Document Type : Research Paper
Authors
Division of Biotechnology, Department of Applied Sciences, University of Technology, Baghdad, Iraq
Abstract
Keywords
INTRODUCTION
Among the pathogens implicated in urinary tract infections (UTIs), Staphylococcus epidermidis and Staphylococcus haemolyticus, both Gram-positive bacteria, are notable. While commonly found on the skin and mucous membranes, these organisms can pose a threat, particularly in healthcare settings, where they may exploit compromised immune defenses or the presence of urinary catheters to cause infections. Conversely, Proteus mirabilis, a Gram-negative bacterium typically inhabiting the gastrointestinal tract, is recognized as a frequent cause of UTIs, often associated with urinary catheterization or structural urinary tract abnormalities. Notoriously resilient, P. mirabilis can instigate stone formation within the urinary tract, necessitating specific antibiotic interventions. In contrast, Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen renowned for its adaptability, thrives in diverse environments, including healthcare facilities. Its propensity for causing UTIs is exacerbated in individuals with weakened immune systems or undergoing invasive medical procedures. Combatting P. aeruginosa infections proves arduous due to its intrinsic and acquired antibiotic resistance, mandating judicious antimicrobial selection guided by susceptibility testing to ensure effective treatment. The global rise in cancer cases is a growing concern, with individuals diagnosed daily. Despite extensive research, new drugs, and numerous studies, exact cancer treatment remains elusive. The continuous increase in cancer cases emphasizes the need for innovative approaches. Nanotechnology is a promising field becoming widely known as a method that shows clear signs of future success in cancer research and treatment, employing nanotechnology in the research of cancer represents a cause of a marked change and approach to address the limitations of treatment. As the need for effective and successful solutions grows and continues to exist, involving nanotechnology in the studies of cancer represents a significant potential frontier [1]. In the sphere of biological sciences: biotechnology and medicine, nanotechnology has gone beyond regular boundaries, venturing into the field of nano-biotechnology. The integration of nanobiotechnology into medicine has been a successful instrument in the current and ongoing improvements of the quality of human life. As a result, nano-medicine has surfaced as a distinct field, allowing researchers and scientists to develop the prevention of disease strategies, and enterprising healthcare measures [2]. It is crucial to embrace the measures of precautions and educate ourselves proactively about the potential in the environment and other pollution issues, as well as the risks of the health-related issues that may come to light from the misemploy of nanotechnology. This is considered of the most importance, stating the growing interest in sustainability worldwide. By combining nanotechnology with the principles of sustainability, the way for a sustainable and promising prospect for this field could be preserved, ensuring that its benefits are realized while minimizing adverse impacts on the environment and living organisms [3]. Nanomaterials is a promising field, offering a wide range of applications especially cancer therapy [4]. Towards the creation of novel anticancer agents, significant investigations have been directed with nanotechnology playing an important role, noting, the majority of interactions between biomaterials and cells transpire at the nanoscale. nanostructures can engage with biomolecules on the cell surface [5]. According to the World Health Organization, there is a projected increase in annual cancer cases, expected to rise from 14 million in 2012 to 22 million by the year 2030 [6]. Most concentrated solution of Ag/TiO2 nanofibers led to the complete cessation of oral cancer cell proliferation and migration. This exceptional performance is likely attributed to the heightened reactivity of the surface and the level of the interaction between titanium dioxide and silver on an atomic level, accompanied by the silver ions release. These mechanisms collectively underpin the remarkable efficacy of the nanofibers generation. The observation of the biological effects underscores the possible utility of silver and titanium nanofibers in both anticancer [7]. Ag/TiO2 nanoparticles (NPs) exhibited significant generation of reactive species of oxygen (ROS), which ultimately led to the complete suppression of cell growth of cancer upon their systemic in vitro application. Furthermore, the Ag/TiO2 NPs efficiently absorbed visible light, thereby enhancing their anticancer sensitivity by inducing cancer cell death and inhibiting cell proliferation, as confirmed through cell viability assay testing [8].
The study aimed to evaluate the effect of TiO2-NPs, obtained through green synthesis, on the viability of HSSCC cells at different concentrations, focusing on concentration-dependent effects over different periods and exploring the MIC properties that reduce biofilm formation isolated from urinary tract infections. TiO2-NPs when presented as a homogeneous mixture showed strong anticancer potential and produced maximum antioxidant activity, demonstrating a strong and necessary anticancer ability against melanoma cell line, which helped to overcome melanoma cells. The observations indicate that TiO2-NPs may become an anti-cancer therapeutic treatment in the future.
MATERIALS AND METHODS
Materials
The media utilized included Mannitol Salt agar , MacConkey agar , Muller-Hinton broth (MHB) utilized to anti-biofilm ,Alcoholic ethanol was used in the extraction process of Q. infectoria plants. In addition to its use titanium tetra isopropoxide (TTIP) in preparing nanoparticle TiO2.
Collection of microbial isolated
The study involved gathering form UTI infection acquired in hospital. The diagnosis was established through the examination of the isolates morphological trait via culture media growth. The identified microbes include s.epidemidis, p.mirabilis ,p.aeruginosa ,s.hamolyticuds.
Preparation of plant extract
Q. infectoria gall underwent a cleaning process by washing with tap water to eliminate the tiny particles of dust from its outer surface. The plants were then cleansed with distilled water and left to dry at room temperature. After complete dryness of the plant at room temperature a fine powder was obtained when transferred and crushed by the grinder. Extraction by Alcohol: by the method described in reference [9] alcoholic extract of the plant was prepared, the extraction was performed by a continuous process in a Soxhlet applying Ethanol as a solvent at eighty percent concentration for seven hours, a further concentration process were carried out using a Rotary Evaporator, then the product were dried using oven at forty Celsius degree and preserved in bottles that were clean and sterilized, the product were kept in refrigerator until utilization.
Green synthesized TiO2 nanoparticles preparation
The Green synthesized process of TiO2 nanoparticles was carried out by applying one point six millimoles of Titanium tetraisopropoxide in fifty milliliters of isopropyl alcohol thereafter the addition of twenty-five milliliters of plant extract took place, under continuous mixing at seventy Celsius degree in two hours period. Afterward, the homogeneous mixture is left to age for over twenty-five hours. The obtained yellowish brown liquid was then transferred over hot air and kept at eighty Celsius degrees for eight hours then a dark and dry homogenous material was obtained and then converted to a powder. The acquired powder was rinsed multiple times with DI and ethanol. The resulting product was dried, calcined at three hundred fifty celsius degrees for three hours, ground, and then stored as TiO2-NPs, a method used with some modifications [10].
Characterisation of TiO2 nanoparticles
The analysis of X-ray diffraction on the produced powder was performed via (XRD Shimadzu-Japan). The characterization of Titanium dioxide nanoparticles’s morphology was performed by Scanning electron microscopy (SEM) to determine the shape and size of these nanoparticles. It was subjected to a Scanning Electron Microscope (Zeiss, Jena, Germany). The spectra over the range of 400 - 4000 cm-1 were recorded employing a Fourier Transform Infrared device (FTIR Shimadzu-Japan). Finally, TiO2 nanoparticles were characterized using a UV-Vis spectrophotometer (Chrom Tech, USA).
Anticancer Activity
Preparation of Cancer Cell Lines
The cytotoxic impact of TiO2-NPs, the experiment was conducted to investigate the toxic effects of test substances at concentrations (0.0, 1.95, 3.9, 7.8, 15.62, 31.25, 62.5, 125, 250, 500, 1000 µg/mL) on a HSSCC line in passage 27. The cells were cultured in RPMI-1640 medium supplemented with 10% Fetal Calf Serum (FCS). The cytotoxic impact of the test substance was examined by culturing cells in multiwell tissue culture plates (96-Microtiter plates) with a flat bottom. The experiment consisted of three stages: Cells Seeding: After activating and proliferating cancer cell lines for 24 hr, the monolayer growth was treated with Trypsin-Versen solution. Subsequently, 25 mL of RPMI-1640 medium, prepared with serum, was added to each well, adjusting the cell count to 1x104 using a counting chamber. A volume of 100 µl of the cell suspension was distributed into the tissue culture wells, which were then incubated at 37 °C for 24 hr to allow cell attachment to the glass [11].
Preparation of different TiO2-NPs concentrations
Different concentrations of the test substance were prepared using a serum-free tissue culture medium. These concentrations were added to the wells containing adherent cancer cells, and solutions were prepared just before use. Six replicates were used for each treatment. The culture medium in the plates was poured out, designating column 1 as the negative control, to which 200 µl of serum-free culture medium was added. Columns 2 to 12 received increasing concentrations (200 µl per well) of the test substance. The plates were covered, incubated at 37°C, and exposed for different durations (24, 48, 72 hrs).
Minimum Inhibitory Concentration
Microtiter Plate Method
The antibacterial activity of was assessed using minimum inhibitory concentration MIC assays against Gram-negative s.haemolyticus, sepidermids, p.mirabilis,and p.aeruginosa [12] Stated that the MIC was determined on a 96-well microtiter plate using the resazurin assisted microdilution technique in Mueller-Hinton broth (MHB) as follows:
1. Test material, which was: Plant extract (Q.infectoria), and TiO2 nanoparticle.
2. Preparation of test materials with triple the final required concentration.
3. 100 μl of broth medium in each well from 1 to 10 were made.
4. 100 μl of diluted test material was transferred to the first well. by transferring 100 μl from the first to the 10th well (the concentrations were 100, 50, 25, 12.5, 6.25, 3.12, 1.56, 0.78, 0.39, and 0.19 μg/ml) in addition to the controls.
5. Each well was inoculated with 100 μl of bacterial suspension equivalent to McFarland standard no 0. 5 (1.5 ×10 8 CFU\ml).
6. The microtiter plate was incubated at 37 C for 24 h. 8- 30 μl of resazurin (0.015%) prepared in ((3.2.5.5) was added to each well (30 μl/well), and incubated for 2 to 4 hrs. for the observation of color change. After completion of the incubation, rows with no color change (blue resazurin color remained unchanged) were scored as above the MIC whereas the last blue well in the row recorded as MIC.
Anti-biofilm
The microtiter plate (MTP) assay is a qualitative technique that uses a microplate reader to determine an agent’s effectiveness against biofilm formation. The minimum inhibition concentrations MIC obtained from the previous experiment were used to study the effect of the test materials on the formation or inhibition of biofilm of the studied s.haemolyticus, sepidermids, p.mirabilis,and p.aeruginosa isolates that produce strong biofilm, the test materials were: Plant extraction (Q.infectoria), and TiO2 NPs.
The same previously mentioned protocol (MTP) was used for the biofilm formation assay. However, 100 μl of test compounds was added. the plate was incubated at 37 °C for 24h. After that, all wells were washed, stained, and read at 600 nm wavelength using a micro-plate reader. percent of biofilm inhibition was calculated by the equation 1 [13].
Cytotoxicity Assay of TiO2-NPs
After the designated incubation duration, the contents of the plates (cell sediment, culture medium) were discarded, and wells were cleaned three times with phosphate-buffered saline to remove any traces of the TiO2-NPs and unattached cells. Then, ten micro-litters of MTT dye solution (final concentration 0.5 mg/ml) were added to each well and left for 4 hours at 37 ℃ in a CO2 incubator. The cells underwent a multiple times cleaning process with phosphate-buffered saline until any surplus dye was eliminated. After complete drying of the plates, 100 µl of DMSO was added to ensure complete dissolution of the violet formazan crystals. The outcomes were interpreted through an ELISA microplate spectrophotometer employed at a wavelength of 500 nanometers for result analysis. The inhibitory rate was determined using the equation 2 [14].
Antioxidant
The method mentioned in (Brand-Williams et al, 1995) was followed in conducting the antioxidant antioxidant test, using the method (2, 2-diphenyl-1-picryl-hydrazylhydrate DPPH), by adding 0.024 grams of DPPH to 50 milliliters of absolute ethyl alcohol. It is dissolved well by mixing it on a magnetic stirrer without heat, then the volume is supplemented to 100 milliliters with absolute ethyl alcohol to give a final concentration equal to 0.024 mg/ml. Then, half a milliliter of serial concentrations of the test substance (0.0, 25, 50, 100, 150, 200) µg/mL were taken and added to a mixture of DPPH (0.5.) mM and mL (3.3) of absolute ethanol. The amount of color change was measured using The spectrophotometer was at a wavelength of 515 nm during 100 minutes of reaction at room temperature. The plank tube contained (3.3) mL of absolute ethanol and (0.5) mL of the sample, and the control tube contained (3.3) mL of absolute ethanol. And (0.5 mL) of DPPH. The removal percentage was calculated according to the equation 3.
Ascorbic acid or vitamin C at a concentration of 1/1 (1 mg/100 ml of distilled water) is used as a positive control due to its high effectiveness as an antioxidant and is considered a standard material for comparison.
Statically analysis
The one-way analysis of variance (ANOVA) and subsequent post-hoc tests were conducted using IBM SPSS Statistics for Windows, version 26 (SPSS Inc. Chicago, Illinois, United States). Variables were expressed as mean ± standard deviation (SD). The level of significance was set at p≤ 0.05.
RESULTS AND DISCUSSION
In the present study, all isolates were examined primarily for colony characterization by culturing on the selective media, Mannitol salt , MacConkey agar incubated for 24 hours at 37ºC. The appeared colonies of s.epidermidis on Mannitol salt results growth of mucoid metallic pink colonies which refer to the presence of s.epidermidis . and appeared yellow colonies on mannitol salt which refer to the presence of s.haemolyticus .On MacConkey agar, red, mucoid, lactose fermented colonies were considered mirabilis. while colonies on Mackonky, brown , mucoid, colonies which refer to the presence of p.aeruginosa Fig. 1. In addition gram staining and microscopic observation were carried out for all isolates to determine the cell size, color and form and as shown in the Fig. 1 below. spherical or sub-spherical shape budding yeast-like cells Gram-positive bacteria stain violet because of the crystal violet assay when seen with a light microscope. This pigmentation is due to the peptidoglycan layer of the cell wall. On the other hand, gram-negative bacteria do not lag behind the crystal violet stain, because they absorb counter stain (safranin) and appear pink or red due to a layer of peptidoglycan layer that is thinner and confined between the outer and inner envelope of the cell.
Fig. 2 (A) depicts the optical absorption spectrum of plant extract with absorption notably centered around 269 nm. On the other hand Figure 1 (B) shows illustrates the optical absorption spectrum of green synthesized TiO2-NPs. Notably, the absorption edge is centered on 355 nm. To assess the suitability of the manufactured materials for various applications, it is crucial to compute the band gaps. The Tauc plot method was employed for the band gap measurements of the synthesized TiO2-NPs. This analytical approach aids in understanding the electronic properties of the nanoparticles and is essential for evaluating their potential utility in diverse applications.
Fig. 3A illustrates the spectrum of FTIR of plant extract, nanoparticles versus green synthesized TiO2 (Fig. 3B). However, in Fig. 3A, the peaks align with 3377.36 cm-1. In spectra, the observed peaks result from the stretching of hydrogen bonds in the hydroxyl functional group, O-H (alcohol) group. Peaks at 2980.02 cm-1 were identified as the C-H (alkane stretching and -CC-(alkynes ,lack of positional isomerism in symmetrical alkynes) functional group functional group. The spectrum of FTIR of TiO2 nanoparticles had peaks matching 3400.24 cm-1, in the spectra because of the stretching of the hydrogen bond of the hydroxyl O - H group (Alcohol), the peaks equivalent to 2926.01 cm-1 was displayed as the C-H Functional group of (alkane stretching and - CC-(alkynes ,variable not present in symmetrical alkynes). Peaks marked at 1448.54 cm-1 are for C = C (medium weak multiple bands). Functional group of (alkane stretching and - CC-(alkynes ,variable not present in symmetrical alkynes).
Fig. 4 illustrates the X-ray diffraction (XRD) patterns of TiO2-NPs. The distinctive creation of rutile-phase TiO2 nanoparticles is evident in the patterns. The diffraction peaks angles (2θ) = 28, 36.14, 38.83, 41.13, 44.55, 54.03, 56.52, 61.81, 63.43 and 69.47 correspond to the lattice planes of (110), (101), (202), (111), (210), (220), (002), (310), and (112), respectively (JCPDS no. 01-087-0920). To determine the crystallite sizes of TiO2 nanoparticles, Scherrer’s equation.Utilizing a Scanning Electron Microscope (SEM) for the examination of nanoparticle morphological characteristics, the Field Effect Scanning Electron Microscopy is employed. In Fig. 5, SEM micrographs of TiO2 nanoparticles at 120,000X magnification are presented for reference. The images showcase sphere-like structures, SEM This visual data enhances understanding of structural characteristics, representations reveal minimal aggregation in TiO2-NPs, providing valuable insights into their morphology distribution validated through comparison with relevant literature on TiO2-NPs
Various concentrations of the extract were employed, and the viability of HSSCC cells was measured after 24, 48, and 72 hours of exposure. The summarized findings are presented in Table 1. Observing Table 1 it could be assumed that the starting concentration point of the concentration was 7.8 µg/mL the HSSCC viability started gradually drooping after 24 and 72h of incubation with plant extract. 24-Hour Exposure: At a concentration of 0 µg/mL, cell viability was (mean ± SD; 1.77 ± 0.01). As the concentration increased, no significant differences were observed within this time frame (p > 0.05). The p-value for the overall effect was 0.22. 48 Hours Exposure: Similar to the 24-hour exposure, no significant differences were observed between concentrations (p > 0.05) at the beginning of the 48-hour exposure. The p-value for the overall effect remained non-significant at 0.21. 72 Hours Exposure: Table 1 represents the effects of plant extract concentrations on HSSCC cells at different time durations 24, 48, and 72 hours, while Fig. 6 represents the inhibition rate of plant extract. Concentration levels were divided into control (0 µg/mL) which serves as the baseline control group with no treatment, low concentrations (1.95 µg/mL, 3.9 µg/mL, and 7.8 µg/mL) representing the lower end of the dose range, intermediate (15.62 µg/mL, 31.25 µg/mL and 62.5 µg/mL) these concentrations cover the mid-range of doses and a higher concentrations (125 µg/mL, 250 µg/mL, 500 µg/mL and 1000 µg/mL) these concentrations represent the higher end of the dose range. At 24 hours of exposure, the inhibition rates (mean ± standard deviation) of plant extract on HSSCC cell viability were observed. assessing the impact of plant extract on HSSCC cell viability. After 72 hours of exposure, significant differences were observed in cell viability at varying concentrations. At 0 µg/mL and 1.95 µg/mL concentrations, the viability was significantly different from the other concentrations. At the highest concentration of 250 µg/mL, cell viability was significantly lower compared to other concentrations. The overall effect was statistically significant with a p-value of 0.01. The impact of the obtained plant extract on HSSCC cell activity and viability changes depending on the duration of exposure and the concentration of the dosage were the main results of this study indicated that after 72 hours of exposure, significant differences in cell viability at various concentrations was observed, higher cell viability was observed with the lower concentrations and vice versa, reduced viability at the high concentrations. Therefore, the dose-dependent effect of the plant extract on the activity of HSSCC in a certain period would be suggested. Furthermore, it could be concluded that TiO2-NPs have concentration-dependent effects on the viability of HSSCC cells. Obtained results provide valuable insights into the possible employment of these nanoparticles and plant extract as a therapeutic agent against cancer and specifically HSSCC, however, the mechanism of action and safety should be further investigated. Torres et al. (2016) conducted a study on the impact of α/β-thujone on glioblastoma, using both models in vitro and in vivo. They found that α/β-thujone possesses the capacity to reduce the feasibility of a cell and exhibits anti-proliferative, pro-apoptotic, and anti-angiogenic characteristics in vitro.
On the other hand, In the studies conducted in living organisms, α/β-thujone was observed to prompt regression of neoplasia and inhibit markers related to angiogenesis such as CD31, Ang-4 and VEGF within the tumor [15]. The antitumoral efficacy of the Q. infectoria extract has been assessed across various cancer cell lines [16-19]. These studies collectively contribute to our understanding of the potential anticancer properties associated with the use of Q. infectorias in homeopathic treatments.
On the other hand, Table 2 and Fig. 7, demonstrate the different concentrations of TiO2-NPs on HSSCC viability: The control (0 µg/mL): This serves as the baseline control group with no treatment. It helps you compare the effects of the treatment groups to the untreated cells. Low Concentrations (7.8 µg/mL, 3.9 µg/mL, and 1.95 µg/mL): These concentrations represent the lower end of your dose range. You might expect minimal or no effects on cell viability or proliferation at these levels. Intermediate Concentrations (62.5 µg/mL, 31.25 µg/mL, and 15.62 µg/mL): These concentrations cover a mid-range of doses. Effects on cell viability and proliferation may become more pronounced at these levels. Higher Concentrations (1000 µg/mL, 500 µg/mL, 250 µg/mL, and 125 µg/mL): These concentrations represent the higher end of your dose range. You may anticipate more significant effects on HSSCC cells at these concentrations, including potential cytotoxicity or inhibition of cell growth. Surprisingly all the examined concentrations were cider to the HSSCC and the viability in almost all concentrations was significantly decreasing.
MIC plant extract the result t of Table 3 the minimum inhibiter concentration plant extract were are identical for three pathogen isolated p.mirabilis however a lower minimum inhibiter contraction were deducted when s.haemolyticus was tested given 3.12 mg/ml. MIC of green synthesis TiO2 were identical the three pathogenic namely s.epidermidis , S.haemolyticus , p.mirabilis given concertation of 50 mg/ml will the concentration 25 was the MIC of the p.aeruginosa and this was the lower MIC among the green synthesis TiO2 and the rest of the all tested plant extract and TiO2 nanopartical. Where the results shown in the Table 3 indicated that the types of interactive oxygen (ROS), along with free radicals, produce the TiO2 which has damaged the wall of bacterial cells and also prevents respiratory enzymes.
Biofilms are accumulations of bacteria that are adhered to the surfaces and embedded in a self-produced matrix. The matrix of biofilm contains substances, such as proteins ,polysaccharides, and extra-cellular DNA, which protect bacteria from harsh environmental conditions and provides resistance to human immunity. The film can also withstand a variety of chemotherapeutic agents. Since infections produced by bacteria that form a biofilm are hard to treat, there is a need to search for new and novel biofilm inhibitors . In this study, the ability of, S.epidermids , S.hoemolyticus ,p mirobilis and P aeruginasa isolates to form biofilm was tested using a Microtiter plate (MTP) and read by Microtiter spectrophotometer. The results showed that there are some isolates that adhere and are able to produce biofilm while others are non-adherent and unable to form biofilm. as shown in the Table 4 in use condition plant extract There is s.epidermidis isolates that are strong for biofilm however ,in the case of used TiO2 (green synthesis) moderate biofilm four isolates of , S.aeruginosa, p mirabilis ,and week product biofilm in S..haemolyticus
Table 5 displays the antioxidant activity of two substances: TiO2-NPs and the plant extract. The antioxidant activity is measured at different concentrations (µg/mL) for each substance. The green synthesized TiO2-NPs generally exhibit higher antioxidant activity compared to plant extract across all concentrations. The antioxidant activity increases with the concentration for both green synthesized TiO2-NPs and plant extract, suggesting a dose-dependent relationship. The TiO2-NPs appear to enhance the antioxidant activity of plant extract, as evidenced by higher percentages at each concentration compared to the plant extract alone. The highest antioxidant activity is observed at the highest concentration (200 µg/mL) for both green synthesized TiO2-NPs and plant extracts. This data suggests that the green synthesized TiO2-NPs may have a synergistic effect on antioxidant activity, providing a potentially more potent antioxidant formulation compared to the plant extract. In addition, it is worth mentioning that many studies have dedicated different activities to nanoparticles derived from microorganisms [20-24], while others dedicated those from plant extract [25-29]. However, among all living organisms, plants exhibit the most promising capacity for nanoparticle biosynthesis, making them well-suited for extensive biosynthetic processes. In contrast to microorganisms, the production of nanoparticles derived from plants is characterized by greater speed and enhanced stability [30].
CONCLUSION
The results of this study demonstrate that the production of TiO2-NPs by the green synthesis approach yielded nanoparticles with consistent stability, and these were characterized by their physicochemical characteristics of chemical compositions, shape and size. Most of the isolates that were used in present study high antibiotic resistance in most of the antibiotic classes that reached 100%.s.haemolyticus,sepidermids,p.mirabilis,and p.aeruginosa isolates the ability to produce biofilm in four levels of intensity: strong, moderate, weak, and non-biofilm-producing. The MIC can reduce the biofilm formation of isolated from urinary tract infections. TiO2-NPs when presented as a homogenized blend exhibited robust anticancer potential and yielded the maximum antioxidant activity, demonstrating robust and necessary anticancer ability against the skin cancer cell line, which helped to overcome the cell cancer cells. The observations suggested that the TiO2-NPs may impaled as a future therapeutic anticancer cure. The effects on both anticancer and antioxidant activity can be associated with both the direct influence of the TiO2-NPs, as well as the indirect effects via the antioxidant capacity of the TiO2-NPs in supporting to combat cancer.
ACKNOWLEDGMENT
The authors wish to express their genuine gratitude to the Department of Applied Science Laboratories, University of Technology, (Baghdad/Iraq) for its experimental aid.
CONFLICT OF INTEREST
The authors declare that there are no conflicts of interest regarding this article.
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