2-ethoxy-2-oxoethyl ethyl methylenemalonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: ISO 10993-10: 2010 Standard, Biological Evaluation of Medical Devices, Part 10: Tests for Irritation and Skin Sensitization, Pages 11-14, GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: ISO 10993-10: 2010 Standard, Biological Evaluation of Medical Devices, Part 10: Tests for Irritation and Skin Sensitization, Pages 11-14

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1-(2-ethoxy-2-oxoethyl) 3-ethyl 2-methylidenepropanedioate

EC Number:

817-217-9

Cas Number:

116280-23-0

Molecular formula:

C10H14O6

IUPAC Name:

1-(2-ethoxy-2-oxoethyl) 3-ethyl 2-methylidenepropanedioate

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Bond.Ease™ Topical Skin Adhesive (polymerized)
- Batch no.: 10312101
- Physical state: Liquid
- Purity: >96%

Test animals

Species:

other: Albino rabbits (Oryctolagus cuniculus)

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Bakkom Rabbitry, Viroqua, WI
- Sex: Female. Females used were nulliparous and non-pregnant.
- Age at study initiation: The rabbits were young adults.
- Weight at study initiation: Each rabbit weighed at least 2.0 kg.
- Housing: Animals were individually housed in stainless steel caging. Housing dimensions complied with NIH and AAALAC International guidelines for this species.
- Diet: Animals were supplied with certified commercial feed, ad libitum. There were no known contaminants present in the feed expected to interfere with the test results.
- Water: Potable water was supplied from the St. Paul municipal water supply, ad libitum. There were no known contaminants present in the water expected to interfere with the test results.
- Acclimation period: The animals were acclimated for a minimum of 5 days under the same conditions as the actual test.

ENVIRONMENTAL CONDITIONS
The environmental conditions in the animal rooms were maintained according to WuXi AppTec SOP: ILS-0018. The temperature and photo-period were set to meet the AAALAC International recommendations for these species. The laboratory and animal rooms were maintained as limited-access facilities.

Test system

Preparation of test site:

clipped

Vehicle:

other: 0.9% Normal Saline (NS) and Cottonseed Oil (CSO)

Controls:

yes, concurrent vehicle

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.2 ml x 5 sites

VEHICLE 1 (Normal Saline)
- Amount(s) applied: 0.2 ml x 5 sites
- Concentration (if solution): 0.9%
- Lot/batch no.: J0S679

VEHICLE 2 (Cottonseed Oil)
- Amount(s) applied: 0.2 ml x 5 sites
- Lot/batch no.: ZN1009

Duration of treatment / exposure:

72-hour

Observation period:

24, 48 and 72-hour

Number of animals:

3

Details on study design:

TEST ARTICLE ADMINISTRATION
The two test article extracts and the two vehicle controls were each injected into 3 rabbits . Each rabbit received five sequential 0.2 ml intracutaneous injections of the test article extract on the right side of the vertebral column and similarly the control vehicle on the left side. The second test and control extract injections were parallel and distal to the first injection sites.

OBSERVATIONS AND SCORING
The animals were observed daily for abnormal clinical signs. The appearance of each injection site was noted at 24 ± 2, 48 ± 2 and 72 ± 2 hours post injection. The tissue reactions were rated for gross evidence of erythema and edema. The skin was lightly swabbed with dilute alcohol to enhance the appearance of any erythema or edema. The intradermal injection of CSO frequently elicits an inflammatory response. CSO erythema scores <= 2 are considered normal. A well-defined positive response is characterized by a score greater than 2.

EVALUATION CRITERIA
According to ISO 10993:10 test criteria, if the difference between the average scores for the extract of the test article and the vehicle control is less than or equal to 1.0, the test article is considered nonirritating.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Remarks:

Control Score, Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

erythema score

Remarks:

Test Score, Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

erythema score

Remarks:

Control Score, Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.91

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

erythema score

Remarks:

Test Score, Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

1.1

Remarks on result:

other: Very slight erythema (barely perceptible)

Results 5

Irritation parameter:

edema score

Remarks:

Control Score, Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

edema score

Remarks:

Test Score, Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

edema score

Remarks:

Control Score, Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.04

Remarks on result:

no indication of irritation

Results 8

Irritation parameter:

edema score

Remarks:

Test Score, Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.4

Remarks on result:

no indication of irritation

Results 9

Irritation parameter:

other: Total Erythema and Edema Control Score

Remarks:

Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 10

Irritation parameter:

other: Total Erythema and Edema Test Score

Remarks:

Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 11

Irritation parameter:

other: Total Erythema and Edema Control Score

Remarks:

Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

1

Remarks on result:

no indication of irritation

Results 12

Irritation parameter:

other: Total Erythema and Edema Test Score

Remarks:

Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

1.5

Remarks on result:

other: slight erythema and edema

Results 13

Irritation parameter:

other: Comparative results (Average Test - Average Control)

Remarks:

Vehicle: NS

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Results 14

Irritation parameter:

other: Comparative results (Average Test - Average Control)

Remarks:

Vehicle: CSO

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.5

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The differences in the mean test and control scores of the extract dermal observations were less than 1, indicating that the requirements of the ISO Intracutaneous Reactivity Test have been met by the test article. Therefore, the test article is considered non-irritating.

Executive summary:

The study was conducted in accordance with ISO 10993 -10:2010 Standard, Biological Evaluation of Medical Devices, Part 10: Tests for Irritation and Skin Sensitization, Pages 11 -14. Each rabbit received five sequential 0.2mL intracutaneous injections along either side of the dorsal mid-line with the test article extract on one side and the concurrent vehicle control on the other. The irritation reaction of the test extracts were compared to vehicle controls and recorded over a 72 -hour period according to the standard ISO irritation scoring system.

Vehicle controls: 0.9% Normal Saline (NS) and Cottonseed Oil (CSO)

Observation period: 24, 48 and 72 hours.

Clinical observations: None of the animals on study showed abnormal clinical signs during the 72 hour test period.

Dermal observations: At the 48 hour observation period, one hematoma was observed at site 1 of CSO test on animal 22031.

There were no other significant dermal reactions observed at the injected test and control sites on the rabbits at the 24, 48 and 72 hour observation periods.

Evalutation criteria:

According to ISO 10993:10 test criteria, if the difference between the average scores for the extract of the test article and the vehicle control is less than or equal to 1, the test article is considered non-irritating.

Results:

- With vehicle: NS:

Total Erythema and Edema Control Score: Average: 0

Total Erythema and Edema Test Score: Average: 0

Comparative results (Average Test - Average Control): 0 - 0 = 0

- With Vehicle: CSO:

Total Erythema and Edema Control Score: Average: 1

Total Erythema and Edema Test Score: Average: 1.5

Comparative results (Average Test - Average Control): 1.5 - 1 = 0.5

The test was considered valid based upon scientific judgment. The differences in the mean test and control scores of the extract dermal observations were less than 1, indicating that the requirements of the ISO Intracutaneous Reactivity Test have been met by the test article. Therefore, the test article is considered non-irritating.