Nanotechnology is a field that is turning twenty-four hours by twenty-four hours, doing an impact in every portion of human life. Biological methods of “ greenish synthesis ” of nanoparticles have proven in assorted experiments to be better methods due to slower dynamicss, they offer better use and control over crystal growing and their stabilisation. Biosynthesis of nanoparticles by works infusions is presently continuously traveling on. Plant infusions are really cost effectual and eco-friendly and therefore can be economic and efficient option for the big scale synthesis of nanoparticles.
In this undertaking, biogenesis of Ag nanoparticles ( AgNPs ) will be carried out utilizing aqueous infusion of Juglans regia fruits outer shell and fruit organic structure of Tribulus terrestris as agribusiness waste and Ag nitrate ( AgNO3 ) at ambient temperature. For word picture of nanoparticles UV/Vis Spectrophotometer, Fourier Transform Infra-Red Spectroscopy ( FTIR ) , Scaning Electron Microscopy ( SEM ) and Energy Dispersive Spectroscopy ( EDS ) will be analyzed. Genotoxicity is a one type of toxicity which is cause of chromosomal aberrance. In this undertaking we will analyse the Ag nanoparticles synthesized by Juglans regia fruits outer shell for genotoxic activity on human leucocyte cell civilization and hence the consequence will follow.
NANOMEDICINE, GREEN SYNTHESIS, UV- ANALYSIS, EDS, SEM, FTIR, GENOTOXICITY
Nanobiotechnology is a term that refers to the interection of nanotechnology and biological science. This subject allows scientists to make systems that can be used in assorted Fieldss of biological science. The medical application of nanotechnology is the nanomedicine. Nanomedicine has a big figure of applications like, it can be used to name and handle malignant neoplastic disease, drug, detoxification, drug bringing, surgery etc. The nanoparticles used for nanomedicine are of three different types: polymeric, lipid-based and metal based. Polymeric nanoparticles are prepared utilizing hydrophilic and hydrophobic bio degradable and biocompatible polymers for drug and cistron bringing, severally. For lipid based oil-in-water nano emulsions are used, where the oil droplets are reduced to less than 100nm in diameter. For metal based assorted metals such as gold, Ag and Fe are used in assorted forms like, domains, rods etc. The nanoparticles which are used for medical intent have toxic effects on the life system. The toxic effects depend mostly on the type of stuffs used and accumulation behavior or the variety meats in which they accumulated. Sometimes the nanoparticles have toxic consequence on cell ‘s chromosome breakage, chromosomal mutant etc. The genotoxic consequence of nanoparticles can be reduced to a big extent by the usage of organic based nanoparticles. so, in the present survey, we concentrated upon nanomedicine synthesis by green synthesis method by utilizing outer shell pulverization of walnut ( Juglans regia ) and the dried fruit organic structures of Tribulus terrestris which are nil but an agricultural waste.
Material and method
Collection of works stuffs
The outer shell of the Juglans regia dried fruit organic structures of Tribulus terrestris was collected from local market for the synthesis of nanoparticles.
Preparation of fruit infusions
The outer shell of Juglans regia was grinded good with howitzer and stamp to do a pulverization. Five gms of pulverization sample was assorted into 100 milliliter of deionized H2O so the mixture was boiled for 10 min. After chilling the fruit infusion was filtered with Whatman No. 1 filter paper. The filtrate was stored for farther usage. The Tribulus terrestris infusion was prepared individually by utilizing the above method.
Synthesis of Ag nanoparticles
The 100 milliliter of aqueous filtrate infusion of Juglans regia was taken into 250 milliliter of conelike flask. Then the infusion was assorted into Silver nitrate ( AgNO3 ) to do the concluding volume concentration of 1 mM solution. The reaction mixture was kept into dark room status until the coloring material alteration was occurred. Then after happening of color word picture was done of the Ag nanoparticles reaction solution coloring material alterations have observed for word picture of Ag nanoparticles.
The above process is repeated for synthesising Ag nanoparticles form Tribulus terrestris works infusion.
Genotoxicity testing of the nanoparticles
5ml of media ( RPMI ) 1640- ready mix was assorted with 0.5ml of blood. Then the RPMI assorted blood is incubated for 72 hour. Every 24 hours the CO2 is released by taking it out of the brooder and assorted good. Now at 71st hr it is treated with antecedently prepared nanoparticle solution and was kept in brooder once more. After incubation for 1 hr 2 beads of colchicines were added. Then it was kept for 15 min in brooder.
After incubation it was centrifuged at 2000 revolutions per minute for 5 min and so the supernatant was discarded. After that 6ml of hypotonic solution ( 0.075M KCl ) was added and it was incubated once more for 6-7 min. The assorted solution was so centrifuged at 2000 revolutions per minute for 5 mins and supernatant was discarded. Then 6ml of newly prepared fixative ( Methanol: Acetic Acid= 3:1 ) was added and assorted well and kept for nightlong in the brooder.
Then it was centrifuged at 2500 revolutions per minute for 7 min. The procedure was repeated until the white pellet was obtained. Then the supernatant was removed and slides were prepared with the white pellet. The slides was so stained with giemsa solution ( 4ml giemsa+ 46ml two-base hit distilled H2O ) for 4 min and once more in dual distilled H2O for 1 min. Then the slides were air dried and observed under fluorescence microscope for any chromosomal aberrance.
Standardizing protocol in advancement. Previously we worked with a specific concentration of Ag nanoparticles for genotoxicity testing, but we did non acquire any consequence out of it. So, we are go oning our work with more silver nanoparticles solutions of different concentrations of and the consequence is expecting.
As a agribusiness waste outer shell of walnut ( Juglans regia ) and dried fruit organic structures of Tribulus terrestris is used for the synthesis of Ag nanoparticles and its genotoxicity testing is done in human leukocyte civilization. Different concentrations of Ag nitrate solution is being used to fix the nanoparticles. Gentoxicity testing of prepared nanoparticles are besides in the procedure to watch out the expected DNA harm and other chromosomal abnormalcies and its ruptures. Previously this genotoxicity testing was done ( Acharya and Panda, 2010 ) . Which provided grounds of terrible DNA damaging potency of Ag nanoparticles in a dose dependent mode.
In decision, the consequence of the above work is still waiting as the standardisation of the protocol is on the procedure. The anticipated consequence of the current survey expected to uncover some of import findings on assorted effects of presenting silver nanoparticle synthesized from agricultural waste as a nanomedicine in human leucocyte cell civilization. Furthermore the consequence will demo the feasibleness of utilizing AgNP-P in a big graduated table mode for bring forthing nanomedicines.