Reaction mass of dodecyl acrylate and tridecyl acrylate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vitro

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

In vitro sensitization: Dendritic Cell Line Activation Assay; Myeloid U 937 Skin Sensitization Test (MUSST).
The myeloid U937 skin sensitization test (MUSST) is a dendritic cell activation test to predict skin sensitizing potential. The test is performed using the human pro-monocytic cell line U937 as surrogate for dendritic cells. As readout, the change in the expression of the cell membrane marker CD86 measured by flow cytometry after 48 hours of the test substance exposure was determined.

GLP compliance:

yes

Remarks:

non-GLP, but study conducted under similar Quality assurance system.

Type of study:

activation of dendritic cells

Test material

Test material information

Constituent 1

Reference substance name:

2-Propenoic acid, C12-14-alkyl esters

EC Number:

282-516-8

EC Name:

2-Propenoic acid, C12-14-alkyl esters

Cas Number:

84238-60-8

Molecular formula:

Unspecified

IUPAC Name:

84238-60-8

Test material form:

other: liquid/clear, colorless

In vitro test system

Details on the study design:

The cytotoxicity of the test substance was evaluated by flow cytometry using propidium iodide staining after 48 hours exposure. For the purpose the CV75 value was derived from the concentration response curve. The CV75 is the estimated concentration that affords 75% cell viability and was determined to be 1297.8 µg/mL for Laurylacrylate 1214 under the chosen exposure conditions on U937 cells. In the main test, test substance was assessed at six final concentrations 2595.6 µg/mL, 1297.8 µg/mL, 648.9 µg/mL, 350.0 µg/mL and 324.0 µg/mL to be 162.2 µg/mL for Laurylacrylate 1214 under the chosen exposure conditions on U937 cells.
After 48 hours of exposure U937 cells were stained with FITC labeled anti-human-CD 86 antibody and propidium iodide and the fluorescence intensity was analyzed using flow cytometry. A test substance was predicted to have a dendritic cell activating potential when the marker expression exceeded the threshold of 1.2 with respect to vehicle treated cells (VC) at any tested sufficiently non-cytotoxic (cell viability >/= 70%) concentration in two experiments.
The strong sensitizer ethylene diamine (EDA, 70 µg/mL) was used as positive and lactic acid (LA, 200 µg/mL) as non-sensitizing negative control.

Results and discussion

In vitro / in chemico

Results

Key result

Run / experiment:

other: treated

Parameter:

other: CD86 expression

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

not applicable

Remarks on result:

no indication of skin sensitisation

Other effects / acceptance of results:

In summary, after 48 hours of exposure to test substance Laurylacrylate 1214 CD 86 expression was not induced in U937 cells at concentration between 162.2 to 2595.6 µg/mL affording at least 70% viability. From this it has to be concluded that test substance Laurylacrylate 1214 does not induce dendritic cell activation.

Applicant's summary and conclusion

Interpretation of results:

other: no induction dendritic cell activation.

Conclusions:

In summary, after 48 hours of exposure to test substance Laurylacrylate 1214 CD 86 expression was not induced in U937 cells at concentration between 162.2 to 2595.6 µg/mL affording at least 70% viability. From this it has to be concluded that test substance Laurylacrylate 1214 does not induce dendritic cell activation.

Executive summary:

The myeloid U937 skin sensitization test (MUSST) is a dendritic cell activation test to predict skin sensitizing potential. The test is performed using the human pro-monocytic cell line U937 as surrogate for dendritic cells. As readout, the change in the expression of the cell membrane marker CD86 measured by flow cytometry after 48 hours of the test substance exposure was determined.

The cytotoxicity of the test substance was evaluated by flow cytometry using propidium iodide staining after 48 hours exposure. For the purpose the CV75 value was derived from the concentration response curve. The CV75 is the estimated concentration that affords 75% cell viability and was determined to be 1297.8 µg/mL for Laurylacrylate 1214 under the chosen exposure conditions on U937 cells. In the main test, test substance was assessed at six final concentrations 2595.6 µg/mL, 1297.8 µg/mL, 648.9 µg/mL, 350.0 µg/mL and 324.0 µg/mL to be 162.2 µg/mL for Laurylacrylate 1214 under the chosen exposure conditions on U937 cells.

After 48 hours of exposure U937 cells were stained with FITC labeled anti-human-CD 86 antibody and propidium iodide and the fluorescence intensity was analyzed using flow cytometry. A test substance was predicted to have a dendritic cell activating potential when the marker expression exceeded the threshold of 1.2 with respect to vehicle treated cells (VC) at any tested sufficiently non-cytotoxic (cell viability >/= 70%) concentration in two experiments. The strong sensitizer ethylene diamine (EDA, 70 µg/mL) was used as positive and lactic acid (LA, 200 µg/mL) as non-sensitizing negative control.

In summary, after 48 hours of exposure to test substance Laurylacrylate 1214 CD 86 expression was not induced in U937 cells at concentration between 162.2 to 2595.6 µg/mL affording at least 70% viability. From this it has to be concluded that test substance Laurylacrylate 1214 does not induce dendritic cell activation.