|Year : 2019 | Volume
| Issue : 1 | Page : 16-20
Synthesis and evaluation of the antibacterial effect of titanium dioxide nanoparticles in comparison with ampicillin, colistin, and ertapenem on Staphylococcus aureus
Mahmoud Bahmani1, Morovat Taherikalani2, Mojtaba Khaksarian3, Mahmoud Rafieian-Kopaei4, Behnam Ashrafi1, Mohammadreza Nazer1, Setareh Soroush2, Naser Abbasi5, Rouhollah Heydari1, Leila Zarei1, Mohsen Alizadeh6
1 Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorrmabad, Iran
2 Department of Microbiology, Lorestan University of Medical Sciences, Khorrmabad, Iran
3 Department of Physiology, Razi Herbal Medicines Research Center, School of Medicine, Lorestan University of Medical Sciences, Khorrmabad, Iran
4 Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
5 Biotechnology and Medicinal Plants Research Center, Ilam University of Medical Sciences, Ilam, Iran
6 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
|Date of Web Publication||22-Aug-2019|
Department of Microbiology, Razi Herbal Medicines Research Center, School of Medicine, Lorestan University of Medical Sciences, Khorramabad
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Objectives: Today, the emergence of antibiotic-resistant strains and the acquisition of antibiotic resistance have caused many problems in the treatment of Staphylococcus aureus infection and is one of the most important health issues. At present, nanotechnology has a significant impact on the various fields including pharmacology, health, medicine, and food. This study aimed to synthesize the titanium dioxide nanoparticles (Tio2NPs) and its antibacterial effect on S. aureus compared with conventional antibiotics. Materials and Methods: In this study, Tio2NPs were tested on S. aureus compared with a number of antibiotics. First, Tio2NPs were synthesized. The shape and size of the particles as well as the synthesis quality were investigated using scanning electron microscopy (SEM), atomic force microscopy (AFM), dynamic light scattering, zeta potential and X-ray diffraction, and Fourier-transform infrared spectroscopy. The antimicrobial effect of Tio2NPs on S. aureus strain (ATCC 12600) was then studied by disc diffusion method. Antibiotics of ampicillin, colistin, and ertapenem were used as control group. Results: Based on the results and given that the polydispersity index of the samples was below 0.5 (0.466), the other sample was estimated at 347.9 nm. The zeta potential of Tio2 sample was estimated at −9.48 indicating the stability of the nanoparticle and its suspension in a suitable amount per unit of time in the solution. The results of the AFM showed that the lower mean value obtained for the Tio2NPs was 0.5 nm, and the growth of the nanoparticles was noticeable in some regions and uniform and low in some others. The result of SEM showed that the size of the nanoparticle was 28.45–34.14 nm. The best inhibition zone diameter was obtained for ampicillin (30.66 mm), followed by ertapenem (15 mm) and Tio2(10 mm). Colistin without inhibition zone was identified as ineffective group. Conclusion: Due to the development of drug resistance to antibiotics, the production and synthesis of antimicrobial agents is one of the requirements of the current time. Due to the excellent synthesis of Tio2 and the presence of very fine nanoparticles, it can be used as a strong antimicrobial compound, especially on S. aureus infection.
Keywords: Antibiotic, disease, infection, nanoparticle, Staphylococcus aureus, titanium dioxide
|How to cite this article:|
Bahmani M, Taherikalani M, Khaksarian M, Rafieian-Kopaei M, Ashrafi B, Nazer M, Soroush S, Abbasi N, Heydari R, Zarei L, Alizadeh M. Synthesis and evaluation of the antibacterial effect of titanium dioxide nanoparticles in comparison with ampicillin, colistin, and ertapenem on Staphylococcus aureus. J Pharm Negative Results 2019;10:16-20
|How to cite this URL:|
Bahmani M, Taherikalani M, Khaksarian M, Rafieian-Kopaei M, Ashrafi B, Nazer M, Soroush S, Abbasi N, Heydari R, Zarei L, Alizadeh M. Synthesis and evaluation of the antibacterial effect of titanium dioxide nanoparticles in comparison with ampicillin, colistin, and ertapenem on Staphylococcus aureus. J Pharm Negative Results [serial online] 2019 [cited 2020 Aug 13];10:16-20. Available from: http://www.pnrjournal.com/text.asp?2019/10/1/16/265148
| Introduction|| |
Staphylococci are gram-positive cocci, whose shape is similar to a bunch of grapes. Organisms may also be seen in isolated clinical specimens as single cells, pairs of cells, or short chains. The majority of staphylococci have a diameter of 0.5–1.5 μm and are immotile and able to grow under various conditions, both aerobic and anaerobic, in high-concentration salts (10% sodium chloride), as well as under the temperature range of 18°C–40°C.,,,,,,, Golden Staphylococcus, in addition to its wide distribution in nature, inhabits in the human body, and Staphylococcus aureus is present everywhere such as the respiratory tract and on the skin of many adults. The emergence of antibiotic-resistant strains and the acquisition of antibiotic resistance have been associated with many problems in the treatment of staphylococcal infections.S. aureus produces extracellular enzymes and materials, which include catalase, coagulase, hyaluronidase, staphylokinase, leukocidin, exotoxins, and enterotoxins.,S. aureus causes the disease through the production of toxins or direct invasion and destruction of the tissue. Clinical manifestations of some staphylococcal diseases are exclusively the result of the activity of toxin (such as staphylococcal scalded skin syndrome, staphylococcal food poisoning, and toxic shock syndrome), while other diseases are due to the rapid growth of the organism and the formation of abscess and tissue destruction (such as skin infections, endocarditis, pneumonia, empyema, osteomyelitis, and septic arthritis. Food poisoning, aphthous ulcers, folliculitis, furuncles, and carbonicles are among other diseases caused by S. aureus.,, Today, nanotechnology has a significant impact on the various fields of industry, health, medicine, and food. Nanotechnology can resolve many biomedical problems and cause change in the fields of health and pharmacology. Titanium dioxide nanoparticles (TiO2 NPs) are more stable, long lasting, safe, and also resistant to a wide range of microbes., Titanium is more useful for its antibacterial properties and also in the environment, water purification, gas sensors, and high-performance solar cells., It also has anticancer effects.,,, Considering its application in different industries, in this study, TiO2 NPs were tested on S. aureus compared to some effective antibiotics. Hence, this study aimed to synthesize the TiO2 NPs and its antibacterial effect on S. aureus compared with conventional antibiotics.
| Materials and Methods|| |
Strain of Staphylococcus
Staphylococcus strain with ATCC 12600 was purchased from the Iranian Research Organization for Science and Technology. Antibiotic disks of ampicillin, colistin, and ertapenem (MAST ® DISC, United Kingdom) were also prepared.
Preparation of titanium dioxide nanoparticles
First, 1 ml of titanium isopropoxide (Sigma-Aldrich, USA) solution was added to 20 ml of sterile distilled water, and the solution was gently stirred. The solution was completely stirred for 5 h on a shaker stirrer at 50°C. The solution was then stirred for 24 h at 50°C and finally converted to a white powder, i.e., Tio2 NPs, in furnace at 550°C.
Determination of antimicrobial effect by disk diffusion method
The bacteria culture from suspension with 0.5 McFarland turbidity was inoculated into the medium on Mueller-Hinton Agar using a sterile swab, and a methicillin disk was placed on the medium. In addition, 40 μL of standard plant stock solutions and active ingredients was added to the blank discs and placed on the medium. The plates were then incubated at 37°C for 24 h. The test was performed in triplicate, and results were expressed as mean ± standard deviation. Inhibition zone was measured by Colis.
| Results|| |
In order to find out more and to investigate the TiO2 crystalline structure used in the experiments, scanning electron microscopy (SEM), density functional theory, zeta potential, and atomic force microscopy (AFM) were used. [Figure 1] is a SEM image of the TiO2 NPs. Based on the results of this microscope, the shape of the NP is round and its size 28.45–34.14 nm. The results of the dynamic light scattering (DLS) that is used to determine the distribution and size of particles in solutions and suspensions based on the motion of particles in a sample are illustrated in [Figure 2]. The DLS image shows the histogram of the distribution of TiO2 particle size, according to which the polydispersity index (PDI) of the samples is below 0.5 (0.466) and therefore that of the other sample was estimated at 347.9 nm [Figure 2]. The zeta potential of TiO2 NP sample was estimated at −9.48 indicating the stability of the nanoparticle and its suspension in a suitable amount per unit of time in the solution [Figure 3]. The results of the AFM showed that the lower mean value obtained for the TiO2 NPs was 0.5 nm, and the growth of the nanoparticles was noticeable in some regions and uniform and low in some others [Figure 4]. Based on the results obtained from X-ray diffraction (XRD), sample synthesis is well done [Figure 5]. Furthermore, the results of Fourier-transform infrared spectroscopy are shown in [Table 1].
|Figure 1: Scanning electron microscopy image of titanium dioxide nanoparticle|
Click here to view
|Figure 2: Dynamic light scattering image of titanium dioxide nanoparticle|
Click here to view
|Figure 4: Atomic force microscopy image of titanium dioxide nanoparticle|
Click here to view
|Table 1: Fourier-transform infrared spectroscopy of titanium dioxide nanoparticles|
Click here to view
Based on the results of XRD, the nanoparticle crystal system was tetragonal and was in the anatase structure. Based on the standard card of the XRD device, the desired nanoparticle was detected in Tio2. The most stable phase is TiO2 at the usual pressure, and temperature is anatase structure.
According to results in [Table 1], Peak at a wavelength of 3421/cm related to the stretching vibration of water molecules link with O-H. Peak in 1623/cm related to the bending vibration of O-H molecules and the peak in 1389/cm also belong to the link titanium oxide.
After preparation of the TiO2 NP solution and selected antibiotics, antibacterial activity of the groups on S. aureus was investigated. The growth inhibition zone diameters are shown in [Table 2]. Accordingly, the most effective groups on S. aureus included antibiotics such as ampicillin and ertapenem, followed by TiO2 NPs. Colistin antibiotic was found to be ineffective on S. aureus [Table 2].
| Discussion|| |
Germs, including S. aureus, are able to bind to the surfaces of wound and ulcer and cause infection, thereby creating a source of contamination of the environment. S. aureus is one of the main microbial agents involved in nosocomial infections. In this study, TiO2 NPs were synthesized, and its antibacterial effect on S. aureus was investigated in comparison to common antibiotics. Based on the results, ampicillin and ertapenem antibiotics formed a higher inhibition zone diameter than TiO2 NPs and therefore were more effective. Colistin antibiotic was also found to be ineffective on S. aureus. The study of Saadat et al. (2012) showed that TiO2 NPs with a diameter of 40–65 nm had a minimum inhibitory concentration (MIC) of 40 μ/ml on Pseudomonas aeruginosa. The study of Rezaei and Kermanshahi (2015) showed that TiO2 NPs with a size of 21 nm and 1% concentration had MIC and minimum bactericidal concentration of 0.5%. The size of the NP is very effective in their antimicrobial effect, and the smaller their size, the more antimicrobial effect they show. The type of bacteria is also effective in the sensitivity of NP. According to the results of our study, the size of NPs in AFM, DLS, and SEM was 0.5 nm, 347.9 nm, and 28.45-34.14 nm, respectively. The reason for its effect is the small size of NPs. The study of Bottero et al. (2011) showed that Tio2 showed antibacterial effect against Bacillus subtilis, Escherichia More Details coli, and Pseudomonas. The NP affect the enzymes and proteins of the respiratory cycle due to the ability to bind to sulfhydryl, carboxylic, and phosphate groups and cause their conformation to change, and subsequently, the cycle is inefficient. In the second part, the effect of NPs on proteins and DNA and enzymes that play an essential role in cell growth was studied. Bacteria bound to NPs due to the presence of a phosphate group and as a result make its conversion from flexible state to hard state, and in the DNA, transformation, rotation, and movement are the results of certain processes such as replication and transcription. Nanoparticles also activate the free radicals and oxygen, which have antimicrobial activity, inside the cell.S. aureus is one of the most common pathogens that cause nosocomial infections, which can also cause infections in any part of the body due to containing various enzymes such as coagulase, hyaluronidase, nuclease, lipase, leukocidin, and hemolysin. Because of its structures, S. aureus leads to the emergence of antibiotic-resistant strains. Because this bacterium is resistant to antibiotics and, in our study, the PDI of TiO2 NPs was also 0.466 and also due to the appropriate dispersion of NPs, in the solution, its antimicrobial effect is intensified.
| Conclusion|| |
Due to the development of drug resistance to antibiotics, the production and synthesis of antimicrobial agents is one of the requirements of the current time. Due to the excellent synthesis of Tio2 and the presence of very fine nanoparticles, it can be used as a strong antimicrobial compound, especially on S. aureus infection.
This article was extracted from PhD thesis of Dr. Mahmoud Bahmani with code A-10-1379-1. The authors would like to express their gratitude for financial support of the Research and Technology Deputy of Lorestan University of Medical Sciences, Khorramabad, Iran.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Blackburn CW, Peter JM. Foodborne Pathogens, Hazard, Risk Analyses and Control. Woodhead Publishing: CRC Press; 2002. p. 385-90.
Hui YH, Smith RA, Spoorke DG. Foodborne Disease Handbook. 2nd
ed. USA: Marcel Dekker Inc.; 2001. p. 345-72, 427-8.
Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG Jr. Staphylococcus aureus
infections: Epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 2015;28:603-61.
Coates R, Moran J, Horsburgh MJ. Staphylococci: Colonizers and pathogens of human skin. Future Microbiol 2014;9:75-91.
Archer NK, Mazaitis MJ, Costerton JW, Leid JG, Powers ME, Shirtliff ME, et al. Staphylococcus aureus
biofilms: Properties, regulation, and roles in human disease. Virulence 2011;2:445-59.
MacNeal WJ, Frisbee FC, McRae MA. Staphylococcemia 1931-1940. Five hundred patients. Am J Clin Pathol 1942;12;281-94.
Moreillon P, Que YA, Glauser MP. Staphylococcus aureus
(including staphylococcal toxic shock). In: Mandell GL, Bennett JE, Dolin R, editors. Mandell, Douglas and Bennett's Principles and Practice of Infectious Disease. 6th
ed. Philadelphia: Elsevier Livingstone; 2005. p. 2321-51.
Mardaneh J. Morai Medical Microbiology, 2016 (General and Specific Bacteriology). 1st
ed. Tehran: Andisheh Rafi Publication; 2016.
Kluytmans J, van Belkum A, Verbrugh H. Nasal carriage of Staphylococcus aureus
: Epidemiology, underlying mechanisms, and associated risks. Clin Microbiol Rev 1997;10:505-20.
Cunha BA. Methicillin-resistant Staphylococcus aureus
: Clinical manifestations and antimicrobial therapy. Clin Microbiol Infect 2005;11 Suppl 4:33-42.
Levinson W, Jawetz E. Medical Microbiology & Immunology Examination and Board Review. 6th
ed. USA: McGraw-Hill; 2000. p. 85-9.
Ananou S, Maqueda M, Martínez-Bueno M, Gálvez A, Valdivia E. Control of Staphylococcus aureus
in sausages by enterocin AS-48. Meat Sci 2005;71:549-56.
Blackburn CV, Peter JM. Foodborne Pathogens, Hazard, Risk Analyses and Control. USA: CRC Press; 2002. p. 385-90.
Normanno G, Firinu A, Virgilio S, Mula G, Dambrosio A, Poggiu A, et al.
Coagulase-positive staphylococci and Staphylococcus aureus
in food products marketed in Italy. Int J Food Microbiol 2005;98:73-9.
Jay MJ. Modern Food Microbiology. 6th
ed. USA: An Aspen Publication; 2000. p. 441-56.
Mritunjai S, Shinjni S, Prasad S, Gambhir SI. Nanothehnology in Medicine and antibacterial effect of silver nanoparticles. J Nanomater Biostrut 2008;3:115-22.
Fujishima A, Zhang X. Titanium dioxide photocatalysis: Present situation and future approaches. C R Chemie 2006;9:750-60.
Kasem KK, Dahn M. Photodissociation of water using colloidal nanoparticles of doped titanium (IV) oxide semiconductors for hydrogen production. Curr Sci 2010;99:1068-73.
Han K, Yu M. Study of the preparation and properties of UV-blocking fabrics of a PET/TiO2 nano composite prepared by in situ
poly condensation. J Appl Polym Sci 2006;100:1588-93.
Fu G, Vary PS, Lin CT. Anatase tiO2 nanocomposites for antimicrobial coatings. J Phys Chem B 2005;109:8889-98.
Wang ML, Tuli R, Manner PA, Sharkey PF, Hall DJ, Tuan RS, et al.
Direct and indirect induction of apoptosis in human mesenchymal stem cells in response to titanium particles. J Orthop Res 2003;21:697-707.
Wang JJ, Sanderson BJ, Wang H. Cyto – And genotoxicity of ultrafine tiO2 particles in cultured human lymphoblastoid cells. Mutat Res 2007;628:99-106.
Lagopati N, Kitsiou PV, Kontos AI, Venieratos P, Kotsopoulou E, Kontos AG, et al
. Photo-induced treatment of breast epithelial cancer cells using nanostructured titanium dioxide solution. J Photochem Photobiol A 2010;214:215-23.
Lee YS, Yoon S, Yoon HJ, Lee K, Yoon HK, Lee JH, et al.
Inhibitor of differentiation 1 (Id1) expression attenuates the degree of tiO2-induced cytotoxicity in H1299 non-small cell lung cancer cells. Toxicol Lett 2009;189:191-9.
Rodbar Mohammadi SH, Mohammadi P, Eskandari M. The evaluation of antifungal activity of titanium dioxide nanoparticles and ethylene diamine tetra acetic acid on growth inhibition of standard strain of Candida albicans
. Yasuj Univ Med Sci J 2010;58:134-41.
Clinical and Laboratory Standards Institute. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically: Approved Standard. 8th
ed. Wayne: Clinical and Laboratory Standards Institute; 2009.
Saadat M, Roudbar Mohammadi SH, Khavarinejad RE, Taghavi A. Evaluation of antimicrobial activity of titanium dioxide nanoparticles, EDTA and garlic on standard strain of Pseudomonas aeruginosa
using standard microdilution method. Comp Pathobiol Sci Res Eighth 2012;10:11-6.
Rezaee P, Kermanshahi R. Investigating the effect of antimicrobial activity of silver nanoparticles, titanium dioxide on two pathogenic bacterial strains with food source. Biotechnol Tarbiat Modares Univ 2015;6:1-6.
Valodkar M, Bhadoria A, Pohnerkar J, Mohan M, Thakore S. Morphology and antibacterial activity of carbohydrate-stabilized silver nanoparticles. Carbohydr Res 2010;345:1767-73.
Bottero J, Auffan M, Mouneyrac C, Botta C, Labille J, Masion A. Manufactured metal and metal-oxide nanoparticles: Properties and perturbing mechanisms of their biological activity in ecosystems. Comptes Rendus Geoscience 2011;343:168-76.
Mirzajani F, Ghassempour A, Aliahmadi A, Esmaeili MA. Antibacterial effect of silver nanoparticles on Staphylococcus aureus
. Res Microbiol 2011;162:542-9.
Panyala NM, Penamendez E, Have MJ. Silver or nanoparticles: A hazardous threat to environment and human health? J Appl Biomed 2008;6:117-22.
Hantson P, Léonard F, Maloteaux JM, Mahieu P. How epileptogenic are the recent antibiotics? Acta Clin Belg 1999;54:80-7.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2]