Uses of nanoparticles (NPs) are increasing day by day almost in every field of science. Due to its vast demanding usage its necessary to assess their adverse impact on organisms. Metal NPs in particular have received implicated interest due to its widespread medical, military, industrial and co
Assessment of genotoxicity of cobalt doped zinc oxide nanoparticles
Uses of nanoparticles (NPs) are increasing day by day almost in every field of science. Due to its vast demanding usage its necessary to assess their adverse impact on organisms. Metal NPs in particular have received implicated interest due to its widespread medical, military, industrial and cosmetic applications.
However as particle size decreases, some metal NPs are observed with increasing toxicity as well. Thus, aim of our project is to assess Genotoxcicity of Cobalt-doped Zinc Oxide NPs on Eukaryotes and Prokaryotes. Which will eventually show us whether these NPs are toxic at DNA level or not.
Prokaryotic model organism will be E.coli, and in case for eukaryotes yeast in unicellular category whereas rats belongs to multicellular eukaryotic organism group.
Recently Nanoparticles have revolutionized in every field of science and technology and have many applications, promises to offer. Nonetheless, their safety and toxicity is still a concern. For this very purpose, the current study is designed to investigate the potential toxicity of Cobalt-doped Zinc Oxide Nanoparticles both on eukaryotic and prokaryotic model organisms.
As per Food and Drug Administration (FDA) USA Comet Assay, Micronucleus Assay and Ames test are three authentic tests for assessments of genotoxicity of any chemical such as drugs, paints, pesticides etc. However at least two of the above mention test are mandatory before approval of any chemical for commercial use.
To achieve this goal following objectives would be followed:
Project Implementation Methods includes in vitro and in vivo testing of organisms as per discussed:
Comet Assay and Gel electrophoresis:
These are some of the most sophisticated techniques to check genotoxicity. Comet Assay and Micronucleus assays are very reliable techniques through which we can analyze DNA damage in current project.
The comet assay, a gel electrophoresis–based method that can be used to measure DNA damage in individual eukaryotic cells. It is versatile, relatively simple to perform and sensitive. Although most investigations make use of its ability to measure DNA single-strand breaks, modifications to the method allow detection of DNA double-strand breaks, cross-links, base damage and apoptotic nuclei.
In this method first step is to lyse cell by using salts and detergents, Followed by electrophoresis from lysed cell DNA released and then electrophoresed in a low melting agarose gel under alkaline PH conditions.
Those cells will migrate quickly towards anode, which contains huge amount of double stranded breaks (DBs). This technique is reliable because it’s easy, less time consuming and cheaper than other techniques. It requires smaller amount of cells and displays results.
Micronuclei (MN) are formed during anaphase from chromosomal fragments or whole chromosomes that are left behind when the nuclei divides over time, the assay has evolved to include a pretreatment with a cytokinesis-blocking agent that inhibits cell division, thereby giving the cells a binucleated appearance. This enables more accurate scoring and the ability to sieve out the dividing cells from the non-dividing ones.
As per Food and Drug Administration (FDA) USA Comet Assay, Micronucleus Assay and Ames test are three authentic tests for assessments of genotoxicity of any chemical such as drugs, paints, pesticides etc. However at least two of the above mention test are mandatory before approval of any chemical for commercial use.
These tests should enable identification of hazards with respect to DNA damage and fixation.
After this study people will keep in row "Genotoxicity of Nanoparticles" as important attribute to deal with futher medicines, cosmetics and vice versa. This will provide a new horizone in study and practical procedure of widely used nanoparticles.
Initially we will evaluate nanoparticles under investigation and analyze their potential genotoxic effect on biotic environment.
The second objective of this study us to optimize internationally accepted standardized tests approved by Food and Drug Administration Authority USA. These tests are mandatory for assessment of genotoxicity of any chemicals of agricultural and industrial importance before their commercial use.
By achieving this, we will become one of the few laboratories in Pakistan, which will be able to perform such commercially important assays for industries like pesticides fertilizers, paints, sprays.
| Item Name | Type | No. of Units | Per Unit Cost (in Rs) | Total (in Rs) |
|---|---|---|---|---|
| Zinc Nitrate. Hexahydrate | Equipment | 1 | 3000 | 3000 |
| Cobalt Nitrate. Hexahydrate | Equipment | 1 | 13000 | 13000 |
| Potassium Hydroxide | Equipment | 2 | 2500 | 5000 |
| Burette 50 ml | Equipment | 2 | 2500 | 5000 |
| Beakers 50, 250 and 500 ml | Equipment | 2 | 1500 | 3000 |
| Conical flask 100, 250 ml | Equipment | 3 | 700 | 2100 |
| Reagent bottles 50, 100 and 250 ml | Equipment | 2 | 1700 | 3400 |
| Graduated cylinders 10, 50, 100 and 250 ml | Equipment | 2 | 2850 | 5700 |
| Spirit lamps 2 550 each | Equipment | 4 | 2500 | 10000 |
| Nickle nitrate | Equipment | 1 | 10000 | 10000 |
| characterization of NPs | Miscellaneous | 1 | 8000 | 8000 |
| Transportation of NPs | Miscellaneous | 1 | 2000 | 2000 |
| Total in (Rs) | 70200 |
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