Toxicity Estimation of Graphene Nanoparticles in Mus MusculusUsing Multiple Biomarkers
Main Article Content
Keywords
Gel electrophoresis, head DNA percentage, tail length, tail moment, hemolysis
Abstract
Graphene, a strong, super elastic, conductive, and crystalline carbon allotrope, is used in gene and drug delivery, osteon and dental implants, tissue grafting, biosensing, bioimaging, and photothermal therapies, etc. In the current study we evaluated the mutagenic potential of graphene by using Ames tests. Genetic damage and Hemolytic potential was also tested using comet assay and hemolysis assay. Male albino mice (n=66) were randomly selected, 5ml fresh blood from two of them were drawn for Hemolysis assay, and the remaining mice were divided into four groups; three treatment groups (dosages of 110mg/Kg, 220mg/kg, and 330mg/kg) and a control group. After treatment, blood was drawn for comet assay. Each group was again subdivided into three sub groups which were treated with L ascorbic acid (0.025 mg), retinol (0.03 mg) and no treatment for 15 days, and at 16th day same procedure was utilized for sample collection for comet assay. Data were recorded in CASP software and analyzed by using one-way ANOVA. The results of the Ames test indicate graphene is non-mutagenic, with a mutagenicity index less than 2 and 1.8 for TA98 and TA100 strains respectively. Dose dependent increase in hemolysis rate was also found in graphene because it can rupture the membrane of red blood cells (erythrocytes). Comet assay results showed Short-term oral exposure to graphene in male albino mice causes DNA damage, which can be repaired if exposure is ceased and some nutritional supplements if added.
References
1. Ames BN, Kammen HO and Yamasaki E, 1975. Hair dyes are mutagenic: identification of a variety of mutagenic ingredients. Proceedings of the National Academy of Sciences. 72: 2423-2427.
2. Akram R, Iqbal R, Hussain R, Jabeen F and Ali M, 2021. Evaluation of oxidative stress, antioxidant enzymes and genotoxic potential of bisphenol A in fresh water bighead carp (Aristichthys nobils) fish at low concentrations. Environmental pollution. 268: 115896.
3. Attri J, Dhawan V, Mahmood, S, Pandhi P, Parwana HK and Nath R, 2003. Effect of vitamin C supplementation on oxidative DNA damage in an experimental model of lead-induced hypertension. Annals of nutrition and metabolism. 47:294-301.
4. Begum P, Ikhtiari R and Fugetsu B, 2011. Graphene phytotoxicity in the seedling stage of cabbage, tomato, red spinach, and lettuce. Carbon. 49: 3907-3919.
5. Bhattacharya K, Gogoi B, Buragohain AK and Deb P, 2014. Fe2O3/C nanocomposites having distinctive antioxidant activity and hemolysis prevention efficiency. Materials Science and Engineering . 42: 595-600.
6. Chen GY, Yang HJ, Lu CH, Chao YC, Hwang SM, Chen CL and Hu YC, 2012. Simultaneous induction of autophagy and toll-like receptor signaling pathways by graphene oxide. Biomaterials. 33: 6559-6569.
7. Dasmahapatra AK, Dasari TP and Tchounwou PB, 2019. Graphene-based nanomaterials toxicity in fish. Reviews of Environmental Contamination and Toxicology Volume 247: 1-58.
8. Demir E and Marcos R, 2018. Toxic and genotoxic effects of graphene and multi-walled carbon nanotubes. Journal of Toxicology and Environmental Health, Part A.81: 645-660.
9. Dahanayaka M, Liu B, Srikanth N and Zhou K, 2020. Ionised graphene oxide membranes for seawater desalination. Desalination. 496: 114637.
10. Dash BS, Jose G, Lu YJ and Chen JP, 2021. Functionalized reduced graphene oxide as a versatile tool for cancer therapy. International journal of molecular sciences. 22: 2989.
11. Foroutan T, Nafar M and Motamedi E, 2020. Intraperitoneal injection of graphene oxide nanoparticle accelerates stem cell therapy effects on acute kidney injury. Stem Cells and Cloning: Advances and Applications, 13: 21.
12. Ghaffar A, Hussain R, Ahmad N, Ghafoor R, Akram MW, Khan I and Khan A, 2021. Evaluation of hemato-biochemical, antioxidant enzymes as biochemical biomarkers and genotoxic potential of glyphosate in freshwater fish (Labeo rohita). Chemistry and Ecology. 37: 646-667.
13. Ghaffar A, Hussain R, Abbas G, Khan R, Akram K, Latif H and Khan A, 2021. Assessment of genotoxic and pathologic potentials of fipronil insecticide in Labeo rohita (Hamilton, 1822). Toxin Reviews. 40:1289-1300.
14. Hussain R, Ali F, Javed MT, Jabeen G, Ghaffar A, Khan I, Liaqat, S, Hussain, T, Abbas RZ and Riaz A, 2021. Clinico-hematological, serum biochemical, genotoxic and histopathological effects of trichlorfon in adult cockerels. Toxin Reviews. 40:1206-1214.
15. Hussain R, Ghaffar A, Abbas G, Jabeen G, Khan I, Abbas RZ, Noreen S, Iqbal Z, Chaudhary I. R, and Ishaq HM, 2022. Thiamethoxam at sublethal concentrations induces histopathological, serum biochemical alterations and DNA damage in fish (Labeo rohita). Toxin Reviews. 41:154-164.
16. Hussain R, Naz S, Alam S, Ali HM, Ali A, Khan SM and Li K, 2024. Temporal and Dosage Impact of Magnesium Oxide Nanoparticles on Grass Carp: Unveiling Oxidative Stress, DNA Damage and Antioxidant Suppression. Toxicology Mechanisms and Methods, (just-accepted). 1-17.
17. Ivask A, Voelcker NH, Seabrook SA, Hor M, Kirby JK, Fenech M and Ke PC2015. DNA melting and genotoxicity induced by silver nanoparticles and graphene. Chemical Research in Toxicology. 28: 1023-1035.
18. Kong Z, Hu W, Jiao F, Zhang P, Shen J, Cui, B and Liang L, 2020. Theoretical evaluation of DNA genotoxicity of graphene quantum dots: A combination of density functional theory and molecular dynamics simulations. The Journal of Physical Chemistry B. 124: 9335-9342.
19. Kaźmierczak-Barańska J, Boguszewska K and Karwowski BT 2020. Nutrition can help DNA repair in the case of aging. Nutrients.12:33-64.
20. Liao KH,Lin YS, Macosko CW and Haynes CL, 2011. Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. ACS applied materials and interfaces. 3: 2607-2615.
21. Li Y, Wang Y, Tu L, Chen D, Luo Z, Liu D and Wang S, 2016. Sub-acute toxicity study of graphene oxide in the Sprague-Dawley rat. International journal of environmental research and public health. 13: 1149.
22. Lamponi S, 2022. Preliminary in vitro cytotoxicity, mutagenicity and antitumoral activity evaluation of graphene flake and aqueous graphene paste. Life, 12: 242.
23. Mortelmans K, and Zeiger E, 2000. The Ames Salmonella/microsome mutagenicity assay. Mutation research/fundamental and molecular mechanisms of mutagenesis, 455: 29-60.
24. Murata M and Kawanishi, S, 2000. Oxidative DNA damage by vitamin A and its derivative via superoxide generation. Journal of Biological Chemistry, 275: 2003-2008.
25. Ma Y, Shen H, Tu X and Zhang Z, 2014. Assessing in vivo toxicity of graphene materials: current methods and future outlook. Nanomedicine. 9: 1565-1580.
26. Mana MA, Niepes RA, Florida MA and Paculba RA, 2023. Growth performance of goats (Capra hircus L.) on forage legumes mixed with guinea grass (Megathyrsus maximus). Journal of Global Innovations in Agricultural Sciences. 11:305-309.
27. Nersesyan A, Fenech M, Bolognesi C, Mišík M, Setayesh T, Wultsch G and Knasmüller S, 2016. Use of the lymphocyte cytokinesis-block micronucleus assay in occupational biomonitoring of genome damage caused by in vivo exposure to chemical genotoxins: Past, present and future. Mutation Research/Reviews in Mutation Research. 770: 1-11.
28. Naz S, Hussain R, Ullah Q, Chatha AMM, Shaheen A and Khan RU, 2021. Toxic effect of some heavy metals on hematology and histopathology of major carp (Catla catla). Environmental science and pollution research. 28: 6533-6539.
29. Nasirzadeh N, Azari MR, Rasoulzadeh Y and Mohammadian Y, 2019. An assessment of the cytotoxic effects of graphene nanoparticles on the epithelial cells of the human lung. Toxicology and Industrial Health. 35:79-87.
30. Ou L, Lv X, Wu Z, Xia W, Huang Y, Chen L and Qi L, 2021. Oxygen content-related DNA damage of graphene oxide on human retinal pigment epithelium cells. Journal of Materials Science: Materials in Medicine. 32: 1-9.
31. Parveen N, Ahmad S and Shadab GHA, 2014. Iron induced genotoxicity: attenuation by vitamin C and its optimization. Interdisciplinary Toxicology. 7:154-158.
32. Pelin M, Sosa S, Prato M and Tubaro A, 2018. Occupational exposure to graphene based nanomaterials: risk assessment. Nanoscale. 10: 15894-15903.
33. Priyadarsini S, Sahoo SK, Sahu S, Mukherjee S, Hota G and Mishra M, 2019. Oral administration of graphene oxide nano-sheets induces oxidative stress, genotoxicity, and behavioral teratogenicity in Drosophila melanogaster. Environmental Science and Pollution Research. 26: 19560-19574.
34. Park W, Shin H, Choi B, Rhim WK, Na K and Han DK, 2020. Advanced hybrid nanomaterials for biomedical applications. Progress in Materials Science.114: 100-686.
35. Purcarea C, Laslo V, Memete AR, Agud E, Miere F and Vicas SI, 2022. Antigenotoxic and antimutagenic potentials of proline in allium cepa exposed to the toxicity of cadmium. Agriculture. 12: 1568.
36. Radadiya TM, 2015. A properties of graphene. European Journal of Material Sciences. 2: 6-18.
37. Shi LQ and Zheng RL, 2006. DNA damage and oxidative stress induced by Helicobacter pylori in gastric epithelial cells: protection by vitamin C and sodium selenite. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 61: 631-637.
38. Soares JC, Pereira TCB, Costa KM, Maraschin T, Basso NR and Bogo MR, 2017. Developmental neurotoxic effects of graphene oxide exposure in zebrafish larvae (Danio rerio). Colloids and Surfaces B. Biointerfaces. 157: 335-346.
39. Singh N, Sahoo SK and Kumar R, 2020. Hemolysis tendency of anticancer nanoparticles changes with type of blood group antigen: An insight into blood nanoparticle interactions. Materials Science and Engineering.109:110645.
40. Serrano-Aroca Á, Takayama K, Tuñón-Molina A, Seyran M, Hassan SS, Pal Choudhury P and Brufsky A, 2021. Carbon-based nanomaterials: promising antiviral agents to combat COVID-19 in the microbial-resistant era. ACS nano. 15: 8069-8086.
41. Saebo IP, Bjørås M, Franzyk H, Helgesen E and Booth JA, 2023. Optimization of the hemolysis assay for the assessment of cytotoxicity. International journal of molecular sciences. 24: 2914.
42. Vallelian F, Buehler PW and Schaer DJ, 2022. Hemolysis, free hemoglobin toxicity, and scavenger protein therapeutics. Blood, The Journal of the American Society of Hematology. 140: 1837-1844.
43. Wang Y, Li Z, Wang J, Li J and Lin Y, 2011. Graphene and graphene oxide: biofunctionalization and applications in biotechnology. Trends in biotechnology. 29:205-212.
44. Wang JQ, Hussain R, Ghaffar A, Afzal G, Saad AQ, Ahmad N and Khan A, 2022. Clinicohematological, mutagenic, and oxidative stress induced by pendimethalin in freshwater fish bighead carp (Hypophthalmichthys nobilis). Oxidative Medicine and Cellular Longevity. 2022: 2093822.
45. Zhang X, Zhang J, Wang Q, Ghimire S, Mei L and Wu C, 2022. Effects of particle size and surface charge on mutagenicity and chicken embryonic toxicity of new silver nanoclusters. ACS omega. 7: 17703-17712.
2. Akram R, Iqbal R, Hussain R, Jabeen F and Ali M, 2021. Evaluation of oxidative stress, antioxidant enzymes and genotoxic potential of bisphenol A in fresh water bighead carp (Aristichthys nobils) fish at low concentrations. Environmental pollution. 268: 115896.
3. Attri J, Dhawan V, Mahmood, S, Pandhi P, Parwana HK and Nath R, 2003. Effect of vitamin C supplementation on oxidative DNA damage in an experimental model of lead-induced hypertension. Annals of nutrition and metabolism. 47:294-301.
4. Begum P, Ikhtiari R and Fugetsu B, 2011. Graphene phytotoxicity in the seedling stage of cabbage, tomato, red spinach, and lettuce. Carbon. 49: 3907-3919.
5. Bhattacharya K, Gogoi B, Buragohain AK and Deb P, 2014. Fe2O3/C nanocomposites having distinctive antioxidant activity and hemolysis prevention efficiency. Materials Science and Engineering . 42: 595-600.
6. Chen GY, Yang HJ, Lu CH, Chao YC, Hwang SM, Chen CL and Hu YC, 2012. Simultaneous induction of autophagy and toll-like receptor signaling pathways by graphene oxide. Biomaterials. 33: 6559-6569.
7. Dasmahapatra AK, Dasari TP and Tchounwou PB, 2019. Graphene-based nanomaterials toxicity in fish. Reviews of Environmental Contamination and Toxicology Volume 247: 1-58.
8. Demir E and Marcos R, 2018. Toxic and genotoxic effects of graphene and multi-walled carbon nanotubes. Journal of Toxicology and Environmental Health, Part A.81: 645-660.
9. Dahanayaka M, Liu B, Srikanth N and Zhou K, 2020. Ionised graphene oxide membranes for seawater desalination. Desalination. 496: 114637.
10. Dash BS, Jose G, Lu YJ and Chen JP, 2021. Functionalized reduced graphene oxide as a versatile tool for cancer therapy. International journal of molecular sciences. 22: 2989.
11. Foroutan T, Nafar M and Motamedi E, 2020. Intraperitoneal injection of graphene oxide nanoparticle accelerates stem cell therapy effects on acute kidney injury. Stem Cells and Cloning: Advances and Applications, 13: 21.
12. Ghaffar A, Hussain R, Ahmad N, Ghafoor R, Akram MW, Khan I and Khan A, 2021. Evaluation of hemato-biochemical, antioxidant enzymes as biochemical biomarkers and genotoxic potential of glyphosate in freshwater fish (Labeo rohita). Chemistry and Ecology. 37: 646-667.
13. Ghaffar A, Hussain R, Abbas G, Khan R, Akram K, Latif H and Khan A, 2021. Assessment of genotoxic and pathologic potentials of fipronil insecticide in Labeo rohita (Hamilton, 1822). Toxin Reviews. 40:1289-1300.
14. Hussain R, Ali F, Javed MT, Jabeen G, Ghaffar A, Khan I, Liaqat, S, Hussain, T, Abbas RZ and Riaz A, 2021. Clinico-hematological, serum biochemical, genotoxic and histopathological effects of trichlorfon in adult cockerels. Toxin Reviews. 40:1206-1214.
15. Hussain R, Ghaffar A, Abbas G, Jabeen G, Khan I, Abbas RZ, Noreen S, Iqbal Z, Chaudhary I. R, and Ishaq HM, 2022. Thiamethoxam at sublethal concentrations induces histopathological, serum biochemical alterations and DNA damage in fish (Labeo rohita). Toxin Reviews. 41:154-164.
16. Hussain R, Naz S, Alam S, Ali HM, Ali A, Khan SM and Li K, 2024. Temporal and Dosage Impact of Magnesium Oxide Nanoparticles on Grass Carp: Unveiling Oxidative Stress, DNA Damage and Antioxidant Suppression. Toxicology Mechanisms and Methods, (just-accepted). 1-17.
17. Ivask A, Voelcker NH, Seabrook SA, Hor M, Kirby JK, Fenech M and Ke PC2015. DNA melting and genotoxicity induced by silver nanoparticles and graphene. Chemical Research in Toxicology. 28: 1023-1035.
18. Kong Z, Hu W, Jiao F, Zhang P, Shen J, Cui, B and Liang L, 2020. Theoretical evaluation of DNA genotoxicity of graphene quantum dots: A combination of density functional theory and molecular dynamics simulations. The Journal of Physical Chemistry B. 124: 9335-9342.
19. Kaźmierczak-Barańska J, Boguszewska K and Karwowski BT 2020. Nutrition can help DNA repair in the case of aging. Nutrients.12:33-64.
20. Liao KH,Lin YS, Macosko CW and Haynes CL, 2011. Cytotoxicity of graphene oxide and graphene in human erythrocytes and skin fibroblasts. ACS applied materials and interfaces. 3: 2607-2615.
21. Li Y, Wang Y, Tu L, Chen D, Luo Z, Liu D and Wang S, 2016. Sub-acute toxicity study of graphene oxide in the Sprague-Dawley rat. International journal of environmental research and public health. 13: 1149.
22. Lamponi S, 2022. Preliminary in vitro cytotoxicity, mutagenicity and antitumoral activity evaluation of graphene flake and aqueous graphene paste. Life, 12: 242.
23. Mortelmans K, and Zeiger E, 2000. The Ames Salmonella/microsome mutagenicity assay. Mutation research/fundamental and molecular mechanisms of mutagenesis, 455: 29-60.
24. Murata M and Kawanishi, S, 2000. Oxidative DNA damage by vitamin A and its derivative via superoxide generation. Journal of Biological Chemistry, 275: 2003-2008.
25. Ma Y, Shen H, Tu X and Zhang Z, 2014. Assessing in vivo toxicity of graphene materials: current methods and future outlook. Nanomedicine. 9: 1565-1580.
26. Mana MA, Niepes RA, Florida MA and Paculba RA, 2023. Growth performance of goats (Capra hircus L.) on forage legumes mixed with guinea grass (Megathyrsus maximus). Journal of Global Innovations in Agricultural Sciences. 11:305-309.
27. Nersesyan A, Fenech M, Bolognesi C, Mišík M, Setayesh T, Wultsch G and Knasmüller S, 2016. Use of the lymphocyte cytokinesis-block micronucleus assay in occupational biomonitoring of genome damage caused by in vivo exposure to chemical genotoxins: Past, present and future. Mutation Research/Reviews in Mutation Research. 770: 1-11.
28. Naz S, Hussain R, Ullah Q, Chatha AMM, Shaheen A and Khan RU, 2021. Toxic effect of some heavy metals on hematology and histopathology of major carp (Catla catla). Environmental science and pollution research. 28: 6533-6539.
29. Nasirzadeh N, Azari MR, Rasoulzadeh Y and Mohammadian Y, 2019. An assessment of the cytotoxic effects of graphene nanoparticles on the epithelial cells of the human lung. Toxicology and Industrial Health. 35:79-87.
30. Ou L, Lv X, Wu Z, Xia W, Huang Y, Chen L and Qi L, 2021. Oxygen content-related DNA damage of graphene oxide on human retinal pigment epithelium cells. Journal of Materials Science: Materials in Medicine. 32: 1-9.
31. Parveen N, Ahmad S and Shadab GHA, 2014. Iron induced genotoxicity: attenuation by vitamin C and its optimization. Interdisciplinary Toxicology. 7:154-158.
32. Pelin M, Sosa S, Prato M and Tubaro A, 2018. Occupational exposure to graphene based nanomaterials: risk assessment. Nanoscale. 10: 15894-15903.
33. Priyadarsini S, Sahoo SK, Sahu S, Mukherjee S, Hota G and Mishra M, 2019. Oral administration of graphene oxide nano-sheets induces oxidative stress, genotoxicity, and behavioral teratogenicity in Drosophila melanogaster. Environmental Science and Pollution Research. 26: 19560-19574.
34. Park W, Shin H, Choi B, Rhim WK, Na K and Han DK, 2020. Advanced hybrid nanomaterials for biomedical applications. Progress in Materials Science.114: 100-686.
35. Purcarea C, Laslo V, Memete AR, Agud E, Miere F and Vicas SI, 2022. Antigenotoxic and antimutagenic potentials of proline in allium cepa exposed to the toxicity of cadmium. Agriculture. 12: 1568.
36. Radadiya TM, 2015. A properties of graphene. European Journal of Material Sciences. 2: 6-18.
37. Shi LQ and Zheng RL, 2006. DNA damage and oxidative stress induced by Helicobacter pylori in gastric epithelial cells: protection by vitamin C and sodium selenite. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 61: 631-637.
38. Soares JC, Pereira TCB, Costa KM, Maraschin T, Basso NR and Bogo MR, 2017. Developmental neurotoxic effects of graphene oxide exposure in zebrafish larvae (Danio rerio). Colloids and Surfaces B. Biointerfaces. 157: 335-346.
39. Singh N, Sahoo SK and Kumar R, 2020. Hemolysis tendency of anticancer nanoparticles changes with type of blood group antigen: An insight into blood nanoparticle interactions. Materials Science and Engineering.109:110645.
40. Serrano-Aroca Á, Takayama K, Tuñón-Molina A, Seyran M, Hassan SS, Pal Choudhury P and Brufsky A, 2021. Carbon-based nanomaterials: promising antiviral agents to combat COVID-19 in the microbial-resistant era. ACS nano. 15: 8069-8086.
41. Saebo IP, Bjørås M, Franzyk H, Helgesen E and Booth JA, 2023. Optimization of the hemolysis assay for the assessment of cytotoxicity. International journal of molecular sciences. 24: 2914.
42. Vallelian F, Buehler PW and Schaer DJ, 2022. Hemolysis, free hemoglobin toxicity, and scavenger protein therapeutics. Blood, The Journal of the American Society of Hematology. 140: 1837-1844.
43. Wang Y, Li Z, Wang J, Li J and Lin Y, 2011. Graphene and graphene oxide: biofunctionalization and applications in biotechnology. Trends in biotechnology. 29:205-212.
44. Wang JQ, Hussain R, Ghaffar A, Afzal G, Saad AQ, Ahmad N and Khan A, 2022. Clinicohematological, mutagenic, and oxidative stress induced by pendimethalin in freshwater fish bighead carp (Hypophthalmichthys nobilis). Oxidative Medicine and Cellular Longevity. 2022: 2093822.
45. Zhang X, Zhang J, Wang Q, Ghimire S, Mei L and Wu C, 2022. Effects of particle size and surface charge on mutagenicity and chicken embryonic toxicity of new silver nanoclusters. ACS omega. 7: 17703-17712.
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Sep 2, 2024
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Toxicity Estimation of Graphene Nanoparticles in Mus MusculusUsing Multiple Biomarkers. (2024). Journal of Population Therapeutics and Clinical Pharmacology, 31(9), 24-40. https://doi.org/10.53555/yzw8y742
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Author Biographies
Aqsa Fayyaz
Government College University Faisalabad.
Tayyaba Ali
Government College University Faisalabad
Farkhanda Asad
Government College University Faisalabad.
Farhat Jabeen
Government College University Faisalabad.
How to Cite
Toxicity Estimation of Graphene Nanoparticles in Mus MusculusUsing Multiple Biomarkers. (2024). Journal of Population Therapeutics and Clinical Pharmacology, 31(9), 24-40. https://doi.org/10.53555/yzw8y742