Pine, Pinus sylvestris, ext.

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Read across from structural analogue

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

Read across from structural analogue

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

reference to other study

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Table 7.6.2/2: In vivo micronucleus test results

Group	Concentration		MNF (%)	PCE (%)
	CP (mg/kg bw)	PNE (mg/kg bw)
Distilled water			2.18 ± 0.58	52.90 ± 4.71
CP	30	-	36.68 ± 1.56*	40.74 ± 4.42*
PNE	-	2000	2.38 ± 1.18	53.95 ± 4.50
PNE + CP	30	500	27.31 ± 1.29*	51.13 ± 4.21*
PNE + CP	30	1000	18.40 ± 2.73*	47.58 ± 3.25***
PNE + CP	30	2000	8.88 ± 1.38*	42.50 ± 4.32***
PNE + CP	30	4000	10.53 ± 2.42*	40.40 ± 3.96***

 

CP: Cyclophosphamide; PNE: Pine needle extract

* p < 0.01, compared to distilled water control; * * p < 0.01, * * * p < 0.05, compared to CP groups.

There was a significant negative correlation (r = -0.9782, p < 0.05) between PNE dose and MNF in the PNE-treated groups in comparison to the CP positive group up to 2000 mg/kg bw of PNE. However, when the dose of PNE reached 4000 mg/kg bw, it exhibited some increase of MNF compared with the 2000 mg/kg bw group. This suggested that PNE inhibited the MNF in certain concentration ranges.

Conclusions:

Based on the read-across approach, the target substance did not induce micronuclei in mice.

Executive summary:

In an in vivo micronucleus test, ten male mice were injected intraperitoneally with Pine needle extract at the dose level of 2000 mg/kg bw and animals were sacrificed 30 h after the test item administration. Negative control (distilled water) and positive controls (cyclophosphamide at 30 mg/kg) were also included in the study. Antimutagenic effect of PNE was also studied with following dose levels at 10 males/group: (PNE + CP): 500 mg/kg bw + 30 mg/kg bw; 1000 mg/kg bw + 30 mg/kg bw; 2000 mg/kg bw + 30 mg/kg bw; 4000 mg/kg bw + 30 mg/kg bw.

Test material did not induce micronucleated PCE when compared with negative control. PNE could decrease the micronucleus frequency (MNF) compared with the cyclophosphamide (positive group) and the PNE + CP groups. Results showed that PNE could decrease the micronucleus induced by CP, and micronucleus frequency decreased markedly at the dose of 2000 mg/kg bw PNE (p < 0.01) demonstrating the antimutagenic effect. Positive control induced a statistically significant increase in micronucleated polychromatic erythrocytes indicating the validity of the study.

 

Based on the read-across approach, the target substance did not induce micronuclei in mice.

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1995

Materials and methods

Principles of method if other than guideline:

In an in vivo micronucleus test, mice injected intraperitoneally with Pine needle extract and animals sacrificed for examination of micronucleated polychromatic erythrocytes.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

90082-72-7

Cas Number:

90082-72-7

IUPAC Name:

90082-72-7

Test material form:

not specified

Details on test material:

- Name of test material (as cited in study report): Pine needle extract (ointment); which is mainly composed of steroidal saponins, triterpenoidal saponins, terpenes and aromatic acids.
- Source: Jiangsu Institute of Cancer Research, China.

Test animals

Species:

mouse

Strain:

Balb/c

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding Center of Nanjing University, Nanjing, China.
- Weight at study initiation: 18-22 g (at receipt)

Administration / exposure

Route of administration:

intraperitoneal

Duration of treatment / exposure:

30 h after test item administration

Frequency of treatment:

Single

Post exposure period:

None

Doses / concentrations

Remarks:

Doses / Concentrations:
2000 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

10 males/dose

Control animals:

yes

Positive control(s):

- Positive control: Cyclophosphamide: 30 mg/kg bw
- Route of administration: Intraperitoneal

Examinations

Tissues and cell types examined:

Micronucleus frequency in polychromatic erythrocytes (PCE) was determined by the routine method of Schmid (1973). 1000 PCEs in each animal were examined for micronucleus.

Evaluation criteria:

No data

Statistics:

The data were statistically evaluated with the t-test.

Results and discussion

Test results

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Table 7.6.2/2: In vivo micronucleus test results

Group	Concentration		MNF (%)	PCE (%)
	CP (mg/kg bw)	PNE (mg/kg bw)
Distilled water			2.18 ± 0.58	52.90 ± 4.71
CP	30	-	36.68 ± 1.56*	40.74 ± 4.42*
PNE	-	2000	2.38 ± 1.18	53.95 ± 4.50
PNE + CP	30	500	27.31 ± 1.29*	51.13 ± 4.21*
PNE + CP	30	1000	18.40 ± 2.73*	47.58 ± 3.25***
PNE + CP	30	2000	8.88 ± 1.38*	42.50 ± 4.32***
PNE + CP	30	4000	10.53 ± 2.42*	40.40 ± 3.96***

 

CP: Cyclophosphamide; PNE: Pine needle extract

* p < 0.01, compared to distilled water control; * * p < 0.01, * * * p < 0.05, compared to CP groups.

There was a significant negative correlation (r = -0.9782, p < 0.05) between PNE dose and MNF in the PNE-treated groups in comparison to the CP positive group up to 2000 mg/kg bw of PNE. However, when the dose of PNE reached 4000 mg/kg bw, it exhibited some increase of MNF compared with the 2000 mg/kg bw group. This suggested that PNE inhibited the MNF in certain concentration ranges.

Applicant's summary and conclusion

Conclusions:

Under the test conditions, Pine needle extract did not induce micronuclei in mice.

Executive summary:

In an in vivo micronucleus test, ten male mice were injected intraperitoneally with Pine needle extract at the dose level of 2000 mg/kg bw and animals were sacrificed 30 h after the test item administration. Negative control (distilled water) and positive controls (cyclophosphamide at 30 mg/kg) were also included in the study. Antimutagenic effect of PNE was also studied with following dose levels at 10 males/group: (PNE + CP): 500 mg/kg bw + 30 mg/kg bw; 1000 mg/kg bw + 30 mg/kg bw; 2000 mg/kg bw + 30 mg/kg bw; 4000 mg/kg bw + 30 mg/kg bw.

Test material did not induce micronucleated PCE when compared with negative control. PNE could decrease the micronucleus frequency (MNF) compared with the cyclophosphamide (positive group) and the PNE + CP groups. Results showed that PNE could decrease the micronucleus induced by CP, and micronucleus frequency decreased markedly at the dose of 2000 mg/kg bw PNE (p < 0.01) demonstrating the antimutagenic effect. Positive control induced a statistically significant increase in micronucleated polychromatic erythrocytes indicating the validity of the study.

 

Under the test conditions, Pine needle extract did not induce micronuclei in mice.