Synthesis of TiO2 NPs by using plant extracts.
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More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
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Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
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This book describes a comprehensive overview of electro-chemical sensors and biosensors for the analyses, investigation, and monitoring of the most significant unsafe analytes in the ecological as well as environmental field in industry, in treatment plants, and in environmental research. The contributed chapters stretch the reader a comprehensive study, state-of-the-art picture of the field of electro-chemical sensors or biosensors appropriate to environmental analytes, from the theoretical principles of their design to their implementation, realization, and potential application. It covers the most recent techniques and nanocomposites/nanomaterials for the preparation, construction, validation, analyses, and design of electro-chemical sensors/biosensors for bio-analytical, clinical, and environmental applications—emphasizing the latest classes of selective, sensitive, robust, fast response, stable, electro-chemical sensors as well as electrochemical biosensors for in vivo/vitro diagnosis.
Development in advanced nanotechnology and the conservatory of innovative chemical sensors, biosensors, ionic sensors with various composites/materials and nanodevices has been a regulating key task in the fabrication and improvement of very precise, perceptive, accurate, sophisticated, sensitive, and consistent efficient chemical sensors [1–3]. The exploration for even tiny electrodes accomplished in nano-level imaging and controlling of doped nanomaterials, doping agents (host-guest), biological, chemical, pathological samples, and chemical sensors has recently extended the attention of awareness of the scientist, mainly for control monitoring, owing to the amplifying essential for environmental safety and health monitoring [4–6]. Recently, great attention is provided for the detection of various unsafe, carcinogenic, toxic, hazardous chemicals, or biomolecules to live safely and as well as to prevent the ecological system from harmful effects of toxins [7–9].
Ronkainen et al. describe the enzyme-based electrochemical glutamate biosensor development, which has been proposed to play a significant role in various neurological and psychiatric disorders. In this contribution, the design, construction, and optimization of enzyme-based electrochemical biosensors for in vivo and in vitro detection of glutamate were discussed in this contribution [10–17]. Various glutamate biosensors have been discussed, including the developed glutamate monitoring dynamic levels of extracellular glutamate in the living brain tissue adding to the current medical knowledge of these complex neurotransmitter systems and ultimately impacting treatment plans [18–21]. More significantly, glutamate biosensors have been used in environmental monitoring, in the fermentation industry, and in the food industry for determination of Monosodium glutamate (MSG), a common flavor-enhancing food additive. With continuous developments in molecular biology, nanofabrication methods, immobilization methods of biomolecules and multiplexing capabilities, the production of sensitive, selective, fast, and easy-to-use biosensors for quantification of glutamate, and other neurotransmitters will be feasible in the not too distant future.
Qijin et al. approached some graphene paper-based electro-chemical sensors to illustrate recent advances in the research and development of 2D graphene papers as new and noble materials for electro-chemical sensors. It covers the design, fabrication, functionalization, and application evaluation of graphene papers. Precise monitoring of chemical or biological processes is of extreme importance for medical and biological applications. Electro-chemical sensors can ideally fulfill that goal by converting a chemical or biological response into a processable and quantifiable signal. In the past two decades, intensive research and development of electro-chemical sensors have enabled to fabricate different types of devices [22–31]. After the development of many successful commercial electro-chemical sensors in the classic configurations, currently, there is a notable transition and increasing demands for the development of flexible and wearable sensors. The development of flexible electro-chemical sensors depends crucially on the discovery and preparation of freestanding and flexible new materials. They first summarized the mainstream methods for fabrication of graphene papers/membranes with the focus on chemical vapor deposition techniques and solution-processing assembly. A large portion of this work is devoted to the highlights of specific functionalization of graphene papers with polymer and nanoscale functional building blocks for electrochemical sensing purposes. In terms of electrochemical sensing applications, the emphasis is on enzyme-graphene and nanoparticle-graphene paper-based systems for detection of glucose. We conclude this chapter with brief remarks and an outlook. In short, worldwide researchers have explored graphene paper–based sensors by exploiting their unique advantages including high sensitivity, conductivity, and in-situ sensing. The recent research advances suggest that graphene paper-based materials could play a significant role in developing flexible sensors and electronic devices due to their intriguing structural and functional features.
Murray et al. focused on managing H2O cross-sensitivity using composite electrolyte “NOx sensors”. They approached NOx sensors composed of PSZ, FSZ, and PSZ–FSZ composite electrolytes which were investigated using impedance spectroscopy under dry and humidified gas conditions. The microstructural properties, NOx sensitivity, oxygen partial pressure and temperature dependence, as well as the response time of the sensors composed of the various electrolytes were characterized in order to interpret the electrochemical response with respect to water cross-sensitivity. In this approach, impedance spectroscopy was used to interpret the electrochemical response of NOx sensors composed of PSZ, FSZ, and PSZ–FSZ composite electrolytes during operation under dry and humidified gas conditions. Analysis of the electrochemical responses of the 50PSZ–50FSZ-based sensors indicated PSZ contributed to lower water cross-sensitivity, while FSZ promoted NOx sensitivity. Finally, sensors composed of the 50PSZ–50FSZ composite electrolyte demonstrated significant sensitivity to NO and low cross-sensitivity to water with negligible temperature dependence [32–35].
Lutic et al. describe electro-chemical sensors for monitoring indoor and outdoor air pollution. They approached a comprehensive presentation of the most common electro-chemical sensors used in real monitoring applications of air purity testing. The air quality monitoring stations based on electro-chemical sensors are nowadays used to determine the global pollution index of the atmospheric air, in order to prevent the risks toward human health and damage of environment, especially in the highly populated and industrialized urban areas. The electro-chemical gas sensors are nowadays indispensable in the monitoring of the atmosphere quality, especially due to pollutants associated with human activities. Carbon monoxide, sulfur oxides, hydrogen sulfide, and nitrogen oxides are only a few of species which can seriously damage the environment equilibrium by smog formation, acid rain, soil deterioration, water contamination as well as some direct damages on the human health [36–38]. The electrochemical gas sensing is based on gas oxidation or reducing reactions on sensing surfaces with catalytic potential, surfaces which suffer noticeable charge changes, that can be amplified and processed in order to generate a signal. The electro-chemical sensors are fast, reliable, small and cheap; therefore, their use covers nowadays the exhaust systems from automotives, domestic/residential gas detection, and leak checkers. Respecting the rated voltage as said by the manufacturer, using the sensor in the right temperature range, avoiding the deterioration due to exposure to humidity, avoiding contamination with various chemicals, lack of sudden exposure to extreme temperatures, and avoiding the mechanical shocks are basic conditions to preserve their work function and accuracy.
Lee et al. have focused on fabrication and characteristics of metal-loaded mixed metal oxides gas sensors for the detection of toxic gases for environmental purposes. They approached developing gas sensors which permit individuals to circumvent poisonous gases that may be produced in spaces with residues of inorganic/organic waste with certain temperature at 50°C or above. The response, sensitivity, and selectivity of these gas sensors to types of carcinogenic gases such as H2S, toluene, and aldehyde were examined. The thick-film semiconductor sensors that detect some toxic gases were fabricated using nano-sized sensing materials powder (SnO2, WO3, and ZnO), and these were prepared via sol-gel and precipitation methods. Response to various lethal gases was measured and is defined as the ratio (Ra/Rg) of the resistance of the sensor film in air to the resistance of the film in toxic gas. Generally, semiconductor metal-oxide gas sensors can be used for diverse applications, ranging from equipment to monitor environmental and occupational safety to facilitating quality assurance through novel measurement. The nature of the gas-sensitive material and the concentration of the target gas (usually a few ppb~ppm) determine the measuring range and limitations of the device [39–41].
Zhang describes the potential application of nanosensors in dissolved gases for the detection in oil-insulated transformers in this contribution. Here, it is approached on the adsorption processes between modified CNTs (CNTs-OH, Ni-CNTs) and dissolved gases in transformers oil including C2H2, C2H4, C2H6, CH4, CO, and H2 which have been simulated based on the first principle theory. Additionally, the density of states, adsorption energy, charge transfer amount, and adsorption distance of adsorption process between CNTs and dissolved gas were also calculated in his chapter. Two kinds of sensors, mixed acid-modified CNTs and NiCl2-modified CNTs, were prepared to conduct the dissolved gases response experiment. Afterward, the gas response mechanisms were investigated. Finally, the results between response experiment and theoretical calculation were compared, reflecting a good coherence with each other. The carbon nanotube (CNT) based gas sensors possess a relatively high sensitivity and fine linearity and could be employed in dissolved gas analysis equipment in the transformer [42–47].
Finally, this book generally reviews the recent and advanced methods and substantial applications of biosensors, gas sensor and chemical sensors. Contributed chapters are scratched by expert scientists and professors in the electrochemical sensor field. This book aims to make a connection between undergraduates, post-graduates, graduates, and scientists on their researches in sensor development based on enzyme-based sensors, graphene-based sensors, NOx sensors, gas sensors, hazardous and toxic gas sensors, and nano-sensors in environmental and biomedical sciences in order to initiate researchers into various sensors study in as straightforward a way as possible and as well as present the scientist the opportunities offered by the health care science and ecological fields. However, each chapter delivers methodological details beyond the level originally in representative journal articles and explores the potential applications of biological and chemical sensors to a substantial level in health care, real clinical, food, industrial, cancer diagnostics, biomedical, environmental science and detection of infectious organisms, also providing a brochure for the future as well as in the safety and security arena. The primary target audience for this book “Electrochemical Sensors Technology” includes students, researchers, technologists, physicists, chemists, biologists, engineers, and professionals who are interested in bio, chemi, and gas sensors and associated topics.
The incidence of microbial attack in different sectors such as food, textiles, medicine, water disinfection, and food packaging leads to a constant trend in the search for new antimicrobial substances. The increased resistance of some bacteria to some antibiotics and the toxicity to the human body of some organic antimicrobial substances has increased the interest in the development of inorganic antimicrobial substances. Among these compounds, metal and metal oxide compounds have attracted significant attention due to their broad-spectrum antibacterial activities. On the other hand, nanoscale materials are well known thanks to their increased properties due to their high surface area-to-volume ratio. Antimicrobial NPs have shown excellent and different activities from their bulk properties [1, 2].
During last decades, metal oxide nanoparticles, such as zinc oxide (ZnO), manganese oxide (MgO), titanium dioxide (TiO2), and iron oxide (Fe2O3), have been extensively applicable thanks to their unique physiochemical properties in biological applications. Among metal oxide antimicrobial agents, TiO2 is a valuable semiconducting transition metal oxide material and shows special features, such as easy control, reduced cost, non-toxicity, and good resistance to chemical erosion, that allow its application in optics, solar cells, chemical sensors, electronics, antibacterial and antifungal agents [3]. In general, TiO2 nanoparticles (TiO2 NPs) present large surface area, excellent surface morphology, and non-toxicity in nature. Several authors have reported that TiO2 NPs have been one of the most studied NPs thanks to their photocatalytic antimicrobial activity, exerting excellent bio-related activity against bacterial contamination [4, 5, 6, 7].
Antimicrobial activity of nanoparticles is highly influenced by several intrinsic factors such as their morphology, size, chemistry, source, and nanostructure [8, 9, 10, 11]. Specifically, antimicrobial activity of TiO2 NPs is greatly dependent on photocatalytic performance of TiO2, which depends strongly on its morphological, structural, and textural properties [12]. Several TiO2 NPs have been developed through different methods of synthesis. Specifically, in this chapter, eco-friendly synthesis based on biological sources, such as natural plant extracts and metabolites from microorganisms, which have resulted in TiO2 NPs with different size, shape, morphology, and crystalline structures will be presented. Titanium dioxide produces amorphous and crystalline forms and primarily can occur in three crystalline polymorphous: anatase, rutile, and brookite. Studies on synthesis have stated that the crystalline structure and morphology of TiO2 NPs is influenced by process parameters such as hydrothermal temperatures, starting concentration of acids, etc. [13]. The crystal structures and the shape of TiO2 NPs are both the most important properties that affect their physicochemical properties, and therefore their antimicrobial properties [14]. Regarding the crystal structures, anatase presents the highest photocatalytic and antimicrobial activity. Some works have shown that anatase structure can produce OH˙ radicals in a photocatalytic reaction, and as it will be clearly explained below, bacteria wall and membranes can be deadly affected [15, 16].
The potential health impact and toxicity to the environment of NPs is currently an important matter to be addressed. Several works have confirmed that metal oxide NPs conventionally synthesized using chemical methods, such as sol–gel synthesis and chemical vapor deposition, have shown different levels of toxicity to test organisms [17, 18, 19, 20]. In recent years, researchers have emphasized on the development of nanoparticles promoted through environmental sustainability and processes characterized by an ecological view, mild reaction conditions, and non-toxic precursors. Due to this growing sensitivity toward green chemistry and biological processes, ecological processes are currently being investigated for the synthesis of non-toxic nanoparticles.
These biological methods are considered safe, cost-effective, biocompatible, non-toxic, sustainable, and environmentally friendly processes [20]. Furthermore, it has been described that chemically synthesized NPs have exhibited less stability and added agglomeration, resulting in biologically synthesized NPs that are more dispersible, stable in size, and the processes consuming less energy [21].
These biosynthetic methods, also called “green synthesis,” use various biological resources available in nature, including live plant [22], plant products, plant extracts, algae, fungi, yeasts [23], bacteria [24], and virus for the synthesis of NPs. Among these methods, the processes that use plant-based materials are considered the most suitable for large-scale green synthesis of NPs with respect to their ease and safety [25]. On the other hand, the reduction rate of metal ions in the presence of the plant extract is much faster compared to microorganisms, and provides stable particles [26]. Plants contain biomolecules that have been highly studied by researchers like phenols, nitrogen compounds, terpenoids, and other metabolites. It is well known that the hydroxyl and carboxylic groups present in these biocompounds act as stabilizers and reducing agents due to their high antioxidant activity [12]. Thus, plant extracts have been studied as one of the best green alternatives for metal oxide nanoparticles synthesis [27]. In recent years, TiO2 nanoparticles have been obtained by using different plant extracts, but not all of them have been studied for their antimicrobial activity. Table 1 presents a compilation of synthesized TiO2 nanoparticles from green synthesis by using plant extracts that were tested against different microorganisms.
Source | Titanium precursor | Size (nm) | Shape/crystal structure | Target microorganism (method) |
---|---|---|---|---|
Azadirachta indica leaves extract [28] | TiO2 | 25–87 (SEM) | Spherical/anatase-rutile | S. typhi, E. coli, and K. pneumoniae (broth micro dilution method) |
Psidium guajava leaves extract [29] | TiO(OH)2 | 32.58 (FESEM) | Spherical shape and clusters/anatase-rutile | S. aureus and E. coli (agar diffusion) |
Vitex negundo Linn leaves extract [30] | Ti{OCH(CH3)2}4 | 26–15 (TEM) | Spherical and rod shaped/tetragonal phase anatase | S. aureus and E. coli (agar diffusion) |
Morinda citrifolia leaves extract [31] | TiCl4 | 15–19 (SEM) | Quasi-spherical shape/rutile | S. aureus, B. subtilis, E. coli, P. aeruginosa, C. albicans, A. niger (agar diffusion) |
Trigonella foenum-graecum leaf extract [21] | TiOSO4 | 20–90 (HR-SEM) | Spherical/anatase | E. faecalis, S. aureus, S. faecalis, B. subtilis., Y. enterocolitica, P. vulgaris, E. coli, P. aeruginosa, K. pneumoniae, and C. albicans (agar diffusion) |
Orange peel extract [32] | TiCl4 | 20–50 (SEM) | Irregular and angular structure with high porous net/anatase | S. aureus, E. coli, and P. aeruginosa (agar diffusion) |
Glycyrrhiza glabra root extracts [33] | TiO2 | 60–140 (FESEM) | Spherical shape/anatase | S. aureus and K. pneumoniae (agar diffusion) |
Synthesis of TiO2 NPs by using plant extracts.
Different factors need to be evaluated in this research field in order to obtain TiO2 NPs with better properties and to maintain their biocompatibility. It has been shown that nanoparticles obtained from green synthesis can have a better morphology and size translated into better antimicrobial activity. Mobeen and Sundaram have obtained TiO2 NPs from titanium tetrachloride precursor through a chemical and a green synthesis method. Sulfuric acid and ammonium hydroxide were used in the chemical-based method and, in the green synthesis, those chemical reagents were replaced by an orange peel extract [32]. The nanoparticles obtained by using the natural extract presented a well-defined and smaller crystalline nature (approx. 17.30 nm) compared to the nanoparticles synthesized through the chemical method (21.61 nm). Both methods resulted in anatase crystalline structures, and, when evaluating the antimicrobial activity, the more eco-friendly NPs revealed higher bactericidal activity against Gram-positive and Gram-negative bacteria compared to the chemically synthesized nanoparticles.
Bavanilatha et al. have also detailed TiO2 NPs green synthesis with Glycyrrhiza glabra root extract. Antibacterial activity against Staphylococcus aureus and Klebsiella pneumonia were investigated and in vivo toxicity tests using the zebrafish embryonic model (Danio rerio) were also carried out [33]. Results have demonstrated their biocompatibility because healthy embryos of adult fish to different variations of NP and no distinctive malformations were observed at every embryonic stage with respect to embryonic controls.
Subhapriya and Gomathipriya have biosynthesized TiO2 NPs by using a Trigonella foenum-graecum leaf extract, obtaining spherical NPs and their size varied between 20 and 90 nm, and their antimicrobial activity was evaluated through the standard method of disc diffusion [21]. The NPs showed significant antimicrobial activity against Yersinia enterocolitica (10.6 mm), Escherichia coli (10.8 mm), Staphylococcus aureus (11.2 mm), Enterococcus faecalis (11.4 mm), and Streptococcus faecalis (11.6 mm). Results confirmed developed TiO2 NPs as an effective antimicrobial drug that can lead to the progression of new antimicrobial drugs.
Spherical TiO2 NPs were synthesized from plants, in particular by applying a Morinda citrifolia leaf extract, and through advanced hydrothermal method [31]. Developed TiO2 NPs showed a size between 15 and 19 nm in an excellent quasispherical shape. In addition, their antimicrobial activity was tested against human pathogens, such as Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. TiO2 NPs exhibited interesting antimicrobial activity, principally against Gram-positive bacteria.
In addition to plants, other organisms can produce inorganic compounds at an intra or extracellular level. The synthesis of TiO2 NPs through microorganisms, including bacteria, fungi, and yeasts, also meets the requirements and the exponentially growing technological demand toward eco-friendly strategies, by avoiding the use of toxic chemicals in the synthesis and protocols [34]. The metabolites generated by microorganism present bioreducing, capping, and stabilizing properties that improve the NPs synthesis performance. Jayaseelan et al. have stated glycyl-L-proline, one of the most abundant metabolite from Aeromonas hydrophilia bacteria, as the main compound that acted as a capping and stabilizing agent during TiO2 NPs green synthesis [35]. Moreover, the interest in fungi in green synthesis of metal oxide nanoparticles has increased over last years. Fungi enzymes and/or metabolites also present intrinsically the potential to obtain elemental or ionic state metals from their corresponding salts [34, 36]. Different works based on the green synthesis of TiO2 NPs from bacteria and fungus are presented in Table 2. Some of them have been synthesized with antimicrobial and antifungal purposes, and their target microorganisms are also declared.
Microorganism | Titanium precursor | Size (nm) | Shape/crystal structure | Target microorganisms (method) |
---|---|---|---|---|
Aeromonas hydrophilia [46] | TiO(OH)2 | 28–54 (SEM) ~ 40.5 (XRD) | Spherical/uneven | S. aureus, S. pyogenes (agar diffusion) |
Aspergillus flavus [34] | TiO2 | 62–74 (TEM) | Spherical/anatase and rutile | E. coli, P. aeruginosa, K. pneumoniae, B. subtilis (agar diffusion and MIC) |
Bacillus mycoides [37] | Titanyl hydroxide | 40–60 (TEM) | Spherical/anatase | E. coli (toxicity) |
Bacillus subtilis [38] | K2TiF6 | 11–32 (TEM) | Spherical | Aquatic biofilm |
Fusarium oxysporum [36] | K2TiF6 | 6–13 (TEM) | Spherical/brookite | — |
Lactobacillus sp. [51] | TiO(OH)2 | ~ 24.6 (TEM) | Spherical/anatase-rutile | — |
Planomicrobium sp. [39] | TiO2 | 100–500 (SEM) | Irregular/pure crystalline | B. subtilis, K. planticola, Aspergillus niger (agar diffusion) |
Propionibacterium jensenii [52] | TiO(OH)2, 300°C | 15–80 (FESEM) | Spherical | — |
Saccharomyces cerevisiae [51] | TiO(OH)2 | ~ 12.6 (TEM) | Spherical/anatase-rutile | — |
Examples of TiO2 NPs synthesis through microorganisms, both bacteria and fungus strains.
Two important factors that affect NPs synthesis are the type of microorganisms and their source. Some microorganisms widely used in the food industry are Lactobacillus, a bacterium used in dairy products and as a probiotic supplement, and Saccharomyces cerevisiae, a yeast commonly used in bakery. Jha et al. have investigated the effectiveness of both microorganisms to synthesize TiO2 NPs. A comparison between synthesis through Lactobacillus from yogurt and probiotic tablets resulted in different NP sizes: a particle size of 15–70 nm for yogurt, and 10–25 nm for tablets. This difference was due to the purity of the bacteria [40]. In general, TiO2 NP synthesis through microorganisms has not provided stable sizes, being not industrially scalable compared to the synthesis of nanoparticles from plants.
Harmful bacteria, such as Staphylococcus aureus, Burkholderia cepacia, Pseudomonas aeruginosa, Clostridium difficile, Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii, Mycobacterium tuberculosis, and Neisseria gonorrhoeae, are responsible for bacterial infections that can cause serious diseases in humans year after year [40]. The principal solution is the use of antibiotics, antimicrobial and antifungal agents. Nevertheless, in recent years there has been an increase in the resistance of several bacterial strains to these substances, and therefore there is currently a great interest in the search for new antimicrobial substances. The antimicrobial nanoparticles have been studied due to their high activity, specifically the metal oxide nanoparticles [41, 42, 43]. In this sense, titanium dioxide nanoparticles are one of the antimicrobial NPs whose study has gained interest during last years.
TiO2 is a thermally stable and biocompatible chemical compound with high photocatalytic activity and has presented good results against bacterial contamination [44]. Table 3 presents some research including the antimicrobial capacity of TiO2 NPs.
Microorganism | NPs | Results |
---|---|---|
Methicillin-resistant Staphylococcus aureus [45] | Fe3O4-TiO2 core/shell magnetic NPs | The survival ratio [%] of bacteria decreased from 82.40 to 7.13%. |
Staphylococcus saprophyticus [45] | Fe3O4-TiO2 core/shell magnetic NPs | The survival ratio [%] of bacteria decreased from 79.15 to 0.51%. |
Streptococcus pyogenes[57] | Fe3O4-TiO2 core/shell magnetic NPs | The survival ratio [%] of bacteria decreased from 82.87 to 4.45%. |
Escherichia coli [46] | TiO2 nanotubes ~ 20 nm | 97.53% of reduction |
Staphylococcus aureus [46] | TiO2 nanotubes ~ 20 nm | 99.94% of reduction |
Bacillus subtilis [47] | TiO2 NPs co-doped with silver (19–39 nm) | 1% Ag-N-TiO2 had the highest antibacterial activity with antibacterial diameter reduction of 22.8 mm |
Mycobacterium smegmatis [48] | Cu-doped TiO2NPs ~20 nm | The percentage of inhibition was around 47% |
Pseudomonas aeruginosa [49] | TiO2 NPs 10–25 nm | Although it was not completely euthanized, their survival was significantly inhibited. |
Shewanella oneidensis MR-1 [48] | Cu-doped TiO2 NPs ~20 nm | The percentage of inhibition was around 11% |
TiO2 nanoparticles against different microorganisms and their antimicrobial activities.
The principal factors differentiating the antimicrobial activity between TiO2 NPs were their morphology, crystal nature, and size. According to López de Dicastillo et al. [11], hollow TiO2 nanotubes presented interesting antimicrobial reduction thanks to the enhancement of specific surface area. This fact can be explained by the nature of titanium dioxide, and one of the main mechanisms of its action is through the generation of reactive oxygen species (ROS) on its surface during the process of photocatalysis when it exposed to light at an appropriate wavelength. It is important to highlight that some research works have evidenced antimicrobial activity of TiO2 NPs increased when they were irradiated with UV-A light due to the photocatalytic nature of this oxide. The time of irradiation varied between 20 min [45] and 3 hours [50].
Titanium dioxide nanoparticles (TiO2 NPs) are one of the most studied materials in the area of antimicrobial applications due to its particular abilities, such as bactericidal photocatalytic activity, safety, and self-cleaning properties. The mechanism referred to the antimicrobial action of TiO2 is commonly associated to reactive oxygen species (ROS) with high oxidative potentials produced under band-gap irradiation photo-induces charge in the presence of O2 [51]. ROS affect bacterial cells by different mechanisms leading to their death. Antimicrobial substances with broad spectrum activity against microorganisms (Gram-negative and Gram-positive bacteria and fungi) are of particular importance to overcome the MDR (multidrug resistance) generated by traditional antibiotic site-specific.
The main photocatalytic characteristic of TiO2 is a wide band gap of 3.2 eV, which can trigger the generation of high-energy electron–hole pair under UV-A light with wavelength of 385 nm or lower [52]. As mentioned above for bulk powder, TiO2 NPs have the same mechanism based on the ROS generation with the advantage of being at nanoscale. This nanoscale nature implies an important increase of surface area-to-volume ratio that provides maximum contact with environment water and oxygen [53] and a minimal size, which can easily penetrate the cell wall and cell membrane, enabling the increase of the intracellular oxidative damage.
Bacteria have enzymatic antioxidant defense systems like catalases and superoxide dismutase, in addition to natural antioxidants like ascorbic acid, carotene, and tocopherol, which inhibit lipid peroxidation or O-singlet and the effects of ROS radicals such as OH2˙− and OH˙. When those systems are exceeded, a set of redox reactions can lead to the death cell by the alteration of different essential structures (cell wall, cell membrane, DNA, etc.) and metabolism routes [54]. In the following sections, several ways that cellular structures were affected in the presence of TiO2 NPs will be described. In order to understand the genome responses of bacteria to TiO2-photocatalysis, some biological approaches related to expression of genes encoding to defense and repair mechanism of microorganism will explained below. Different mechanisms and processes of antimicrobial activity of TiO2 NPs are represented as a global scheme in Figure 1.
Scheme of main antimicrobial activity-based processes.
ROS are responsible for the damage by oxidation of many organic structures of microorganisms. One of them is the cell wall, which is the first defense barrier against any injury from the environment, thus being the first affected by oxidative damage. Depending on the type of microorganism, the cell wall will have different composition; that is, in fungi and yeast, cell walls are mainly composed of chitin and polysaccharides [55], Gram-positive bacteria contain many layers of peptidoglycan and teichoic acid, and Gram-negative bacteria present a thin layer of peptidoglycan surrounded by a secondary lipid membrane reinforced with transmembrane lipopolysaccharides and lipoproteins [56]. Thus, the effect of TiO2 NPs will be slightly different depending type of microorganism.
It has been studied that the composition of the cell wall in Pichia pastoris (yeast) changed in the presence of TiO2, increasing the chitin content in response to the ROS effects [57]. The cell wall of Escherichia coli (Gram-negative) composed of lipo-polysaccharide, phosphatidyl-ethanolamine, and peptidoglycan has been reported to be sensitive to the peroxidation caused by TiO2 [58]. The damage can be quantified by assessing the production of malondialdehyde (MDA), which is a biomarker of lipid peroxidation, or through ATR-FTIR of the supernatant of cell culture, which evidenced the way that porins and proteins on the outer membrane were affected, probably as a result of greater exposure to the surface of TiO2 [59]. In fungi, the release of OH˙ captured hydrogen atoms from sugar subunits of polysaccharides, which composed the cell wall, leading to the cleavage of polysaccharide chain and the exposition of cell membrane [60].
In terms of genetic issues, there is evidence that the bacteria change the level expression of certain genes encoding for proteins involved in lipopolysaccharide and peptidoglycan metabolism, pilus biosynthesis, and protein insertion related to the cell wall which values were lower-expressed after exposition to TiO2 NPs [61].
The second usual cellular target of most of antibiotics is the cell membrane mainly composed by phospholipids, which grant the cell a non-rigid cover, permeability, and protection. Most of the studies with TiO2 NPs have been focused to the loss of membrane integrity caused by oxidation of phospholipids due to ROS such hydroxyl radicals and hydrogen peroxide [62, 63], which led to an increase in the membrane fluidity, leakage of cellular content, and eventually cell lysis.
Gram-positive bacteria present only one membrane protected by many layers of peptidoglycan, whereas Gram-negative bacteria are composed by two membranes, inner and outer, and a thin layer of peptidoglycan between them. The outer membrane is exposed, thus, more liable to mechanical breakage due to the lack of peptidoglycan protective cover, like in Gram-positive bacteria [64]. Some studies have demonstrated a better antimicrobial performance of TiO2 NPs against Gram-positive bacteria [65] while others reported that Gram-negative bacteria were more resistant [66, 67]. It can be concluded that the bacterial inactivation effectiveness depends mainly on the resistant capacity of cell wall structures and the damage level of ROS generation [68].
In contrast with the lower expression of genes related to the cell wall seen before, the level expression of genes encoding for enzymes involved in metabolism of lipid essential for the cell membrane structure, are over-expressed [61]. It would be concluded that cells compensate the initial cell wall damage by reinforcing the second defense barrier, the cell membrane, in a way to provide support against the oxidation produced by ROS.
In fungi, the biocidal effect is not quite different. In the presence of TiO2 NPs and UV light, hydroxyl radicals, hydrogen peroxide, and superoxide anions initially promote oxidation of the membrane, leading to an unbalance in the cell permeability, even decomposition of cell walls [69]. This oxidation can inhibit cell respiration by affecting intracellular membranes in mitochondria. Studies have demonstrated biocidal effects on Penicillium expansum [70], but there is still research on other strains.
Beyond the relatively well-studied initial lipoperoxidation attack of TiO2 NPs on the outer/inner cell membrane of the microorganism, specific mechanisms are still aimed of being solved.
As the oxidative damage generates lipoperoxidation of cell membranes due to their lipid nature, the respiratory chain, which takes place in the double-membrane mitochondria, is also affected. This organelle is a natural source of ROS in aerobic metabolism because superoxide anions are produced in the electron transfer respiratory chain process. Mitochondria can control this fact by converting them into H2O2 by superoxide dismutase (SOD), and finally into water by glutathione peroxidase and catalase [71]. The presence of TiO2 NPs increases the production of ROS at levels that this enzymatic defense mechanism cannot attenuate the damage, even a dysregulation in electron transfer through the mitochondrial respiratory chain implies an increase in ROS generation [72].
The genetic approaches have indicated that changes in level expression in genes related to the energy production in mitochondria prioritize the most efficient pathway to uptake oxygen, which is through ubiquinol coenzyme [61]. This coenzyme presented a higher capacity to exchange electrons, while the coenzyme-independent oxygen uptake pathways were expressed at lower level.
Damage at molecular level in DNA affects all regulatory microorganism metabolism, replication, transcription, and cell division. DNA is particularly sensitive to oxidative damage because oxygen radicals, specially OH˙ produced by Fenton reaction [73], may attack the sugar-phosphate or the nucleobases and cause saccharide fragmentation aimed to the strand break [74].
DNA strand modifications are more lethal than base modifications (punctual mutation). Mitochondrial DNA is more vulnerable to oxidative damage than nuclear DNA because it is closer to a major cellular ROS source [75].
Besides the enzymatic detoxification system (SOD, glutathione and catalase), DNA injuries are covered by a set of structures related to post-translational modification, protein turnover, chaperones (related to folding), DNA replication and repair, which are significantly over-expressed in the presence of TiO2 NPs [61].
Iron is an essential ion for cell growth and survival, but it can turn potentially toxic if some malfunction in homeostatic regulation occurs (i.e., Fenton reaction that produces ROS). Bacteria are able to regulate iron concentration in order to maintain it in a physiological range [76]. This regulation involves directly siderophores to active transport of iron in cell [77], whose coding genes related to siderophore synthesis and iron transport protein are significantly lower-expressed in the presence of TiO2 NPs, decreasing the ability to assimilate and transport it, leading to cell death [61]. The loss of homeostasis regulation was confirmed by ICP-MS analysis, which revealed that the presence of TiO2 NPs significantly reduced the cellular iron level in Pseudomonas brassicacearum, directly proportional to the cell viability [78].
Regarding the functions related to Pi group (PO43−) uptake, major differences were found in the expression of set of genes contained in Pho regulon, which were significantly lower when compared to the control [61]. The Pho regulon is a regulatory network in bacteria, yeast, plants, and animals, related to assimilation of inorganic phosphate, merely available in nature, and essential to nutritional cross-talk, secondary metabolite production, and pathogenesis [79].
This suggested that the microorganisms were highly deficient in phosphorus uptake and metabolism in the presence of TiO2 NPs. It should be also noted that the Pho regulon has been reported to regulate biofilm synthesis capacity and pathogenicity [80].
TiO2 NPs can directly oxidize components of cell signaling pathways and even change the gene expression by interfering with transcription factors [81]. There is evidence to confirm the interference of TiO2 NPs in biosynthesis pathways of signaling molecules that bind lipopolysaccharide, stabilize and protect the cell wall against oxidative damage [82]. Moreover, a significant decrease in the synthesis of quorum-sensing signal molecule related to functions like pathogenesis and biofilm development was observed. This was corroborated through Scanning Electron Microscopy (SEM) images of bacteria (P. aeruginosa) growth in the presence of TiO2 NPs without UV irradiation. Cells appeared mainly non-aggregated and dispersed in the substratum, compared with controls without NPs where cells were mainly aggregated by lateral contact. This suggested that TiO2 NPs not only affected microorganisms by oxidative damage, but also bacteria aggregation and biofilm formation, which directly influenced in pathogenicity [83].
In plants and algae, ROS can act as signaling intermediates in the process of transcription factor controlling stress response by H2O2, which is activated by a GSH peroxidase, and not by peroxides directly. But there is still lack of research in this area [84].
The control of morphology and crystal structure of TiO2 NPs is the most important factor to enhance their antimicrobial activity. The appropriate design based on desirable surface properties given by shaped nanoparticles can improve effectiveness that is also dependent on the type of bacteria. The route of synthesis of TiO2 NPs is also a key factor. Recent works have revealed more eco-friendly synthesis methods, principally based on plant-based compounds and microorganisms, such as bacteria and fungus. Antimicrobial activity of different TiO2 NPs against Gram-positive and Gram-negative bacteria including antibiotic-resistant strains has been confirmed in different works.
Specific studies on antimicrobial mechanisms have evidenced that microorganism exposed to photocatalytic TiO2 NPs exhibited cell inactivation at regulatory network and signaling levels, an important decrease in the activity of respiratory chain, and inhibition in the ability to assimilate and transport iron and phosphorous. These processes with the extensive cell wall and membrane alterations were the main factors that explain the biocidal activity of TiO2 NPs.
The authors acknowledge the financial support of CONICYT through the Project Fondecyt Regular 1170624 and “Programa de Financiamiento Basal para Centros Científicos y Tecnológicos de Excelencia” Project FB0807, and CORFO Project 17CONTEC-8367.
The authors declare no conflict of interest.
IntechOpen implements a robust policy to minimize and deal with instances of fraud or misconduct. As part of our general commitment to transparency and openness, and in order to maintain high scientific standards, we have a well-defined editorial policy regarding Retractions and Corrections.
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\\n\\nA Correction will be issued by the Academic Editor when:
\\n\\n3.1. ERRATUM
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\\n\\n3.2. CORRIGENDUM
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\\n\\n4. FINAL REMARKS
\\n\\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\\n\\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\\n\\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
\\n\\nAny suggestions or comments on this Policy are welcome and may be sent to permissions@intechopen.com.
\\n\\nPolicy last updated: 2017-09-11
\\n"}]'},components:[{type:"htmlEditorComponent",content:'IntechOpen’s Retraction and Correction Policy has been developed in accordance with the Committee on Publication Ethics (COPE) publication guidelines relating to scientific misconduct and research ethics:
\n\n1. RETRACTIONS
\n\nA Retraction of a Chapter will be issued by the Academic Editor, either following an Author’s request to do so or when there is a 3rd party report of scientific misconduct. Upon receipt of a report by a 3rd party, the Academic Editor will investigate any allegations of scientific misconduct, working in cooperation with the Author(s) and their institution(s).
\n\nA formal Retraction will be issued when there is clear and conclusive evidence of any of the following:
\n\nPublishing of a Retraction Notice will adhere to the following guidelines:
\n\n1.2. REMOVALS AND CANCELLATIONS
\n\n2. STATEMENTS OF CONCERN
\n\nA Statement of Concern detailing alleged misconduct will be issued by the Academic Editor or publisher following a 3rd party report of scientific misconduct when:
\n\nIntechOpen believes that the number of occasions on which a Statement of Concern is issued will be very few in number. In all cases when such a decision has been taken by the Academic Editor the decision will be reviewed by another editor to whom the author can make representations.
\n\n3. CORRECTIONS
\n\nA Correction will be issued by the Academic Editor when:
\n\n3.1. ERRATUM
\n\nAn Erratum will be issued by the Academic Editor when it is determined that a mistake in a Chapter originates from the production process handled by the publisher.
\n\nA published Erratum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n3.2. CORRIGENDUM
\n\nA Corrigendum will be issued by the Academic Editor when it is determined that a mistake in a Chapter is a result of an Author’s miscalculation or oversight. A published Corrigendum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n4. FINAL REMARKS
\n\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\n\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\n\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
\n\nAny suggestions or comments on this Policy are welcome and may be sent to permissions@intechopen.com.
\n\nPolicy last updated: 2017-09-11
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