Cytogenetic damage by vanadium(IV) and vanadium(III) on the bone marrow of mice

Abstract

Vanadium is a strategic metal that has many important industrial applications and is generated by the use of burning fossil fuels, which inevitably leads to their release into the environment, mainly in the form of oxides. The wastes generated by their use represent a major health hazard. Furthermore, it has attracted attention because several genotoxicity studies have shown that some vanadium compounds can affect DNA; among the most studied compounds is vanadium pentoxide, but studies in vivo with oxidation states IV and III are scarce and controversial. In this study, the genotoxic and cytotoxic potential of vanadium oxides was investigated in mouse bone marrow cells using structural chromosomal aberration (SCA) and mitotic index (MI) test systems. Three groups were administered vanadium(IV) tetraoxide (V₂O₄) intraperitoneally at 4.7, 9.4 or 18.8 mg/kg, and three groups were administered vanadium(III) trioxide (V₂O₃) at 4.22, 8.46 or 16.93 mg/kg body weight. The control group was treated with sterile water, and the positive control group was treated with cadmium(II) chloride (CdCl₂). After 24 h, all doses of vanadium compounds increased the percentage of cells with SCA and decreased the MI. Our results demonstrated that under the present experimental conditions and doses, treatment with V₂O₄ and V₂O₃ induces chromosomal aberrations and alters cell division in the bone marrow of mice.

Keywords:

• Metals
• bone marrow
• vanadium oxides
• ; mitotic index
• structural chromosomal aberrations
• genotoxic effects in vivo

I. Introduction

Vanadium (V) is a strategic metal that has many important industrial applications, used for producing high-quality specialized metal alloys and strengthening steel in the aerospace industry and in the chemical industry, for the production of catalysts, electronics and batteries, e.g., vanadium redox batteries (Huang et al. 2015, Hu et al. 2018). Among the compounds that are employed in diverse industrial processes are vanadium(III) trioxide (V₂O₃) and vanadium(IV) tetraoxide (V₂O₄), which are used in electro-optical switches, aerospace technology and semiconductor manufacturing (Rodríguez-Mercado et al. 2010, Zhang et al. 2021, Vanitec 2023) New uses and applications are continually being discovered for this metal (SCIREA 2019).

Vanadium is released into the atmosphere as a consequence of burning fossil fuels and metallurgic industry activities. Measured concentrations in coals worldwide vary from 7 to 100 mg/kg, with an estimated mean of 25 mg/kg. Schlesinger et al. (2017) estimated that extraction associated with coal mining had increased from approximately 100 × 10⁹ g V/year in 2000 to 180–190 x 10⁹ V/year in 2014–16, and their release caused residual oil ash. Levels in air vary by season and location (urban, industrial and rural), and volcanic eruptions, weathering of V-rich rocks and wildfires can lead to their introduction into the ambient environment. The main compounds released consist of V oxides, which accumulate in the soil and water, resulting in constant exposure to organisms (ATSDR 2012, Huang et al. 2015, Cohen 2022).

The most common nonoccupational sources of V exposure are contaminated food (WHO 1996, Anke 2004, Mehmood et al. 2021), because soils contain high concentrations, and from there, they pass to food (Chen et al. 2021). For example, following a diesel spill in Oaxaca, Mexico, V levels in sediments were found to range from 95 to 149 μg V/g compared to levels of < 5 μg V/g at distanced reference sites (Salazar-Coria et al. 2007). Similarly, Hernandez and Rodríguez (2012) found that levels of V in urban soils of Salamanca, Mexico, reached values of > 600 mg V/kg; these were far higher than in rural soils, but even those were relatively high (i.e., 126 mg V/kg) due partly to the region’s volcanic nature that contains many basalts and rhyolites.

The total ingested in the diet is varied, with an average V intake of 13 μg/day/person in the United Kingdom and an estimated dietary intake between 6 and 18 µg/day in the USA (EFSA 2004). In Germany and México, V intake is 10–20 µg/day for women and > 20 µg/day for men (Anke 2004, Rodríguez-Mercado et al. 2011). Another report indicated that typical daily doses consumed by humans have been estimated at 0.01–0.03 mg V/day (Willsky et al. 2011, Pessoa et al. 2015). Whole grains, seafood and meats generally contain 5–30 ng/kg of V (EFSA 2004), and parsley, mushrooms, black pepper, cereals and shellfish are some of the main sources of V, with concentrations ranging from 0.05 to 2 mg/kg (Tripathi et al. 2018). Another important source of V exposure is the use of dietary mineral supplements, which can lead to the consumption of more than 10 mg V/day (Barceloux and Barceloux 1999, EFSA 2004, Leopardi et al. 2005).

Workers occupationally exposed to V are at risk of developing respiratory diseases (IARC 2006) because the particulates are small, breathable and may penetrate deep into the pulmonary tract (Fortoul et al. 2011, Rojas-Lemus et al. 2021) and increase heart rates (Magari et al. 2002). Some epidemiological studies have indicated that a correlation exists between exposure to airborne V particles and the incidence of cancer in residents of metropolitan areas (Kraus et al. 1989). In leukocytes from 52 exposed workers, an increase in the plasma concentration of V and no difference in DNA migration, with comet assay, were reported under standard conditions, but when the authors used a modified technique that detects DNA damage due to the oxidation of DNA bases, they found increased tail lengths, and those workers also had a 2.5-fold higher micronuclei frequency (Ehrlich et al. 2008). Vanadium(V) pentoxide (V₂O₅) has been designated as possibly carcinogenic to humans (Group 2B) by the International Agency for Research on Cancer (IARC 2006), based on evidence of lung cancer in mice published by the NTP (2002), and this chemical and other vanadium compounds have been designated as category 2 by the German Commissions for the Investigation of Health Hazards of Chemical Compounds in the Work Area (Beyersmann and Hartwing 2008).

Due to the above, genotoxicity studies of V have attracted attention, and several studies have shown that some V compounds can affect DNA, among other toxic effects. The most studied compound at the cytogenetic level is V₂O₅, both in vitro and in vivo, which is known to produce DNA single-strand breaks in whole blood leukocytes in vitro (Rojas et al. 1996). The results from in vivo studies reported an increased number of cells with damage, with a single-cell gel electrophoresis assay, in liver, kidney, lung, spleen, and heart cells but not in bone marrow (Altamirano-Lozano et al. 1999). Additionally, in male mice injected intraperitoneally with V₂O₅ every 3^rd day for 60 days, a DNA damage-dependent dose was reported in testis cells (Altamirano-Lozano et al. 1996), in the bone marrow, and no differences were observed in sister chromatid exchanges or in average generational time (Altamirano-Lozano et al. 1993). Rojas-Lemus et al. (2014) reported that inhaled V₂O₅ (0.02 M) 2 hours twice a week increased the frequency of micronuclei in exposed males compared with the control, and no cytotoxic effect was observed.

Studies with other V oxides showed that V₂O₄ increases the frequency of sister chromatid exchanges and the structural chromosomal aberration (SCA) and induces cytostatic and cytotoxic effects in human cells cultured in vitro (Rodríguez-Mercado et al. 2003, 2010, Rodríguez-Mercado and Altamirano-Lozano 2006), and some authors consider that some vanadium(III) compounds can induce structural chromosome damage (Lloyd et al. 1998, Caicedo et al. 2008). Nevertheless, we did not find significant differences in the SCA when human lymphocyte cultures were treated with vanadium(III) trioxide (Rodríguez-Mercado et al. 2010). However, in vivo studies with vanadium oxides III and IV are scarce and inconclusive. Therefore, the aim of this study was to evaluate the genotoxic and cytotoxic potential of the in vivo intraperitoneal administration of two vanadium oxides, V₂O₄ and V₂O₃, in mouse bone marrow cells using the SCA and mitotic index (MI) cytogenetics assay.

II. Materials and methods

2.1. Testing drug

The treatment was performed with vanadium oxides obtained from Aldrich Chemical Company (Milwaukee, Wis., USA), V₂O₃ (V = 67.98%, O = 32.02%; CAS no. 1314–34-7, 99.6% pure), and V₂O₄ (V = 61.43%, O = 38.57%; CAS no. 12036–21-4, 99.6% pure), which were macerated and dissolved in injectable water. Cadmium(II) chloride (CdCl₂•2.5 H₂O CAS 7790–78-5) and KCl were obtained from Sigma–Aldrich (St. Louis, MO, USA). Giemsa was obtained from Hycel of Mexico. Absolute methanol and acetic acid were obtained from JT Baker Mexico.

2.2. Animals

Male CD-1 mice, six to seven weeks old, weighing 25–35 g, were obtained from Bioterium of the FES-Zaragoza, UNAM. The animals were handled in accordance with official regulations in Mexico (NOM-062-ZOO-1999). The animals were housed in polypropylene cages under a controlled temperature of 22 ± 1 °C, and free access to water and food (Harlan 2018S with 18% protein rodent diet) was permitted. The photoperiod consisted of 12 h of artificial light and 12 h of darkness, with a relative humidity of 31%.

2.3. Experimental procedure

The mice were separated and housed in groups of six animals for a total of 8 groups. Three groups were administered 4.7, 9.4 and 18.8 mg/kg body weight (bw) of V₂O₄, and these values correspond to 1/32, 1/16, and 1/8 of the lethal dose 50 (LD₅₀= 150 mg/kg Aragón and Altamirano-Lozano 2001). Three groups were administered 4.22, 8.46 or 16.93 mg/kg bw of V₂O₃; these doses of V₂O₃ were calculated based on the amount of vanadium present in V₂O₄. The vanadium concentration was calculated in both compounds, which were 2.87, 5.75 and 11.5 mg of V in each dose of compounds, respectively. The doses of both compounds contained the same amount of elemental V. The control group was treated with sterile water, and the positive control group was treated with 7 mg/kg bw CdCl₂ (Ali 2012, El-Habit and Abdel-Moneim 2014)

All treatments were injected intraperitoneally (ip) with a volume of 0.1 mL/10 g of mouse body weight. The intraperitoneal route of administration generally offers the second most rapid absorption among the parenteral routes, conferred by the large surface area of the lining of the peritoneal cavity, and the rich blood supply to that area process (Stump et al. 1999, Gad 2014). Bone marrow was collected 24 h after the treatment. The experimental protocol was approved by the Committee of Ethics and Biosecurity of FES-Zaragoza UNAM (registration number FESZ-CE/21–118-01).

2.4. Cytogenetic assay

Twenty-two hours after vanadium compound treatment, the mice were injected with 0.2% colchicine (0.1 mL/10 g bw), to arrest proliferating cells in metaphase (Florian and Mitchison 2016), and 2 h later, the animals were sacrificed by cervical dislocation. Bone marrow tissue was obtained from each femur with hypotonic solution (KCl 0.075 M), and samples remained for 1 h at 37 °C. The cells were then fixed with methanol-acetic acid (3:1). To prepare the slides for cytogenetic analysis, 5 drops of the fixed cell suspension were placed on clean glass slides, which were then stained for 30 min with a 5% Giemsa solution.

The MI was calculated as the proportion of cells undergoing cell division. A total of 4000 cells per mouse were assessed, and the MI was calculated using the following formula: MI= (Number of dividing cells, metaphases/total number of bone marrow cells counted) x 100.

The SCA analysis was based on the recommendations of Adler (1984), Savage (2004) and the OECD TG 475 (2013) with slight modifications. The type of aberration scored included chromatid and chromosome breaks, fragments and pulverizations. At least one hundred cells in metaphase per mouse were reviewed, and only metaphases containing 40 ± 2 centromeres were analyzed. The total number and type of aberrations, as well as the percentage of aberrant cells per treatment, were evaluated. All slides were coded and evaluated in a double-blind manner.

2.5. Statistical analysis

The results are expressed as the means ± standard deviations (SD). The Z test was used to assess significant MI differences. Student’s t test was used to compare the group means for SCA. Tukey’s ANOVA was also applied to the difference between groups of the percentage of cells with SCA.

3. Results

The administration of V compounds to mice resulted in no changes in the weight of exposed or control mice. It is important to highlight that the animals treated did not present other symptoms of toxicity.

The SCA and MI results are shown in Table 1. The data show a reduction in the MI in the treated groups compared to the negative control group. The control group had an MI of 4.1, while the groups treated with V₂O₄ showed reductions of 44, 28.5 and 32.6%, respectively; in the groups treated with V₂O₃, the reductions in the MI were 35.6, 31.4 and 26.8%, respectively.

Table 1. Mitotic index (MI) and structural chromosomal aberrations (SCA) in male mouse bone marrow cells following exposure ip to vanadium(IV) tetraoxide (V₂O₄) or vanadium(III) trioxide (V₂O₃).

Dose (mg/kg)	MI (%) ± SD (Inhibition %)	SCA					Total of SCA excluding Pu	Cells with SCA	% of cells with SCA
		Total of cells scored	Fg	Ct	Cs	Pu
Control group
0.0	4.1 ± 0.90	635	40	1	1	0	42	26	4.0 ± 2.4
Vanadium(IV) tetraoxide (V2O4)
4.7	2.2 ± 0.20^a (44.0)	614	127	9	0	0	136	78	12.6 ± 7.6^a
9.4	2.9 ± 0.52^a (28.5)	625	112	31	3	6	146	104	16.6 ± 4.4^a
18.8	2.7 ± 0.53^a (32.6)	583	101	37	1	0	139	104	18.0 ± 7.0^a
Vanadium(III) trioxide (V2O3)
4.22	2.6 ± 0.72^a (35.6)	670	88	36	1	2	125	98	14.6 ± 7.1^a
8.46	2.8 ± 0.40^a (31.4)	650	116	35	6	1	157	102	15.6 ± 3.7^a
16.9	3.0 ± 0.68^a (26.8)	649	154	44	0	0	198	124	19.0 ± 5.6^a
Positive control group (CdCl2)
7	2.6 ± 0.27^a (36.0)	631	61	36	4	1	101	78	12.3 ± 7.5^a

SD: standard deviation; ip; intraperitoneally; Ct; chromatid type breaks; Fg; fragments; Cs; chromosomal type breaks; Pu; pulverizations; CdCl₂; cadmium(II) chloride. Six mice were treated per dose. Differences between total chromosome aberrations and the number of cells with rations SCA is due to some cells containing or having more than one aberration.

^ap < 0.05, compared vs. control.

The percentage of cells with SCA increased in the treated groups. The control group had 4.03% of cells with SCA, while the groups treated with V₂O₄ showed increases of approximately 3-, 4- and 4.5-fold, respectively; in the groups treated with V₂O_3, the increases were approximately 3.7-, 3.9- and 4.8-fold, respectively. The most frequent aberration types were fragments and chromatid breaks, and in some concentration cells with 10 or more aberrations, pulverization was observed (Figure 1). Differences between total chromosome aberrations and the number of cells with aberrations are due to some cells containing or having more than one aberration.

Figure 1. Some of chromosome aberrations in bone marrow cells from CD-1 male mice treated ip with vanadium(IV) or vanadium(III). A) Normal chromosome (control group), B) fragments (dose of 16.9 mg/Kg of V₂O₃), C) chromatid breaks (dose of 18.8 mg/Kg of V₂O₄), D) chromosomal breaks (dose of 8.46 mg/Kg of V₂O₃), E) chromatid breaks, and F) pulverizations (dose of 9.4 mg/Kg of V₂O₄).

As expected, when mice were treated with cadmium(II) chloride at a dose of 7 mg/kg, a clear reduction in MI and an increase in SCA were observed.

4. Discussion

The results of this work show that there were no differences in mouse weight after intraperitoneal administration of the compounds V₂O₄ or V₂O₃. In another work where V₂O₄ was administered for 60 days to male mice at similar doses (4.7, 9.4 or 18.8 mg/kg bw), no changes in animal weight were observed (Aragón and Altamirano-Lozano 2001). However, a reduction in the weight of male CD-1 mice was found when sodium orthovanadate (Na₃VO₄) was administered orally for 5 weeks at concentrations of 7.5, 75, 750 and 1500 mg/L (Leopardi et al. 2005).

Therefore, we can say that the effect of V on the weight of mice depends on the compound, time and the dose administered. Considering that the weight of the whole organism is one of the parameters that provide evidence of toxicity of the different compounds administered (OECD 2013), we can conclude that under the present experimental conditions and doses of V₂O₄ and V₂O₃ administered in this study do not produce toxicity in mice at this treatment time (24 h).

The data show a reduction in the MI in the groups treated with both compounds compared to the negative control group. MI is a parameter that shows the percentage of cells that reached the metaphase stage of the total of cells that are in mitosis, indicating that the cells are in division, and the reduction of this parameter shows that V₂O₄ and V₂O₃ produce an effect on cell proliferation.

Few in vivo studies have evaluated the cytotoxic effect of V oxides. Ciranni et al. (1995) reported that SVO₅ (vanadyl sulfate), a compound that contains vanadium(IV), produced an appreciable cytotoxic effect, using polychromatic erythrocytes (PCEs) and normochromatic erythrocytes (NCEs) ratio data after administration of 100 mg/kg intragastrically to male mice.

Previous studies performed with vanadium in oxidation estate +5, which is the main compound studied, show that the administration of 23 mg/kg bw ip of V₂O₅ to mice decreases the MI, but with the administration of doses of 5.75 and 11.5 mg/kg, MI does not decrease (Altamirano-Lozano et al. 1993). Rojas-Lemus et al. (2014) reported a decrease in the reticulocyte count in mice that inhaled V₂O₅ (0.02 M) 2 h/twice a week, and the PCE/NCE ratio is indicative of cytotoxic effects.

Rodríguez-Mercado et al. (2010) reported that in human leukocytes treated in vitro with three oxides, V₂O₃, V₂O₄ or V₂O_5, decreased MI in all groups treated, so they concluded that the three compounds were capable of inducing cytotoxicity. In another study with the same model, V₂O₃ caused a decrease in the MI and replication index, which may suggest that V₂O₃ can alter cell cycle progression (Mateos-Nava et al. 2017, 2021).

The decrease in MI may be related to the ability of V compounds to alter the activity of different molecules, such as ATPases, protein kinases, ribonucleases and phosphatases, and it has also been shown that V oxides could modify the levels of some proteins responsible for regulating the cell cycle (Attia et al. 2005, Mateos-Nava et al. 2021, Rojas-Lemus et al. 2021). Furthermore, V oxides delay the cell cycle via a mechanism that does not necessarily involve DNA damage, since it has been observed that disrupting cytoskeletal microtubules, hyperosmotic stress and inhibiting histone deacetylases induce the modification of Cdc25C through the p38 MAPK pathway (Zhang et al. 2003, Liu et al. 2012, Mateos-Nava et al. 2021).

Although generally the reduction of MI is a consequence of a reduced rate of cell proliferation, there could also be a contribution from cells that have permanently lost their proliferative capacity since it has been reported that some V compounds have a cytotoxic effect in vitro and that this effect is closely related to its chemical form (Rodriguez-Mercado et al. 2010), which increases with the oxidation state and produces this effect only in certain cell types (Rojas-Lemus et al. 2014). Guerrero-Palomo et al. (2019) reported that classical apoptotic mechanisms are not related to the cell death induced by some V compounds and showed that higher reactive oxygen species production was induced by SVO₅ in the human lung adenocarcinoma cell Line A549.

The MI data do not present adose–response relationship, which can be explained on the one hand by the difference between the concentrations used, which is not large enough to cause more cytotoxicity, but, if SCA is produced, the cell cycle will stop. Alternatively, because metals can have multiple biological effects on cellular signaling pathways, such as adaptation/tolerance mechanisms to low doses (Giani et al. 2021, Shi et al. 2022), in this work, a single injection of V compounds was administered at doses of 1/32, 1/16 and 1/8 of the LD₅₀. Nevertheless, currently, there is still a gap in knowledge regarding the dose–response relationship, especially for metal compounds (Shahid et al. 2020).

Chromosome damage is a very important indicator of genetic damage relevant to environmental studies. In somatic cells, chromosomal aberrations and mutations play a key role in processes leading to malignancies if mutations occur in proto-oncogenes, tumor suppressor genes, and/or DNA damage response genes. The accumulation of DNA damage in somatic cells has also been proposed to play a role in degenerative conditions such as accelerated aging, immune dysfunction and cardiovascular and neurodegenerative diseases (Niida and Nakanishi 2006, Doherty 2012, Valko et al. 2016, Mosesso and Cinelli 2019).

The increase in the percentage of cells with SCA in the groups treated with V₂O₄ found in this study is in accordance with one previous study reported by Ciranni et al. (1995), in which they administered single intragastric doses of three vanadium salts, 100 mg/kg bw SVO₅, 75 mg/kg bw sodium orthovanadate (Na₃VO₄) and 50 mg/kg bw ammonium metavanadate (NH₄VO₃) (-vanadate in oxidation state V) to male mice. They found an increased frequency of micronucleus by all compounds tested, but only the group treated with SVO₅ presented increased chromosome aberrations in bone marrow cells, and the highest effect was observed at 24 h. This chromosome damage was due mainly to breaks and fragments.

Villani et al. (2007) reported that there was no significant difference in comet assays and micronuclei in the PCE ratio in male CD1 mice exposed to doses of 2–1000 mg/L SVO₅ in drinking water for 5 weeks, both in the bone marrow and peripheral erythrocytes. The authors concluded that the data indicate scarce bioavailability for orally administered vanadium(IV) and lack of significant genotoxic potential in vivo.

On the other hand, in in vitro studies with human isolated lymphocytes, Wozniak and Blasiak (2004) found that DNA damage was induced with 0.5 or 1 mM SVO₅ within one hour of exposure. Studies conducted by our working group reported that treatment of human peripheral lymphocyte cultures with 1, 2, 4 or 8 µg/mL V₂O₄ produced an increase in chromatid breaks and acentric fragments as well as dicentrics (Rodríguez-Mercado et al. 2010). Additionally, vanadium(IV) induces DNA double-strand breaks (Rodríguez-Mercado et al. 2011), and this damage to the DNA can lead to chromosome lesions since it is generally accepted that mechanisms involved in repairing double-strand breaks and the genetic processes of recombination are mainly responsible for forming chromosomal aberrations (Obe and Durante 2010, Yamamura et al. 2018). It is likely that the increase in the SCA that we observed in this work is caused by this type of damage. Our results confirm that V₂O₄ causes DNA damage in both in vivo and in vitro cytogenetic tests and in different cell types.

Few studies have been carried out to assess the genotoxic potential of V₂O₃. Owusu-Yaw et al. (1990) reported an increase in SCA frequencies induced by V₂O₃ in CHO cells. In human blood leukocytes treated in vitro with the same compound produced DNA-single-strand breaks at all of the concentrations and treatment times tested (Rodríguez-Mercado et al. 2011) but showed no significant increase in the frequency of SCA (Rodríguez-Mercado et al. 2010).

In this study, we found that both V oxides produce SCA in non-dose dependent manner. The most frequent aberration types were fragments and chromatid breaks in both compounds, although it should be noted that some chromosome-type breaks and pulverisation were also observed. The increase in the percentage of cells with SCA with treatment with V₂O₄ and V₂O₃ may be based on different mechanisms. One of the modes of action in the induction of genotoxic effects of V compounds is the generation of ROS and/or DNA–protein cross-links because vanadium(IV) has the capability to bind DNA through interactions with guanine and adenine N-7 atoms and backbone PO₂ groups, resulting in the formation of 8-hydroxy-2′-deoxyguanosine, which is a common oxidative DNA adduct, and multiple and accumulative DNA single- and double-strand breaks cause clastogenicity (Rodríguez-Mercado et al. 2011, Espinosa-Zurutuza et al. 2018) or its ability to react with hydrogen peroxide and generate hydroxyl radicals (•OH) (Prousek 2007, He et al. 2022).

Unrepaired or misrepaired breaks may very well result in chromosomal damage, a lesion in the DNA, which is converted to a single-strand break during repair when a section of DNA is removed. If DNA synthesis occurs before the repair is complete, this single-strand break can be converted to a double-strand break, leading to the production of a visible aberration. For this reason, the production of chromosome aberrations requires cells to pass through S-phase for aberration formation. In addition, these aberrations will be chromatid type at the first metaphase after development (Danford 2012). Consequently, most of these aberrations may contribute to mutagenesis and carcinogenesis. Chromosome-type and chromatid-type aberrations and micronuclei can predict cancer risk (Doherty 2012). Therefore, the possibility that V oxides may produce cancer or chronic degenerative disease should not be ruled out.

The results show that both oxides produce a similar increase in SCA, despite having different oxidation states, which can be explained because we administered the same amount of elemental V, as stated in the material and methods section, and the applied doses of both compounds contain the same amount of elemental V. However, there is also the possibility that part of the vanadium(III) administered could be oxidized to vanadium(IV). Since V compounds that enter the bloodstream are subjected to speciation, it is known that vanadyl bound to transferrin is distributed to the body tissues and bones; vanadate (V) and oxidovanadium (IV) bound to transferrin can enter the intracellular space by endocytosis, analogous to Fe³⁺, and vanadium (IV) is not easily reduced to vanadium (III) under physiological conditions, but vanadium (III) can be oxidized to vanadium (IV) (Rehder 2013, 2015, López-Valdez et al. 2022).

Many environmental factors can damage DNA, and understanding the mechanisms associated with the outcomes of this process will provide researchers with tools to understand DNA environmental insults and ways to prevent or avoid them. Our results contribute to a better understanding of the mechanisms by which metals induce genotoxic effects. Finally, we want to note that the cytogenetic damage that V compounds produce is similar to that produced by other metals, such as Cd (Fahmy et al. 2000, Celik et al. 2005, 2009, El-Habit and Abdel-Moneim 2014).

5. Conclusions

In conclusion, our results showed that V₂O₄ and V₂O₃ treatment in mice reduced the MI, so it altered cell proliferation and increased the proportion of cells with SCA in bone marrow.

Acknowledgements

We thank MVZ. Adriana Altamirano Bautista and MVZ. Roman Hernández Meza for their technical support in the care and handling of the animals.

Disclosure statement

The authors declare no conflicts of interest.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article.

Additional information

Funding

During the development of this work, partial support was provided by Dirección General de Asuntos del Personal Académico, Universidad Nacional Autónoma de México PAPIIT IA201123 and IN210324.