Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 December 2017 to 15 December 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: paste

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
other: reconstructed human skin
Cell source:
other: EpiDerm TM (EPI-200-MatTek Corporation)
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm TM (EPI-200-MatTek Corporation)
- Tissue batch number(s): (Lot 25864)
- Delivery date: 12 December 2017
- Date of initiation of testing: no data

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)
- Temperature of post-treatment incubation: 42 hour post-dose incubation at 37°C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: not indicated (according to MatTEek SOP)

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE: no data (according to MatTEek SOP)
- MTT concentration: not specified
- Incubation time: no data
- Spectrophotometer: not identified measured at 570 nm
- Wavelength: 570 nm
- Filter: none
- Linear OD range of spectrophotometer: no data

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: yes (1.601 ± 0.007 (mean and SD of MTT value of 3 tissues exposed to H2O) )
- Barrier function: yes (ET 50% of tissues exposed to 100 µL Triton X-100 1% (n=3): 5.84 h)
- Morphology: yes ( Histological examination demonstrates human epidermis-like structure: including multiple layers (at least 4) of viable epithelial cells (basal layer, stratum spinosum, stratum granulosum) which are present under multilayered stratum corneum ; Tissue thickness: 128 µm )
- Contamination: No contamination reported
- Reproducibility: yes (The mean OD of the 6 wells containing extraction solvent alone (blanks) should be ≤ 0.1., but was 0.149 evaluated not to have affected the measurements, based upon historical system data for blanks; < 0.244)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µl

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl PBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl SDS
- Concentration (if solution): 5% in water
Duration of treatment / exposure:
60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)
Duration of post-treatment incubation (if applicable):
42 hour post-dose incubation at 37°C, 5% CO2
Number of replicates:
3/treatment

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
106.9
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
mean viability 4.5%
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
No colour interference or interference with MTT reduction as tested in a pre-test

Any other information on results incl. tables

Name

Code

Mean of OD

SD of OD

Mean of viability (%)

SD of viability (%)

CV %

Classification

DPBS

NC

1.768

0.210

100.000

11.904

11.904

Non-Irritant

SDS 5%

PC

0.080

0.016

4.535

0.909

20.046

Irritant

Substance

TA2

1.889

0.142

106.873

8.008

7.493

Non-Irritant

 

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is non-irritant in the EpiDermTM model
Executive summary:

The substance was tested for skin irritation in the EpiDermTM model according to OECD 439. 30 uL of the substance, positive control (5% SDS) and negative control (PBS) was applied to the tissue for 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2) and thereafter incubated for 42 hour post-dose at 37°C, 5% CO2. The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 106.9% of negative controls, while validity criteria were evaluated to be met. The substance is concluded to be non-irritant.