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Toxicological information

Repeated dose toxicity: other routes

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Administrative data

Endpoint:
repeated dose toxicity: other route
Remarks:
other: mice IP injection on days -5, 2, 9, and 16 of 18 day study
Type of information:
experimental study
Adequacy of study:
disregarded due to major methodological deficiencies
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Peer reviewed publication using non-guideline and non-GLP study design to examine the effects of intraperitoneal injection of DINP on atopic dermatitis-like skin lesions in NC/Nga mice.

Data source

Reference
Reference Type:
publication
Title:
Effects of diisononyl phthalate on atopic dermatitis in vivo and immunologic responses in vitro.
Author:
Koike E, Yanagisawa R, Sadakane K, Inoue K, Ichinose T, Takano H.
Year:
2010
Bibliographic source:
Environ Health Perspect

Materials and methods

Principles of method if other than guideline:
NC/Nga mice aged 7 weeks were treated with 0.15, 1.5, 15 or 150 mg/kg/day DINP on days -5, 2, 9, and 16 of the study. Five days following the first IP injection of DINP the mice were exposed to the allergen Dermatophagoides pteronyssinus via intradermal injection in the ventral side of the right ear every 2-3 days. The purpose of the experiment was to determine the effects of IP injection of DINP on atopic dermatitis-like skin lesions. The effects of DINP in vitro exposure on bone-marrow derived dendritic cells or splenocytes was evaluated.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
DINP purchased from Wako Pure Chemical Industries, Osaka, Japan. Dermatophagoides pteronyssinus (Dp) purchased from Cosmo Bio LSL, Tokyo Japan. CAS or catalog number was not provided.

Test animals

Species:
mouse
Strain:
other: NC/Nga
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 7 weeks and 11-15 weeks
- Weight at study initiation: 20-23 g at 7 weeks and 24-27 g at 11-15 weeks
- Fasting period before study: no
- Housing: no information
- Diet ad libitum: CE-2; Clea Japan Inc., Tokyo Japan)
- Water ad libitum: distilled
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-26
- Humidity (%): 40-69
- Air changes (per hr): no information
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To: no information

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
olive oil
Details on exposure:
NC/Nga mice aged 7 weeks were IP injected with 0.15, 1.5, 15 or 150 mg/kg/day DINP dissolved in 0.1 ml olive oil on days -5, 2, 9, and 16 of the study. Five days following the first IP injection of DINP the mice were exposed to 5 ug of the allergen Dermatophagoides pteronyssinus dissolved in 10 ul saline via intradermal injection in the ventral side of the right ear on days 0, 3, 5, 8, 10, 12, 15, and 17 of the study.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
Study lasted 18 days.
Frequency of treatment:
DINP IP injection on days -5, 2, 9, and 16 of the study.
Dp injection on days 0, 3, 5, 8, 10, 12, 15, and 17 of the study.
In vitro exposure of DINP to differentiated bone marrow cells and splenocytes continuous for 24 hours.
In vitro exposure of DINP + Dp to differentiated bone marrow cells continuous for 72 hours.
Doses / concentrations
Remarks:
Doses / Concentrations:
In vivo DINP concentrations 0.15, 1.5, 15 or 150 mg/kg/day.
Dp concentration 5 ug.
In vitro DINP concentration 0.1, 1, 10, and 100 uM.
No. of animals per sex per dose:
n=12 for ear thickness.
n=5 for histological changes, eosinophil infiltration, mast cell degranulation
n=3 for bone marrow derived cultures
n=3 for splenocytes
Control animals:
yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: No data

FOOD CONSUMPTION: No data

FOOD EFFICIENCY: No data


WATER CONSUMPTION: No data


OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: No data

CLINICAL CHEMISTRY: Limited data
- Time schedule for collection of blood: Sampled by cardiac puncture 24 hours after last injection of Dp on day 18
- Animals fasted: No
- IgG, IgE and histamine levels were examined

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Histologic evaluation of the right ears
Ear thickness
Number of infiltrated eosinophils and mast cells
Degranulation of mast cells
Quantitation of cytokines and chemokines the ear tissue
Cytokine production of bone marrow cells
Cytokine and antigen stimulated proliferation of splenocytes
Statistics:
Significance of variation was determined by one-way analysis of variance or Kruskal-Wallis analysis. Differences were analyzed using Dunnett's or Steel multiple comparison test. p<0.05 was considered significant.

Results and discussion

Effect levels

open allclose all
Dose descriptor:
LOEL
Effect level:
15 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: Ear thinkness significantly increased. The authors noted that it was not dose dependent. The high dose 150 mg/kg/day was not significantly different.
Dose descriptor:
LOEL
Effect level:
0.15 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: Decrease in expression of Eeotaxin and eotaxin-2
Dose descriptor:
NOEL
Effect level:
>= 150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: infiltration of eosinophils
Dose descriptor:
NOEL
Effect level:
>= 150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: Mast cell degranulation
Dose descriptor:
NOEL
Effect level:
>= 150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: No effect on expression of IL-4, IL-5, IL-13, INF-gamma
Dose descriptor:
NOEL
Effect level:
>= 150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
DINP + Dp
Sex:
male
Basis for effect level:
other: IgG and IgE levels
Dose descriptor:
NOEL
Effect level:
>= 100 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: No effect on INF-gamma and IL-17 production in the cocultures of T cells and bone marrow cells.
Dose descriptor:
LOEL
Effect level:
100 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in Th2 chemokines TARC/CCL17 and MDC/CCL22 from bone marrow cells and an increase in CCR7 positive cells following in vitro exposure
Dose descriptor:
LOEL
Effect level:
10 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in CXCR4 positive bone marrow cells following in vitro exposure
Dose descriptor:
LOEL
Effect level:
0.1 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: MHCII positive bone marrow cells following in vitro exposure
Dose descriptor:
LOEL
Effect level:
1 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in CD86 positive bone marrow cells following in vitro exposure.
Dose descriptor:
LOEL
Effect level:
1 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in Dp-specific antigen presenting activity measured as cell proliferation.
Dose descriptor:
LOEL
Effect level:
10 other: uM
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in IL-4 production in splenocytes following in vitro exposure
Dose descriptor:
LOEL
Effect level:
0.001 other:
Based on:
test mat.
Remarks:
DINP
Sex:
not specified
Basis for effect level:
other: Increase in proliferation of splenocytes following in vitro exposure to DINP in the presence of Dp

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
DINP did not cause a dose dependent aggravation of AD-like skin lesions related to Do in NC/Nga mice. In vitro exposure to DINP enhanced the Th2 responses through activation of bone marrow cells and splenocytes although, this response occured at very high doses (100uM) or did not follow a clear dose response.