Z-109

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 09 June 2009 and 11 June 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The assay has undergone inter-laboratory validation and has been shown to reliably detect test materials that are negligible or moderate to severe ocular irritants. The strain of rabbit used in these laboratories has been shown to produce satisfactory responses using known ocular-irritants and non-ocular irritants during in house validation. Study conducted to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

The purpose of this study was to determine the eye irritation potential of the test material using the SkinEthic Reconstituted Human Corneal 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.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Details on test material:

- Substance type: UVCB
- Physical state: liquid
- Analytical purity: ~90%
- Lot/batch No.: CM71217681
- Expiration date of the lot/batch: 27-Apr-2010
- Stability under test conditions: Unknown; is excluded from the statement of compliance.
- Storage condition of test material: At room temperature (range of 20 ± 5 °C), light protected.

Test animals / tissue source

Species:

other: SkinEthic Reconstituted Human Corneal model

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The SkinEthic RHC model consists of transformed human keratinocytes of the cell line HCE (LSU EYE Center, New Orleans, USA) that form a corneal epithelial tissue (mucosa), devoid of stratum corneum, resembling, histologically, the mucosa of the human eye.
On arrival, the SkinEthic HCE tissues (Day 6 cultures), were stored at room temperature prior to transferring into 24-well plates designated ‘arrival plates’ containing 300 μl of maintenance medium. It was important to ensure that there were no air bubbles present under the tissue inserts. The tissues were incubated overnight at 37°C, 5% CO2 in air.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Triplicate tissues for negative control group; triplicate tissues for positive control group

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
- Concentration (if solution): Undiluted


VEHICLE
- Amount(s) applied (volume or weight with unit): N/A
- Concentration (if solution): N/A
- Lot/batch no. (if required): N/A
- Purity: N/A

Duration of treatment / exposure:

10 min

Observation period (in vivo):

All the tissues were rinsed after 10 min treatment, and each tissue was placed into a pre-labelled 24-well plate containing 300 μl of maintenance medium (at room temperature for 3 h) .

Number of animals or in vitro replicates:

Triplicate tissues for negative control group;
Triplicate tissues for test material treated group;
Triplicate tissues for positive control group

Details on study design:

See information presented in the below section "Any ohter information on materials and methods incl. tables"

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

OD540

Run / experiment:

Relative mean viability of test material treated tissues (%)

Value:

81.7

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

The relative mean viability of the test material treated tissues after a 10 minute exposure was 81.7%. According to the protocol followed the test material was considered to be a Non-Irritant (NI).

Other effects:

The test material was not able to directly reduce MTT.
Control Tissue: The positive control result of 70.5% fell within the historical range of 12.4 to 75.0% for the positive control for this study type in this testing facility.

Any other information on results incl. tables

The test material was classified according to the following criteria:

i) If the percentage relative mean tissue viability was ≥ 60% the test material was considered to be non-irritant.

ii) If the percentage relative mean tissue viability was < 60% the test material was considered to be an irritant.

Results.The relative mean viability of the test material treated tissues after a 10 minute exposure was 81.7%.

It was considered unnecessary to proceed with tissue histopathology.

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

The test material was considered to be non-mutagenic under the conditions of this test.