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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:
23 May 2016 - 17 June 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

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

In vitro test system

Test system:
human skin model
Remarks:
reconstructed human epidermis
Source species:
human
Cell type:
other: human epidermal keratinocytes
Cell source:
foreskin from multiple donors
Vehicle:
unchanged (no vehicle)
Details on test system:
EpiskinTM model consists of an airlifted, living, multilayered epidermal tissue construction (surface 0.38 cm2) supplied by SkinEthic Laboratories, Lyon, France.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL- Amount(s) applied (volume or weight with unit): 10 µL of the test item.NEGATIVE CONTROL- Amount(s) applied (volume or weight): 10 µLPOSITIVE CONTROL- Amount(s) applied (volume or weight): 10 µL- Concentration (if solution): SDS at 5% (w/v).
Duration of treatment / exposure:
Exposure period of 15 minutes, followed by rinsing.
Duration of post-treatment incubation (if applicable):
42-hour recovery period.
Number of replicates:
Triplicate tissues for each tested substance (test item, negative control, positive control).

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
treated
Value:
109
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: mean cOD of the three negative controls should be = 0.6 and the Standard Deviation (SD) value of the % viability should be = 18%,
- Acceptance criteria met for positive control: relative mean viability of the positive control should be = 40% of the relative mean viability of the negative control and the SD value of the % viability should be = 18%.

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 42 hours of recovery period, the relative mean viability of the tissues treated with the test item was 109% with a standard deviation of 2%.
As the mean viability was > 50% after the MTT reduction, theIL-1a concentration in culture media samples retained from the three negative controls and the three test item-treated tissues were analysed by ELISA.
The IL-1a concentration value of one tissue was found Below the Limit of Quantification (< 5.00 pg/mL). Consequently, the mean IL-1a concentration from the three test item-treated tissues was not calculated. The IL-1a concentration values of the two other test item-treated tissues were found < 60 pg/mL (5.04 and 20.0 pg/mL).
Therefore, the results met the criteria for an in vitro classification as non-irritant to skin.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the test item 1,10-decanediyl bis methacrylate is considered to be non-irritant to skin. According to the results of this study, the classification of the test item should be:- no Category (UN GHS and Regulation (EC) No. 1272/2008).
Executive summary:

The objective of this study was to evaluate the skin irritation potential of the test item,1,10-decanediyl bis methacrylate, using the Episkin 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).

 

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 +37°C, 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).

In addition, the concentration of the inflammatory mediator IL-1a was evaluated in the culture medium retained following the 42-hour recovery period. This quantification, based on an ELISA assay, was performed since the mean relative viability of the test item-treated tissues was > 50% following the MTT reduction assay.

 

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 42 hours of recovery period, the relative mean viability of the tissues treated with the test item was 109% with a standard deviation of 2%.

As the mean viability was > 50% after the MTT reduction, the IL-1a concentration in culture media samples retained from the three negative controls and the three test item-treated tissues were analysed by ELISA.

The IL-1a concentration value of one tissue was found Below the Limit of Quantification (< 5.00 pg/mL). Consequently, the mean IL-1a concentration from the three test item-treated tissues was not calculated. The IL-1a concentration values of the two other test item-treated tissues were found < 60 pg/mL (5.04 and 20.0 pg/mL).

Therefore, the results met the criteria for an in vitro classification as non-irritant to skin.

Conclusion

Under the experimental conditions of this study, the test item 1,10-decanediyl bis methacrylate is considered to be non-irritant to skin.