Also, the Mycobacterium tuberculosis (Mtb) escapes from immune system with different tricks and survive. However, with the discovery of amazing nano-drugs in ...
Mixed metal oxide nanoparticles; The inhibitor of growth of Mycobacterium tuberculosis into the THP-1 cells Jafari AR1*, Mosavi T1, Mosavari N2, Majid pour A1, Movahedzade F3, Tebyaniyan M2, Kamalzadeh M2, Dehgan pour M2, Jafari S4 1
Rasoul-e-Akram Hospital, Antimicrobial Resistance Research Center, Iran University of medical Sciences, Tehran, Iran. 2
3
Razi Vaccine and Serum Research Institute, Karaj, Iran
Department of Medicinal Chemistry and Pharmacology, Institute for Tuberculosis Research, University of Illinois at Chicago 4
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran, Iran
Aims and objectives. Humans have always been in constant battle with tuberculosis. In today's world, the use of various antibiotics, has caused the spread of Multidrug-Resistant Mycobacterium tuberculosis (MDR). This shows that currency antibiotics are ineffective in controlling the spread of TB infection in the host cells, especially Macrophage. Also, the Mycobacterium tuberculosis (Mtb) escapes from immune system with different tricks and survive. However, with the discovery of amazing nano-drugs in medicine, we should not unaware the anti-mycobacterial properties and the bactericidal mechanism of nanoparticles (NPs). The purpose of this study were synthesis of mixture metal oxide NPs for inhibit the growth of Mtb into the Macrophages, and in the next stage, investigation of the cytotoxicity effects of NPs against of THP-1 cells. Methods Ag NPs was synthesized by chemical reduction in aqueous solution. The ZnO NPs was synthesized by chemical deposition in aqueous solution. The DLS, SEM, TEM and UV-Vis absorption spectra and ICP-MS were used to identify the properties of NPs. Ag and ZnO NPs were mixed together at a ratio of 8ZnO / 2Ag. Next, The NPs were dilution into the LJ medium. Bacteria were added to each falcon tubes. Falcons, for a period of 28 days were incubated at 37 ° C. After this period, the MIC was determined. Separately, the MIC for Ag and ZnO NPs were done. Atomic Force Microscopy (AFM) was showed the interaction of NPs and Mtb. In the next phase, the toxicity of NPs to THP-1 was conducted with MTT test. Then, the THP-1 cells infected accordance with the protocol of “W.R. Jacobs lab, 120707” was conducted in laboratory cell cultures. Macrophage was infected four hours at 37 ° C and %5 CO2 with Mtb. THP-1 cells were lysed. And its contents for bacterial culture, as well as counting the colonies CFU and preparation for TEM imaging was used. Results Nano-sized particles according to the results of the DLS, SEM, TEM and UV-Vis absorption spectra were estimated at around 30 to 80 nm. The initial concentration of Ag NPs was 20 ppm and also ZnO NPs was estimated about 60 ppm. The MIC ratio of 8ZnO/2Ag NPs against Mtb were detected about 1/32 of initial concentration. Ag NPs in the range of concentrations had no antimycobacterial effects on Mtb. While ZnO NPs showed potent antibacterial activity at 1/128 of initial concentrations. ZnO NPs in all concentration showed cytotoxic activity. And about 100% of THP-1 cells were alive in the presence of Ag NPs in the concentration of 1/32 and 1/64. However, in the ratio of 8ZnO/2Ag, about 39.94% of the cells in concentration of 1/16 were still alive. And also, about 100% of THP-1 in concentrations of 1/32 were still alive. The CFU after lysing of the THP-1, equivalent of the concentration of initial dilution; 1/32, the number was zero. Conclusions Although the cytotoxicity of Ag NPs was low, But was not able to inhibit the growth of Mtb in vitro. Meanwhile, super strong anti-mycobacterial agent “ZnO NPs” was able to inhibit bacterial growth, but had highly toxic to the human Macrophage cells. By mixing Ag and ZnO NPs, the ratio 8ZnO/2Ag, we could find a new formulation that in addition to having a potent antibacterial activity against Mtb, had no cytotoxic effects on THP-1. And also was capable of both in vitro and Ex-vivo to inhibit Mtb. Keywords: Nanoparticles, THP-1 cells, Mycobacterium tuberculosis
