Registration Dossier

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

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:
liquid

In vitro test system

Test system:
human skin model
Vehicle:
unchanged (no vehicle)
Details on test system:
Three-dimensional human epidermis model
The EpiDermTM model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multi layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs ®, 10 mm ø) and commercially available as kits (EpiDerm™ 200) containing 24 tissues on shipping agarose.
Tissue model: EPI-200
Tissue Lot Number: 25800
Origin: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia
Irritation test:
On the day of arrival in the laboratory, the tissues were transferred to sterile 6-well plates with 0.9 mL assay medium and preconditioned in the incubator at 37°C. After 1 hour, the pre-incubation medium was replaced by fresh medium and reconditioning continued for 18 ± 3 hours. Three tissues were treated with the test substance, the PC and the NC, respectively. Thirty microliters (30 μL) undiluted liquid test substance were applied by using a pipette. Control tissues were concurrently treated with 30 μL sterile PBS (NC) or with 30 μL 5% SDS (PC). A nylon mesh was carefully placed onto the tissue surface afterwards. The tissues were kept under the laminar flow hood at room temperature for 25 minutes overall and for 35 minutes in the incubator.
The tissues were washed with sterile PBS to remove residual test material 1 hour after start of application. Rinsed tissues were blotted on sterile absorbent paper and transferred into new 6-well plates pre-filled with 0.9 mL fresh medium. When all tissues were rinsed the surface of each tissue was carefully dried with a sterile cotton swab. Subsequently, the tissues were placed into the incubator at 37°C for 24 ± 2 hours. After 24 ± 2 hours, the tissues were transferred into new 6-well plates pre-filled with
0.9 mL fresh medium and placed into the incubator for an additional 18 ± 2-hour post-incubation period.
After the post-incubation period, the assay medium was replaced by 0.3 mL MTT solution and the tissues were incubated in the incubator for 3 hours. After incubation, the tissues were washed with PBS to stop the MTT incubation. The formazan that was metabolically produced by the tissues was extracted by incubation of the tissues in isopropanol. The optical density at a wavelength of 570 nm (OD570) of the extracts was spectrophotometrically determined. Blank values were established of 4 microtiter wells filled with isopropanol for each microtiter plate.


Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
irritation test
Value:
110

Any other information on results incl. tables

Test substance

 

tissue 1

tissue 2

tissue 3

mean

SD

CV[%]

NC

mean OD570

1.756

1.639

1.739

1.711

 

 

viability

[% of NC]

102.6

95.8

101.6

100.0

3.7

3.7

16/0068-1

mean OD570

1.914

1.837

1.896

1.882

 

 

viability

[% of NC]

111.9

107.3

110.8

110.0

2.4

2.2

PC

mean OD570

0.016

0.038

0.044

0.043

 

 

viability

[% of NC]

2.7

2.2

2.5

2.5

0.2

9.6

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results observed and by applying the evaluation criteria, it was concluded that C9-11 Methacrylate does not show a skin irritation potential in the EpiDerm™ in vitro skin irritation and corrosion test strategy under the test conditions chosen.