MnFe2O4@L-Cysteine as a drug delivery system, in vitro cytotoxicity evaluation on human Breast cancer cell (MCF7) and DFT calculation

MnFe2O4@L-Cysteine as a drug delivery system, in vitro cytotoxicity evaluation on human Breast cancer cell (MCF7) and DFT calculation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article MnFe2O4@L-Cysteine as a drug delivery system, in vitro cytotoxicity evaluation on human Breast cancer cell (MCF7) and DFT calculation Neda hasankhani, Sharieh Hosseini, Elham Askarizadeh, Bita mehravi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4396900/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Thanks to their high hydrophilic and magnetic properties, MnFe 2 O 4 nanoparticles (NP) are recognized as favorable drug carriers. In this study, L-Cysteine-modified MnFe 2 O 4 nanoparticles (MnFe 2 O 4 @L-Cysteine) were prepared and characterized. Their cytotoxicity against human breast cancer cell lines (MCF7) was also evaluated by MTT assay. To simulate drug delivery systems, the interaction between modified NP and 5-fluorouracil (5-FU) was examined as a breast cancer drug. The MTT results showed the applicability of MnFe 2 O 4 @L-Cysteine nanoparticles as a potential cytotoxic agent in breast cancer treatment. Based on the theoretical calculations, the adsorption energy between L-Cysteine and 5-FU was − 12.029 KJ/mol and their interaction was spontaneous and exothermic at the temperature range of 278.15 to 288.15 K. Also, the drug release thermodynamically is feasible at body temperature. The calculated electronic descriptors indicated that the electrons were transferred from L-Cysteine to 5- FU. Overall, MNFe2O4@L-Cysteine, in addition to being non-toxic has the potential to deliver 5-FU anticancer drug. Modified magnetic nanoparticles L-Cysteine MCF7 DFT Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 1-Introduction Nanocarriers, such as nanoparticles, polymers, dendrimers, micelles, and liposomes are widely used for drug delivery in biological systems for targeted drug delivery, controlled release, and impressive loading of drugs[ 1 – 3 ]. Inorganic nanoparticles have attracted the attention of researchers due to their promising properties[ 4 , 5 ]. The most serious property of inorganic nanoparticles is their high surface-to-volume ratio[ 5 – 7 ], Surface modification improves the targeted drug delivery and release [ 8 ]. Among the inorganic nanoparticles, spinel structures, ferrites, and special MnFe 2 O 4 nanoparticles have found extensive applications in magnetic resonance imaging [ 9 – 16 ], magnetic hyperthermia[ 17 – 20 ], lithium-ion batteries[ 21 – 24 ], supercapacitor[ 25 – 27 ] ,sensors[ 28 – 31 ], catalyst[ 32 – 35 ], hydrogen production[ 36 – 38 ], heavy metal removal[ 39 – 41 ] and drug delivery systems[ 24 , 42 – 45 ]. Regarding the higher compatibility of MnFe 2 O 4 compared to hematite, cobalt ferrite, magnetite, and nickel ferrite, it is could be a favorable candidate for different biomedical applications especially magnetic resonance imaging (MRI) and drug delivery [ 46 , 47 ]. The MnFe 2 O 4 nanoparticles are favorable carrier drug due to their high hydrophilic and magnetic properties. Furthermore, they have shown high capacity for loading and controlled release of drugs. MnFe 2 O 4 nanoparticles have high deposition tendency due to their high surface energy and magnetic properties, necessitating the modification of MnFe2O4 NPs [ 48 – 51 ]. One of these modifiers is L-Cysteine, which includes SH, NH2, and COOH functional groups. L-Cysteine is a nontoxic α-amino acid that can be synthesized in the body. It has found several applications in biomedicine, catalysis, water treatment, and cancer treatment[ 52 – 56 ]. Furthermore, hydrophilic groups of L-Cysteine such as NH 2 and COOH are easily soluble in water. Thus, L-Cysteine is a proper candidate for surface modification of magnetic nanoparticles to increase their adsorption efficiency [ 57 – 60 ]. Other components (e.g. carboxylic acids, amines [ 61 ], and enediol [ 62 ], are also used to modify metallic nanoparticles whose complex preparation have limited their biomedical applications [ 63 ]. Naderi and coworkers loaded curcumin on MnFe 2 O 4 Np/carboxymethyl chitosan hydrogel. Their studies showed that 67.2% of curcumin was loaded by hydrogen bonding interaction between NH 2 groups of the chitosan and hydroxyl groups of the drug [ 42 ]. Wang et al.[ 43 ] investigated loading of doxorubicin by MnFe 2 O 4 /graphene oxide nanocomposite at various pH levels. Based on their reports, the highest release rate was related to acidic pH followed by neutral and alkaline pH, respectively. The interaction of hydroxyl and carboxylic groups of graphene oxide and doxorubicin functional groups (NH 2 and OH) was stronger in neutral and alkaline media than in acidic media. Fahmi et al[ 24 ] investigated the release rate of naproxen from MnFe 2 O 4 Np/cellulose acetate nanofibers. They reported the higher release rate of the drug in neutral pH due to low electrostatic interactions. In this study, MnFe 2 O 4 nanoparticles were synthesized and modified with L-Cysteine, and their cytotoxicity was examined against human breast cancer cell lines (MCF7). To simulate drug delivery systems, the interaction between modified NP and a breast cancer drug (5-fluorouracil (5-FU)) was investigated. 2-Materials and Characterizations Analytical grade MnCl 2 . 4 H 2 O, FeCl 3 . 6H 2 O, L-Cysteine, and NaOH were purchased from Merk and applied without further purification. Fourier transforms spectra were recorded using a Perkin Elmer FT-IR spectrometer. X-ray diffraction (XRD) was applied to investigate the phase structures of the nanoparticles with Cu Kα radiation. The morphology of nanoparticles was characterized by SEM, EDX mapping, and EDAX using a Zeiss scanning electron microscope equipped with an energy-dispersive X-ray spectrometer. The vibration sample magnetometer (VSM) was measured using a Vibrating-sample magnetometer (Magnates 120 Daghigh Kavir Company, Kashan, Iran). 2-1-Preparation of MnFeO@L-Cysteine MNPs The MnFe 2 O 4 NPs were synthesized by the solvothermal method reported previously [ 64 ]. The surface modification with L-Cysteine was carried out as follows: MnFe 2 O 4 NPs (0.1 g) were dispersed in 25 mL of deionized water. L-Cysteine (0.05 g) was dissolved in 10 mL deionized water and added dropwise to the suspension of MnFe 2 O 4 under vigorous stirring at room temperature. After 2 h the modified NPs were collected with the external magnet, washed several times with deionized water and ethanol, and dried under vacuum at 60 ᵒC for 24h. 2-2-Cytotoxicity studied by MTT test The Human breast carcinoma cells were prepared by the National Cell Bank of Iran at the Pasteur Institute. The Human breast carcinoma cells (MCF7) were cultured in PRMI-1640 medium, containing 10% fetal bovine serum (FBS), 5% horse serum, 2mM L-glutamine, 1% penicillin/streptomycin and 2 gr/L sodium bicarbonate and preserved at 37° C and pH = 7.2 in a humidified atmosphere containing 5% CO 2 . At this stage, 10 4 cells were cultured in 1L culture media. Then, cell viability and proliferation were measured using the MTT microculture colorimetric test. The stock solution was prepared by dissolving 5 mg of tetrazolium in 1 ml of phosphate-buffered saline. Cells were then exposed to nanoparticles with different concentrations (1, 5, 10, 25, 50, 100, and 250 µg/ml) for 24 hours. In the next step, 10 microliters of MTT stock were added to the cells. After keeping the stock for 4 hours in incubation, 10 microliters of DMSO were added and absorbance was checked at 570 nm using an Eliezer reader. The cell viability was calculated according to the following equation: %viability= \(\frac{ \text{m}\text{e}\text{a}\text{n} \text{e}\text{x}\text{p}\text{e}\text{r}\text{i}\text{m}\text{e}\text{n}\text{t}\text{a}\text{l} \text{a}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e}}{\text{m}\text{e}\text{a}\text{n} \text{n}\text{e}\text{g}\text{a}\text{t}\text{i}\text{v}\text{e} \text{c}\text{o}\text{n}\text{t}\text{r}\text{o}\text{l} \text{a}\text{b}\text{s}\text{o}\text{r}\text{b}\text{a}\text{n}\text{c}\text{e}}\) ×100 (1) The cell viability was investigated by reading the optimal density at 570 nm. The viability (mean ± SD) was plotted in the form of histograms, using the Microsoft Excel program. 2-3-Computational details : All geometry optimization and energy calculation were carried out by the Gaussian 09 software [ 65 ]. Quantum mechanical calculations were performed by the Density Functional Theory (DFT) method at the Lee-Yong-Parr Exchange-correlation hybrid functional (B3LYP) level of theory and 6-31G(d) basis set. Vibrational analysis was carried out after structural optimization at B3LYP/6-31G*. The absence of imaginary vibration frequencies indicated the stability of structures. Furthermore, the highest occupied energy (HOMO), lowest unoccupied energy (LUMO), and energy gap (Eg) were determined for the considered structures. Eg value can be used to check the sensitivity of L-Cysteine toward 5-fU. The reactivity parameters including electronegativity (A), chemicalpotential(µ), chemical hardness(η), electrophilicity(ω) and softness (S)were calculated following equations[ 66 ] : A= -HOMO (2) $$\mu =\frac{ (\text{E} \text{H}\text{O}\text{M}\text{O} +\text{E} \text{L}\text{U}\text{M}\text{O})}{2}$$ 3 η = \(\frac{\text{E} \text{L}\text{U}\text{M}\text{O}- \text{E} \text{H}\text{O}\text{M}\text{O}}{2}\) ( 4) ω= \(\frac{{\mu } 2}{2{\eta } }\) (5) S= \(\frac{1}{2{\eta }}\) (6) The binding energies (E ads ) of 5-FU molecule and L-Cysteine were determined as the difference between the total energy of complexes and the sum of the energies of the isolated L-Cysteine and 5-FU as follows: E ads = E 5FU/Cys – (E Cys + E 5−FU ) (7) The negative adsorption energy demonstrated a thermodynamically favorable adsorption process. The zero-point energy (ZPE) was also considered in the calculation. Negative adsorption energy shows the stability of the 5-FU/L-Cysteine complex. The adsorption Gibbs free energies and adsorption enthalpy were computed by the following equations (at T = 298/15 K and P = 1 atm): Where G complex, G drug, G L−Cys, H complex, H drug, and H L−Cys are the Gibbs free energy and enthalpy of the species specified in the equation. ∆G ads = G complex – (G drug + G L−Cys ) (8) ∆H ads = H complex – (H drug + H L−Cys ) (9) Where G complex, G drug, G L−Cys, H complex, H drug, and H L−Cys are the Gibbs free energy and enthalpy of the species specified in the equation. All of the calculation was performed in gas and water phases. 3-Results and Discussion The bare MnFe 2 O 4 and MnFe 2 O 4 @L-Cysteine were examined by FT-IR method. For bare MnFe 2 O 4 NP (Fig. 1 ), the bands at ~ 580–650 cm − 1 correspond to the Mn-O and Fe-O bonds. The peak at 3400 cm − 1 can be assigned to the hydroxyl group on the surface of MnFe 2 O 4 NP. After the functionalization of MnFe 2 O 4 NP with L-Cysteine (MnFe 2 O 4 @L-cyc), some changes emerged in the FT-IR spectrum of MnFe 2 O 4 NP (Fig. 1 ). The strong peaks at ~ 580–650 cm − 1 can be attributed to Mn-O and Fe-O in the structure of MnFe 2 O 4 NP. The new bands at 3440 cm − 1 and the peak at 1510 cm − 1 are assigned to the N-H group. The peaks at 2857 and 2935 cm − 1 are related to the CH 2 group. The peaks at 1411 and 1633 cm − 1 can be ascribed to the carbonyl group. The peaks at 1308 and 1071 cm − 1 can be assigned to C-O and C-N bonds, respectively. All of these new bands (in comparison with bare MnFe 2 O 4 ) are related to L-Cysteine, implying the bonding of L-Cysteine to the surface of MnFe 2 O 4 NP. Because of the low concentration of the thiol group, the S-H band was not observed in the spectrum of MnFe 2 O 4 @L-Cysteine [ 67 ]. The XRD patterns of the bare MnFe 2 O 4 NP and MnFe 2 O 4 @L-Cysteine are shown in Fig. 2 . The diffraction peaks of both compounds are the same and show the phase composition of MnFe 2 O 4 NP. The diffraction peaks correspond to the crystal planes (111), (220), (311), (222), (400), (422) (511), and (440) with 2θ values of 18.08°, 29.74°, 35.02°, 36.66°, 42.57°, 52.82°, 56.26°, and 61.74°, respectively which is correspond to the spinel structure of MnFe 2 O 4 according to the JCPDS standard card No. 88-1965. These data indicate that the functionalization of bare MnFe 2 O 4 with L-Cysteine did not alter the phase composition of MnFe 2 O 4 NP. The vibration sample magnetometer (VSM) was utilized for the investigation of the magnetic properties of MnFe 2 O 4 NP and MnFe 2 O 4 @L-Cysteine(Fig. 3 ) in a magnetic field range of -15000 to 15000 Oe. Both samples exhibited narrow hysteresis loops with superparamagnetic behavior. The saturation magnetization (Ms) values of bare MnFe 2 O 4 and MnFe 2 O 4 @L-Cysteine are 29.64 emu/g and 18.36 emu/g, respectively, indicating that the coating of MnFe 2 O 4 with non-magnetic L-Cysteine reduced the magnetic properties of bare MnFe 2 O 4 NPs. MnFe 2 O 4 @L-Cysteine NPs still showed acceptable magnetic properties and could be separated by an external magnet. SEM images of MnFe 2 O 4 @L-Cysteine can be seen in Fig. 4 which shows non-uniform semi spherical particles with some aggregations. The diameter of nanoparticles ranged in 70–110 nm. The Energy Dispersive X-ray spectrum and EDX mapping are shown in Figs. 5 and 6 , respectively. These data confirm the presence of manganese, iron, oxygen, and sulfur groups, confirming the coverage of MnFe 2 O 4 with L-Cysteine. 3-1-Cytotoxicity study by MTT assay: Nowadays amino acid-based nanocarriers are applied as a multipurpose tool for disease treatment due to their low immunogenicity, good biocompatibility, tunable structure, and ease of chemical modification. As an amino acid, cysteine has shown interesting applications in chemical modification, drug delivery, bioimaging, and in vivo long blood circulation. Thanks to its thiol (SH) groups, it has found new properties such as tumor proliferation[ 68 ] Mohammad Zaki Fahmi and coworkers synthesized MnFe 2 O 4 composite nanofibers including cellulose and collagen. The MTT test against Hella cells indicated the low toxicity of this nanofiber [ 24 ]. In another study, Kanagesan et al. prepared MnFe 2 O 4 NP and evaluated their biocompatibility with murine breast cancer cells (4T1). Their results showed the dose-dependent cytotoxic effect of MnFe 2 O 4 NP against 4T1[ 69 ]. In this study, cytotoxicity of MnFe 2 O 4 NP, L-Cysteine, and MnFe 2 O 4 @L-Cysteine was in vitro examined against MCF7 cells through the MTT test. In this method, the viability percentage of MCF7 cells was calculated by measuring the reduction of tetrazolium salt to purple formazan in living cells. The breast cancer cells were treated with different concentrations (1, 5, 10, 25, 50, 100 and 250 µg/L) of nanoparticles, modifier, and modified nanoparticles within the incubation period of 24 h. Based on the results, the cell viability percentage decreased with increasing the concentration of nanoparticles. Although there is no regular trend in the case of cysteine alone, a decreasing trend was observed for viability percentage at higher concentrations. This indicates the significant dose-dependence of antiproliferative activity of MnFe 2 O 4 and L-Cysteine. However, the cell viability of MnFe 2 O 4 @ L-Cysteine was not dose-dependent (Fig. 7 ). Moreover, MnFe 2 O 4 , L-Cysteine, and MnFe 2 O 4 @L-Cysteine have low toxicity with the cell viability above 95%. MTT test results are in agreement with the earlier findings on the good biocompatibility of MnFe 2 O 4 NP and L-Cysteine [ 68 , 70 , 71 ], which makes them appropriate for biomedical purposes. 3-2Theoretical study: In this section investigates the loading of 5-FUas an anti-breast cancer drug by MnFe 2 O 4 @L-Cysteine nanocarrier with Quantum mechanical calculations, considering the interaction between 5FU and L-cysteine. First, the structures of 5-FU and L-Cysteine were optimized at the DFT/B3LYP/6-31G(d) level. Vibrational analysis was carried out after the optimization of structures at mentioned theoretical level. The absence of imaginary vibration frequencies illustrated the stability of structures. The optimized structures are shown in Fig. 8 . The theoretical bond lengths and angles of the optimized structures well agree with previous research. Then, 5-FU was placed on various positions of L-Cysteine (including NH 2 , COOH, and S) through its nitrogen, fluorine, and carbon atoms both vertically and horizontally in both media (gas and aqueous). The results showed no favorable interaction between L-Cysteine and 5-FU in the gas phase. The most stable complex was obtained after optimization in the aqueous phase (Fig. 9 ). The negative adsorption energy demonstrated a thermodynamically favorable adsorption process. Moreover, the 5-FU molecule were adsorbed through the nitrogen atom on the sulfur atom of L-Cysteine with the adsorption energy of -12.029 KJ.mol − 1 . The value of adsorption energy illustrated physical adsorption. Changes of enthalpy (∆H), free Gibbs energy (∆G), and entropy were calculated for interaction between L-Cysteine and 5-FU. The results indicated the negative variation of adsorption enthalpy, implying exothermic interaction of 5FU and L-Cysteine at ambient temperature. The Gibbs free energy change of binding was obtained positive. As a result, this interaction does not happen spontaneously at room temperature. It seems that the interaction temperature must be lowered to bind the 5-FU to the L-Cysteine as it reduces the vibrations of the bonds in the complex [ 72 ]. Temperature is an important factor in adsorption and release reactions. Therefore, the interaction between L-Cysteine and 5FU was examined at different temperatures (T = 278.15-305.15K) in aqueous media. The results in Table 1 show that the interaction between 5-FU and L-Cysteine was spontaneous at temperature range of 278.15 to 288.15K. With increasing the temperature, the values of adsorption-free Gibbs energy, enthalpy, and entropy got more positive but the equilibrium constant decreased especially at 308.15 K (body temperature). Therefore, drug release seems to be thermodynamically feasible at physiological conditions. The diagram of changes of ∆G, ∆H, ∆S, and K versus temperature is shown in Fig. 10 . Table 1 The calculated binding energy (E b Kcal/mol), ∆G(Kcal/mol), ∆H(Kcal/mol), ∆S(cal/molK), and K in 278.15-308.15K T ∆H ads ∆Gads ∆Sads Kth 278.15 -3.49 -3.205 -264.988 3.37*10 2 283.15 -3.336 -2.025 -263.589 3.708*10 288.15 -2.961 -0.842 -262.183 4.376 293.15 -2.586 0.344 -260.770 5.530*10 − 1 298.15 -2.211 1.532 -259.351 7.456*10 − 2 303.15 -1.836 2.730 -257.948 1.058*10 − 2 308.15 -1.461 3.939 -256.516 1.57*10 − 3 As an intermolecular interaction or a chemical reaction occurs between the frontier orbitals of the involved species, the frontier molecular including the highest occupied molecular orbital (HOMO), lowest unoccupied molecular orbital (LUMO) and H-L gap (Eg) were calculated as listed in Table 2 . The HOMO/LUMO contour is also depicted in Fig. 11 . The HOMO energy of 5-FU is smaller (less negative) than L-Cysteine, implying that this structure has higher tendency to react with electrophilic species. The high HOMO energy in this system indicates that the electrons in this orbital can be more easily given to an electrophile species. L-Cysteine is more electrophile compared with 5FU. Figure 12 shows the HOMO and the map of molecular electrostatic potential (MEP) of the L-Cysteine. As depicted in Table 2 , Eg(L-H) of the L-Cysteine /5FU (5.410 eV) is lower than L-Cysteine(6.250), suggesting a notable rise in the reactivity and conductivity of the complex. The ω index approved the tendency of a molecule to absorb electrons and is closely related to the electron affinity of the molecule. It also gives useful information about the electron transfer in the electrostatic interaction. Since the formation of the complex involves non-covalent interactions and the electron transfer from the nucleophilic to the electrophilic species, according to the values of ω in the Table 2 , the electron is transferred from L-Cysteine to 5 FU. Also, the softness is a degree of interactivity of molecule. As seen in Table 2 , L-cysteine is more reactive to 5Fu. Table 2 The calculated Energy gap (Eg eV), HOMO(eV), LUMO(eV), Chemical potential (µ eV), Chemical hardness(η eV), Electrophilicity index(ω eV), Softness (S ev − 1 )and Electronegativity(A eV) of the drug, L-cysteine and their complex Compound E HOMO E LUMO ∆Eg η µ ω S A 5Fu -6.390 -0.96 5.430 2.715 -3.675 2.487 0.184 0.96 L-cysteine -6.600 -0.350 6.250 3.125 -3.475 1.932 0.16 0.35 L-cysteine /5F -6.420 -1.010 5.410 2.705 -3.715 2.551 0.184 1.01 The natural bond orbital (NBO) analysis was carried out by the B3LYP functional and 6-31G (d) basis set. The NBO was used to analyze the interaction between the donor and the acceptor orbitals. The Lewis and non-Lewis orbitals illustrate the bonding and antibonding orbital NBOs, respectively. The energy of charge transfer between donor and acceptor orbitals was determined using second order perturbation theory. Eq. 10 estimates the stabilization energy (E2). E 2 = q i \(\frac{\left(Fij\right)2}{\mathcal{E}j-\mathcal{E}i}\) (10) Where qi shows the donor orbital, Ei denotes the diagonal element, Ej is orbital energy, and F(ij) represents diagonal NBO Fock matrix element. The stabilization energy (E2) determined the interaction between the donor and acceptor orbitals, the larger the stabilization energy the stronger the interaction. Stabilization energy of the interacting donor and acceptor orbitalis are depicted in Table 3 . We considered the most probable bonding to antibonding interactions, which could be due to charge transfer between l-cysteine and 5FU. The major interactions arise from the charge transfer between lone pair S and ơ* antibonding orbital (N20-H25) with energy of 8.86Kcal/mol. This result agrees with the FMO evaluation. Table 3 The second −order perturbation theory analysis of the most interacting of the natural bond orbitals Charge transfer E2(Kcal/mol) Ej-Ei(a.u) F(i,j) a.u Lp S͢͢ → ơ* N20-H25 8.86 0.71 0.071 Lp O23→ơ* N4-H12 1.91 1.18 0.043 Lp O23→ơ*C5-C7 0.15 0.63 0.009 Conclusions The MnFe 2 O 4 @L-Cysteine drug delivery system was synthesized and characterized in this research. Then, the cytotoxicity of MnFe 2 O 4 , L-Cysteine and MnFe 2 O 4 @L-Cysteine was determined on the MCF7 cells line by MTT assay. The MTT results illustrated that the MnFe 2 O 4 @L-Cysteine does not have cytotoxicity against the MCF7 cells line while increasing the MnFe 2 O 4 @L-Cysteine concentration does not have a significant effect on the cytotoxicity of this carrier. Also, theoretical studies evaluated the power of L-cysteine as a modifying agent of MnFe 2 O 4 and drug delivery system for 5-FU drugs. DFT calculation indicated that the 5-FU anticancer drug was physically adsorbed on L-Cysteine in aqueous media. Thermodynamic analysis revealed that this interaction is exothermic. The drug binding is not favorable at ambient temperature, whereas the drug release was feasible at body temperature. Also, the electronic calculations showed a decline in the gap energy (Eg) of the complex by -13.4%. The obtained results from the hybridization of experimental and theoretical studies show that the MnFe 2 O 4 @L-Cysteine nanocarrier can be a promising candidate for drug delivery systems and these results might be effective in nanomedicine scope. Declarations Ethic approval and consent to participate We had no participant in our study and the study was in vitro which had no need ethic approval Consent for publication Not applicable Availability of data and material All raw data and materials on which this study is based are included in the manuscript. Competing interests The authors declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this paper Funding We have no funding Authors contributions Neda. Hasankhani : experimental and laboratory test Sharieh. Hosseini : wrote the main manuscript text and supervision Elham askarizadeh .: wrote the manuscript text Bita.M ehravi : laboratory test All authors reviewed the manuscript References E. Abbasi, M. Milani, S. Fekri Aval, M. Kouhi, A. Akbarzadeh, H. Tayefi Nasrabadi, P. Nikasa, S.W. Joo, Y. Hanifehpour, K. 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Massabni, Chemical and spectroscopic studies of a new palladium (II) complex with N-acetyl-L-cysteine, Journal of Coordination Chemistry 61(22) (2008) 3666-3673. Y. Meng, S. Han, Z. Gu, J. Wu, Cysteine‐Based Biomaterials as Drug Nanocarriers, Advanced Therapeutics 3(5) (2020) 1900142. S. Kanagesan, S.B.A. Aziz, M. Hashim, I. Ismail, S. Tamilselvan, N.B.B.M. Alitheen, M.K. Swamy, B. Purna Chandra Rao, Synthesis, characterization and in vitro evaluation of manganese ferrite (MnFe2O4) nanoparticles for their biocompatibility with murine breast cancer cells (4T1), Molecules 21(3) (2016) 312. M. Aghajanzadeh, E. Naderi, M. Zamani, A. Sharafi, M. Naseri, H. Danafar, In vivo and in vitro biocompatibility study of MnFe2O4 and Cr2Fe6O12 as photosensitizer for photodynamic therapy and drug delivery of anti-cancer drugs, Drug Development and Industrial Pharmacy 46(5) (2020) 846-851. R. Kavkhani, A. Hajalilou, E. Abouzari-Lotf, L.P. Ferreira, M.M. Cruz, M. Yusefi, E. Parvini, A.B. Ogholbeyg, U.N. Ismail, CTAB assisted synthesis of MnFe2O4@ SiO2 nanoparticles for magnetic hyperthermia and MRI application, Materials Today Communications 31 (2022) 103412. F. Ghazali, S. Hosseini, S. Ketabi, DFT and Molecular Simulation Study of Gold Clusters as Effective Drug Delivery Systems for 5-Fluorouracil Anticancer Drug, Journal of Cluster Science (2022) 1-11. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4396900","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":303889739,"identity":"238d228e-d12f-4b7f-aad4-9cba93d63011","order_by":0,"name":"Neda hasankhani","email":"","orcid":"","institution":"Tehran medical sciences university, Islamic Azad University","correspondingAuthor":false,"prefix":"","firstName":"Neda","middleName":"","lastName":"hasankhani","suffix":""},{"id":303889740,"identity":"76f563b9-14ae-4ae6-ad49-c0c42d8a34f1","order_by":1,"name":"Sharieh Hosseini","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAklEQVRIiWNgGAWjYDACCSBOYDiQAOYkMEjIMTDwkKjFmDgtDDAtQJDYQEiL/OzmZx8eVNzJ4+c//vDBwx0W6RuOnz344AODnZxuA3YtBneOGc9IOPOsWHJGjrFB4hmJ3A1n8pINZzAkG5sdwKFFIsGYIbHtcOKGGzxsEoltQC0HcsykeRgOJG7DoUV+RvpnhsR/QC3njz//AdSSbnD+DX4tDDdygLY0ALUcSDADWieRYHCDgC1ABcUMCccOJ84E+gXkMMOZN94YG84wwO0XoMM2M/6oOZzYDwyxjz/b6uT5zucYPvhQYSeHSwsmUACrNCBWOdjeBlJUj4JRMApGwUgAAI/eZg8QpBEzAAAAAElFTkSuQmCC","orcid":"","institution":"Tehran medical sciences university, Islamic Azad University","correspondingAuthor":true,"prefix":"","firstName":"Sharieh","middleName":"","lastName":"Hosseini","suffix":""},{"id":303889741,"identity":"fa2737c9-0707-49c1-9e0e-1e7469748429","order_by":2,"name":"Elham Askarizadeh","email":"","orcid":"","institution":"Tehran medical sciences university, Islamic Azad University","correspondingAuthor":false,"prefix":"","firstName":"Elham","middleName":"","lastName":"Askarizadeh","suffix":""},{"id":303889742,"identity":"9a97721b-8fb6-4e3c-8ca1-5b3b17f3f147","order_by":3,"name":"Bita mehravi","email":"","orcid":"","institution":"Iran University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Bita","middleName":"","lastName":"mehravi","suffix":""}],"badges":[],"createdAt":"2024-05-09 19:23:46","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4396900/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4396900/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":56887719,"identity":"10c8c3dd-9740-4731-a1b7-28aff1793860","added_by":"auto","created_at":"2024-05-21 18:57:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":97278,"visible":true,"origin":"","legend":"\u003cp\u003eIR spectra of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-cysteine NPs\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/1794468339c78a21a2f51d89.png"},{"id":56887715,"identity":"c3a3f0e7-836d-43fd-909e-b4dc71bed6d8","added_by":"auto","created_at":"2024-05-21 18:57:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":21214,"visible":true,"origin":"","legend":"\u003cp\u003eThe XRD patterns of the bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/ffe1bc1ba0310cce91794c08.png"},{"id":56887714,"identity":"dc357fe3-ccdb-4c03-ab04-b4c23a1a737c","added_by":"auto","created_at":"2024-05-21 18:57:55","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":115151,"visible":true,"origin":"","legend":"\u003cp\u003eHysteresis curves of: a) bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and b) MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/6d557cf4f81715e486e2f350.png"},{"id":56887722,"identity":"7463afaa-71f1-4fe6-8cd9-95261d001957","added_by":"auto","created_at":"2024-05-21 18:57:59","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":700452,"visible":true,"origin":"","legend":"\u003cp\u003eThe SEM image of MnFe2O4@L-Cysteine\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/92feecdf08e96f6a9e6df676.png"},{"id":56887686,"identity":"43b85734-b3f7-4734-b10b-7785ef762df4","added_by":"auto","created_at":"2024-05-21 18:57:51","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":59001,"visible":true,"origin":"","legend":"\u003cp\u003eEDAX analysis of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/de591cf9a29b699b19209aec.png"},{"id":56887730,"identity":"9cdcb322-fbf2-4f04-8731-e3f3eaae6e7f","added_by":"auto","created_at":"2024-05-21 18:58:01","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":438905,"visible":true,"origin":"","legend":"\u003cp\u003eSEM-EDX mapping of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/f01cf6898c5f92a0d81d82e4.png"},{"id":56887716,"identity":"82bb15e6-153a-407d-8739-d09f4a1d0c63","added_by":"auto","created_at":"2024-05-21 18:57:56","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":95981,"visible":true,"origin":"","legend":"\u003cp\u003eThe viability percentage of the MCF7 cell line versus the log of various concentrations of a) MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e b) L-Cysteine and c) MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine with a 24 h exposure. Each point represents a mean of three experiments. Cell survival in all of experimental concentration (1, 5, 10, 25, 50, 100 and 250 μg/L)\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/e5bc94347c07c12ae89bdf80.png"},{"id":56887717,"identity":"14e6a48e-0ff4-48ef-af3b-0fd98536ed5d","added_by":"auto","created_at":"2024-05-21 18:57:56","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":38333,"visible":true,"origin":"","legend":"\u003cp\u003eThe optimized structure of a)5-FU and b)L-Cysteine(N=blue, O=red, S=yellow, F=cyan)\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/49b8195a540f86ec14eedb33.png"},{"id":56887713,"identity":"f2dd2298-aa5f-44c8-b279-86c4ba6d2298","added_by":"auto","created_at":"2024-05-21 18:57:54","extension":"jpg","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":30496,"visible":true,"origin":"","legend":"\u003cp\u003eThe more stable complex in the water phase\u003c/p\u003e","description":"","filename":"9.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/80f2a296ab33c78c8bf81060.jpg"},{"id":56887721,"identity":"fe7e8978-a49a-4ec1-bcde-1001b4436725","added_by":"auto","created_at":"2024-05-21 18:57:58","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":41639,"visible":true,"origin":"","legend":"\u003cp\u003eThe changes of a)∆G ads b)∆H c)∆S and d) K versus temperature(208.15-308.15K)\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/18cc77a90a728a45873d1362.png"},{"id":56887720,"identity":"b0b79a14-1e29-421c-93e7-6c4bacb3f88b","added_by":"auto","created_at":"2024-05-21 18:57:58","extension":"png","order_by":11,"title":"Figure 11","display":"","copyAsset":false,"role":"figure","size":343036,"visible":true,"origin":"","legend":"\u003cp\u003eHOMO/LUMO contoure of a)5FU b)L-Cysteine c05FU/L-Cysteine complex\u003c/p\u003e","description":"","filename":"11.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/541a2764ae75eab467730ad7.png"},{"id":56887724,"identity":"5ed5b71f-868c-41fc-997c-b48836f4c0ac","added_by":"auto","created_at":"2024-05-21 18:57:59","extension":"png","order_by":12,"title":"Figure 12","display":"","copyAsset":false,"role":"figure","size":213309,"visible":true,"origin":"","legend":"\u003cp\u003eThe HOMO and The map of molecular electrostatic potential (MEP) of the L-Cysteine\u003c/p\u003e","description":"","filename":"12.png","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/98455af37989a526ddd49568.png"},{"id":57245954,"identity":"05cf29ba-0efc-4625-8c14-d75cb3f3d097","added_by":"auto","created_at":"2024-05-28 05:45:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2980319,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4396900/v1/1398aec4-d9fd-4ffe-b5db-e366a17d092c.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"MnFe2O4@L-Cysteine as a drug delivery system, in vitro cytotoxicity evaluation on human Breast cancer cell (MCF7) and DFT calculation","fulltext":[{"header":"1-Introduction","content":"\u003cp\u003eNanocarriers, such as nanoparticles, polymers, dendrimers, micelles, and liposomes are widely used for drug delivery in biological systems for targeted drug delivery, controlled release, and impressive loading of drugs[\u003cspan additionalcitationids=\"CR2\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Inorganic nanoparticles have attracted the attention of researchers due to their promising properties[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The most serious property of inorganic nanoparticles is their high surface-to-volume ratio[\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], Surface modification improves the targeted drug delivery and release [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Among the inorganic nanoparticles, spinel structures, ferrites, and special MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles have found extensive applications in magnetic resonance imaging [\u003cspan additionalcitationids=\"CR10 CR11 CR12 CR13 CR14 CR15\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], magnetic hyperthermia[\u003cspan additionalcitationids=\"CR18 CR19\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], lithium-ion batteries[\u003cspan additionalcitationids=\"CR22 CR23\" citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], supercapacitor[\u003cspan additionalcitationids=\"CR26\" citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] ,sensors[\u003cspan additionalcitationids=\"CR29 CR30\" citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], catalyst[\u003cspan additionalcitationids=\"CR33 CR34\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e], hydrogen production[\u003cspan additionalcitationids=\"CR37\" citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e], heavy metal removal[\u003cspan additionalcitationids=\"CR40\" citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] and drug delivery systems[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan additionalcitationids=\"CR43 CR44\" citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Regarding the higher compatibility of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e compared to hematite, cobalt ferrite, magnetite, and nickel ferrite, it is could be a favorable candidate for different biomedical applications especially magnetic resonance imaging (MRI) and drug delivery [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. The MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles are favorable carrier drug due to their high hydrophilic and magnetic properties. Furthermore, they have shown high capacity for loading and controlled release of drugs. MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles have high deposition tendency due to their high surface energy and magnetic properties, necessitating the modification of MnFe2O4 NPs [\u003cspan additionalcitationids=\"CR49 CR50\" citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. One of these modifiers is L-Cysteine, which includes SH, NH2, and COOH functional groups. L-Cysteine is a nontoxic α-amino acid that can be synthesized in the body. It has found several applications in biomedicine, catalysis, water treatment, and cancer treatment[\u003cspan additionalcitationids=\"CR53 CR54 CR55\" citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. Furthermore, hydrophilic groups of L-Cysteine such as NH\u003csub\u003e2\u003c/sub\u003e and COOH are easily soluble in water. Thus, L-Cysteine is a proper candidate for surface modification of magnetic nanoparticles to increase their adsorption efficiency [\u003cspan additionalcitationids=\"CR58 CR59\" citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOther components (e.g. carboxylic acids, amines [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e], and enediol [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e], are also used to modify metallic nanoparticles whose complex preparation have limited their biomedical applications [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eNaderi and coworkers loaded curcumin on MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eNp/carboxymethyl chitosan hydrogel. Their studies showed that 67.2% of curcumin was loaded by hydrogen bonding interaction between NH\u003csub\u003e2\u003c/sub\u003e groups of the chitosan and hydroxyl groups of the drug [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Wang et al.[\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e] investigated loading of doxorubicin by MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e/graphene oxide nanocomposite at various pH levels. Based on their reports, the highest release rate was related to acidic pH followed by neutral and alkaline pH, respectively. The interaction of hydroxyl and carboxylic groups of graphene oxide and doxorubicin functional groups (NH\u003csub\u003e2\u003c/sub\u003e and OH) was stronger in neutral and alkaline media than in acidic media. Fahmi et al[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] investigated the release rate of naproxen from MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003eNp/cellulose acetate nanofibers. They reported the higher release rate of the drug in neutral pH due to low electrostatic interactions.\u003c/p\u003e \u003cp\u003eIn this study, MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles were synthesized and modified with L-Cysteine, and their cytotoxicity was examined against human breast cancer cell lines (MCF7). To simulate drug delivery systems, the interaction between modified NP and a breast cancer drug (5-fluorouracil (5-FU)) was investigated.\u003c/p\u003e"},{"header":"2-Materials and Characterizations","content":"\u003cp\u003eAnalytical grade MnCl\u003csub\u003e2\u003c/sub\u003e. 4 H\u003csub\u003e2\u003c/sub\u003eO, FeCl\u003csub\u003e3\u003c/sub\u003e. 6H\u003csub\u003e2\u003c/sub\u003eO, L-Cysteine, and NaOH were purchased from Merk and applied without further purification.\u003c/p\u003e\n\u003cp\u003eFourier transforms spectra were recorded using a Perkin Elmer FT-IR spectrometer. X-ray diffraction (XRD) was applied to investigate the phase structures of the nanoparticles with Cu K\u0026alpha; radiation. The morphology of nanoparticles was characterized by SEM, EDX mapping, and EDAX using a Zeiss scanning electron microscope equipped with an energy-dispersive X-ray spectrometer. The vibration sample magnetometer (VSM) was measured using a Vibrating-sample magnetometer (Magnates 120 Daghigh Kavir Company, Kashan, Iran).\u003c/p\u003e\n\u003ch3\u003e2-1-Preparation of MnFeO@L-Cysteine MNPs\u003c/h3\u003e\n\u003cp\u003eThe MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs were synthesized by the solvothermal method reported previously [\u003cspan class=\"CitationRef\"\u003e64\u003c/span\u003e]. The surface modification with L-Cysteine was carried out as follows: MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs (0.1 g) were dispersed in 25 mL of deionized water. L-Cysteine (0.05 g) was dissolved in 10 mL deionized water and added dropwise to the suspension of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e under vigorous stirring at room temperature. After 2 h the modified NPs were collected with the external magnet, washed several times with deionized water and ethanol, and dried under vacuum at 60 ᵒC for 24h.\u003c/p\u003e\n\u003ch3\u003e2-2-Cytotoxicity studied by MTT test\u003c/h3\u003e\n\u003cp\u003eThe Human breast carcinoma cells were prepared by the National Cell Bank of Iran at the Pasteur Institute. The Human breast carcinoma cells (MCF7) were cultured in PRMI-1640 medium, containing 10% fetal bovine serum (FBS), 5% horse serum, 2mM L-glutamine, 1% penicillin/streptomycin and 2 gr/L sodium bicarbonate and preserved at 37\u0026deg; C and pH\u0026thinsp;=\u0026thinsp;7.2 in a humidified atmosphere containing 5% CO\u003csub\u003e2\u003c/sub\u003e. At this stage, 10 \u003csup\u003e4\u003c/sup\u003e cells were cultured in 1L culture media. Then, cell viability and proliferation were measured using the MTT microculture colorimetric test. The stock solution was prepared by dissolving 5 mg of tetrazolium in 1 ml of phosphate-buffered saline. Cells were then exposed to nanoparticles with different concentrations (1, 5, 10, 25, 50, 100, and 250 \u0026micro;g/ml) for 24 hours. In the next step, 10 microliters of MTT stock were added to the cells. After keeping the stock for 4 hours in incubation, 10 microliters of DMSO were added and absorbance was checked at 570 nm using an Eliezer reader. The cell viability was calculated according to the following equation:\u003c/p\u003e\n\u003cp\u003e%viability=\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{ \\text{m}\\text{e}\\text{a}\\text{n} \\text{e}\\text{x}\\text{p}\\text{e}\\text{r}\\text{i}\\text{m}\\text{e}\\text{n}\\text{t}\\text{a}\\text{l} \\text{a}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e}}{\\text{m}\\text{e}\\text{a}\\text{n} \\text{n}\\text{e}\\text{g}\\text{a}\\text{t}\\text{i}\\text{v}\\text{e} \\text{c}\\text{o}\\text{n}\\text{t}\\text{r}\\text{o}\\text{l} \\text{a}\\text{b}\\text{s}\\text{o}\\text{r}\\text{b}\\text{a}\\text{n}\\text{c}\\text{e}}\\)\u003c/span\u003e\u003c/span\u003e \u0026times;100 (1)\u003c/p\u003e\n\u003cp\u003eThe cell viability was investigated by reading the optimal density at 570 nm. The viability (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) was plotted in the form of histograms, using the Microsoft Excel program.\u003c/p\u003e\n\u003cdiv class=\"Heading\"\u003e\u003cstrong\u003e2-3-Computational details\u003c/strong\u003e:\u003c/div\u003e\n\u003cp\u003eAll geometry optimization and energy calculation were carried out by the Gaussian 09 software [\u003cspan class=\"CitationRef\"\u003e65\u003c/span\u003e]. Quantum mechanical calculations were performed by the Density Functional Theory (DFT) method at the Lee-Yong-Parr Exchange-correlation hybrid functional (B3LYP) level of theory and 6-31G(d) basis set. Vibrational analysis was carried out after structural optimization at B3LYP/6-31G*. The absence of imaginary vibration frequencies indicated the stability of structures. Furthermore, the highest occupied energy (HOMO), lowest unoccupied energy (LUMO), and energy gap (Eg) were determined for the considered structures. Eg value can be used to check the sensitivity of L-Cysteine toward 5-fU. The reactivity parameters including electronegativity (A), chemicalpotential(\u0026micro;), chemical hardness(\u0026eta;), electrophilicity(\u0026omega;) and softness (S)were calculated following equations[\u003cspan class=\"CitationRef\"\u003e66\u003c/span\u003e] :\u003c/p\u003e\n\u003cp\u003eA= -HOMO (2)\u003c/p\u003e\n\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equ1\" class=\"mathdisplay\"\u003e$$\\mu =\\frac{ (\\text{E} \\text{H}\\text{O}\\text{M}\\text{O} +\\text{E} \\text{L}\\text{U}\\text{M}\\text{O})}{2}$$\u003c/div\u003e\n\u003cdiv class=\"EquationNumber\"\u003e3\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003e\u0026eta; = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{\\text{E} \\text{L}\\text{U}\\text{M}\\text{O}- \\text{E} \\text{H}\\text{O}\\text{M}\\text{O}}{2}\\)\u003c/span\u003e\u003c/span\u003e ( 4)\u003c/p\u003e\n\u003cp\u003e\u0026omega;= \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{{\\mu } 2}{2{\\eta } }\\)\u003c/span\u003e\u003c/span\u003e (5)\u003c/p\u003e\n\u003cp\u003eS= \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{1}{2{\\eta }}\\)\u003c/span\u003e\u003c/span\u003e (6)\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eThe binding energies (E\u003csub\u003eads\u003c/sub\u003e) of 5-FU molecule and L-Cysteine were determined as the difference between the total energy of complexes and the sum of the energies of the isolated L-Cysteine and 5-FU as follows:\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eE\u003csub\u003eads\u003c/sub\u003e = E\u003csub\u003e5FU/Cys\u003c/sub\u003e\u0026ndash; (E \u003csub\u003eCys\u003c/sub\u003e + E \u003csub\u003e5\u0026minus;FU\u003c/sub\u003e) (7)\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eThe negative adsorption energy demonstrated a thermodynamically favorable adsorption process.\u003c/p\u003e\n\u003cp\u003eThe zero-point energy (ZPE) was also considered in the calculation. Negative adsorption energy shows the stability of the 5-FU/L-Cysteine complex. The adsorption Gibbs free energies and adsorption enthalpy were computed by the following equations (at T\u0026thinsp;=\u0026thinsp;298/15 K and P\u0026thinsp;=\u0026thinsp;1 atm):\u003c/p\u003e\n\u003cp\u003eWhere G \u003csub\u003ecomplex,\u003c/sub\u003e G \u003csub\u003edrug,\u003c/sub\u003e G\u003csub\u003eL\u0026minus;Cys,\u003c/sub\u003e H \u003csub\u003ecomplex,\u003c/sub\u003e H \u003csub\u003edrug,\u003c/sub\u003e and H\u003csub\u003eL\u0026minus;Cys\u003c/sub\u003e are the Gibbs free energy and enthalpy of the species specified in the equation.\u003c/p\u003e\n\u003cp\u003e∆G \u003csub\u003eads\u003c/sub\u003e = G \u003csub\u003ecomplex\u003c/sub\u003e\u0026ndash; (G \u003csub\u003edrug\u003c/sub\u003e + G\u003csub\u003eL\u0026minus;Cys\u003c/sub\u003e) (8)\u003c/p\u003e\n\u003cp\u003e∆H \u003csub\u003eads\u003c/sub\u003e = H \u003csub\u003ecomplex\u003c/sub\u003e\u0026ndash; (H \u003csub\u003edrug\u003c/sub\u003e + H\u003csub\u003eL\u0026minus;Cys\u003c/sub\u003e) (9)\u003c/p\u003e\n\u003cp\u003eWhere G \u003csub\u003ecomplex,\u003c/sub\u003e G \u003csub\u003edrug,\u003c/sub\u003e G\u003csub\u003eL\u0026minus;Cys,\u003c/sub\u003e H \u003csub\u003ecomplex,\u003c/sub\u003e H \u003csub\u003edrug,\u003c/sub\u003e and H\u003csub\u003eL\u0026minus;Cys\u003c/sub\u003e are the Gibbs free energy and enthalpy of the species specified in the equation.\u003c/p\u003e\n\u003cp\u003eAll of the calculation was performed in gas and water phases.\u003c/p\u003e"},{"header":"3-Results and Discussion","content":"\u003cp\u003eThe bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine were examined by FT-IR method. For bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e), the bands at \u003cem\u003e~\u003c/em\u003e\u0026thinsp;580\u0026ndash;650 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e correspond to the Mn-O and Fe-O bonds. The peak at 3400 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e can be assigned to the hydroxyl group on the surface of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP. After the functionalization of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP with L-Cysteine (MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-cyc), some changes emerged in the FT-IR spectrum of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The strong peaks at \u003cem\u003e~\u003c/em\u003e\u0026thinsp;580\u0026ndash;650 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e can be attributed to Mn-O and Fe-O in the structure of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP. The new bands at 3440 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and the peak at 1510 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are assigned to the N-H group. The peaks at 2857 and 2935 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e are related to the CH\u003csub\u003e2\u003c/sub\u003e group. The peaks at 1411 and 1633 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e can be ascribed to the carbonyl group. The peaks at 1308 and 1071 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e can be assigned to C-O and C-N bonds, respectively. All of these new bands (in comparison with bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e) are related to L-Cysteine, implying the bonding of L-Cysteine to the surface of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP. Because of the low concentration of the thiol group, the S-H band was not observed in the spectrum of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine [\u003cspan class=\"CitationRef\"\u003e67\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eThe XRD patterns of the bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The diffraction peaks of both compounds are the same and show the phase composition of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP. The diffraction peaks correspond to the crystal planes (111), (220), (311), (222), (400), (422) (511), and (440) with 2\u0026theta; values of 18.08\u0026deg;, 29.74\u0026deg;, 35.02\u0026deg;, 36.66\u0026deg;, 42.57\u0026deg;, 52.82\u0026deg;, 56.26\u0026deg;, and 61.74\u0026deg;, respectively which is correspond to the spinel structure of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e according to the JCPDS standard card No. 88-1965. These data indicate that the functionalization of bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e with L-Cysteine did not alter the phase composition of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP.\u003c/p\u003e\n\u003cp\u003eThe vibration sample magnetometer (VSM) was utilized for the investigation of the magnetic properties of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine(Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e) in a magnetic field range of -15000 to 15000 Oe. Both samples exhibited narrow hysteresis loops with superparamagnetic behavior. The saturation magnetization (Ms) values of bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine are 29.64 emu/g and 18.36 emu/g, respectively, indicating that the coating of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e with non-magnetic L-Cysteine reduced the magnetic properties of bare MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NPs. MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine NPs still showed acceptable magnetic properties and could be separated by an external magnet.\u003c/p\u003e\n\u003cp\u003eSEM images of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine can be seen in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e which shows non-uniform semi spherical particles with some aggregations. The diameter of nanoparticles ranged in 70\u0026ndash;110 nm. The Energy Dispersive X-ray spectrum and EDX mapping are shown in Figs.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e, respectively. These data confirm the presence of manganese, iron, oxygen, and sulfur groups, confirming the coverage of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e with L-Cysteine.\u003c/p\u003e\n\u003ch3\u003e3-1-Cytotoxicity study by MTT assay:\u003c/h3\u003e\n\u003cp\u003eNowadays amino acid-based nanocarriers are applied as a multipurpose tool for disease treatment due to their low immunogenicity, good biocompatibility, tunable structure, and ease of chemical modification. As an amino acid, cysteine has shown interesting applications in chemical modification, drug delivery, bioimaging, and in vivo long blood circulation. Thanks to its thiol (SH) groups, it has found new properties such as tumor proliferation[\u003cspan class=\"CitationRef\"\u003e68\u003c/span\u003e]\u003c/p\u003e\n\u003cp\u003eMohammad Zaki Fahmi and coworkers synthesized MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e composite nanofibers including cellulose and collagen. The MTT test against Hella cells indicated the low toxicity of this nanofiber [\u003cspan class=\"CitationRef\"\u003e24\u003c/span\u003e]. In another study, Kanagesan et al. prepared MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP and evaluated their biocompatibility with murine breast cancer cells (4T1). Their results showed the dose-dependent cytotoxic effect of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP against 4T1[\u003cspan class=\"CitationRef\"\u003e69\u003c/span\u003e].\u003c/p\u003e\n\u003cp\u003eIn this study, cytotoxicity of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP, L-Cysteine, and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine was in vitro examined against MCF7 cells through the MTT test. In this method, the viability percentage of MCF7 cells was calculated by measuring the reduction of tetrazolium salt to purple formazan in living cells. The breast cancer cells were treated with different concentrations (1, 5, 10, 25, 50, 100 and 250 \u0026micro;g/L) of nanoparticles, modifier, and modified nanoparticles within the incubation period of 24 h. Based on the results, the cell viability percentage decreased with increasing the concentration of nanoparticles. Although there is no regular trend in the case of cysteine alone, a decreasing trend was observed for viability percentage at higher concentrations. This indicates the significant dose-dependence of antiproliferative activity of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and L-Cysteine. However, the cell viability of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@ L-Cysteine was not dose-dependent (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e). Moreover, MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, L-Cysteine, and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine have low toxicity with the cell viability above 95%. MTT test results are in agreement with the earlier findings on the good biocompatibility of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e NP and L-Cysteine [\u003cspan class=\"CitationRef\"\u003e68\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e70\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e71\u003c/span\u003e], which makes them appropriate for biomedical purposes.\u003c/p\u003e\n\u003ch3\u003e3-2Theoretical study:\u003c/h3\u003e\n\u003cp\u003eIn this section investigates the loading of 5-FUas an anti-breast cancer drug by MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine nanocarrier with Quantum mechanical calculations, considering the interaction between 5FU and L-cysteine.\u003c/p\u003e\n\u003cp\u003eFirst, the structures of 5-FU and L-Cysteine were optimized at the DFT/B3LYP/6-31G(d) level. Vibrational analysis was carried out after the optimization of structures at mentioned theoretical level. The absence of imaginary vibration frequencies illustrated the stability of structures. The optimized structures are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e. The theoretical bond lengths and angles of the optimized structures well agree with previous research. Then, 5-FU was placed on various positions of L-Cysteine (including NH\u003csub\u003e2\u003c/sub\u003e, COOH, and S) through its nitrogen, fluorine, and carbon atoms both vertically and horizontally in both media (gas and aqueous). The results showed no favorable interaction between L-Cysteine and 5-FU in the gas phase. The most stable complex was obtained after optimization in the aqueous phase (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e9\u003c/span\u003e). The negative adsorption energy demonstrated a thermodynamically favorable adsorption process. Moreover, the 5-FU molecule were adsorbed through the nitrogen atom on the sulfur atom of L-Cysteine with the adsorption energy of -12.029 KJ.mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The value of adsorption energy illustrated physical adsorption. Changes of enthalpy (∆H), free Gibbs energy (∆G), and entropy were calculated for interaction between L-Cysteine and 5-FU. The results indicated the negative variation of adsorption enthalpy, implying exothermic interaction of 5FU and L-Cysteine at ambient temperature. The Gibbs free energy change of binding was obtained positive. As a result, this interaction does not happen spontaneously at room temperature.\u003c/p\u003e\n\u003cp\u003eIt seems that the interaction temperature must be lowered to bind the 5-FU to the L-Cysteine as it reduces the vibrations of the bonds in the complex [\u003cspan class=\"CitationRef\"\u003e72\u003c/span\u003e]. Temperature is an important factor in adsorption and release reactions.\u003c/p\u003e\n\u003cp\u003eTherefore, the interaction between L-Cysteine and 5FU was examined at different temperatures (T\u0026thinsp;=\u0026thinsp;278.15-305.15K) in aqueous media. The results in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e show that the interaction between 5-FU and L-Cysteine was spontaneous at temperature range of 278.15 to 288.15K. With increasing the temperature, the values of adsorption-free Gibbs energy, enthalpy, and entropy got more positive but the equilibrium constant decreased especially at 308.15 K (body temperature). Therefore, drug release seems to be thermodynamically feasible at physiological conditions. The diagram of changes of ∆G, ∆H, ∆S, and K versus temperature is shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e10\u003c/span\u003e.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe calculated binding energy (E \u003csub\u003eb\u003c/sub\u003e Kcal/mol), ∆G(Kcal/mol), ∆H(Kcal/mol), ∆S(cal/molK), and K in 278.15-308.15K\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eT\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e∆H ads\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e∆Gads\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e∆Sads\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eKth\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e278.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.205\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-264.988\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.37*10 2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e283.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.336\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.025\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-263.589\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.708*10\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e288.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.961\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.842\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-262.183\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e4.376\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e293.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.586\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.344\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-260.770\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.530*10\u0026thinsp;\u0026minus;\u0026thinsp;1\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e298.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-2.211\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.532\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-259.351\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e7.456*10\u0026thinsp;\u0026minus;\u0026thinsp;2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e303.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.836\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.730\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-257.948\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.058*10\u0026thinsp;\u0026minus;\u0026thinsp;2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e308.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.461\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.939\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-256.516\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.57*10\u0026thinsp;\u0026minus;\u0026thinsp;3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eAs an intermolecular interaction or a chemical reaction occurs between the frontier orbitals of the involved species, the frontier molecular including the highest occupied molecular orbital (HOMO), lowest unoccupied molecular orbital (LUMO) and H-L gap (Eg) were calculated as listed in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The HOMO/LUMO contour is also depicted in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e11\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eThe HOMO energy of 5-FU is smaller (less negative) than L-Cysteine, implying that this structure has higher tendency to react with electrophilic species. The high HOMO energy in this system indicates that the electrons in this orbital can be more easily given to an electrophile species. L-Cysteine is more electrophile compared with 5FU. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e12\u003c/span\u003e shows the HOMO and the map of molecular electrostatic potential (MEP) of the L-Cysteine. As depicted in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, Eg(L-H) of the L-Cysteine /5FU (5.410 eV) is lower than L-Cysteine(6.250), suggesting a notable rise in the reactivity and conductivity of the complex.\u003c/p\u003e\n\u003cp\u003eThe \u0026omega; index approved the tendency of a molecule to absorb electrons and is closely related to the electron affinity of the molecule. It also gives useful information about the electron transfer in the electrostatic interaction. Since the formation of the complex involves non-covalent interactions and the electron transfer from the nucleophilic to the electrophilic species, according to the values of \u0026omega; in the Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, the electron is transferred from L-Cysteine to 5 FU. Also, the softness is a degree of interactivity of molecule. As seen in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, L-cysteine is more reactive to 5Fu.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe calculated Energy gap (Eg eV), HOMO(eV), LUMO(eV), Chemical potential (\u0026micro; eV), Chemical hardness(\u0026eta; eV), Electrophilicity index(\u0026omega; eV), Softness (S ev\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e)and Electronegativity(A eV) of the drug, L-cysteine and their complex\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCompound\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eE HOMO\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eE LUMO\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e∆Eg\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u0026eta;\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u0026micro;\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u0026omega;\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eS\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5Fu\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-6.390\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.430\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.715\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.675\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.487\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.184\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.96\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-cysteine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-6.600\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.350\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e6.250\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e3.125\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.475\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.932\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.35\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-cysteine /5F\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-6.420\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-1.010\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e5.410\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.705\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-3.715\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e2.551\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.184\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.01\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe natural bond orbital (NBO) analysis was carried out by the B3LYP functional and 6-31G (d) basis set. The NBO was used to analyze the interaction between the donor and the acceptor orbitals. The Lewis and non-Lewis orbitals illustrate the bonding and antibonding orbital NBOs, respectively. The energy of charge transfer between donor and acceptor orbitals was determined using second order perturbation theory. Eq.\u0026nbsp;10 estimates the stabilization energy (E2).\u003c/p\u003e\n\u003cp\u003eE\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;q\u003csub\u003ei\u003c/sub\u003e \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\frac{\\left(Fij\\right)2}{\\mathcal{E}j-\\mathcal{E}i}\\)\u003c/span\u003e\u003c/span\u003e (10)\u003c/p\u003e\n\u003cp\u003eWhere qi shows the donor orbital, Ei denotes the diagonal element, Ej is orbital energy, and F(ij) represents diagonal NBO Fock matrix element. The stabilization energy (E2) determined the interaction between the donor and acceptor orbitals, the larger the stabilization energy the stronger the interaction. Stabilization energy of the interacting donor and acceptor orbitalis are depicted in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. We considered the most probable bonding to antibonding interactions, which could be due to charge transfer between l-cysteine and 5FU. The major interactions arise from the charge transfer between lone pair S and ơ* antibonding orbital (N20-H25) with energy of 8.86Kcal/mol. This result agrees with the FMO evaluation.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eThe second \u0026minus;order perturbation theory analysis of the most interacting of the natural bond orbitals\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCharge transfer\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eE2(Kcal/mol)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eEj-Ei(a.u)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eF(i,j) a.u\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLp S͢͢ \u0026rarr; ơ* N20-H25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e8.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.71\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.071\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLp O23\u0026rarr;ơ* N4-H12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.91\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.18\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.043\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLp O23\u0026rarr;ơ*C5-C7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.63\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.009\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine drug delivery system was synthesized and characterized in this research. Then, the cytotoxicity of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e, L-Cysteine and MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine was determined on the MCF7 cells line by MTT assay. The MTT results illustrated that the MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine does not have cytotoxicity against the MCF7 cells line while increasing the MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine concentration does not have a significant effect on the cytotoxicity of this carrier. Also, theoretical studies evaluated the power of L-cysteine as a modifying agent of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e and drug delivery system for 5-FU drugs. DFT calculation indicated that the 5-FU anticancer drug was physically adsorbed on L-Cysteine in aqueous media. Thermodynamic analysis revealed that this interaction is exothermic. The drug binding is not favorable at ambient temperature, whereas the drug release was feasible at body temperature. Also, the electronic calculations showed a decline in the gap energy (Eg) of the complex by -13.4%. The obtained results from the hybridization of experimental and theoretical studies show that the MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine nanocarrier can be a promising candidate for drug delivery systems and these results might be effective in nanomedicine scope.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthic approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe had no participant in our study and the study was in vitro which had no need ethic approval\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll raw data and materials on which this study is based are included in the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this paper\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe have no funding\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNeda. Hasankhani\u003c/strong\u003e: experimental and laboratory test\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSharieh. Hosseini\u003c/strong\u003e: wrote the main manuscript text and supervision\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eElham askarizadeh\u003c/strong\u003e.: wrote the manuscript text\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBita.M ehravi\u003c/strong\u003e: laboratory test\u003c/p\u003e\n\u003cp\u003eAll authors reviewed the manuscript\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eE. Abbasi, M. Milani, S. Fekri Aval, M. Kouhi, A. Akbarzadeh, H. Tayefi Nasrabadi, P. Nikasa, S.W. Joo, Y. Hanifehpour, K. Nejati-Koshki, Silver nanoparticles: synthesis methods, bio-applications and properties, Critical reviews in microbiology 42(2) (2016) 173-180.\u003c/li\u003e\n\u003cli\u003eA. Akbarzadeh, M. Samiei, S. 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Ketabi, DFT and Molecular Simulation Study of Gold Clusters as Effective Drug Delivery Systems for 5-Fluorouracil Anticancer Drug, Journal of Cluster Science (2022) 1-11.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Modified magnetic nanoparticles, L-Cysteine, MCF7, DFT","lastPublishedDoi":"10.21203/rs.3.rs-4396900/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4396900/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThanks to their high hydrophilic and magnetic properties, MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles (NP) are recognized as favorable drug carriers. In this study, L-Cysteine-modified MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e nanoparticles (MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine) were prepared and characterized. Their cytotoxicity against human breast cancer cell lines (MCF7) was also evaluated by MTT assay. To simulate drug delivery systems, the interaction between modified NP and 5-fluorouracil (5-FU) was examined as a breast cancer drug. The MTT results showed the applicability of MnFe\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e4\u003c/sub\u003e@L-Cysteine nanoparticles as a potential cytotoxic agent in breast cancer treatment. Based on the theoretical calculations, the adsorption energy between L-Cysteine and 5-FU was \u0026minus;\u0026thinsp;12.029 KJ/mol and their interaction was spontaneous and exothermic at the temperature range of 278.15 to 288.15 K. Also, the drug release thermodynamically is feasible at body temperature. The calculated electronic descriptors indicated that the electrons were transferred from L-Cysteine to 5- FU. Overall, MNFe2O4@L-Cysteine, in addition to being non-toxic has the potential to deliver 5-FU anticancer drug.\u003c/p\u003e","manuscriptTitle":"MnFe2O4@L-Cysteine as a drug delivery system, in vitro cytotoxicity evaluation on human Breast cancer cell (MCF7) and DFT calculation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-21 18:57:43","doi":"10.21203/rs.3.rs-4396900/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"daa9c688-1652-48f9-8d7e-6c5a2a3b45d8","owner":[],"postedDate":"May 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-05-28T05:37:11+00:00","versionOfRecord":[],"versionCreatedAt":"2024-05-21 18:57:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4396900","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4396900","identity":"rs-4396900","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}