Registration Dossier

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name: 3,4-Epoxycyclohexylmethyl methacrylate
- Chemical Name: 2-Propenoic acid, 2-methyl-7-oxabicyclo[4.1.0]hept-3-ylmethyl ester
- CAS No.: 82428-30-6
- Physical State / Colour: liquid / light yellow, transparent
- Density: 1.079 g/mL at 20 °C
- Molecular Weight: 196.2 g/mol

Test animals / tissue source

Species:
other: Bovine Corneas
Strain:
other: not applicable

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
750 µL
Duration of treatment / exposure:
10 min
Duration of post- treatment incubation (in vitro):
- 2 h (opacity measurement)
- 3.5 h total (determination of optical density)
Number of animals or in vitro replicates:
3 corneas
Details on study design:
- 3 corneas for the test item
- 3 corneas as negative controls treated with physiological saline 0.9% NaCl
- 3 corneas as positive control treated with ethanol 100%
- The BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Run / experiment:
mean test item
Value:
10.16
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
IVIS 2.08
Positive controls validity:
valid
Remarks:
IVIS 86.02

Any other information on results incl. tables

RESULTS

 Cornea No.  Test Item  Corr. Opacity  Corr. OD490  IVIS
 1  Neg. Control  0.00  0.027  
 2  Neg. Control  4.00  0.021  
 3  Neg. Control  1.00  0.035  
 mean  Neg. Control  1.67  0.028  2.08
 4  Pos. Control  50.33  1.919  
 5  Pos. Control  64.33  2.053  
 6  Pos. Control  53.33  2.031  
 mean  Pos. Control  56.00  2.001  86.02
 7  Test Item  13.33  -0.001  
 8  Test Item  9.33  -0.022  
 9  Test Item  8.33  -0.012  
 mean  Test Item  10.33  -0.011  10.16

The following mean in vitro irritation score was calculated: 10.16

No prediction can be made regarding the classification of the test substance 3,4-Epoxycyclohexylmethyl methacrylate according to the evaluation criteria. Further testing in another suitable method is required.

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

HISTOPATHOLOGY

Negative control samples did not show any histological lesions.

In positive control samples, the main lesions consisted of vacuolated corneal epithelia and single cell apoptosis. In one sample (PK1), there was a marked epithelial flattening/height reduction associated with a focal chronic injury. This corneal sample is deemed to be affected before study start by an inflammatory/degenerative that is supported by the presence of brownish pigment (hemosiderin). In sample PK 3, there was a slight laceration of the cuboidal basal cell layer.

In test item treated samples, there was slight multifocal apoptosis/necrosis in the epithelium of all specimens.

Applicant's summary and conclusion

Interpretation of results:
other: Irritation score of 10.16 and histopathological findings indicate slight signs of irritation
Remarks:
Criteria used for interpretation of results: BCOP standard evaluation criteria
Conclusions:
No prediction can be made regarding the classification of the test substance 3,4-Epoxycyclohexylmethyl methacrylate according to the evaluation criteria. Further testing in another suitable method is required.
Under the conditions of this study, 3,4-Epoxycyclohexylmethyl methacrylate caused indicators for irritation consisting of slight multifocal apoptosis/necrosis associated with multifocal vacuolation in the epithelium of all specimen.
Executive summary:

The eye irritancy potential of 3,4-Epoxycyclohexylmethyl methacrylate was investigated in the bovine corneal opacity and permeability assay.

- Preparation of the test item: tested as provided by the sponsor

- Mean in vitro irritation score: 10.16

- Classification: No prediction can be made

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.