers. To establish a cell line, human leukocyt
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