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In this project, five different nanoparticles based on different ratios of iron, zinc, and manganese were synthesized to form Znx Mn1-X Fe2O4. Then, one of them was selected and polyethylene glycol was conjugated to its surface using tartaric acid to improve its biocompatibility. Also, to improve the target ability of the prepared polyethylene glycol conjugated Znx Mn1-X Fe2O4 as a nano-platform, folic acid was used as a targeting agent. The nano-platform was characterized using different techniques such as X-ray diffractometey, vibrating-sample magnetometer, ultraviolet–visible spectroscopy, field emission-scanning electron microscopy, Fourier-transform infrared spectroscopy and thermogravimetric analysis. In vivo and in vitro tests including hemolysis, blood aggregation and lethal dose showed that Zn0.5Mn0.5 Fe2O4 were the most suitable photosensitizers among all. Also, in vivo and in vitro biocompatibility analysis revealed that incorporation of polyethylene glycol can significantly improve the biocompatibility of the nano-platforms. The cytotoxicity results revealed that cell viability of cancerous cells was lower when the nano-platform was coated with folic acid and poly ethylene glycol under irradiation of visible light while whiteout shining the light, the cell viability was almost 100% for polyethylene coated nano-platform. These results can prove the great potential of folic acid to improve the photodynamic therapy performance of Zn0.5Mn0.5 Fe2O4.
Introduction
Nowadays, photodynamic therapy (PDT) has become one of the new approaches for the treatment of various cancers [1, 2]. PDT consists a photosensitizer (PS) which can be excited by irradiation of light (visible and infra-red lights are more appropriate) [3]. PSs usually do not accumulate in the nuclei of the normal cells which can remove concerns about them being carcinogenic materials, a property which makes them appropriate candidates for the treatment of cancer [4]. Moreover, the side effects in chemotherapy or radiotherapy have not been observed either [5]. Transition metals based photosensitizer are an important category as they can be excited using visible or infrared lights which are less harmful than ultra violet lights for human body [6, 7, 8, 9–10]. In 2024, Fatemeh Javani Jouni, and coworkers used zinc oxide quantum dots and visible light (blue light) on MDA-MB-231 cancer cells to inhibit cancer markers and induce apoptosis [11]. In 2023, Chandran Murugan a ferrite nanoparticle based on cerium and molybdenum disulfide and utilized it as dual-performance cancer therapies for phtotothermal therapy (PTT) and reactive singlet oxygen (ROS) generation [12]. Also, in 2023, Yang Shuai utilized a modified nanoplatform of SnFe2O4 nanozyme. The nanoplatform could act as a photoabsorber to convert light energy into heat for PTT while as a photosensitizer it could transfer the photon energy to generate ROS for PDT [13]. Therefore, several transition metals such as manganese (Mn), iron (Fe), and zinc (Zn) were selected to combine with one another to prepare nano-platforms for using in PDT. For example, Fe can be merged with oxygen and other metals to form spinel ferrites which are usually demonstrated by MFe2O4 structure in which M can be a third metal. Fe and Zn, and Mn-based nanostructures have attracted significant attraction in biomedical applications. In tumor microenvironment glutathione (GSH) acts as a neutralization agent which is not suitable for PDT, and It has been found that MnO2 could reduce the level of the GSH by its consumption [14]. Hence, in this work, a series of ferrite nano-platforms consist of different ratios of Zn and Mn were synthesized to utilize anticancer potential of Zn and Mn while using the whole Znx Mn1-x Fe2O4 platform for PDT. Then, polyethylene glycol (PEG) was added to one of the prepared nano-platforms to improve its hydrophilicity and reduce probable toxicity. Also, a chemical targeting agent (folic acid (FA)) was attached to the surface of one of the prepared platforms to enhance its efficiency of treatment [15, 16, 17–18].
Materials and Methods
Material
Zinc (II) chloride (Sigma, ≥ 98%), manganese (II) chloride tetrahydrate (Sigma, ≥ 99.0%), Iron (III) chloride hexahydrate (Sigma, ≥ 98%), sodium hydroxide (NaOH) (Merck, ≥ 99%) PEG (Merck, Mn = 6000), FA (Sigma, > 97%), tartaric acid (Tar) (Sigma, ≥ 99.5%), N-hydroxysuccinimide (NHS)(Sigma, 98%), N–N′-ethylcarbodiimide hydrochloride (EDC.HCl)(Sigma), and 1,3-Diphenylisobenzofuran (DPBF)(Sigma, 97%) were purchased from Sigma-Aldrich chemical company. Other chemicals were from chemical lab in purity grades.
Preparation of Zinc-based Magnetic Nano-platform
Here all nano-platforms were synthesized via co-precipitation [19]. In this method (Fig. 1A), Znx Mn1-x Fe2O4, where X = 0, 0.25, 0.5, 0.75, and 1 were synthesized by dissolving the appropriate combination of Zinc (II) chloride, manganese (II) chloride tetrahydrate, Iron (III) chloride hexahydrate salts at various molar ratios in 20 mL of distilled. The mixture was then heated and agitated until its temperature reached 70 °C. In addition, 4 g of NaOH was dissolved in 30 mL of distilled water, which was then stirred and elevated to 70 °C. Then, two solutions were combined and agitated at 100 °C for 30 min. A magnet was then utilized to isolate the desired product. The solid product was then rinsed three times with distilled water to eliminate impurities. All of the subsequent tests were conducted on Znx Mn1-x Fe2O4 (X = 0, 0.25, 0.5, 0.75, and 1) to determine the most suitable composition, which was Zn 0.5. It possesses the optimal balance of all the characteristics required for the intended PDT.
[See PDF for image]
Fig. 1
A preparation of Zn based nano-platforms by co-precipitation method, B preparation of PEG-Zn 0.5 nano-platforms, C Preparation of FA-PEG-Zn 0.5 nano-platforms
Preparation of PEG Conjugated Tar Coated Zn 0.5 (PEG-Zn 0.5)
After evaluating and determining the anti-cancer potential of each of the prepared photosensitizers, the best one (Zn 0.5) was chosen for this step. In first step, 300 mg (two mmol) of Tar and 6 g (one mmol) of PEG were dissolved in 50 mL of distilled water (Fig. 1B); then, one mmol of EDC and one mmol of NHS were added to the solution. Tar solution was magnetically agitated at cold temperature (using ice bath) for 48 h (hrs). After this time period, 236 mg of Zn 0.5 (1 mmol) was added to the PEG-Tar solution, followed by the addition of one mmol of EDC and one mmol of NHS; then, the mixture was stirred for another 48 h at cold temperature. The product was then separated with a magnet and rinsed three times to remove unreacted substances. PEG-Zn 0.5 was then desiccated at room temperature.
Preparation of FA-PEG-Zn 0.5
500 mg of PEG-Zn 0.5, FA (0.5 mmol), NHS (0.5 mmol), and EDC (0.5 mmol) were added to a 0.125 M of NaOH solution (20 mL). The solution was then agitated for 48 h at cold temperature in a dark condition. After 48 h, the FA-PEG-Zn 0.5 was magnetically separated and at least three times rinsed with distilled water to remove impurities. Then, it was dried at room temperature (Fig. 1C).
Characterization
The prepared photosensitizers were characterized using an X-ray diffractometer (XRD) (PW1730 Philips, Netherlands, = 1.54056 A) to measure their crystal structure. In addition, the most prominent peak in the XRD pattern was used to determine the crystallite dimension of each sample. All elements in the prepared samples were determined using energy dispersive X-ray analysis (EDX) (MIRA II (SAMX detector, France). Ultraviolet–visible (UV–Vis) (Biochrom WPA Biowave II, United Kingdom) spectroscopy was conducted to confirm the preparation of various products, the morphology of samples was determined using field emission-scanning electron microscopy (FE-SEM; Mira III; Tescan; Czech Republic). Chemical composition of each sample was determined using Fourier-transform infrared spectroscopy (FT-IR) (VERTEX 70, Brucker, Germany). The magnetic properties of samples were calculated using a vibrating-sample magnetometer (VSM) (Magnet Kavir Company, 1.5 T). Thermal characteristics were determined by performing thermogravimetric analysis (TGA) (Q600, Ta, USA) under argon gas at a heating rate of 15 ºC per minute.
Determination of the Band Gap of the Nano-platforms
The nano-platforms' band gap was determined using the Tauc and Devis Mott method. Briefly, the UV–Vis spectrum of each nano-platform was recorded between 200 and 750 nm, a curve was drawn for each sample according to Eq. 1, and the band gap (Eg) was calculated by extrapolating the curve.
1
where, α, h, C, λ, K, and Eg represent, the absorption coefficient, Planck constant, velocity of light, wave length, equation constant, and band gap, respectively.Biocompatibility Test
All procedures conducted in this study were in accordance with the ethical standards of the University of Guilan's research committee, as evidenced by the letter with the identifier 12078/p15.
Hemolysis
The hemolysis ability of all samples in this endeavor was evaluated to determine their biomedical safety and potential. One of our co-authors voluntarily donated blood sample for this purpose (5 mL). The red blood cells (RBCs) were then extracted via centrifuge at 4000 rpm for 5 min. The RBCs were rinsed with phosphate buffer saline (PBS) at least three times and diluted to a volume of 50 mL. The samples were then weighed at 5 mg, and 1 mL of PBS was added to each. In microtubes, samples and diluted RBCs were then combined in a 1:1 ratio (each sample was performed three times). RBCs were combined with fresh PBS and distilled water as negative and positive controls. The samples were incubated at 37 °C for four hrs [20]. Then, samples were centrifuged, and the supernatants were collected in order to ascertain the percentage of hemolysis using a microplate reader at 540 nm.
Aggregation Test
Separated RBCs were also evaluated by aggregating assay [21]. In the initial step and for this purpose, RBCs were diluted with phosphate buffer saline (1:10). 1 mL of diluted RBCs were then combined with 1 mg of each sample. All samples were incubated for one hour at 37 °C. Finally, a 40 × optical microscope (Nikon eclipse E100) was utilized to capture micrographs of samples.
Lethal Dose
To assess the safety of each sample in a living organism, a lethal dose test was conducted. The lethal dose test began by selecting three adult rodents per sample. Mice weighed approximately 30 g and were housed in the laboratory for at least seven days to acclimate to the test's conditions. One mouse was then administered 35 mg/kg of each sample, which is 20 times greater than our practical dose. If the treated rodent survived after 24 h, the same dose would be administered to two additional mice. The sample lethal dose can then be determined if all three rodents survive. Alternatively, if one of the three mice did not survive, three more mice would be selected and administered lesser doses to determine the lethal dose for each sample.
Determination of Singlet Oxygen Generation Using DPBF Assay
The generation of ROS by Zn 0.5 and PEG@ Zn 0.5 was measured by observing the photooxidation of DPBF [22, 23]. DPBF is an acceptor as it can absorb singlet oxygen to produce colorless compounds. To minimize the possibility of ROS quenching by samples, the test was conducted at modest sample concentrations, with DPBF and sample concentrations of 10 µM and 10 µg/mL, respectively. The prepared samples were exposed for one minute to visible light. Then, the absorbance of the supernatant was measured with a UV–Vis spectrophotometer at 425 nm.
In Vitro Photo-cytotoxicity Assay
To evaluate the efficacy of the prepared photosensitizers for treatment of breast cancer, human adenocarcinoma breast cell lines (MDA) were cultured. In a 96-well plate containing 100µL of Dulbecco's Modified Eagles Medium (DMEM) including 10% fetal bovine serum (FBS), and 1% penicillin/streptomycin, MDA cells were seeded. Various sample concentrations were introduced to the cells. The cells were exposed for 20 min to visible light (250W, wavelength 400–700 nm). Wells were incubated for an additional 72 h. Then, 20µL of a solution containing 5-dimethylthiazol-z-yl)-2,5-diphenyltetrazolium bromide (MTT) was added to each well. After a few hours, 100 µL of dimethyl sulfoxide was added to this solution to dissolve the formazan crystals, and the absorbance was measured using a microplate reader at 570 nm. For comparison, a second group was treated with nano-platforms without exposure to light.
Statistical Analysis
The significance of the difference between the results was determined using the ANOVA test.
Results and Discussion
Characterization of Photosensitizer
Figure 2A illustrates the XRD patterns of every nano-platform. Both Zn 0 and Zn 1 exhibited the characteristic diffraction pattern of cubic phase with the space group Fd3m. Zn 0 (8.5110 A°) and Zn 1 (8.4300 A°) possess the reference codes 01–074-2403 (MnFe2O4) and 00–001-1108 (ZnFe2O4), and all three crystallographic parameters were identical [24]. In addition to Zn 0 and Zn 1, the characteristic peaks of all doped samples were located in accordance with their component. For instance, the characteristic peaks of Zn 0.25 and Zn 0.75 are closer to Zn 0 and Zn 1, respectively. Debye-Scherer-equation was used to determine the crystallite size (D) of metal photosensitizers as follows:
2
where λ is the X-ray wavelength, K is the shape factor, θ is the Bragg angle, and β is the half maximum intensity line broadening (FWHM). Table 1 presents the crystallite dimension of metal photosensitizers. As can be seen, the size of all photosensitizers falls between 10 and 30 nm, which is ideal for delivery system in cancer.[See PDF for image]
Fig. 2
A X-ray pattern, B The EDX analysis, and C VSM analysis of Zn 1, Zn 0.75, Zn 0.5, Zn 0.25, and Zn 0; D UV–Vis of Zn 0.5, PEG, PEG@ Zn 0.5, and FA-PEG@ Zn 0.5; FE-SEM image of (E) Zn 0.5 and (F) FA-PEG@ Zn 0.5 (scale 200 nm); G FTIR spectrum of Zn 0.5, PEG, PEG@ Zn 0.5, FA, and FA-PEG@ Zn 0.5; H TGA analysis of Zn 0.5 and FA-PEG@ Zn 0.5
Table 1. The crystallite size of all metal photosensitizers (nm)
Zn 0 | Zn 0.25 | Zn 0.5 | Zn 0.75 | Zn 1 |
|---|---|---|---|---|
25 | 15 | 22 | 24 | 10 |
Figure 2B depicts the EDX analysis of Zn-based nano-platforms. As can be seen, all samples contain iron and oxygen. According to our preparation, the intensity of two characteristic peaks at 5.93 and 1.11 keV, which were attributed to manganese and zinc, decreased or increased based on the amounts of these metals in samples, confirming the successful preparation of zinc-based nano-platforms containing varying amounts of zinc and manganese.
In Table 2, magnetic parameters of various nano-platforms are listed. According to the table, the Ms and Mr were progressively increased by the addition of manganese. This can be explained by the fact that Zn is a nonmagnetic metal and the addition of manganese which is inverse spinel, can increase the Ms. When Zn ions were substituted into manganese ferrite, as their affinity toward tetrahedral sites is greater than that of ferric ions, the amounts of ferric ions on A sites decreased, and as a result of antiferromagnetic coupling, the magnetic moment on the B lattice increased, as did the saturation magnetization [25]. However, the magnetic properties of each photosensitizer, even after the addition of FA-PEG to Zn 0.5, were adequate for our needs. In addition, the parameter R can demonstrate the various forms of group inter-grain exchange, and, according to the references, a magnetostatic interaction can be determined in all nano-platforms because the value of R was less than 0.5 for all of them [26, 27]. Figure 2C illustrates the hysteresis loop for every photosensitizer.
Table 2. Magnetic parameters for all synthesized photosensitizers (emu/g)
Samples | Saturation magnetization Ms | Remanent magnetization Mr | Remanence ratio (R = Mr/Ms) |
|---|---|---|---|
Zn 0 | 24.2 | 3.2 | 0.1 |
Zn 0.25 | 24.8 | 1.9 | 0.1 |
Zn 0.5 | 18.0 | 0.1 | 0.0 |
Zn 0.75 | < 1 | 0 | - |
Zn 1 | 10.0 | 1.6 | 0.2 |
FA-PEG@ Zn 0.5 | 15.0 | 0.0 | 0.0 |
Characterization of PEG@ Zn 0.5 and FA-PEG@ Zn 0.5
After preparing PEG@ Zn 0.5 and FA-PEG@ Zn 0.5, the UV–Vis spectrophotometer was utilized as a characterization technique to demonstrate the successful deposition of PEG on Zn 0.5. As shown in Fig. 2D, PEG exhibited an absorption peak in the range of 200 to 300 nm, whereas Zn 0.5 exhibited no absorption peak in the same range. However, when PEG and FA-PEG were incorporated onto the surface of Zn 0.5, the characteristic peak was observed at the same wavelength range (200 to 400 nm) and the general trend of the curve was similar to that of Zn 0.5. This indicates that PEG and FA-PEG were successfully incorporated into the surface of Zn 0.5.
Figure 2E and F illustrates that all of the manufactured nanoparticles are spherical and homogenous. In addition, the addition of PEG to Zn 0.5 reduced the size of nanoparticles from approximately 45 nm to 30 nm, demonstrating the ability of PEG to disperse and stabilize nanoparticles, particularly in an aqueous environment. As a result, the FA-PEG@ Zn 0.5 particle size became highly suitable for passive targeting. When tumor tissue reaches a volume of 22 mm3 or more, its permeability typically decreases, impairing its ability to excrete waste, assimilate nutrients, and oxygenate cells. To circumvent this issue, tumor tissues initiate angiogenesis, resulting in abnormalities in the membrane and the formation of vessels with pores larger than 100 nm. Moreover, tumor tissues typically lack an efficient lymphatic drainage system. All of these factors result in anomalous transport dynamics, particularly for macromolecular materials such as PEG. In solid tumors, this phenomenon is known as the enhanced permeability and retention effect (EPR) of macromolecules and lipids [28, 29]. Therefore, the small size of our prepared nano-platforms, coupled with the presence of PEG (EPR effect), can enhance drug delivery to tumor cells through passive targeting.
Figure 2G illustrates the FTIR spectra of the samples. The bands at 452 and 569 cm−1 in Zn 0.5 are attributed to bending and stretching vibrations of octahedral and tetrahedral spinel sites. The remaining peaks correspond to hydroxyl group bonds on metals [30, 31]. According to the structure of folic acid, its FTIR curve displayed two characteristic peaks at 1730 and 3427 cm−1 that correspond to carboxylic acid and amine bonds, respectively [32]. In addition, PEG has a characteristic peak near 1100 cm−1 that corresponds to its ether groups [33, 34]. After incorporation of PEG and FA onto Zn 0.5, the corresponding peaks of these two materials appeared on the FA-PEG@ Zn 0.5 curve. These observations demonstrate that FA-PEG@ Zn 0.5 was successfully prepared.
Figure 2H compares the thermogravimetric analyses of Zn 0.5 and FA-PEG@ Zn 0.5. Zn 0.5 demonstrated a weight loss of 10.4% up to 700 ºC, whereas FA-PEG@ Zn 0.5 demonstrated a weight loss of approximately 30.9% under the same conditions. Since the difference between the two trends can be attributed to the presence of FA-PEG, it can be concluded that approximately 20.5% of nanocarriers are coated with FA-PEG on Zn 0.5. This can demonstrate the successful deposition of FA-PEG on the nano-platform, which can decrease its potential toxicity and increase the sample's efficiency as a DDS.
Determining the Band Gap
As described in the materials and methods section, the most effective nano-platform was chosen for modification with PEG and FA. Table 3 displays band gaps of nano-platforms before and after incorporation of PEG and FA. According to the explanations in the references, a material is a semiconductor if its band gap is less than 4.0 eV [35]. Furthermore, if the excitation energy of a material is between 1.6 eV and 3.2 eV, this particular material can excited by visible light which makes it a great candidate for photodynamic therapy. Therefore, one of our goals in this project was to keep band gap of prepared nano-platforms in the visible range especially after incorporation of PEG and FA. Hence, as it was shown in Table 3, all nano-platforms' band gap were in the visible light range.
Table 3. Band gaps of various metal nano-platforms (electron volt)
Zn 0 | Zn 0.25 | Zn 0.5 | Zn 0.75 | Zn 1 | PEG@ Zn 0.5 | FA-PEG@ Zn0.5 |
|---|---|---|---|---|---|---|
2.4 | 2.1 | 2.2 | 2.3 | 1.9 | 2.2 | 2.1 |
Biocompatibility Test
Hemolysis
Table 4 displays the percentage of hemolysis activity of Zn-based nano-platforms. According to the data presented, the toxicity of Zn-based nano-platforms increased as their Zn content rises. This may be due to the fact that when ZnFe2O4 is reduced to nanoscale form, it possesses innate cytotoxicity [36, 37]. However, after coating Zn-based nano-platform with PEG, the metal-based nano-platforms' hemolysis activity decreased to a lower range. This observation can be ascribed to the biocompatibility, hydrophilicity, and immunological resistance of PEG [38]. Figure 3 shows a schematic illustration of various samples.
Table 4. Percent of hemolysis for all of the prepared nano-platforms
Sample | Hemolysis % (5 mg/mL) |
|---|---|
Zn 0 | 4.2 ± 4.1 |
Zn 0.25 | 3.9 ± 1.7 |
Zn 0.5 | 2.1 ± 3.6 |
Zn 0.75 | 6.3 ± 2.8 |
Zn 1 | 2.8 ± 1.7 |
FA-PEG@ Zn 0.5 | 2.1 ± 2.0 |
[See PDF for image]
Fig. 3
The schematic illustration of hemolytic activity of (A) Zn 0, (B) Zn 0.25, (C) Zn 0.5, (D) Zn 0.75, (E) Zn 1, and (F) FA-PEG@ Zn 0.5
Aggregation Test
Analyses of RBC aggregation were conducted on all of the prepared nano-platforms. As shown in Fig. 4, all bare nano-platforms (Zn 0, Zn 0.25, Zn 0.5, Zn 0.75, and Zn 1) exhibited a moderate to high level of aggregation relative to positive and negative controls, with the level of aggregation increasing as the quantity of Zn in them increased. This finding may also be associated with the capacity of nanoscale ZnFe2O4 to induce intrinsic cytotoxicity against cells [36]. The successful preparation of PEG@ Zn 0.5 and FA-PEG@ Zn 0.5 can further be proved by the fact that the RBC aggregation level decreased substantially after PEG was used as a coating agent. In addition, it demonstrates that PEG can reduce the toxicity of metal photosensitizers and may increase their blood circulation time.
[See PDF for image]
Fig. 4
RBCs aggregation images of (A) FA-PEG@ Zn 0.5, (B) PEG@ Zn 0.5, (C) Zn 0, (D) Zn 0.25, (E) Zn 0.5, (F) Zn 0.75, (G) Zn 1, (H) Negative control, and (I) Positive control
Lethal Dose
According to the test, all rodents survived after receiving a dose of 35 mg/kg. Therefore, it can be concluded that the lethal doses of the prepared nano-platforms are greater than 20 times the practical dosages used in this study, therefore, the prepared nano-platforms can be considered safe when not exposed to direct light. After receiving 35 mg/kg doses of the samples, the weight changes of mice were monitored for one week, and as shown in Fig. 5, all mice exhibited similar trends in comparison to the control group, indicating that they all continued their normal activities [39].
[See PDF for image]
Fig. 5
The weight change in mice after treatment using 35 mg/kg of each sample
ROS Generation
DPBF, an acceptor that can absorb and rapidly scavenge singlet oxygen to form an endoperoxide that decomposes into 1,2 dibenzoylbenzene, was used to measure ROS production. As shown in Fig. 6, the absorbance of DPBF significantly decreased more than the control with time, confirming the production of ROS by Zn 0.5. In addition, the results demonstrated that the prepared nano-platform retains its PDT potential to produce ROS even after coating.
[See PDF for image]
Fig. 6
DPBF assay to measure ROS generation of Zn 0.5, PEG@ Zn 0.5
In Vitro Photo-cytotoxicity Assay
Figure 7A depicts the anticancer effects of all bare nano-platforms (Zn 0—Zn 1) on MDA. As shown in the figure, Zn 0.5 and Zn 0.75 exhibited the greatest performance, reducing cell viability the most when they were exposed to light. In the presence of light, the viability of MDA cells treated with Zn 0.5 and Zn 0.75 was reduced to 64.1% and 60.4%, respectively, at a concentration of 4 mM, whereas in the absence of light, the viability of the cells was 83.1% and 89.0% for the same concentration. These outcomes can be explained by two factors: a) Mn inhibits GSH activity in tumor microenvironment [14], and b) ability of Zn to cause inherent cytotoxicity against cells when it is reduced from bulk to nanoscale [36]. Therefore, the combination of these two properties could enhance the efficacy of PDT. In contrast to the common core–shell structure, our synthesized nano-platforms have a cubic spinel structure, which makes them a suitable candidate for PDT because all metals in this structure are simultaneously in contact with the microenvironment and perform their function. As previously explained, PEG was used as a coating agent to enhance the biocompatibility of the synthesized nano-platforms. As described in the previous section, PEG can reduce the toxicity of PEG@Zn 0.5 in the absence of light. The cytotoxicity results of PEG @Zn 0.5 at both concentrations (3 and 4 mM) demonstrated an equal reduction in cell viability, proving that the addition of PEG enhanced the activity of nano-platforms (Fig. 7B). At 3 and 4 mM, the addition of FA to PEG@ Zn 0.5 decreased cell viability from 76.7% and 64.1% to 57.9% and 40.0%, respectively. This demonstrates the potential of FA to enhance the PDT performance of Zn 0.5 by improving the targeting ability of the prepared nano-platforms [40].
[See PDF for image]
Fig. 7
A Cell viability of bare photosensitizers on MDA cancer cells after 72 h (under visible light and without visible light) (P Value < 0.001 ***) (P Value < 0.05 *); B Cell viability of FA-PEG@ Zn 0.5, PEG@ Zn 0.5, and Zn 0.5 on MDA cancer cells after 72 h (under visible light and without visible light) (P Value < 0.001 ***) (P Value < 0.01 **)
Conclusions
In this study, nano-platforms with the basic structure of Znx Mn1-X Fe2O4 were synthesized using co-precipitation method. Then, one of the prepared nano-platforms was selected, and was modified with PEG and FA to improve nano-platforms biocompatibility and target ability. The in vitro photo-cytotoxicity assay showed the potential of the prepared nano-platform in PDT under visible light, minimizing the potential damage caused by UV light exposure. The addition of PEG and FA to the nano-platforms improved their dispersibility and targeting ability enhancing the antitumor efficiency of Znx Mn1-X Fe2O4. Also, the small size of the prepared nano-platforms is advantageous for the accumulation of drugs at the tumor site, increasing the efficacy of treatment. All the results showed that the unique properties of the Znx Mn1-X Fe2O4-based nano-platform make it a promising candidate for further development in the field of photodynamic therapy.
Acknowledgements
We are thankful to the Research Council of the University of Guilan for the partial support of this work. The collaboration of the Iran National Science Foundation (Grant No. 99023183) is also acknowledged.
Author Contributions
Mostafa Zamani-Roudbaraki and Mozhgan Aghajanzadeh-Kiyaseh wrote the main manuscript text.
Setareh Jashnani, Hossein Khoramabadi and S. Shirin Shahangian prepared the data and validated them. Farhad Shirini Superved the research program.
Funding
The authors have not disclosed any funding.
Data Availability
No datasets were generated or analysed during the current study.
Declarations
Competing Interests
The authors declare no competing interests.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
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