Veterynarna biotehnologija – Veterinary biotechnology, 2016, 28, 225-232 [in Ukrainian].
Institute of Veterinary Medicineof NAAS
F.D.Ovcharenko Institute of Biocolloidal Chemistry
IN VIVO EVALUATION OF GENOTOXICITY AND MUTAGENICITY OF MULTICOMPONENT VACCINE“MULTIBOVISAN” MODIFED BIOSAFETY GOLD NANOPARTICLES
Introduction. Vaccine genotoxicity and mutagenicityestimation in vivo is highly prognostic because of the possibility to predict the malignant degeneration of the eukaryotic cells as well as the level of risk for the posterity health in the case of essential changes in the DNA of animal cells.
The goal of the work was in vivoestimation of genotoxicity and mutagenicity parameters for multicomponent vaccine “Multibovisan” modifed biosafety gold nanoparticles.
Materials and methods. For the in vivoestimation of genotoxicity and mutagenicity of “Multibovisan”,“Multibovisan+ AuNP (0,5%) ” and “Multibovisan+ AuNP (1%) ” vaccines, rabbit males have been used.
All experiments with laboratory animals have been carried out in compliance with “Guide for the Care and Use of Laboratory Animals”. Rabbits were injected subcutaneously with 2 ml of vaccinse. Vaccination was performed twice, according to the specification of the vaccine. Experimental group consisted of 3 animals. The animals were kept in the vivarium in accordance with the appropriate sanitary regulations on a standard diet with 12-hour light regime and free access to food and water.
The level of DNA damage has been estimated by the Comet assay (alkaline gel-electrophoresis of isolated eukaryotic cells).
Cell isolation from liver, kidneys, spleen, bone marrow, heart, lungs, testicles and muscles of the hind limbs has been performed according to standard protocols.
Micronucleus test in vivo has been performed according to standard protocols.
The experiments have been done in two parallels. Statistical analysis of obtained results has been performed comparing the indexes of DNA damage in experimental and control groups. The data of two replications have been combined and average parameter for each group has been calculated. Statistically significant high indexes of DNA damage (data close to positive control) serve as criteria of positive result. The differences p<0.05 were considered as significant.
Results of research and discussion. When using the Comet assay method in alkaline conditions for testing “Multibovisan”, “Multibovisan+ AuNP (0.5 %)” and “Multibovisan+ AuNP (1.0 %)” vaccines genotoxicity, electrophoretic tracks of the DNA comet typical of the genotoxic influence on the eukariotic cell have not been observed.
The obtained genotoxicity indexes ІDNA are close to those of the “Multibovisan” control . Studied veterinary vaccines “Multibovisan”, “Multibovisan + AuNP (0.5%)” and “Multibovisan + AuNP (1.0%)”are not genotoxic in the invivo experiments. Modification ofthe vaccine “Multybovisan”withgold nanoparticles does not decrease its genotoxiceffect. Investigated veterinary vaccines are biosafe according to the genotoxicity parameter.
By the micronucleus test in vivo, absence of the mutagenic effect of “Multibovisan”, “Multibovisan+ AuNP (0.5%)” and “Multibovisan+ AuNP (1.0%)” vaccines, has been shown. In the micronucleus test vaccines “Multibovisan” and “Multibovisan+ AuNP (0.5 %)” “Multibovisan+ AuNP (1.0 %)” were characterized by similar valuesofmutagenicity.
Conclusions and prospects for further research.The full set of experimental investigations of veterinary vaccines“Multibovisan”, “Multibovisan+ AuNP (0.5%)” and “Multibovisan+ AuNP (1.0%)” showed the absence of the genotoxic and mutagenic effects. Investigated veterinary vaccines are biosafe to the genotoxicity and mutagenicity parameters.
Fulfilled investigations open perspectives for using of gold nanoparicles for modification of veterinary vaccines.
Keywords: “Multibovisan”, gold nanoparicles, genotoxicity, mutagenicity, Comet assay method, micronucleus test, biosafety.