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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 28/07/2011 to 14/09/2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Remarks:
"ISO 10993-10: 2010 Standard, Biological Evaluation of Medical Devices, Part 10: Tests for Irritation and Skin Sensitization, Pages 18-26, GLP" comparable to OECD Guideline 406

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Guinea pig maximisation test already available from 2011 in compliance with U.S. Food and Drug Administration Good Laboratory Practice (GLP) regulations set forth in 21 CFR part 58.

Test material

Reference
Name:
Unnamed
Type:
Constituent
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%

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories
- Age: Healthy young adults were used.
- Weight Range: All animals weighed between 300 and 500 g upon assignment to the test.
- Housing: Animals were housed in solid bottom cages with contact bedding and up to two guinea pigs per cage. Housing density complied with AAALAC International recommendations. The test and negative control animals were housed separately.
- Diet: Animals were supplied with certified commercial guinea pig feed, ad libitum. No known contaminants present in the feed were expected to interfere with the test results.
- Water: Potable water was obtained from the St. Paul municipal water supply. No known contaminants present in the water were expected to interfere with the test results.
- Receipt: Animals were received on 07/27/11 according to WuXi AppTec SOP: ILS-0092. Each animal was examined for signs of disease and injury. 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 set to be 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.

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
other: 0.9% Normal Saline / Cottonseed Oil
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: 0.9% Normal Saline / Cottonseed Oil
No. of animals per dose:
11 (test group)
6 (control group)
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: one intradermal exposure and one epidermal exposure
- Exposure period: First induction week: intradermal induction/ Second induction week: epidermal induction
- Test groups:
Intradermal Induction:
1) Syringe 1: 0.1 ml of FCA + 0.9% Sterile Saline - Ratio: 1:1 (v/v)
2) Syringe 2: 0.1 mL of Test Extract
3) Syringe 3: 0.1 mL of FCA + 0.9% Sterile Saline (1:1) + Test Extract - Ratio: 1:1 (v/v)
Epidermal Induction: 0.3 ml of freshly prepared test article extract.
- Control group:
Intradermal Induction:
1) Syringe 1: 0.1 ml of FCA + 0.9% Sterile Saline - Ratio: 1:1 (v/v)
2) Syringe 2: 0.1 mL of Control Vehicle
3) Syringe 3: 0.1 mL of FCA + 0.9% Sterile Saline (1:1) + Control Vehicle - Ratio: 1:1 (v/v)
Epidermal Induction: 0.3 ml of freshly prepared control vehicle.
- Site: The intradermal induction was performed on the shaved shoulder region of the guinea pigs (on each side of the dorsal mid-line). The epidermal induction was performed over the injection sites.
- Frequency of applications: One week after the intradermal induction injections the epidermal induction application was performed.
- Duration:
Three pairs of intradermal injections made simultaneously for the intradermal induction.
Closed patch exposure for 48 hours for the epidermal induction.
- Concentrations:
100% test article extract
100% control vehicle

B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day(s) of challenge: 14 days after completion of epidermal induction.
- Exposure period: 24 hours
- Test groups: 0.3 ml of freshly prepared test article extract
- Control group: 0.3 ml of freshly prepared control vehicle
- Site: right flank (test article extract) / left flank (Control vehicle)
- Concentrations:
100% test article extract
100% control vehicle
- Evaluation (hr after challenge): 24 hours and 48 hours after removal of the dressings.
Positive control substance(s):
yes
Remarks:
Dinitrochlorobenzene (DNCB)

Results and discussion

Positive control results:
For the Induction I and Induction II phases, 0.3% dinitrochlorobenzene (DNCB), a known sensitizer, in ethanol is used. For the challenge phase, 0.15% DNCB in acetone is used. The negative control animals are exposed to the appropriate vehicle (acetone is used for the challenge and ethanol is used for the Inductions I and II) only.
Animals in the positive control test group exhibited moderate and confluent erythema to intense erythema and swelling at the challenge sites (scores of 2-3 during 48 hour score) and all of the animals in the negative control group exhibited discrete or patchy erythema at the challenge sites (score of 1 during 48 hour score) when treated with the 0.15% w/v mixture of DNCB in acetone. Since the sensitization results are based on the scores of the test group subtracted by the scores of the negative control group, the score of 1 for the negative control animals are subtracted from that of the
scores of 2-3, observed for the positive control test group. Therefore, all reactions in the positive control test group are considered to be sensitization reactions. Based on the results obtained, this test methodology demonstrated dermal sensitization in guinea pigs using DNCB, a known sensitizer.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100% test substance
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
No visible change - No erythema and edema
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100% test substance
No. with + reactions:
0
Total no. in group:
11
Clinical observations:
No visible change - No erythema and edema
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100% control vehicle
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
No visible change - No erythema and edema
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100% control vehicle
No. with + reactions:
0
Total no. in group:
6
Clinical observations:
No visible change - No erythema and edema
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.15% DNCB in acetone
No. with + reactions:
11
Total no. in group:
11
Clinical observations:
Discrete or patchy erythema to intense erythema and swelling (scores of 1, 2, 3)
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.15% DNCB in acetone
No. with + reactions:
11
Total no. in group:
11
Clinical observations:
Moderate and confluent erythema to intense erythema and swelling (scores 2-3)
Remarks on result:
positive indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
None of the negative control animals challenged with the control vehicle were observed with a sensitization response greater than '0'. None of the animals challenged with the test article extracts were observed with a sensitization response greater than '0'. The normal saline extract of the test material had a sensitization response of '0' under valid test conditions. The cottonseed oil extract of the test material had a sensitization response of '0' under valid test conditions. Under the conditions of this protocol, the test article did not elicit a sensitization response.
Executive summary:

This study was conducted in compliance with U.S. Food and Drug Administration Good Laboratory Practice (GLP) regulations set forth in 21 CFR, Part 58, in accordance with ISO 10993 -10:2010 Biological Evaluation of Medical Devices, Part 10- Tests for Irritation and Skin Sensitization, pages 10 -26 (Comparable to OECD Guideline 406).

Method used: Guinea pig maximisation test.

This test was designed to evaluate the allergenic potential or sensitizing capacity of the test article.

Summary design: Eleven test guinea pigs (per extract) were injected with the test article extract and FCA, and six guinea pigs (per extract) were injected with the corresponding control blank and FCA. On Day 6, the dorsal site was reshaved and SLS in mineral oil was applied. The day after the SLS application, the test animals were topically patched with the appropriate test extract and the control animals were patched with the corresponding control blank. The patches were removed after 48 ± 2 hours of exposure. Following an approximate two-week rest period, the animals were topically patched with the appropriate test extract and corresponding control blank. The patches were removed after 24 ± 2 hours of exposure. The dermal patch sites were observed for erythema and edema 24 ± 2 and 48 ± 2 hours after patch removal. Each anim al was assessed for a sensitization response based upon the dermal scores. The test results were based upon the percentage of animals exhibiting a sensitization response.

Species/Strain: Albino guinea pigs (Cavia porcellus), Hartley strain (specific pathogen free), male.

Results/Conclusion:

None of the negative control animals challenged with the control vehicle were observed with a sensitization response greater than '0'. None of the animals challenged with the test article extracts were observed with a sensitization response greater than '0'. The normal saline extract of the test material had a sensitization response of '0' under valid test conditions. The cottonseed oil extract of the test material had a sensitization response of '0' under val id test conditions. Under the conditions of this protocol, the test article did not elicit a sensitization response.