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Developmental toxicity / teratogenicity

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developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not reported
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets generally accepted scientific standards, well documented and acceptable for publication.

Data source

Reference Type:
Embryotoxic and Teratogenic Effects of Nickel in Swiss Albino Mice during Organogenetic Period
Saini S, Nair N, Saini MR.
Bibliographic source:
BioMed Research International Volume 2013, Article ID 701439, 9 pages

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
Ni was administered orally on body weight base durign the organogenesis period from days 6 to 13 of gestation period and developmental endpoints were assessed on gestnational day 18.
GLP compliance:
not specified
GLP compliance not specified in manuscript
Limit test:

Test material

Constituent 1
Reference substance name:
Nickel chloride
EC Number:
EC Name:
Nickel chloride
Cas Number:
nickel(2+) dichloride
Details on test material:
Nickel chloride hexahydrate procured fromHi-Media Laboratories Pvt. Ltd., Mumbai, (Purity: 97.0%) was used for the study.

Test animals

Details on test animals or test system and environmental conditions:
Swiss albino mice (7–9 weeks of age, 24 ± 2 gm) selected from an inbred colony were maintained on standard mice feed (Aashirwad Ltd., Chandigarh) and tap water ad libitum.

Administration / exposure

Route of administration:
oral: gavage
Details on exposure:
Animals were divided into four groups of ten mice each. Group I was given tap water and served as a control, while groups II, III, and IV were given 46.125, 92.25, and 184.5mgNi/kg b.wt. as NiCl2 ⋅6H2O orally from days 6 to 13 of gestation, that is, organogenetic period. The doses were selected below LD50, that is, 369mgNi/kg b.wt. Dams were sacrificed by cervical dislocation on day 18 of gestation, and uteri of all the sacrificed dams were examined.
Analytical verification of doses or concentrations:
Details on mating procedure:
Female and male mice were housed formating in the ratio of 3 : 1 and examined every morning for vaginal plug. The day on which vaginal plug were detected was considered as day zero of pregnancy.
Duration of treatment / exposure:
From days 6 to 13 of gestation
Frequency of treatment:
Duration of test:
Until gestational day 18.
No. of animals per sex per dose:
10 females
Control animals:
yes, concurrent vehicle


Maternal examinations:
Diet consumption, water intake and body weight of all the groups were recorded daily.
Ovaries and uterine content:
The uteri of all the sacrificed dams were examined.
Fetal examinations:
The implant sites and live fetuses per dam were counted, and the conceptus at each site was classified as being alive, resorbed, or dead. The live fetuses were sexed, weighed, and examined for morphological alterations. Randomly selected 75% of fetuses were fixed in 95% alcohol for double staining (alizarin red S and alcian blue) to observe the skeletal anomalies, and the remaining fetuses were fixed in Bouin’s fixative to study the brain.
Thestatistical analysis of the data was evaluated by using the Microsoft Office Excel 2003 software, and the significance of the data was determined either by using one way analysis of variance (ANOVA) or one way Mann-Whitney U test. The levels of significance were p < 0.05 (almost significant) and p < 0.01 (significant).
Average number of implant sites/dam, Average number of live fetuses/dam
Historical control data:
None reported

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
The pregnant females administered with Ni+2 during organogenetic period revealed an almost significant (p < 0.05) decrease in diet consumption after 92.25 mgNi/kg b.wt. However, after 184.5 mgNi/kg b.wt., the decrease was significant (p < 0.01). Similar pattern of decrease was evident in intake of water. Maternal weight decreased significantly (p < 0.01) after 92.25 and 184.5 mgNi/kg b.wt.

Effect levels (maternal animals)

open allclose all
Dose descriptor:
Effect level:
184.5 mg/kg bw/day (nominal)
Based on:
Basis for effect level:
other: developmental toxicity
Dose descriptor:
Effect level:
92.25 mg/kg bw/day (nominal)
Based on:
Basis for effect level:
other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
The average implant sites per dam decreased after 92.25 and 184.5 mg Ni/kg b.wt. when compared with the control group. Average number of live fetuses/dam were significantly reduced (p < 0.01) in group IV (184.5 mg Ni/kg bw/d). Further, there was a concomitant rise in the percentage of resorbed, dead, and macerated fetuses in group IV. However, in groups II and III, the dead and macerated fetuses were not observed. At 184.5mgNi/kg b.wt. dose level, the incidence of postimplantation death (35.29%) was evident. No change was evident in the sex ratio (M: F) after oral administration of Ni at all the three dose levels. The average fetal weight significantly reduced (p < 0.01) in a dose-dependent manner. The placental weight decreased nonsignificantly in all the treated groups.

Fetuses with macroscopic anomalies such as open eye lids (10% and 12.50% in groups III and IV), club foot (6.25% in group IV) (Figure 13), and umbilical hernia (5% and 6.25% in groups III and IV) were evident. Ophthalmic anomalies, namely, microphthalmia (5%, 5%, and 6.25% in all the treated groups) and exophthalmia (5% in group III) were observed. The occurrence of hydrocephaly in groups III and IV was 5% and 12.50%, and microcephaly in group III was 5%, respectively. No gross anomalies were seen in group II). The double stained (alizarin red S and alcian blue) skeleton of fetuses showed numerous anomalies such as reduced ossification of nasal, frontal, parietal, intraparietal, and supra-occipital bones of skull, absence or gap between the ribs, reduced or fused sternebrae, reduced/absence/displaced vertebral centra in thoracic and lumbar regions, reduced/ absence of caudal vertebrae, reduced pelvic elements, and absence of carpals,metacarpals, tarsals, metatarsals, and phalanges. The percentage of skeletal anomalies was found to be dose dependent.

Effect levels (fetuses)

Dose descriptor:
Effect level:
92.25 mg/kg bw/day (nominal)
Based on:
not specified
Basis for effect level:
reduction in number of live offspring

Fetal abnormalities

not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Executive summary:

Study rated by an independent reviewer