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Article

Titanium Dioxide Nanoparticles-Induced Genotoxic Effects in Mosquito Culex quinquefaciatus

by
Aastha Saini
1,
Kanwaljit Kaur Ahluwalia
1,*,
Amrik Singh Ahluwalia
2,
Neelam Thakur
1,*,
Puneet Negi
3,
Abeer Hashem
4,
Khalid F. Almutairi
5 and
Elsayed Fathi Abd_Allah
5
1
Department of Zoology, Akal College of Basic Sciences, Eternal University, Baru Sahib, Sirmaur 173101, Himachal Pradesh, India
2
Department of Botany, Akal College of Basic Sciences, Eternal University, Baru Sahib, Sirmaur 173101, Himachal Pradesh, India
3
Department of Physics, Akal College of Basic Sciences, Eternal University, Baru Sahib, Sirmaur 173101, Himachal Pradesh, India
4
Botany and Microbiology Department, College of Science, King Saud University, P.O. Box. 2460, Riyadh 11451, Saudi Arabia
5
Plant Production Department, College of Food and Agricultural Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia
*
Authors to whom correspondence should be addressed.
Toxics 2024, 12(12), 871; https://doi.org/10.3390/toxics12120871
Submission received: 17 October 2024 / Revised: 25 November 2024 / Accepted: 26 November 2024 / Published: 29 November 2024
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Abstract

Titanium dioxide (TiO2) nanoparticles are being extensively used in a wide range of industrial applications for producing a variety of different consumer products, including medicines and even food items. The consumption of these products is increasing at an alarming rate, and this results in the release of these nanoparticles in the environment, causing a threat to organisms thriving in aquatic as well as terrestrial ecosystems. That is why screening such materials for their genotoxic effects, if any, becomes essential. A toxicity assay was performed to determine the LD20 of these nanoparticles for the mosquito Culex quinquefaciatus by Probit analysis. Early fourth instar larvae were exposed to the selected dose of 50 µg/mL, which is <LD20 value, for 24 h treatment. Chromosomal slides were prepared from lacto-aceto-orcein-stained gonads of adult mosquitoes developed from treated and control larvae. These nanoparticles were reported cytotoxic as a statistically significant decline in mitotic index in treated mosquitoes than controls were observed. The nanoparticles were also found to induce various structural and numerical chromosomal aberrations in the treated lot. In the end, it can be concluded that these nanoparticles do have a genotoxic effect. The present study provides a caution against further use of these nanoparticles. There must be the development of strategies for the safe, sustainable use as well as proper disposal of these nanoparticles so as to protect both biotic and non-biotic components of the environment.

Graphical Abstract

1. Introduction

Nanomaterials have been implemented in various industrial and agricultural sectors for decades. Due to their ever-increasing commercial applications and presence in the majority of consumer products, these are being extensively investigated for their safety. Genotoxicity, which is the tendency of physical and chemical agents to alter genetic information, is one of the major concerns with respect to nanomaterials [1].
TiO2, often known as titania, was first mass-produced in the early 20th century as a non-toxic alternative to a white dye for paint. Later, it and its nano-forms use were extended to diverse commercial products, and now it can be found in all kinds of toothpaste, ceramics, printing inks, paper, plastics, rubber, synthetic fibers, condensers, paints and crayons, electronic components along with food and cosmetics [2,3,4,5,6,7,8,9]. In addition to these utilities, more and more photodynamic implementation trials are using TiO2 NPs with the main focus on their significance as photosensitizing agents in the treatment of malignant tumors as well as in the photodynamic inactivation of antibiotic-resistant bacteria [10]. These have also been used in photoelectrochemical bio-sensing applications that greatly improve target detection. That is why these are often loaded with various medications and employed as local drug delivery systems for various diseases such as osteoporosis, cancer, and microbial inflammations [11].
Various experimental and epidemiological studies have reported severe effects of titanium dioxide nanoparticles in the liver, brain, kidney and spleen, causing adverse impacts on human health [12,13,14,15]. These are also reported to be neurotoxic, resulting in memory loss and compromised learning capabilities in rodents [16,17,18]. These particles are classified as group 2B carcinogens (suspected carcinogens) by the International Agency for Research on Cancer. In 2021, the European Food Safety Authority (EFSA) declared TiO2-NPs unsafe for their use as food additives as their long-time exposure and high dosage could affect the function of various major organs and recommended even their ban for further use. A ban on titanium dioxide (E171) as a food additive was implemented in the European Union in 2022 as its use is simply to enhance the aesthetic value of food only and does not serve any important purpose. On the one hand, these nanoparticles are being used in diverse applications; on the other hand, these are reported to confer toxicity because of their undesired interactions with many biological components and cellular processes [19,20,21]. In the present scenario, the genotoxicity of TiO2 NPs to living organisms is a topic of high concern and needs proper assessment. Different researchers have already obtained controversial results as far as the genotoxicity of these NPs is concerned using different endpoints in various in vitro and in vivo test models [22,23,24]. In view of these contrasting results, the present piece of research work is carried out particularly at a dose that may not impact the general mortality of the individuals, maintaining the population size. This study is intended to ensure the safety of non-target organisms, including humans, exposed to such environmental pollutants unintentionally.
Because of the time-consuming nature of long-term experiments, a number of quick tests using bacteria, plants, rodents, and mammals as test models are in practice. In vivo studies are preferred over in vitro ones as later results are frequently found falsely positive. However, nowadays, in vivo research on large animals like monkeys, rats, guinea pigs, etc., is viewed as unethical. In light of this, insects have been suggested as useful alternative experimental material for such studies. Among insects, mosquitoes are selected, recommended and proved to be a good test model for assessing the clastogenic, mutagenic, and genotoxic potential of different environmental pollutants [25,26,27,28,29,30,31]. A few of the benefits of their selection in these studies include their short life span, easy rearing and maintenance in the laboratory, and the presence of a low diploid number of only six relatively large-sized chromosomes. Recently, these have already been used as test models for determining the genotoxicity of even nanoparticles [32]. The most dangerous metallic compounds have particle sizes in the submicron range of 0.25–5.0 and have been shown to be the most harmful to human health. Nanoparticles, being still smaller in size, are expected to be more hazardous than their larger counterparts due to their enhanced reactivity and penetrability in cells [33]. So, it becomes essential to analyze their genotoxic impact on living organisms and categorize them as safe or dangerous for biodiversity and human health. The main purpose of this study is to assess the genotoxic potential of these nanoparticles by observing their impact on mitotic index and genetic material.

2. Materials and Methods

A colony of the mosquito Culex quinquefasciatus Say belonging to the family Culicidae and order Diptera was raised and maintained in special mosquito-rearing cages at a constant temperature of 27 ± 2 °C and relative humidity of 75 ± 5% for sufficient and constant availability of the material to carry out various experiments.

2.1. Preparation of TiO2 Nanoparticles

TiO2 nanoparticles used in the present study were synthesized by sol–gel auto-combustion route [34]. Initially, two different solutions, X and Y, were prepared.
  • Solution X: Prepared by mixing titanium (IV) isopropoxide (TTIP, C12H28O4Ti, 97% Sigma Aldrich, Saint Louis, MO, USA) and ethanol (C2H5OH, Himedia 99.8%) in a ratio of 1:3 for 30 min on a magnetic stirrer (Ambala, Haryana, India).
  • Solution Y: Prepared by mixing double-distilled water (DDW), ethanol and acetic acid glacial AR (CH3 COOH) in a ratio of 1:2:7 by stirring for the same time on a magnetic stirrer.
  • Synthesis technique: A milky solution (sol) was obtained by adding, drop by drop, solution X to solution Y. Then, for the formation of gel, this sol was stirred at 25 °C for 10–12 h, maintaining the pH value of approximately 7 by adding ammonium hydro-oxide (NH4OH, Nice 30% NH3). The gel was dried in an oven maintained at 100 °C for 48–72 h to obtain the crystals. A fine powder was prepared by grinding these crystals using pastel mortar for 45 min. This fine powder was calcined at 450 °C for 2 h in the furnace to obtain titanium dioxide nanoparticles. These nanoparticles’ crystalline form is anatase, having a tetragonal structure, and its average crystallite size is about 10 nm.

2.2. Toxicity Assay for Determining LD20 of TiO2 NPs for Mosquito Culex quinquefasciatus

The toxicity assay was conducted by treating 40 early fourth instar larvae of the mosquito to six different doses of these nanoparticles, and a control with an equal number of larvae in an equal volume of distilled water was also maintained. These doses (TiO2 NPs suspensions) were prepared by adding 30, 20, 10, 2, 1, and 0.2 mg of the nanoparticles to 20 mL of distilled water in separate sterilized glass vials, and these concentrations were properly vortexed before their application. After 24 h treatment, the larvae were transferred to distilled water and were fed on larval feed made by mixing powdered dog biscuits and yeast in a 3:1 ratio. The mortality data of the treated larvae were recorded (Table 1), and from the percent mortality data, LD20 was determined by Probit analysis [35]. From the regression equation y = 1.313x + 1.766 and the regression line so obtained, the value of LD20 was calculated, and it came out to be 66.53 µg/mL (Figure 1).

2.3. Selection of Dose for Genotoxic Assays

The selection of a dose is important for quantitative assessment and a more precise evaluation of genotoxic effects because a dose that is too high or too low can give pseudo-positive or negative results, respectively. The dose that does not reduce the population size drastically but does possess some toxicity is usually selected for such assays. Therefore, a toxicity assay was performed to determine the LD20 of the nanoparticles for the mosquito Culex quinquefaciatus. Out of all the doses used in the toxicity assay, 50 µg/mL dose, the nearest < LD20 value, was selected for the various assays to be performed for assessing the genotoxic potential of TiO2 NPs.

2.4. Genotoxic Assays

Different assays were performed to assess the genotoxic potential of TiO2 nanoparticles and for each assay, 20 early fourth instar mosquito larvae were kept in freshly vortexed 10 mL of the selected dose of TiO2 NPs suspension for 24 h. After the treatment, the larvae were shifted to a nutrient medium, which is distilled water with larval feed added to it, so that they could grow and mature into adults. These adults were used to carry out various assays performed to assess the genotoxic potential of these nanoparticles. Three replicates of this treatment were performed, and a control of 20 larvae in 10 mL of distilled water was also maintained.

2.4.1. Mitotic Index Assay

Mitotic slides were prepared from the gonads of 12–15 h old adult mosquitoes from treated as well as control stocks using the method of Crozier [36] with some modifications. After dissecting out gonads in a dissecting medium of 1% sodium citrate solution, these were fixed for 3–5 min in Carnoy’s fixative (ethanol and glacial acetic acid in a 3:1 ratio) and thereafter stained in 2% lacto-aceto-orcein stain for 10–15 min to obtain proper color. A squash preparation of the stained material was made, and the slides were observed under a microscope to study the frequency of dividing cells.

2.4.2. Chromosomal Aberration Assay

For this assay, chromosomal slides were prepared using the same method but after arresting cell division at the metaphase stage in the gonads by treating the adult mosquitoes with 0.1% colchicine (C22H28NO8, MW 399.44, 98.6% pure procured from Loba Chemie Pvt. Ltd., Mumbai, India) solution about 3–4 h prior to dissection. The slides were screened for chromosomal aberrations, if any, so as to determine genetic damage induced by the nanoparticles.

2.4.3. Statistical Analysis

Student’s t-test was applied to compare the mean percent frequency data of at least three replicates obtained for mitotic indices and chromosomal aberrations induced in the gonadial cells of treated and control mosquitoes.

3. Results

3.1. Mitotic Index

About 600–800 cells were studied in each of the three replicates, and the percentage frequency of dividing cells (MI) was calculated. The mean MI values came out to be 52.05 ± 6.22% for treated mosquitoes against 83.63 ± 5.55% in the control (Table 2, Figure 2). The comparison of mean values revealed a statistically significant decline in mitotic index in treated mosquitoes compared to the controls (p < 0.05).

3.2. Chromosomal Aberrations

In the present study, both structural as well as numerical changes in the chromosomes after treatment of TiO2 nanoparticles were observed. Structural changes include clumping, fragments, terminal fusions, breaks, and translocations of chromosomes (Figure 3), while numerical alterations recorded were aneuploid cells. The hyperploid cells, such as trisomic (2n + 1) and tetrasomic (2n + 2), as well as hypoploid cells with monosomic (2n − 1) and nullisomic (2n − 2) conditions, were observed. An average of 105 cells were studied for chromosomal aberrations in each of the treated and control slots. The percent frequency of total aberrations recorded was 19.32 ± 2.61% in TiO2 NPs treated mosquitoes, and this was only 2.52 ± 0.21% in the controls (Table 3). The outcome is statistically significant (p < 0.05). Out of the total aberrations induced, the frequency of structural aberrations was found to be 15.20 ± 1.21%, and this value is 1.90 ± 0.06% in controls. The most prevalent structural aberration found was cells with clumped chromosomes, having a frequency of 4.38 ± 0.84%, and the least observed was breaks in chromosomes with a frequency of 1.55 ± 0.47.

4. Discussion

The release of TiO2 NPs nanoparticles in the environment, due to their wide range of industrial applications, is increasing and causing a threat to organisms thriving in aquatic as well as terrestrial ecosystems. In order to assess the environmental danger that a particular species is exposed to, it is crucial to understand the effects caused by TiO2 NPs as well as the mechanisms underlying their activity, which may be ROS- or non-ROS-mediated [37,38]. The genotoxic potential of TiO2 NPs using short-term in vivo assays, viz. mitotic index and chromosomal aberration assays, are studied. Both the assays are performed at a selected dose of <LD20. This idea of using <LD20 is endorsed by the recommendation of the cell-inhibiting concentration IC25 for in vitro cytotoxic assays by Santos-Aguilar and others [39]. Further, El Yamani and associates, in 2017 [40], found these nanoparticles genotoxic at non-cytotoxic concentrations in cultured human lymphoblastoid cells by comet assay. This study also emphasizes the importance of the selection of doses for such studies.
Different researchers reached positive as well as negative outcomes in both in vivo and in vitro experiments performed by different test models after TiO2 NPs exposures [21,23,24,37,41,42,43,44]. Contrary results obtained in different studies are obvious because of the use of different concentrations, sizes, and forms of the nanoparticles to assess the genetic damage induced. Donner and others [45] could not find any micronuclei induction in the reticulocytes of rats fed with different doses of these nanoparticles. Lack of absorption from the alimentary tract was explained as a reason for the negative effects observed. However, Gao and coworkers [46] observed an adverse impact on ovarian cells in mice after intragastric infusion of TiO2 NPs, and this observation indicates absorption through the digestive tract wall. Migration across cell membranes leading to the uptake and internalization of these particles into cells was reported by others [43,47]. Dorier and associates [48] stressed the role of some carriers and efflux pumps in the uptake of these nanoparticles across the gut epithelium. Most of these nanoparticles are reported to migrate and accumulate in vital organs, induce toxic effects in their tissues, and disturb the metabolic processes of the organism’s body [49,50,51]. In the present study, these particles were found inside the haemocoel of mosquitoes, and the internalization of these particles is further endorsed by positive results obtained in both the parameters carried out to test the genotoxicity of the nanoparticles in question. A statistical increase in chromosomal aberrations over control was observed. Both structural alterations, including chromosomal clumping, fragments, breaks, terminal fusions, and translocations, as well as numerical alterations comprising hyperaneurploids (2n + 1 and 2n + 2) and hypoaneuploids (2n − 1 and 2n − 2) were encountered. The breaks near the terminal ends of chromosomes make them sticky, leading to clumping and terminal fusions. The reason for the occurrence of aneuploids may be due to the abnormal disjunction of chromosomes during their segregation at anaphase stages of mitosis [29]. These results are in conformity with similar observations on germ and bone marrow cells in mice [52,53,54,55]. An increase in chromosomal abnormalities was also shown in cultured human peripheral blood cells after 24 h exposure to these nanoparticles by Patel and others [56]. Further, they also observed double-strand breaks in cultured bone marrow cells of rats, indicating the genetic damage caused by these nanoparticles. The genetic damage induced by NPs may be due to oxidative stress and the generation of cytokines, which harm DNA, lipids, proteins, and carbohydrates [57,58,59,60].
An EFSA [61] panel chaired by Younes in 2021 published a report regarding the safety assessment of titanium dioxide (E171) and concluded that E171 cannot be considered safe as a food additive, even after considering the limited relevance of genotoxic studies with TiO2 NPs with size < 30 nm. Although NPs present in E171 food additives are bigger (size > 30 nm) and constitute only a minor proportion of the additive, recently, in 2024, Warheit [62] put forth a view that labeling these particles risky may be an error, as its genotoxicity depends on diverse factors and positive results shown in various such studies are controversial. However, as per the report of Weir et al. [2] in the year 2012, these NPs constitute 36% of this additive, which is used in most common food items like chewing gum, candies, white icing on cakes, etc., repeatedly consumed in large amounts, especially by children, its safety assessment is very crucial as future generations of human beings are at stake. If any agent induces chromosomal aberrations, which are direct evidence of genetic damage, that agent cannot be considered safe. In the present study, too, statistically significant chromosomal aberrations are reported to be induced by these NPs, although having a size < 30 nm. The EFSA panel seems to be right or genuine in labeling E171 as unsafe, especially for its repeated exposures.
A statistically significant decline in mitotic index in treated mosquitoes compared to the controls (p < 0.05) was observed in the present research work. A delay in the G2 to M phase of the cell cycle and a slowdown in the mitotic division were observed [63,64]. A decrease in cell division is also reported by Ferrante and team [65]. The reason for this decline may be the impact of these nanoparticles on the expression of genes involved in the regulation of mitosis [22,66,67]. The cytotoxic effects of nanomaterials have been reported to be due to their interaction with enzymes involved in the regulation of cell division [68]. Sarikhani and others, in the year 2022 [69], suggested a decrease in stem cells due to the induction of apoptosis during the cell cycle as the cause of retardation of cell proliferation leading to a decline in the mitotic index of cells. The induction of cellular apoptosis by TiO2 NPs was also shown by other research workers [39,70]. Over the past few decades, the majority of data on the photocatalytic activity of TiO2 NPs and their associated phototoxic effects have been acquired at the cellular level [37,71].
Crystalline form and particle size have been shown to influence the genotoxic potential of TiO2 nanoparticles. The anatase form of the nanoparticles was found to be more genotoxic than its other forms [72,73]. Gurr and others [74] reported 10–20 nm-sized TiO2 nanoparticles-induced genotoxicity in the form of DNA damage and micronuclei formation in human bronchial epithelial cell lines. The reason for this toxicity was found to be oxidative stress, which is evident from their observation of the increased production of hydrogen peroxide in the absence of photo-activation. However, these results were not reported after these cell lines were exposed to 200 to > 200 nm-sized particles under similar conditions. In murine MC3T3-E1 pre-osteoblast cells, the toxicity of the cells treated with 5 nm TiO2 NPs exhibited higher levels of lactate dehydrogenase (LDH) release, mitochondrial damage, and apoptosis when compared to those treated with 32 nm TiO2 NPs [75]. Some other findings also implied that the smaller the particle size, the more genetic damage is caused [76,77,78]. In the present study, the anatase form of these particles measuring 10 nm was used, and it gave statistically significant positive results, thus confirming the above-mentioned observations. The possible mechanism for genetic damage by titanium dioxide nanoparticles was indicated to be due to the release of reactive oxygen species (ROS), which in turn causes lipid peroxidation [3,44,53,79,80]. Mancuso and team [81] reported that the anatase crystal, the form of the nanoparticles used in the present study, is also able to induce the production of ROS in cells, and this could be the plausible reason for their genotoxic potential.

5. Conclusions

From the present study, it is quite evident that TiO2 NPs, particularly having a size of about 10 nm, do possess genotoxic potential, as these nanoparticles have been found to induce statistically significant (p < 0.05) decline in mitotic index and increase in various structural and numerical chromosomal aberrations over control at a dose less than LD20 and are not absolutely safe for use in different consumer products. The present study warrants their careful use and disposal in future. Further, experiments can be conducted to mitigate the harmful impacts of these nanoparticles.

Author Contributions

Conceptualization, K.K.A.; methodology, K.K.A., P.N. and A.S.; software, A.S., N.T. and K.K.A.; validation, K.K.A.; formal analysis, A.S. and K.K.A.; investigation, A.S.; resources, K.K.A.; data curation, A.S. and K.K.A.; writing—original draft preparation, A.S. and K.K.A.; writing—review and editing, K.K.A., A.S.A., N.T., K.F.A., A.H. and E.F.A.; visualization, A.S. and K.K.A.; supervision, K.K.A.; funding acquisition, A.H. and E.F.A. All authors have read and agreed to the published version of the manuscript.

Funding

The authors would like to extend their sincere appreciation to the Researchers Supporting Project Number (RSP2025R356), King Saud University, Riyadh, Saudi Arabia.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The data presented in this study are available on request from the corresponding authors.

Acknowledgments

The authors express their gratitude to Hon’ble Chancellor Eternal University Baru Sahib, Himachal Pradesh, India, for providing the necessary infrastructure to carry out the research. The authors would also like to extend their sincere appreciation to the Researchers Supporting Project Number (RSP2025R356), King Saud University, Riyadh, Saudi Arabia.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Kohl, Y.E.; Rundén-Pran, E.; Mariussen, M.; Hesler, N.; El Yamani, E.M.; Longhin, M.; Dusinska, M. Genotoxicity of nanomaterials: Advanced in vitro models and high throughput methods for human hazard assessment—A review. Nanomaterials 2020, 10, 1911. [Google Scholar] [CrossRef] [PubMed]
  2. Weir, A.P.; Westerhoff, L.; Fabricius, K.; Hristovski, N.; Von Goetz, N. Titanium dioxide nanoparticles in food and personal care products. Environ. Sci. Technol. 2012, 46, 2242–2250. [Google Scholar] [CrossRef] [PubMed]
  3. Luo, Z.; Li, Z.; Xie, I.M.; Sokolova, L.; Song, W.J.; Peijnenburg, M.; Hu, M.; Wang, Y. Rethinking nano-TiO2 safety: Overview of toxic effects in humans and aquatic animals. Nano Micro Small 2020, 16, 2002019. [Google Scholar] [CrossRef] [PubMed]
  4. Contado, C. Nanomaterials in consumer products: A challenging analytical problem. Front. Chem. 2015, 3, 48. [Google Scholar] [CrossRef] [PubMed]
  5. Bogdan, J.; Pławińska-Czarnak, J.; Zarzyńska, J. Nanoparticles of titanium and zinc oxides as novel agents in tumor treatment: A review. Nanoscale Res. Lett. 2017, 12, 1–15. [Google Scholar] [CrossRef]
  6. Sharma, S.; Sharma, R.K.; Gaur, K.; Cátala Torres, J.F.; Loza-Rosas, S.A.; Torres, A.; Saxena, M.; Julin, M.; Tinoco, A.D. Fueling a hot debate on the application of TiO2 nanoparticles in sunscreen. Materials 2019, 12, 2317. [Google Scholar] [CrossRef]
  7. Haider, A.J.; Jameel, Z.N.; Al-Hussaini, I.H. Review on: Titanium dioxide applications. Energy Procedia 2019, 157, 17–29. [Google Scholar] [CrossRef]
  8. Wang, S.; Ding, Z.; Chang, X.; Xu, J.; Wang, D.H. Modified nano-TiO2 based composites for environmental photocatalytic applications. Catalysts 2020, 10, 759. [Google Scholar] [CrossRef]
  9. Cornu, R.; Béduneau, A.; Martin, H. Ingestion of titanium dioxide nanoparticles: A definite health risk for consumers and their progeny. Arch. Toxicol. 2022, 96, 2655–2686. [Google Scholar] [CrossRef]
  10. Ziental, D.; Czarczynska-Goslinska, B.; Mlynarczyk, D.T.; Glowacka-Sobotta, A.; Stanisz, B.; Goslinski, T.; Sobotta, L. Titanium dioxide nanoparticles: Prospects and applications in medicine. Nanomaterials 2020, 10, 387. [Google Scholar] [CrossRef]
  11. Jafari, S.; Mahyad, B.; Hashemzadeh, H.; Janfaza, S.; Gholikhani, T.; Tayebi, L. Biomedical applications of TiO2 nanostructures: Recent advances. Int. J. Nanomed. 2020, 14, 3447–3470. [Google Scholar] [CrossRef] [PubMed]
  12. Minetto, D.; Ghirardini, A.V.; Libralato, G. Saltwater ecotoxicology of Ag, Au, CuO, TiO2, ZnO and C60 engineered nanoparticles: An overview. Environ. Int. 2016, 92, 189–201. [Google Scholar] [CrossRef] [PubMed]
  13. Iavicoli, I.; Leso, V.; Fontana, L.; Bergamaschi, A. Toxicological effects of titanium dioxide nanoparticles: A review of in vitro mammalian studies. Eur. Rev. Med. Pharmacol. Sci. 2011, 15, 481–508. [Google Scholar] [PubMed]
  14. Khosravi-Katuli, K.; Prato, E.; Lofrano, G.; Guida, M.; Vale, G.; Libralato, G. Effects of nanoparticles in species of aquaculture interest. Environ. Sci. Pollut. Res. 2017, 24, 17326–17346. [Google Scholar] [CrossRef]
  15. Dar, G.I.; Saeed, M.; Wu, A. Toxicity of TiO2 nanoparticles. In TiO2 Nanoparticles; Wu, A., Ren, W., Eds.; Wiley-VCH Verlag GmbH and Co. KGaA: Hoboken, NJ, USA, 2020; pp. 67–103. [Google Scholar]
  16. Hu, R.; Gong, X.; Duan, Y.; Li, N.; Che, Y.; Cui, Y.; Hong, F. Neurotoxicological effects and the impairment of spatial recognition memory in mice caused by exposure to TiO2 nanoparticles. Biomaterials 2010, 31, 8043–8050. [Google Scholar] [CrossRef]
  17. Ze, Y.; Hu, R.; Wang, X.; Sang, X.; Ze, X.; Li, B.; Su, J.; Wang, Y.; Guan, N.; Zhao, X.; et al. Neurotoxicity and gene-expressed profile in brain-injured mice caused by exposure to titanium dioxide nanoparticles. J. Biomed. Mater. Res. 2014, 102, 470–478. [Google Scholar] [CrossRef]
  18. Aijie, C.; Huimin, L.; Jia, L.; Lingling, O.; Limin, W.; Junrong, W.; Xuan, L.; Xue, H.; Longquan, S. Central neurotoxicity induced by the instillation of ZnO and TiO2 nanoparticles through the taste nerve pathway. Nanomedicine 2017, 12, 2453–2470. [Google Scholar] [CrossRef]
  19. Yang, Y.; Qin, Z.; Zeng, W.; Yang, T.; Cao, Y.; Mei, C.; Kuang, Y. Toxicity assessment of nanoparticles in various systems and organs. Nanotechnol. Rev. 2017, 6, 279–289. [Google Scholar] [CrossRef]
  20. Gupta, R.; Xie, H. Nanoparticles in daily life: Applications, toxicity and regulations. J. Environ. Pathol. Toxicol. Oncol. 2018, 37, 209–230. [Google Scholar] [CrossRef]
  21. Garcia-Rodriguez, A.; Rubio, L.; Vila, L.; Xamena, N.; Velázquez, A.; Marcos, R.; Hernández, A. The comet assay as a tool to detect the genotoxic potential of nanomaterials. Nanomaterials 2019, 9, 1385. [Google Scholar] [CrossRef]
  22. Carriere, M.; Arnal, M.E.; Douki, T. TiO2 genotoxicity: An update of the results published over the last six years. Mutat. Res. Genet. Toxicol. Environ. Mutatgen. 2020, 854, 503198. [Google Scholar] [CrossRef] [PubMed]
  23. Ling, C.; An, H.; Li, L.; Wang, J.; Lu, T.; Wang, H.; Hu, Y.; Song, G.; Liu, S. Genotoxicity evaluation of titanium dioxide nanoparticles in vitro: A systematic review of the literature and meta-analysis. Biol. Trace Elem. Res. 2021, 199, 2057–2076. [Google Scholar] [CrossRef] [PubMed]
  24. Shi, J.; Han, S.; Zhang, J.; Liu, Y.; Chen, Z.; Jia, G. Advances in genotoxicity of titanium dioxide nanoparticles in vivo and in vitro. NanoImpact 2022, 25, 100377. [Google Scholar] [CrossRef]
  25. Sharma, G.P.; Sobti, R.C.; Chaudhry, A.; Ahluwalia, K.K. Genotoxicity of two heavy metal compounds- lead acetate and mercuric chloride in the mosquito Anopheles stephensi Liston (Culicidae: Diptera). Cytologia 1988, 53, 263–267. [Google Scholar] [CrossRef]
  26. Sharma, G.P.; Sobti, R.C.; Chaudhry, A.; Ahluwalia, K.K. Chromosomal aberrations and dominant lethals in Culex fatigans due to mercuric chloride. Cytobios 1989, 59, 131–135. [Google Scholar] [PubMed]
  27. Sobti, R.; Sobti, A. An introduction to recent approaches underlying mechanistic insights harboring oncobiology. In Handbook of Oncobiology: From Basic to Clinical Sciences; Sobti, R.C., Ganguly, N.K., Kumar, R., Eds.; Springer: Singapore, 2024; pp. 3–47. [Google Scholar]
  28. Ahluwalia, K.K.; Chaudhary, S. Genotoxic potential of mercuric nitrate in Culex quinquefaciatus. Res. Bull. Panjab Univ. 1999, 49, 9–16. [Google Scholar]
  29. Sharma, G.P.; Chaudhry, A.; Sobti, R.C.; Dhar, M.; Ahluwalia, K.K. Effect of mitomycin-C on the genetics of Culex fatigans (Diptera: Culicidae). La Kromosomo II 1990, 58, 1949–1954. [Google Scholar]
  30. Marwaha, L. In vivo genotoxicity evaluation of carbaryl pesticides using polytene chromosomes of Anopheles culicifacies (Diptera: Culicidae). Toxicol. Int. 2016, 23, 4–11. [Google Scholar] [CrossRef]
  31. Mishra, P.; Balaji, A.P.B.; Dhal, P.K.; Kumar, R.S.; Magolassi, S.; Margulis, K.; Tyagi, B.K.; Mukherjee, A.; Chandrasekaran, N. Stability of nano-sized permethrin in its colloidal state and its effect on the physiological and biochemical profile of Culex tritaeniorhynchus larvae. Bull. Entomol. Res. 2017, 107, 676–688. [Google Scholar] [CrossRef]
  32. Ahluwalia, K.K.; Thakur, K.; Ahluwalia, A.S.; Hashem, A.; Avila-Quezada, G.D.; Abd_Allah, E.F.; Thakur, N. Assessment of genotoxicity of zinc oxide nanoparticles using mosquito as test model. Toxics 2023, 11, 887. [Google Scholar] [CrossRef]
  33. Foroozandeh, P.; Aziz, A.A. Insight into cellular uptake and intracellular trafficking of nanoparticles. Nanoscale Res. Lett. 2018, 13, 339. [Google Scholar] [CrossRef] [PubMed]
  34. Kaur, G.; Negi, P.; Konwar, R.J.; Kumar, H.; Devi, N.; Kaur, G.; Boruah, R.; Ranjan, M.; Sooraj, K.P.; Raval, N.; et al. Tweaking the structural, micro-structural, optical and dielectric properties of anatase titanium dioxide via doping of nitrogen ions. Ceram. Int. 2024, 50, 48138–48163. [Google Scholar] [CrossRef]
  35. Finney, D.J. A Statistical Treatment of the Sigmoid Response Curve, 3rd ed.; Cambridge University Press: London, UK, 1971; p. 333. [Google Scholar]
  36. Crozier, R.H. An acetic acid dissociation, air-drying technique for insect chromosomes, with aceto-lactic orcein staining. Stain Technol. 1968, 43, 171–173. [Google Scholar] [CrossRef] [PubMed]
  37. Wang, Y.; Ding, L.; Yao, C.; Li, C.; Xing, X.; Huang, Y.; Gu, T.; Wu, M. Toxic effects of metal oxide nanoparticles and their underlying mechanisms. Sci. China Mater. 2017, 60, 93–108. [Google Scholar] [CrossRef]
  38. Hou, J.; Wang, L.; Wang, C.; Zhang, S.; Liu, H.; Li, S.; Wang, X. Toxicity and mechanisms of action of titanium dioxide nanoparticles in living organisms. J. Environ. Sci. 2019, 75, 40–53. [Google Scholar] [CrossRef]
  39. Santos-Aguilar, P.; Bernal-Ramírez, J.; Vázquez-Garza, E.; Vélez-Escamilla, L.Y.; Lozano, O.; García-Rivas, G.D.J.; Contreras-Torres, F.F. Synthesis and Characterization of Rutile TiO2 Nanoparticles for the Toxicological Effect on the H9c2 Cell Line from Rats. ACS Omega 2023, 8, 19024–19036. [Google Scholar] [CrossRef]
  40. El Yamani, N.; Collins, A.R.; Rundén-Pran, E.; Fjellsbø, L.M.; Shaposhnikov, S.; Zienolddiny, S.; Dusinska, M. In vitro genotoxicity testing of four reference metal nanomaterials, titanium dioxide, zinc oxide, cerium oxide and silver: Towards reliable hazard assessment. Mutagenesis 2017, 32, 117–126. [Google Scholar] [CrossRef]
  41. Chen, Z.; Wang, Y.; Ba, T.; Li, Y.; Pu, J.; Chen, T.; Song, Y.; Jia, G. Genotoxic evaluation of titanium dioxide nanoparticles in vivo and in vitro. Toxicol. Lett. 2014, 226, 314–319. [Google Scholar] [CrossRef]
  42. Demir, E.; Akça, H.; Turna, F.; Aksakal, S.; Burgucu, D.; Kaya, B.; Tokgun, O.; Vales, G.; Creus, S.; Marcos, R. Genotoxic and cell-transforming effects of titanium dioxide nanoparticles. Environ. Res. 2015, 136, 300–308. [Google Scholar] [CrossRef]
  43. Kazimirova, A.; El Yamani, N.; Rubio, L.; García-Rodríguez, A.; Barancokova, M.; Marcos, R.; Dusinska, M. Effects of titanium dioxide nanoparticles on the Hprt gene mutations in V79 hamster cells. Nanomaterials 2020, 10, 465. [Google Scholar] [CrossRef]
  44. Cao, Y.; Chen, J.; Bian, Q.; Ning, J.; Yang, L.; Ou, T.; Sang, Y.; Wei, S. Genotoxicity evaluation of titanium dioxide nanoparticles in vivo and in vitro: A meta-analysis. Toxics 2023, 11, 882. [Google Scholar] [CrossRef] [PubMed]
  45. Donner, E.M.; Myhre, A.; Brown, S.C.; Boatman, R.; Warheit, D.B. In vivo micronucleus studies with 6 titanium dioxide materials (3 pigment-grade and 3 nanoscale) in orally-exposed rats. Regul. Toxicol. Pharmacol. 2016, 74, 64–74. [Google Scholar] [CrossRef] [PubMed]
  46. Gao, G.; Ze, Y.; Li, B.; Zhao, X.; Sang, X.; Zhang, T.; Sheng, L.; Hu, R.; Gui, S.; Sang, X.; et al. Ovarian dysfunction and gene-expressed characteristics of female mice caused by long term exposure to titanium dioxide nanoparticles. J. Hazard. Mater. 2012, 243, 9–27. [Google Scholar] [CrossRef] [PubMed]
  47. Ye, L.; Hong, F.; Ze, X.; Li, L.; Zhou, Y.; Ze, Y. Toxic effects of TiO2 nanoparticles in primary cultured rat sertoli cells are mediated via a dysregulated C a2+/PKC/p38 MAPK/NF-κ B cascade. J. Biomed. Mater. Res. A 2017, 105, 1374–1382. [Google Scholar] [CrossRef] [PubMed]
  48. Dorier, M.; Brun, E.; Veronesi, G.; Barreau, F.; Pernet-Gallay, K.; Desvergne, C.; Rabilloud, T.; Carapito, C.; Herlin-Boime, N.; Carrière, M. Impact of anatase and rutile titanium dioxide nanoparticles on uptake carriers and efflux pumps in Caco-2 gut epithelial cells. Nanoscale 2015, 7, 7352–7360. [Google Scholar] [CrossRef]
  49. Morgan, A.M.; Abd El-Hamid, M.I.; Noshy, P.A. Reproductive toxicity investigation of titanium dioxide nanoparticles in male albino rats. World J. Pharm. Pharm. Sci. 2015, 4, 34–49. [Google Scholar]
  50. Herrera-Rodriguez, M.A.; del Pilar Ramos-Godinez, M.; Cano-Martínez, A.; Segura, F.C.; Ruiz-Ramírez, A.; Pavón, N.; Lira-Silva, E.; Bautista-Pérez, R.; Thomas, R.S.; Delgado-Buenrostro, N.L.; et al. Food-grade titanium dioxide and zinc oxide nanoparticles induce toxicity and cardiac damage after oral exposure in rats. Part. Fibre Toxicol. 2023, 20, 43. [Google Scholar] [CrossRef]
  51. Bautista-Pérez, R.; Cano-Martínez, A.; Herrera-Rodriguez, M.A.; Ramos-Godinez, M.D.P.; Pérez Reyes, O.L.; Chirino, Y.I.; Serrano, Z.J.R.; López-Marure, R. Oral exposure to titanium dioxide E171 and zinc oxide nanoparticles induces multi-organ damage in rats: Role of Ceramide. Int. J. Mol. Sci. 2024, 25, 5881. [Google Scholar] [CrossRef]
  52. Rad, J.S.; Alfatemi, M.H.; Rad, M.S.; Rad, M.S.; Sen, D.J.; Mohsenzadeh, S. In-vivo titanium dioxide (TiO2) nanoparticles effects on chromosomal abnormalities and lactate dehydrogenase activity. Am. J. Adv. Drug Deliv. 2013, 1, 232–237. [Google Scholar]
  53. Rizk, M.Z.; Ali, S.A.; Hamed, M.A.; El-Rigal, N.S.; Aly, H.F.; Salah, H.H. Toxicity of titanium dioxide nanoparticles: Effect of dose and time on biochemical disturbance, oxidative stress and genotoxicity in mice. Biomed. Pharmacother. 2017, 90, 466–472. [Google Scholar] [CrossRef]
  54. Ali, S.A.; Rizk, M.Z.; Hamed, M.A.; Aboul-Ela, E.I.; El-Rigal, N.S.; Aly, H.F.; Abdel-Hamid, A.H.Z. Assessment of titanium dioxide nanoparticles toxicity via oral exposure in mice: Effect of dose and particle size. Biomarkers 2019, 24, 492–498. [Google Scholar] [CrossRef] [PubMed]
  55. Salman, A.S.; Al-Shaikh, T.M.; Hamza, Z.K.; El-Nekeety, A.A.; Bawazir, S.S.; Hassan, N.S.; Abdel-Wahhab, M.A. Matlodextrin-cinnamon essential oil nanoformulation as a potent protective against titanium nanoparticles-induced oxidative stress, genotoxicity, and reproductive disturbances in male mice. Environ. Sci. Pollut. Res. 2021, 28, 39035–39051. [Google Scholar] [CrossRef] [PubMed]
  56. Patel, S.; Patel, P.; Bakshi, S.R. Titanium dioxide nanoparticles: An in vitro study of DNA binding, chromosome aberration assay and comet assay. Cytotechnology 2017, 69, 245–263. [Google Scholar] [CrossRef] [PubMed]
  57. Kelly, S.A.; Havrilla, C.M.; Brady, T.C.; Abramo, K.H.; Levin, E.D. Oxidative stress in toxicology: Established mammalian and emerging piscine model systems. Environ. Health Perspect. 1998, 106, 375–384. [Google Scholar] [CrossRef] [PubMed]
  58. Jia, X.; Wang, S.; Zhou, L.; Sun, L. The potential liver, brain, and embryo toxicity of titanium dioxide nanoparticles on mice. Nanoscale Res. Lett. 2017, 12, 1–14. [Google Scholar] [CrossRef] [PubMed]
  59. Nagakannan, P.; Tabeshmehr, P.; Eftekharpour, E. Oxidative damage of lysosomes in regulated cell death systems: Pathophysiology and pharmacologic interventions. Free Radic. Biol. Med. 2020, 157, 94–127. [Google Scholar] [CrossRef]
  60. Sallam, M.F.; Ahmed, H.M.; Diab, K.A.; El-Nekeety, A.A.; Abdel-Aziem, S.H.; Sharaf, H.A.; Abdel-Wahhab, M.A. Improvement of the antioxidant activity of thyme essential oil against biosynthesized titanium dioxide nanoparticles-induced oxidative stress, DNA damage, and disturbances in gene expression in vivo. J. Trace Elem. Med. Biol. 2022, 73, 127024. [Google Scholar] [CrossRef]
  61. EFSA Panel on Food Additives and Flavourings (FAF); Younes, M.; Aquilina, G.; Castle, L.; Engel, K.H.; Fowler, P.; Frutos Fernandez, M.J.; Fürst, P.; Gundert-Remy, U.; Gürtler, R.; et al. Safety assessment of titanium dioxide (E171) as a food additive. EFSA J. 2021, 19, e06585. [Google Scholar]
  62. Warheit, D.B. Safety of titanium dioxide (E171) as a food additive for humans. Front. Toxicol. 2024, 6, 1333746. [Google Scholar] [CrossRef]
  63. Chang, H.; Wang, Q.; Meng, X.; Chen, X.; Deng, Y.; Li, L.; Yang, Y.; Song, G.; Jia, H. Effect of titanium dioxide nanoparticles on mammalian cell cycle in vitro: A systematic review and meta-analysis. Chem. Res. Toxicol. 2022, 35, 1435–1456. [Google Scholar] [CrossRef]
  64. Huang, S.; Chueh, P.J.; Lin, Y.W.; Shih, T.S.; Chuang, S.M. Disturbed mitotic progression and genome segregation are involved in cell transformation mediated by nano-TiO2 long-term exposure. Toxicol. Appl. Pharmacol. 2009, 241, 182–194. [Google Scholar] [CrossRef] [PubMed]
  65. Ferrante, M.; Grasso, A.; Salemi, R.; Libra, M.; Tomasello, B.; Fiore, M.; Copat, C. DNA damage and apoptosis as in-vitro effect biomarkers of titanium dioxide nanoparticles (TiO2 NPs) and the food additive E171 toxicity in colon cancer cells: HCT-116 and Caco-2. Int. J. Environ. Res. Public Health 2023, 20, 2002. [Google Scholar] [CrossRef] [PubMed]
  66. Cui, Y.; Liu, H.; Ze, Y.; Zengli, Z.; Hu, Y.; Cheng, Z.; Cheng, J.; Hu, R.; Gao, G.; Wang, L.; et al. Gene expression in liver injury caused by long-term exposure to titanium dioxide nanoparticles in mice. Toxicol. Sci. 2012, 128, 171–185. [Google Scholar] [CrossRef] [PubMed]
  67. Leroux, M.M.; Doumandji, Z.; Chézeau, L.; Gaté, L.; Nahle, S.; Hocquel, R.; Zhernovkov, V.; Migot, S.; Ghanbaja, J.; Bonnet, C.; et al. Toxicity of TiO2 nanoparticles: Validation of alternative models. Int. J. Mol. Sci. 2020, 21, 4855. [Google Scholar] [CrossRef]
  68. Magdolenova, Z.; Collins, A.; Kumar, A.; Dhawan, A.; Stone, V.; Dusinska, M. Mechanisms of genotoxicity. A review of in vitro and in vivo studies with engineered nanoparticles. Nanotoxicology 2014, 8, 233–278. [Google Scholar] [CrossRef]
  69. Sarikhani, M.; Vaghefi Moghaddam, S.; Firouzamandi, M.; Hejazy, M.; Rahimi, B.; Moeini, H.; Alizadeh, E. Harnessing rat derived model cells to assess the toxicity of TiO2 nanoparticles. J. Mater. Sci. Mater. Med. 2022, 33, 41. [Google Scholar] [CrossRef]
  70. Li, N.; Duan, Y.; Hong, M.; Zheng, L.; Fei, M.; Zhao, X.; Wang, J.; Cui, Y.; Liu, H.; Cai, J.; et al. Spleen injury and apoptotic pathway in mice caused by titanium dioxide nanoparticules. Toxicol. Lett. 2010, 195, 161–168. [Google Scholar] [CrossRef]
  71. Tang, Y.; Cai, R.; Cao, D.; Kong, X.; Lu, Y. Photocatalytic production of hydroxyl radicals by commercial TiO2 nanoparticles and phototoxic hazard identification. Toxicology 2018, 406, 1–8. [Google Scholar] [CrossRef]
  72. Khan, M.; Naqvi, A.H.; Ahmad, M. Comparative study of the cytotoxic and genotoxic potentials of zinc oxide and titanium dioxide nanoparticles. Toxicol. Rep. 2015, 2, 765–774. [Google Scholar] [CrossRef]
  73. Ghosh, M.; Öner, D.; Duca, R.C.; Cokic, S.M.; Seys, S.; Kerkhofs, S.; Landuyt, K.V.; Hoet, P.; Godderis, L. Cyto-genotoxic and DNA methylation changes induced by different crystal phases of TiO2-np in bronchial epithelial (16-HBE) cells. Mutat. Res. Mol. Mech. Mutagen. 2017, 796, 1–12. [Google Scholar] [CrossRef]
  74. Gurr, J.R.; Wang, A.S.; Chen, C.H.; Jan, K.Y. Ultrafine titanium dioxide particles in the absence of photoactivation can induce oxidative damage to human bronchial epithelial cells. Toxicology 2005, 213, 66–73. [Google Scholar] [CrossRef] [PubMed]
  75. Zhang, Y.; Yu, W.; Jiang, X.; Lv, K.; Sun, S.; Zhang, F. Analysis of the cytotoxicity of differentially sized titanium dioxide nanoparticles in murine MC3T3-E1 preosteoblasts. J. Mater. Sci. Mater. Med. 2011, 22, 1933–1945. [Google Scholar] [CrossRef] [PubMed]
  76. Sycheva, L.P.; Zhurkov, V.S.; Iurchenko, V.V.; Daugel-Dauge, N.O.; Kovalenko, M.A.; Krivtsova, E.K.; Durnev, A.D. Investigation of genotoxic and cytotoxic effects of micro-and nanosized titanium dioxide in six organs of mice in vivo. Mutat. Res. Toxicol. Environ. Mutagen. 2011, 726, 8–14. [Google Scholar] [CrossRef] [PubMed]
  77. Proquin, H.; Rodriguez-Ibarra, C.; Moonen, C.G.J.; Urrutia Ortega, I.M.; Briede, J.J.; deKok, T.M.; van Loveren, H.; Chirino, Y.I. Titanium dioxide food additive (E171) induces ROS formation and genotoxicity: Contribution of micro and nano-sized fractions. Mutagenesis 2017, 32, 139–149. [Google Scholar] [CrossRef] [PubMed]
  78. Wang, J.; Wang, J.; Liu, Y.; Nie, Y.; Si, B.; Wang, T.; Waqas, A.; Zhao, G.; Wang, M.; Xu, A. Aging-independent and size-dependent genotoxic response induced by titanium dioxide nanoparticles in mammalian cells. J. Environ. Sci. 2019, 85, 94–106. [Google Scholar] [CrossRef]
  79. Hanot-Roy, M.; Tubeuf, E.; Guilbert, A.; Bado-Nilles, A.; Vigneron, P.; Trouiller, B.; Lacroix, G. Oxidative stress pathways involved in cytotoxicity and genotoxicity of titanium dioxide (TiO2) nanoparticles on cells constitutive of alveolo-capillary barrier in vitro. Toxicol. In Vitro 2016, 33, 125–135. [Google Scholar] [CrossRef]
  80. Nichols, C.E.; Shepherd, D.L.; Hathaway, Q.A.; Durr, A.J.; Thapa, D.; Abukabda, A.; Hollander, J.M. Reactive oxygen species damage drives cardiac and mitochondrial dysfunction following acute nano-titanium dioxide inhalation exposure. Nanotoxicology 2019, 12, 32–48. [Google Scholar] [CrossRef]
  81. Mancuso, F.; Arato, I.; Di Michele, A.; Antognelli, C.; Angelini, L.; Bellucci, C.; Lilli, C.; Boncompagni, S.; Fusella, A.; Bartolini, D.; et al. Effects of titanium dioxide nanoparticles on porcine prepubertal sertoli cells: An “in vitro” study. Front. Endocrinol. 2022, 12, 751915. [Google Scholar] [CrossRef]
Toxics 12 00871 g001
Figure 1. Linear regression line and equation, obtained from mortality data, representing the relationship of Probit of kill and log concentrations of TiO2 NPs.
Figure 1. Linear regression line and equation, obtained from mortality data, representing the relationship of Probit of kill and log concentrations of TiO2 NPs.
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Toxics 12 00871 g002
Figure 2. Gonadial cells of adult mosquito Culex quinquefaciatus: (a) control mosquito with more dividing cells and (b) treated mosquito with fewer dividing cells.
Figure 2. Gonadial cells of adult mosquito Culex quinquefaciatus: (a) control mosquito with more dividing cells and (b) treated mosquito with fewer dividing cells.
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Toxics 12 00871 g003
Figure 3. Chromosomal aberrations, structural (b–f) and numerical (g–i) ones induced by TiO2 NPs: (a) normal chromosomal complement (2n = 6), (b) chromosomal fragments, (c) chromosomal break, (d) clumped chromosomes, (e) terminal fusion, (f) translocation, (g) 2n − 1, (h) 2n − 2, and (i) 2n + 2.
Figure 3. Chromosomal aberrations, structural (b–f) and numerical (g–i) ones induced by TiO2 NPs: (a) normal chromosomal complement (2n = 6), (b) chromosomal fragments, (c) chromosomal break, (d) clumped chromosomes, (e) terminal fusion, (f) translocation, (g) 2n − 1, (h) 2n − 2, and (i) 2n + 2.
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Table 1. The percent mortality data of Culex quinquefaciatus mosquito after TiO2 NPs treatment for 24 h.
Table 1. The percent mortality data of Culex quinquefaciatus mosquito after TiO2 NPs treatment for 24 h.
DosesNumber of Larvae Treated (n)Number of Larvae Killed (r)Percentage Mortality (r/n × 100)
(mg/mL)(µg/mL)
1.51500403690%
1.01000403075%
0.5500402255%
0.1100401640%
0.055040615%
0.01104037.5%
Control4025%
Table 2. Percent frequency of dividing cells in the gonads of TiO2 NPs treated and control adult mosquito Culex quinquefaciatus.
Table 2. Percent frequency of dividing cells in the gonads of TiO2 NPs treated and control adult mosquito Culex quinquefaciatus.
TreatmentReplicatesTotal No. of Cells ObservedNo. Of Dividing Cells% Frequency of Dividing CellsPercent Mitotic Index
(Mean ± S.E.)		‘t’-Value
TiO2 NPsR178829036.8052.05 ± 6.22−3.11 *
R266139559.75
R378546859.61
ControlR181472088.4583.63 ± 5.55
R261957192.24
R371550270.20
* The results are statistically significant at p < 0.05. The negative value of ‘t’ indicates decline in mitotic index from control values.
Table 3. Percent frequency of mitotic chromosomal aberrations induced by TiO2 NPs in gonadial cells of mosquito Culex quinquefaciatus.
Table 3. Percent frequency of mitotic chromosomal aberrations induced by TiO2 NPs in gonadial cells of mosquito Culex quinquefaciatus.
TreatmentChromosomal Aberrations
Structural Aberrations (SA)Numerical Aberrations (NA)Total Structural AberrationsTotal Numerical AberrationsTotal Aberrations‘t’-Value
Aneuploids
Clumped
Chromosomes		FragmentsTerminal FusionsBreaksTranslocations(2n + 1)(2n + 2)(2n − 1)(2n − 2)(SA)(NA)(SA + NA)
TiO2 NPs4.38 ± 0.842.55 ± 0.553.52 ± 0.991.55 ± 0.473.17 ± 0.270.95 ± 0.461.90 ± 0.460.62 ± 0.250.62 ± 0.2515.20 ± 1.214.12 ± 1.4019.32 ± 2.612.63 *
Control0.61 ± 0.250.32 ± 0.260.67 ± 0.540.000.28 ± 0.230.000.000.61 ± 0.250.001.90 ± 0.060.61 ± 0.252.52 ± 0.21
* The data are mean ± S.E. of three replicates and are statistically significant at p < 0.05.
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Saini, A.; Ahluwalia, K.K.; Ahluwalia, A.S.; Thakur, N.; Negi, P.; Hashem, A.; Almutairi, K.F.; Abd_Allah, E.F. Titanium Dioxide Nanoparticles-Induced Genotoxic Effects in Mosquito Culex quinquefaciatus. Toxics 2024, 12, 871. https://doi.org/10.3390/toxics12120871

AMA Style

Saini A, Ahluwalia KK, Ahluwalia AS, Thakur N, Negi P, Hashem A, Almutairi KF, Abd_Allah EF. Titanium Dioxide Nanoparticles-Induced Genotoxic Effects in Mosquito Culex quinquefaciatus. Toxics. 2024; 12(12):871. https://doi.org/10.3390/toxics12120871

Chicago/Turabian Style

Saini, Aastha, Kanwaljit Kaur Ahluwalia, Amrik Singh Ahluwalia, Neelam Thakur, Puneet Negi, Abeer Hashem, Khalid F. Almutairi, and Elsayed Fathi Abd_Allah. 2024. "Titanium Dioxide Nanoparticles-Induced Genotoxic Effects in Mosquito Culex quinquefaciatus" Toxics 12, no. 12: 871. https://doi.org/10.3390/toxics12120871

APA Style

Saini, A., Ahluwalia, K. K., Ahluwalia, A. S., Thakur, N., Negi, P., Hashem, A., Almutairi, K. F., & Abd_Allah, E. F. (2024). Titanium Dioxide Nanoparticles-Induced Genotoxic Effects in Mosquito Culex quinquefaciatus. Toxics, 12(12), 871. https://doi.org/10.3390/toxics12120871

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