2,2,3,3,8,8,9,9-octafluorotricyclo[8.2.2.2⁴,⁷]hexadeca-1(12),4,6,10,13,15-hexaene

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vitro

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:

2022

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 442E (In Vitro Skin Sensitisation assays addressing the key event on activation of dendritic cells on the Adverse Outcome Pathway for skin sensitisation)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

other: Human Cell Line Activation Test (h-CLAT) for Skin Sensitisation, DB-ALM Protocol n°158, July 23rd, 2015

GLP compliance:

yes

Type of study:

human Cell Line Activation Test (h-CLAT)

Test material

Test material information

1

Reference substance name:

2,2,3,3,8,8,9,9-octafluorotricyclo[8.2.2.2⁴,⁷]hexadeca-1(12),4,6,10,13,15-hexaene

EC Number:

639-791-8

Cas Number:

3345-29-7

Molecular formula:

C16H8F8

IUPAC Name:

2,2,3,3,8,8,9,9-octafluorotricyclo[8.2.2.2⁴,⁷]hexadeca-1(12),4,6,10,13,15-hexaene

Specific details on test material used for the study:

Batch No.: 340421
Purity: 98.5%

In vitro test system

Details of test system:

THP-1 cell line [442E]

Details on the study design:

Prior to the main study the cell batch was checked for its reactivity towards known positive and negative controls and was found to be acceptable for further testing.
In the present study PARYLENE HT® was dissolved in acetone. For the dose finding assay stock solutions with concentrations ranging from 140.63 mg/mL to 1.10 mg/mL were prepared by a serial dilution of 1:2. Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained with propidium iodide and cell viability was measured by FACS analysis.
Due to a lack of cytotoxicity, no CV75 could be derived. Therefore, the main experiment was performed covering the following concentration steps:
281.26, 234.38, 195.32, 162.77, 135.64, 113.03, 94.19, 78.49 µg/mL
Turbidity of the test item was observed in experiment 2 for all concentration steps when mixing the test item stock solutions with cell culture medium.
Cells were incubated with the test item for 24 h at 37°C. After exposure, cells were stained and cell surface markers CD54 and CD86 were measured by FACS analysis. Cell viability was assessed in parallel using propidium iodide staining.

Vehicle / solvent control:

other: acetone

Positive control:

dinitrochlorobenzene (DNCB) [442E]

Results and discussion

In vitro / in chemico

Results

Results 1

Key result

Group:

test chemical

Run / experiment:

run/experiment 2

Parameter:

EC150, CD86 [442E]

Value:

170 %

Cell viability:

Relative cell viability at the highest test item concentration was reduced to 92.2% (CD86), 90.9% (CD54) and 91.6% (isotype IgG1 control) in the first experiment and to 90.3% (CD86), 88.0% (CD54) and 90.0% (isotype IgG1 control) in the second experiment

Vehicle controls validity:

valid

Results 2

Key result

Group:

test chemical

Run / experiment:

run/experiment 1

Parameter:

EC150, CD86 [442E]

Value:

167 %

Cell viability:

Relative cell viability at the highest test item concentration was reduced to 92.2% (CD86), 90.9% (CD54) and 91.6% (isotype IgG1 control) in the first experiment and to 90.3% (CD86), 88.0% (CD54) and 90.0% (isotype IgG1 control) in the second experiment.

Vehicle controls validity:

valid

Any other information on results incl. tables

Prior to the main study the cell batch was checked for its reactivity towards known positive and negative controls and was found to be acceptable for further testing.

In the present study PARYLENE HT® was dissolved in acetone. For the dose finding assay stock solutions with concentrations ranging from 140.63 mg/mL to 1.10 mg/mL were prepared by a serial dilution of 1:2. Cells were incubated with the test item for 24 h at 37°C. After exposure cells were stained with propidium iodide and cell viability was measured by FACS analysis.

Due to a lack of cytotoxicity, no CV75 could be derived. Therefore, the main experiment was performed covering the following concentration steps:

281.26, 234.38, 195.32, 162.77, 135.64, 113.03, 94.19, 78.49 µg/mL

Turbidity of the test item was observed in experiment 2 for all concentration steps when mixing the test item stock solutions with cell culture medium.

Cells were incubated with the test item for 24 h at 37°C. After exposure, cells were stained and cell surface markers CD54 and CD86 were measured by FACS analysis. Cell viability was assessed in parallel using propidium iodide staining.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

In this study under the given conditions the test item did upregulate the cell surface marker in at least two independent experiment runs. Therefore, the test item can be considered as a skin sensitiser