Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro cytogenicity / micronucleus study
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
no data available
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Reasonbably well described study with minor reporting deficiencies: no information on cytotoxicity, no individual results, purity of the test substance not reported.

Data source

Reference
Reference Type:
publication
Title:
Induction of micronuclei in Syrian hamster embryo cells: comparison to results in the SHE cell transformation assay for national toxicology programm test chemicals
Author:
Gibson DP; Brauninger R; Hussain SS; Kerckaert GA; LeBoeuf RA; Isfort RJ; Aardema MJ
Year:
1997
Bibliographic source:
Mutation Research 392 (1997) 61-70, pp. 61-70

Materials and methods

Test guideline
Guideline:
other: no information available if test was conducted according to guideline
Principles of method if other than guideline:
Testing of 16 chemicals in the in vitro micronucleus assay in SHE cells.
GLP compliance:
no
Type of assay:
in vitro mammalian cell micronucleus test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: solid
Details on test material:
- Name of test material (as cited in study report): Vanadium pentoxide
- Physical state: solid
No further details are given.

Method

Target gene:
not applicable
Species / strain
Species / strain:
other: Syrian hamster embryo (SHE) cells
Details on mammalian cell lines (if applicable):
For the test, the cells were seeded at 1x10^6 cells/T-25 flask for control, and chemically-treated cultures. Afetr approximately 24 hours, the cells were exposed to the test chemical and cytochalasin B (3 µg/mL in DMSO) for 24 hours.
Additional strain characteristics:
not specified
Metabolic activation:
without
Test concentrations with justification for top dose:
10, 15, 20, and 25 µg/mL
Dose levels were selected based on solubility and/or toxicity limits.
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Negative controls:
no
Solvent controls:
not specified
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: colchicine; 0.25 or 0.5 µg/mL
Details on test system and conditions:
METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 24 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 24 hours and 10 minutes: After determination of the toxicity (see below) remaining cells were suspended in 37°c 0.075M KCl for 5-10 minutes, before the cells were collected by centrifugation and fixed in at least two changes of cold 25:1 methanol/acetic acid.

STAIN (for cytogenetic assays): Cells were stained for 1-5 minutes in a 10% Giemsa solution in Gurr buffer.

NUMBER OF REPLICATIONS: no data

NUMBER OF CELLS EVALUATED: In each treatment group, 500 cells were analysed to determine the percentage of binucleated cells and 1000 binucleated cells were analysed to determine the number of micronucleated cells.

DETERMINATION OF CYTOTOXICITY
- Method: other: number of binucleated cells after 24-hour exposure
After a 24-hour treatment period the media was aspirated off and the cells were collected by trypsinisation. An aliquot of cells was counted to determine the number of live cells (determined by trypan blue exclusion) as a measure of toxicity.

OTHER EXAMINATIONS:
Only cells with distinct cytoplasm and distinct binucleation were analysed for the presence of micronuclei. Only micronuclei that were entirely inside the cytoplasm, separate from the main nucleus, less than approximately one-third the size of the main nuclei, and non-refractile were recorded.
Evaluation criteria:
no data
Statistics:
The number of micronucleated binucleated cells (MNBC) in the treated group was compared to the number of MNBC in the vehicle control group using a one-sided Fisher's exact test where p<0.05 was considered significant.

Results and discussion

Test results
Species / strain:
other: SHE cells
Metabolic activation:
without
Genotoxicity:
negative
Remarks:
No statistically significant increase in micronucleated binucleated cells (MNBC) was observed.
Cytotoxicity:
no, but tested up to limit concentrations
Remarks:
(as evidenced by at least a 50% reduction in the relative cell number and/or in the percentage of binucleated cells)
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
No details are reported.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under the test conditions described the test substance, vanadium pentoxide, has no mutagenic potential in the in vitro micronuceus test in Syrian hamster embryo (SHE) cells.
Executive summary:

Vanadium pentoxide as one of 15 other chemical substances was in the in vitro micronucleus assay in SHE at the following concentrations: 10, 15, 20, and 25 µg/mL. Dose levels were selected based on solubility and/or toxicity limits. After a 24-hour treatment period an aliquot of cells was counted to determine the number of live cells (determined by trypan blue exclusion) as a measure of toxicity. Remaining cells were collected by centrifugation, fixed and stained for analysis. In each treatment group, 500 cells were analysed to determine the percentage of binucleated cells and 1000 binucleated cells were analysed to determine the number of micronucleated cells.

Under the test conditions described the test substance, vanadium pentoxide, has no mutagenic potential in the in vitro micronuceus test in Syrian hamster embryo (SHE) cells.