AVITERA ROSE SE

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Remarks:

I

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental Starting Date 22 March 2021 Experimental Completion Date 26 March 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 18 June 2019 Corrected 26 June 2020

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Identification FAT 40879/B TE
Appearance: Dark red powder
Batch Number: BOP 05-20 (MC- 0065114700/800 1:1)
Degree of Purity (% (w/w) 80.5 % all colored organic constituents; main constituent 66.9 %
Manufacture Date: June 26, 2020
Expiry Date: September 08, 2025
Storage Conditions: Freezer (-15 to -20 °C)

In vitro test system

Test system:

human skin model

Remarks:

Commercially available EPISKINTM (RhE) epidermal model was used as test system.

Source species:

human

Cell type:

other:

Cell source:

other: EPISKINTM epidermal tissues were procured from Skin ethic Laboratories, LYON - FRANCE

Vehicle:

water

Details on test system:

EPISKINTM epidermal is a 0.38 cm² Reconstructed human epidermis model. Adult human-derived epidermal keratinocytes were seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model was obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum. It was used for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability.

Control samples:

yes, concurrent negative control

Amount/concentration applied:

10 mg for each insert

Duration of treatment / exposure:

15 minutes

Duration of post-treatment incubation (if applicable):

42 hours after treatment

Number of replicates:

3 replicates

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

100.794

Negative controls validity:

valid

Remarks:

Distilled Water

Positive controls validity:

valid

Remarks:

Sodium Dodecyl Sulphate

Other effects / acceptance of results:

The percent viability of the tissue treated with test item was 100.794 % and it was greater than 50 % of the negative control. Hence, under the conditions of the study the test item was found to be ‘Non Irritant’ to skin in accordance with UN GHS as specified in the OECD Guideline for the Testing of Chemicals.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the results, the % viability of the epidermal tissue treated with FAT 40879/B was 100.794 % and it was greater than 50% of the negative control. Hence, under the conditions of the study the test item was found to be ‘Non Irritant’ to skin in accordance with UN GHS as specified in the OECD Guideline for the Testing of Chemicals.

Executive summary:

In GLP-compliant study conducted according to OECD guideline 439, FAT 40879/B was applied topically to the EPISKINTM epidermal model (three epidermis units were used per test item, positive and negative control) and exposed for 15 minutes. Treatment of epidermal units were terminated by rinsing with Dulbecco’s Phosphate Buffered Saline (DPBS). Epidermal units were then incubated at 37°C for 42 hours. The viability was assessed by incubating the epidermal tissues for 3 additional hours with MTT solution in a 12 well plate (0.3 mg/mL in assay medium; 2 mL per well). The precipitated formazan was then extracted using acidified Isopropanol (0.5 mL) and incubated for 4 hours at room temperature and quantified spectrophotometrically at 560 nm using 96 well plates (200 μL /well) with its respective test item, positive and negative controls. Sodium Dodecyl Sulphate (SDS) 5% and Distilled water treated epidermis were used as Positive (PC) and Negative Controls (NC), respectively. For each treated epidermal tissue, the viability was expressed in percentage (%) with  mean negative control tissues. The Mean Optical Density (OD) of negative controls was found to be 0.588, which is well within the acceptability range. The variation within the replicates was found less than 18 %. The percent viability of the tissue treated with positive control was 17.324 % and it was less than 40 % of the negative control, which reflects the irritant nature of positive control and the sensitivity of the tissues used in the study. The percent viability of the tissue treated with test item was 100.794 % and it was greater than 50 % of the negative control.

Based on the results, the % viability of the epidermal tissue treated with FAT 40879/B was 100.794 % and it was greater than 50 % of the negative control. Hence, under the conditions of the study the test item was found to be ‘Non Irritant’ to skin in accordance with UN GHS as specified in the OECD Guideline for the Testing of Chemicals.