Tris(isopropenyloxy)vinylsilane

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

tris(isopropenyloxy)(vinyl)silane

IUPAC Name:

tris(isopropenyloxy)(vinyl)silane

Constituent 2

Reference substance name:

Tris(isopropenyloxy)vinylsilane

EC Number:

239-362-1

EC Name:

Tris(isopropenyloxy)vinylsilane

Cas Number:

15332-99-7

Molecular formula:

C11H18O3Si

IUPAC Name:

[tris(isopropenyloxy)vinyl]silane

Test material form:

other: liquid

Test animals

Species:

other: EpiSkin™

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 μl
- Concentration (if solution): undiluted

CONTROLS
Negative control: Dulbecco's Phosphate Buffering Saline (DPBS; Gibco, Cat. No. 14040-091, Lot No.: 1581502)..
Positive control: 5% sodium dodecyl sulfate in distilled water (SDS; AppliChem, Art.-No. A1112,0500, CAS No.: 151-21-3, Lot No.: 1V010396).

Details on study design:

TEST SYSTEM: reconstituted three-dimensional human skin model EPISKIN-SM™ (SkinEthic). This skin model consists of normal (non-cancerous), adult human-derived epidermal keratinocytes (NHEK) which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. The NHEK are cultured on chemically modified, collagen-coated cell culture inserts. A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.

PRE-EXPERIMENT
To check the non-specific MTT-reducing capability of the test item 10 μl of the test item were mixed per 2 ml MTT medium and incubated for 3 h at 37 ± 1 °C in the dark. If the mixture turns blue/purple, the test item is presumed to have reduced MTT. [MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. To check the colouring potential of the test item 10 µL of the test item were mixed per 90 µL water in a transparent recipient for 15 min.

EXPERIMENTAL PROCEDURE
Tissues were pre-incubated for at least 24 h at 37 ± 1 °C, 5.0% CO₂, then each group (negative control (first), test item and positive control) was treated in triplicate. After incubation for 15 ± 0.5 minutes, the tissues were washed with DPBS to remove residual test item, then excess DPFS removed with blotting paper. The tissues were then placed in MTT medium and incubated for a further 3 hours.

After incubation the tissues were dried on blotting paper and a biopsy punch was used and the epidermis was separated from the collagen matrix. Epidermis and collagen matrix were extracted with 500 μl of acidic isopropanol. Extraction was carried out protected from light over the weekend at 2 - 8°C, then the tubes were mixed by vortexing. Any visible fragments were removed by centrifugation.

2 x 200 μl aliquots of the extract form each tissue were transferred into a 96-well plate and optical density was measured at 550 nm without reference wavelength in a plate spectrophotometer.

DPBS Dulbecco's Phosphate Buffering Saline
MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: tissue viability

Value:

99

Remarks on result:

other:

Remarks:

Remarks: The tissue viability was assessed by optical density at 550 nm, expressed as a percentage. (migrated information)

In vivo

Other effects:

PRE-EXPERIMENT-test item showed no reduction of MTT relative to negative control. No colouring was detected by unaided eye when 10 μl of the test item were mixed with 90 μl distilled water.

EXPERIMENT: Total mean OD550 of 3 replicate tissues (blank-corrected): negative control: 1.037*; positive control: 0.191; test item: 1.026.
Mean relative tissue viability [%]: negative control: 100.0; positive control: 18.5 test item: 99.0

QUALITY CRITERIA:
Mean absolute OD550 nm NC: pass
Relative % viability PC: pass
SD viability (%): pass

* Corrected mean OD550 of the negative control corresponds to 100% absolute tissue viability.
NC negative control
PC positive control

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Tris(isopropenyloxy)(vinyl)silane has been tested in a reliable in vitro study for skin irritation conducted according to OECD 439 and in compliance with GLP using EPISKIN-SM™ tissue. The relative tissue viability of the test-item treated tissues, assessed by mean optical density at 550 nm relative to solvent control, was 99.0% which is greater than the 50% threshold for skin irritation. The positive control produced the expected reduction in viability. Therefore it is concluded that the test substance is not irritating to the skin.