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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2014-02-17 to 2014-07-07
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well documented GLP study, no OECD Guideline available

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
A reconstructed three dimensional human cornea model is used in this test for determination of tissue distruction/viability using a colorimetric test (measurement of formazan production after incubation with a tetrazolium salt (MTT) as indication of the reduction of mitochondrial dehydrogenase activity).
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals / tissue source

Species:
other: an in vitro test system was used (human derived epidermal keratinocytes)
Details on test animals or tissues and environmental conditions:
The EpiOcularTM model (OCL-200) is a three-dimensional non-keratinized tissue construct composed of normal human derived epidermal keratinocytes used to model the human corneal epithelium. The EpiOcularTM tissues (surface 0.6 cm2) are cultured on especially prepared cell culture inserts and are commercially available as kits (EpiOculaTM 200). Supplier: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Positive (Methyl acetate), and negative controls (deionized water were applied to two tissues each.
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 50 µL


Duration of treatment / exposure:
6 hours, after removal of test substance another 18 hours postincubation period
Observation period (in vivo):
Observations were recorded after removal of test substance.
Number of animals or in vitro replicates:
Two tissues were treated with each the test substance, the PC and the NC.
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: To remove the test substance, the tissues were washed with sterile PBS. For this purpose the tissues were immersed and swiveled three times in each of three beakers filled with PBS. Washed tissues were immediately immersed into 12-well plates, pre-filled with 5 mL/well prewarmed medium (post-soak immersion) in order to remove residual test substance. After 25 minutes of post-soak immersion, each tissue was dried on absorbent paper and transferred to fresh 6-well plates filled with 1 mL/well pre-warmed medium.

TOOL USED TO ASSESS SCORE:
The optical density at a wavelength of 570 nm (OD570) of the extracts containing formazan was determined spectrophotometrically.

Results and discussion

In vivo

Results
Irritation parameter:
other: viability of cells
Basis:
mean
Time point:
other: after removal of test substance and incubation with MTT.
Remarks on result:
other: The mean viability of the test-substance treated tissues was 112 %.
Irritant / corrosive response data:
The EpiOcularTM eye irritation test showed the following results:
The test substance is not able to reduce MTT directly.
The mean viability of the test-substance treated tissues was 112 %.
Generally, the test substance is considered as irritant if the mean tissue viability is ≤ 60 % of the negative control and as non irritant if the viability lies above 60 % of the negative control. Therefore, the test substance is considered to be non-irritating to eyes.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU