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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 December 2013 -- 03 February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: EU Method B.46 (In vitro dermal irritation)
Deviations:
no
Qualifier:
according to
Guideline:
other: OECD Guideline 439 (In vitro dermal irritation)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: colourless liquid
Details on test material:
- Name of test material: Tricyclodecane dimethanol diacrylate
- Physical state: colourless liquid
- Storage conditions: at room temperature and protected from light

Test animals

Species:
other: reconstructed human epidermis
Strain:
other: not applicable
Details on test animals and environmental conditions:
EpiskinTM Model Kit (0.38 cm2 tissues) supplied by SkinEthic Laboratories, Lyon, France.
Medium and Incubation T°C: 37°C
Dates of experimental phase: from 22 January 2014 to 03 February 2014

Test system

Type of coverage:
other: not applicable (in vitro)
Preparation of test site:
other: not applicable (in vitro)
Vehicle:
unchanged (no vehicle)
Controls:
other: in vitro negative and positive controls
Amount / concentration applied:
Amount applied per tissue: 10 ± 2 µL.
Duration of treatment / exposure:
Exposure period of 15 minutes, followed by rinsing
Observation period:
MTT-loading after a 42h-incubation period following rinsing. Observation of MTT-> formazan transformation by viable cells.
Number of animals:
Not applicable
Triplicate for tested substance (test item, negative control, positive control).
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Rinsing: At the end of the treatment period, each tissue was removed from the well of the treatment plate, and rinsed with D-PBS. Rinsing was achieved by gently filling and emptying several times each tissue with D PBS to gently remove any residual test or control items.
The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well and the plates were incubated at +37°C, 5% CO2 in a humidified incubator for 42 hours.

POSITIVE CONTROL
Name: Sodium Dodecyl Sulphate (SDS) at a 5% (w/v) aqueous solution.

NEGATIVE CONTROL
Name: Dulbecco’s Phosphate-Buffered Saline (D-PBS).

SCORING SYSTEM:
- Optical density (OD) was measured at 570 nm:
Relative mean viability (%) = 100 x mean cOD(test item) / mean cOD(negative control)
where:
- mean cOD Negative Control = mean ODNC – mean ODblank
- mean cOD Test Item = mean ODTI – mean ODblank

Interpretation: see below

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
other: other: relative mean viability
Value:
109
Remarks on result:
other:
Remarks:
Basis: mean. Time point: 15 min exposure + 42h expression. Max. score: 100.0. Reversibility: other: not applicable. Remarks: 100% = control. (migrated information)

In vivo

Irritant / corrosive response data:

Following a 15 minutes exposure and a 42 hours recovery periods, the relative mean viability of the tissues treated with the test item was 109% with a standard deviation of 17% as assessed by the MTT assay. As the mean viability was > 50% after the MTT reduction, the results met the criteria for a non-irritant response.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: EU
Conclusions:
Under the experimental conditions of this study, the test item is considered to be non-irritant to skin.
Executive summary:

The objective of this study was to evaluate the skin irritation potential of the test item using the EpiskinTM reconstructed human epidermis model.

The study design was based upon international guidelines (OECD Guideline No. 439 and Commission Regulation (EC) No. 761/2009, B.46). The study was conducted in compliance with CiToxLAB’s standard operating procedures and the principles of Good Laboratory Practice.

 

Methods

 

Preliminary tests were performed to detect the ability of the test item to directly reduce MTT as well as its colouring potential.

Following the preliminary tests, the skin irritation potential of the test item was tested in the main test. The test item and both the negative and positive controls were applied topically on triplicate tissues and incubated at room temperature for 15 minutes. At the end of the treatment period, each tissue was rinsed with D-PBS and incubated for 42 hours at, 5% CO2 in a humidified incubator. The cell viability was then assessed by means of the colourimetric MTT reduction assay.

Relative viability values were calculated for each tissue and expressed as a percentage of the mean viability of the negative control tissues which was set at 100% (reference viability).

 

Results

Preliminary tests

In the preliminary tests, the test item was found not to have direct MTT reducing properties or colouring potential.

 

Main test

All acceptance criteria for the negative and positive controls were fulfilled. The study was therefore considered to be valid.

Following a 15 minutes exposure and a 42 hours recovery periods, the relative mean viability of the tissues treated with the test item was 109% with a standard deviation of 17% as assessed by the MTT assay. As the mean viability was > 50% after the MTT reduction, the results met the criteria for a non-irritant response.

Conclusion

 

Under the experimental conditions of this study, the test item is considered to be non-irritant to skin.

 

According to the results of this study, the classification of the test item should be:

- not classified (Directive 67/548/EEC) and no category (Regulation (EC) No. 1272/2008).