Impact of Chitosan and Calcium phosphate nanoparticles upon VERO cells and their adapted PPRV

Document Type : Original Article

Authors

1 Department of virology, Faculty of Veterinary Medicine, Benha University

2 Veterinary serum and vaccine Research institute, Abbasia, cairo.

Abstract

Nanotechnology that created nanoparticles (NP) having superior properties compared to the original ones offers a vast era for PPR vaccine improvement. African green monkey kidney VERO cell lines were widely used for isolation and propagation of Peste des petits ruminants virus (PPRV) instead of primary cell lines because of the problematic quality and considerable variations in primary cultures. In this study, investigations for the impact of Chitosan nanoparticles (CSN) and Calcium phosphate nanoparticles (CaPN) upon the principle biological elements, VERO cells (in terms of cytotoxicity) and PPRV performance (in terms of cytopathic effect {CPE} and virus titer) were carried out using primary  concentrations of both nanoparticles as  1%(w/v), 5%(w/v), 10%(w/v) then, further concentrations of 0.5%(w/v), 0.25%(w/v) and 0.125%(w/v) were also used. These concentrations were used as supplements to the minimum essential media (MEM) that were regularly used in PPR vaccine production. The results of the study revealed that along of a successive four days of microscopically examined VERO cells that were overlaid with either MEM only or MEM supplemented with different concentrations of CSN and CaPN, the lowest concentrations of 0.125%(w/v) for CSN and CaPN were the minimally cytotoxic  for VERO cells in terms of the ratio among the number of microscopic fields of affected cells comparing to the overall cultured cells sheet, However, the higher the concentration of CSN/CaPN, the higher the VERO cells cytotoxicity. Also, the impact of CSN/CaPN different concentrations supplied MEM upon PPRV performance after inoculating VERO cells revealed that, along seven days of microscopically examined and PPRV inoculated VERO cells that were overlaid with aforementioned concentrations from both of CSN and CaPN did not show any significant impact regarding CPE onset and intensity of eruption or as a PPRV titer (expressed by log 10 TCID50/ml) and may need  further investigations. So, the obtained results from this study recommend the concentration to be used as a MEM supplement during the process of preparing a CSN/CaPN based PPR vaccine was 0.125%(w/v) or less for keeping a considerable VERO cells viability.
 

Keywords