Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: There is no OECD Test Guideline for this study, but it was conducted according to GLP and sufficient information is available for interpretation of results.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Assessment of ocular irritation potential done using the SkinEthic reconstructed human corneal epithelium model (10 minute exposure)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report)
- Physical state: White powder
- Analytical purity: 99.5%
- Storage condition of test material: Room temperature in the dark.

Test animals / tissue source

Species:
other: SkinEthic reconstructed human corneal epithelium model
Strain:
other: SkinEthic reconstructed human corneal epithelium model
Details on test animals or tissues and environmental conditions:
The SkinEthic HCE model consists of transformed human corneal epithelial cells of the cell line HCE (LSU EYE Center, New Orleans, USA) that form a corneal epithelial tissue (mucosa), devoid of stratum, resembling, histologically, the mucosa of the human eye.
The test item is applied directly to the culture surface, at the air interface, so that undiluted and/or end use dilutions can be tested directly. The model consists of an airlifted, living, corneal tissue construct, produced in polycarbonate inserts in serum-free and chemically defined medium.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Negative and positive controls maintained
Amount / concentration applied:
30 mg
Duration of treatment / exposure:
10 minute
Observation period (in vivo):
3 hours
Number of animals or in vitro replicates:
Reconstructed human corneal epithelium
Details on study design:
The experimental design of the study consists of a test for direct reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) by the test item followed by the main test.
For the main test, triplicate SkinEthic tissues were treated with 30mg of the test item for 10 minutes. Triplicate tissues treated with 30 microliter of solution A served as the negative control and triplicate tissues treated with 30 microliter of 2% sodium dodecyl sulphate (SDS) served as a positive control. At the end of the exposure period each SkinEthic tissue was rinsed. The rinsed tissues (two per group) were taken for MTT loading. The remaining tissues were retained for possible histopathology. Following MTT loading the reduced MTT was extracted from the tissues. After extraction the absorbency of triplicate aliquots of the extracted MTT solution for each SkinEthic tissue was measured. The optical density was measured at 540 nm. Data are presented in the form of percentage viability (MTT conversion relative to negative controls).

Results and discussion

In vivo

Results
Irritation parameter:
other: relative mean tissue viability
Basis:
mean
Time point:
other: 10 minute exposure period
Max. score:
100.7
Reversibility:
other: Not applicable - In vitro testing
Remarks on result:
other: End point expressed as %
Irritant / corrosive response data:
The relative mean tissue viability of the test item treated tissues was 100.7% after a 10-minute exposure period.

The test item was classified according to the following criteria:
(1)If the percentage relative mean tissue viability was >60% the test item was considered to be non-irritant (NI)
(2)If the percentage relative mean tissue viability was <60% the test item was considered to be irritant (I)
Other effects:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The relative mean viability of the test item treated tissues after 10-minute exposure period was 100.7%.
The test item was considered to be Non-Irritant.
Executive summary:

The purpose of this test was to determine the eye irritation potential of the test item using the SkinEthic reconstructed Human Corneal Epithelium model (HCE, SkinEthic Laboratories, Nice, France) after a treatment period of 10 minutes. The test is based on the hypothesis that irritant chemicals are able to penetrate the corneal epithelial tissue and are sufficiently cytotoxic to cause cell death.

The experimental design of the study consists of a test for direct reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) by the test item followed by the main test.

For the main test, triplicate SkinEthic tissues were treated with 30mg of the test item for 10 minutes. Triplicate tissues treated with 30 microliter of solution A served as the negative control and triplicate tissues treated with 30 microliter of 2% sodium dodecyl sulphate (SDS) served as a positive control. At the end of the exposure period each SkinEthic tissue was rinsed. The rinsed tissues (two per group) were taken for MTT loading. The remaining tissues were retained for possible histopathology. Following MTT loading the reduced MTT was extracted from the tissues. After extraction the absorbency of triplicate aliquots of the extracted MTT solution for each SkinEthic tissue was measured. The optical density was measured at 540 nm. Data are presented in the form of percentage viability (MTT conversion relative to negative controls).

The test item was classified according to the following criteria:

(1)If the percentage relative mean tissue viability was >60% the test item was considered to be non-irritant (NI)

(2)If the percentage relative mean tissue viability was <60% the test item was considered to be irritant (I)

RESULTS : The relative mean viability of the test item treated tissues after 10-minute exposure period was 100.7%.

CONCLUSION: The test item was considered to be Non-Irritant.