Anti-fibrillation Effect of Gold Nanoparticles Conjugated with Boswellic Acid on α-synuclein

Anti-fibrillation Effect of Gold Nanoparticles Conjugated with Boswellic Acid on α-synuclein | 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 Article Anti-fibrillation Effect of Gold Nanoparticles Conjugated with Boswellic Acid on α-synuclein Masoumeh Gharb, Farima Mozafari, Payam Arghavani, Ali Akbar Saboury, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5383385/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 Jul, 2025 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract Neurodegenerative diseases such as Alzheimer’s and Parkinson’s are characterized by the death of neurons in specific brains. α-synuclein (α-Syn) is a key factor in Parkinson’s disease (PD), forming toxic fibrils when misfolded. Natural products, such as Boswellia serrata , have shown promise in treating neurodegenerative diseases. However, the poor pharmacological performance of Boswellia acids (BAs) limits their effectiveness. Enhancing the bioavailability of BAs through nanocarriers could be a solution. This study explores the potential of β-Boswellic acid conjugated to gold nanoparticles (GNPs) as a novel PD treatment. Covalent and noncovalent conjugations of β-Boswellic acid to GNPs (GNP-BA) were developed to study their impact on α-Syn fibrillation in vitro . The successful synthesis of spherical GNPs (< 32 nm) was confirmed using high-resolution transmission electron microscopy (HR-TEM) and field emission scanning electron microscopy (FESEM). UV-visible and Fourier-transform infrared (FTIR) spectroscopies confirmed the conjugation of BA to GNPs. Specific interactions between α-Syn and GNP-BA conjugates were observed, with GNPs noncovalently bound to BA effectively inhibiting fibril formation. Thioflavin T (ThT) assay and atomic force microscopy (AFM) further supported the inhibitory effect of designed GNPs on α-Syn fibrillation, suggesting a potential therapeutic approach for PD treatment. α-synuclein Boswellic acids gold nanoparticle protein fibrillation Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 INTRODUCTION The role of α-synuclein (α-Syn) in Parkinson's disease (PD) has led to new understandings of the mechanisms behind this debilitating disorder [ 1 ]. The major pathological hallmarks of PD are degeneration of dopamine neurons in the substantia nigra and abnormal deposition of α-Syn into fibrillar Lewy bodies [ 2 ]. Although the physiological function of α-Syn remains unresolved, neuron degeneration in the affected brain regions is associated with the formation of α-Syn oligomers during its fibrillation process [ 3 ]. During this self-aggregation, α-Syn undergoes a structural transformation from intrinsically disordered monomers to form small cytotoxic soluble oligomers, which further assemble into protofibrils and eventually insoluble β-sheet rich fibrils [ 4 – 5 ]. These intermediate oligomers have been identified as highly damaging to cellular function, and are considered the most neurotoxic forms of α-Syn [ 5 – 6 ]. The toxicity of certain α-Syn oligomers is attributed to specific structural features, which lead to a range of detrimental effects including disrupting cell membranes especially mitochondrial, and synaptic impairment [ 7 – 8 ]. As people age, certain small protein fragments and peptides become more prone to aggregation, which can lead to the formation of harmful species. This process is especially apparent in progressive amyloidosis disorders such as PD. The increasing incidence of such diseases in an aging population has led to significant societal costs [ 9 – 10 ]. Therefore, the identification of compounds capable of inhibiting or delaying the aggregation mechanisms associated with neurotoxicity could be a promising approach to preventing or treating PD. Natural products, with their complex molecular frameworks, offer a diverse array of chemical species for medicinal chemists to explore in discovering chemical probes and drugs. For a long time, these natural compounds have been a valuable resource in drug development and treating various diseases. Recently, certain natural products, such as Boswellic acid, have gained significant attention for their potential in treating neurodegenerative complications [ 10 – 11 ]. β-Boswellic acid (BA), a pentacyclic triterpene derived from frankincense resin obtained from the Boswellia serrata tree, has been studied extensively. BA has demonstrated potent efficacy in treating various neurological diseases [ 12 – 13 ]. However, bioavailability has been a major hurdle in translating the preclinical potential of BA into therapeutic effects [ 14 ]. Various approaches have been employed to improve BA's bioavailability, including lecithin formulation, standardized meal administration, and oral co-administration, which have shown enhanced bioavailability and therapeutic effects [ 15 ]. Notably, novel drug delivery systems like nanoparticles (NPs) introducing chemically active surfaces and high surface-to-volume ratios have been proven effective in increasing the bioavailability and pharmacokinetic properties of BA [ 16 ]. Gold nanoparticles (GNPs) are considered one of the most biocompatible nanocarriers for drug delivery due to their physicochemical properties, ease of synthesis in various sizes and shapes, minimal toxicity, and excellent penetration into the blood-brain barrier. Due to their significant surface area, GNPs can carry high drug payloads and enhance drug efficacy in a controlled manner while minimizing the potential for adverse effects [ 16 – 17 ]. Consequently, there has been growing interest in combining BA with GNPs, leveraging the unique physical, chemical, and biological properties of the resulting hybrid materials. In addition to the valuable pharmaceutical effects of BA, conjugation of GNPs with BA is a functionalization approach of NPs in nature, a promising strategy to achieve significant biocompatibility and improved colloidal stability as well as minimizing unwanted biological responses such as the challenge protein corona formation [ 18 – 19 ]. The inhibition of α-Syn fibrillation using metal NPs has been studied broadly [ 20 – 21 ]. suggesting that GNPs potentially may interact and disassemble amyloid fibrils. Therefore, in this investigation, we aim to examine the inhibitory effect of GNP-BA on the fibrillation of α-Syn in vitro. MATERIALS AND METHODS Materials. Tetrachloroauric acid (HAuCl4), 1-ethyl-3-(3dime-thylaminopropyl) carbodiimide (EDC), N-hydroxy succinic-mide (NHS), 2-(N-111Morpholino) ethane sulfonic acid (MES), trisodium citrate, dimethyl sulfoxide (DMSO), thioflavin T (ThT), isopropyl-D-1-thiogalactopyranoside (IPTG) from Sigma-Aldrich (Munich, Germany). PT7-7 α-Syn WT plasmid containing the α-Syn gene was obtained from Addgene. The HiTrap Q FF anion exchange chromatography column was from GE Healthcare. Tween 20, and Triton X-100 (TX-100) were purchased from Merck (Darmstadt, Germany). Boswellic acid was a gift from Kondor Pharma Inc. (Canada). Deionized water was used for making all solutions. Preparation of GNP-BAs Using the Chemical Method. GNPs were synthesized using the Turkevich method (C-G). Trisodium citrate (1%) was added to boiling chloroauric acid (0.8 mM), changing its color to red in 15 min. After the solution cooled, BA was conjugated to the GNPs using EDC/NHS chemistry. The GNPs were first mixed with Tween 20 and cysteamine in an MES buffer. EDC (2 µM) was then added to BA, followed by NHS (4 µM), and the mixture was added to the cysteamine-coated GNPs. This reaction was gently shaken for 48 h. The GNPs were then centrifuged at 10,000 × g for 30 minutes to remove excess reagents. The final GNP-BA conjugates (C-G-BA) were stored at 4°C [ 22 ]. Preparation of GNP-BAs Using the Physical Method. GNPs were synthesized by subjecting a solution of HAuCl₄ and TX-100 to UV irradiation for 10 minutes (U-G). For conjugation, the GNPs were mixed with BA (5 mM) in DMSO and incubated for 48 h. After incubation, the mixture was centrifuged to remove excess BA, and the GNP-BA conjugates (U-G-BA) were stored at 4°C [ 22 ]. Physicochemical Properties of GNP and GNP-BA . The spectra of GNPs and GNP-BA were measured using a UV − visible spectrophotometer (Carry 100 Bio Varian) over a wavelength range of 200–800 nm. A Fourier transform infrared spectrometer (Irprestige-21, Shimadzu) was employed to assess the chemical interactions of different functional groups, with spectra recorded in the range of 4000 to 400 cm⁻¹ at a resolution of 4 cm⁻¹. The particle size, size distribution, and surface charge were analyzed using dynamic light scattering (DLS) and ζ potential measurements (Brookhaven ZetaPlus ζ Potential Analyzer). The surface morphology, size distribution, and crystallinity of the GNPs were examined using a high-resolution transmission electron microscope (FEI Tecnai G2 F20 SuperTwin) operating at an accelerating voltage of 200 kV. A drop of each sonicated and monodispersed GNP sample in deionized water was placed on a transparent carbon-coated copper grid. Elemental composition analysis was performed on a field emission scanning electron microscope (Zeiss 436 Sigma VP) with a carbon-coated copper tape grid. Images were analyzed using ImageJ software. Expression and purification of αSyn. α-Syn was expressed in Escherichia coli BL21(DE3) cells transfected with pT7-7 α-Syn wild type (WT) plasmid, following the method of Hoyer et al. with some modifications [ 23 ]. Briefly, the transfected E. coli cells were grown overnight in Luria broth (LB) containing 100 µg/mL ampicillin. The next day, the pre-culture was used to inoculate fresh LB medium. When the culture reached an OD 600 of 0.6, α-Syn expression was induced by adding 1 mM IPTG, then incubated at 37°C while shaking at 180 rpm for 4 h. Cells were then harvested by centrifugation at 6000 rpm for 5 minutes at 4°C to obtain the cell pellet. The pellet was resuspended in lysis buffer (20 mM Tris base pH 8.0, 1 mM EDTA, and 1 mM PMSF), and the cells were lysed by sonication (50 W, 10 seconds on and 10 seconds off). The lysed cells were then incubated in boiling water for 20 minutes, followed by centrifugation at 18,000 g for 30 minutes at 4°C. The supernatant was collected, and ammonium sulfate was slowly added to a final concentration of 0.36 g/mL. After stirring for 30 minutes at 4°C, the mixture was centrifuged at 18,000 g for 20 minutes at 4°C. The resulting pellet was resuspended in 20 mM Tris buffer (pH 8.0) and loaded onto a HiTrap Q FF anion exchange chromatography column. α-Syn was eluted using 300 mM NaCl, and its purity (≥ 95%) was confirmed by SDS-PAGE. The purified α-Syn was dialyzed against 20 mM Tris base buffer (pH 7.5), and its concentration was determined by measuring absorbance at 275 nm (ε 275 = 5600 M − 1 .cm − 1 ). Notably, the purified α-Syn used in our studies didn’t contain any additional motifs, such as a His-tag, that could interact with the metal ions used in our experiments [ 23 ]. Aggregation of αSyn . To study the effect of BA, GNP, and GNP-BA on α-Syn fibril formation, 90 µM of α-Syn in 20 mM Tris buffer (pH 7.5) was incubated with each sample at 37°C while stirring at 1000 rpm for 60 h. As a control, α-Syn was incubated in Tris buffer alone. Kinetic studies were conducted during the incubation period, followed by characterization of the fibrils after the completion of fibril formation [ 24 ]. Thioflavin (ThT) fluorescence assay. A steady-state ThT kinetic assay was conducted to study α-Syn aggregation, with excitation at 440 nm and emission at 482 nm, using a Cary Eclipse fluorescence spectrophotometer (Varian). Cuvette wells were filled with a final sample volume of 200 µL, containing 90 µM α-Syn and 20 µM ThT. Circular dichroism spectropolarimetry (CD). Far-UV circular dichroism (CD) spectra were recorded using an AVIV 215 spectropolarimeter to analyze the secondary structural changes of α-Syn at the end of fibril formation. The measurements were conducted at room temperature in the presence of C-G-BA and U-G-BA, covering the wavelength range of 190 − 260 nm. Atomic force microscopy (AFM). α-Syn fibrils formed in the presence and absence of C-G-BA and U-G-BA were analyzed by AFM. Images were obtained in semi-contact mode using an AFM (NTEGRA, NT-MDT, Russia) and processed with Nova software (version 1.26.0.1443). RESULT AND DISCUSSION In this study, we have meticulously synthesized GNPs through two distinct methods. The physicochemical attributes of the synthesis process play a pivotal role in the size, morphology, and functional properties of these NPs [ 25 – 26 ]. Subsequently, we employed BA, known for its therapeutic potential in addressing neurodegenerative disorders, as a conjugation agent with GNPs. Our objective was to enhance potential GNP efficacy in neurodegenerative treatment alongside improving their stability, biocompatibility, and bioavailability. The results presented in this article explore the intricacies of this innovative strategy. The Turkevich method, the most common chemical method for the synthesis of GNPs in biological applications, involves the reduction of tetrachloroauric acid (HAuCl4) with trisodium citrate to produce GNPs (C-G) [ 27 ]. Over the past two decades, the development of covalent conjugation techniques has significantly advanced research in biomedicine, materials science, and nanotechnology [ 28 ]. In this study, GNPs were conjugated with BA using cysteamine (CysA) as a linker and EDC/NHS as a cross-linker to establish covalent bonds (Fig. 1 a) [ 28 ]. Another effective method for producing GNPs (U-G) is photochemical synthesis [ 29 ]. While each method for synthesizing GNPs has its advantages and disadvantages, chemical methods are efficient but often involve toxic reducing agents, which pose biological risks. In contrast, photochemical synthesis is a non-toxic, eco-friendly alternative that is increasingly recognized for producing GNPs suitable for various biological applications. This method utilizes UV irradiation in the presence of Triton X-100 micelles, which play a crucial role in stabilizing the GNPs and influencing their morphology (U-G) [ 30 ]. Covalent interactions lead to the creation of new molecules with distinct properties and are generally stronger than noncovalent interactions [ 31 – 32 ]. However, for smaller drug molecules bound to NP surfaces via covalent bonds, these bonds must be broken for the drug release to function effectively. Therefore, for targeted therapy, using non-covalent interactions to create nanoconjugates may be more effective. Such nanoconjugates are stable enough to reach the intended site in the body after administration [ 33 ]. This approach ensures both stability and functionality, thereby enhancing drug delivery efficacy. Accordingly, in this study, the GNPs were also coated with BA using noncovalent electrostatic interactions (U-G-BA) (Fig. 2 a). Fabrication and characteristics of C-G-BA and U-G-BA. The chemical and physical properties of the synthesized GNPs were characterized using a combination of UV-Vis spectroscopy, DLS, ζ potential, FT-IR, and HR-TEM. The formation of GNPs was further confirmed by EDX analysis via FE-SEM. Visual inspection was used to track the formation of C-G-BA and U-G-BA, with a change from light-yellow color to wine red confirming the synthesis. This color transformation corresponds to the surface plasmon resonance (SPR) feature of the produced GNPs [ 34 ]. After three rounds of centrifugation and washing with deionized water, the GNP samples were examined with UV–Vis spectroscopy. SPR band of C-G was observed at 522 nm, and after conjugation with BA (C-G-BA), a redshift to 524 nm was noted. To optimize BA concentration for conjugation with C-G, various concentrations (1, 5, 10, 15, 20, and 25) were tested, with the results indicating that a BA concentration of 5 was the most effective. Similarly, the surface plasmon band of U-G-BA exhibited a slight redshift, shifting from 530 nm to 536 nm compared to U-G (Figure S1 a). This redshift is due to a change in the dielectric environment of the GNPs, confirming the effective conjugation of BA to GNPs in both cases. DLS measurements were carried out to measure the hydrodynamic diameter of GNPs before and after they were conjugated with BA. The results showed an increase in particle size for GNPs-BA compared to naked GNPs (Figure S1 b and Table 1 ). This increase aligns with the expected rise in light scattering due to the adsorption of larger molecules on the particle surface [ 35 ]. U-G-BA had a bigger diameter than C-G-BA, suggesting a higher number of BA equivalents. Nonetheless, the DLS data indicated a monodispersed system for all GNPs and GNP-BA samples (Figure S1 b). The higher ξ potential value indicated that C-G and C-G-BA carry a negative charge, while the U-G exhibits a positive ξ potential. After interacting with BA, the ξ potential of U-G-BA shifts to a negative value (Table 1 ). When particles in a suspension have a high negative or positive ξ potential, they repel one another, which enhances their stability by preventing aggregation. In contrast, if particles have a low ξ potential, there is insufficient repulsive force to prevent interaction, leading to flocculation [ 36 ]. The phase plots for C-G and C-G-BA exhibited a similar pattern, while those for U-G and U-G-BA were inverted due to the electrostatic interaction between BA and U-G. This suggests that U-G had a positive surface charge, and since BA is negatively charged, electrostatic interactions between them were favored. Consequently, the size and charge of the particles play a crucial role in effective conjugation ( Figure S1 c) [ 37 ]. HR-TEM analyses were conducted to evaluate the size and morphology of the GNPs. The images depicted in Fig. 1 b and Fig. 2 b show spherical-shaped GNPs with an average diameter of 16.48 ± 2.13 and 31.23 ± 6.21 for C-G and U-G respectively. The selected area electron diffraction (SAED) pattern confirmed the crystalline nature of the GNPs, revealing bright circular rings (Fig. 1 d and Fig. 2 d). These rings corresponded to reflections from the standard Bragg planes (111), (200), (220), and (311), indicating that the GNPs had a cubic crystal structure (Fig. 1 c and Fig. 2 c) [ 38 ]. Energy-dispersive X-ray spectroscopy (EDX) revealed strong peaks at 2.15 keV, typical of metallic gold nanocrystallites, due to surface plasmon resonance (Fig. 1 e and Fig. 2 e) [ 36 ]. Table 1 Surface charge and size of synthesized GNPs. Nanoparticles ξ potential (mV) particle size (nm) DLS (nm) HR-TEM (nm) C-G −25.59 ± 2.02 23.4 16.48 ± 2.13 C-G-BA −15.74 ± 0.76 27.4 - U-G + 8.16 ± 0.86 48.2 31.23 ± 6.21 U-G-BA −22.56 ± 1.51 55.5 - The FTIR spectrum of BA displayed characteristic absorption bands at 3673, 2925, 1697, 1453, 1380, and 1242 \(\:{\text{c}\text{m}}^{-1}\) corresponding to the following: O–H stretching vibrations, C − H stretching vibrations, C = O stretching vibrations of aryl acid, C − H bending, \(\:-\text{C}\text{O}\text{O}-\) symmetric stretching vibrations of carboxylates and C − COC stretching vibrations of aryl ketone, respectively (Figure S2 a) [ 39 ]. The synthesized C-G was analyzed by FTIR, revealing peaks at 3302 \(\:{\text{c}\text{m}}^{-1}\) , 1635 \(\:{\text{c}\text{m}}^{-1}\) , 1024 \(\:{\text{c}\text{m}}^{-1}\) , and 871 \(\:{\text{c}\text{m}}^{-1}\) ascribed to the presence of O–H, C = C, C–O, and C–C groups in the sample. The O–H peak is linked to water molecules and O–H stretching of citrate molecules, while the C = C, C–O, and C–C peaks confirmed the presence of citrate molecules along with the C-G (Figure S2 a) [ 40 ]. The lack of characteristic peaks of S–H stretching peak around 2601 \(\:{\text{c}\text{m}}^{-1}\) , combined with the presence of N–H bending vibrations at 1603 \(\:{\text{c}\text{m}}^{-1}\:\) and 824 \(\:{\text{c}\text{m}}^{-1}\) , suggests that Cy is linked to AuNPs via Au–S bonds (Figure S2 a). The intense peak at 1641 \(\:{\text{c}\text{m}}^{-1}\) (Figure S2 a) is associated with the carbonyl stretching vibration of the carboxyl group present in BA [ 41 ]. In the case of U-G, the spectrum underwent an increase in the intensity of a band around 1099 \(\:{\text{c}\text{m}}^{-1}\) which corresponds to C-O stretching in the TX-100 molecule. This increase in intensity is due to the coordination of AuNPs with the O-H group of TX-100, leading to a polarity change in the C-O bond and a boost in the intensity of the C = O stretching band (Figure S2 b) [ 42 ]. TX-100 contains hydroxyl and oxyethylene groups that may interact with the BA molecules through hydrogen bonding. This suggests that TX-100 (non-ionic surfactant) with a hydrophilic chain of O–H groups interacts with the N − H groups of BA molecules. The bands at 1404–1387 cm-1 are attributed to the C-H group vibrations found in phenyl rings of TX-100 [ 43 ]. U-G-BA had changes associated with decreased C − O stretching vibration and increased C-O stretching vibration, respectively [ 44 ]. These spectra confirmed that both types of GNPs have incorporated BA, forming GNPs-BA (Figure S2 b). NPs hydrophobicity, size, and surface charge are physicochemical properties that are known to affect protein aggregation [ 45 ]. It is well documented that surface modifications of NPs can alter protein aggregation pathways, either slowing down or accelerating aggregation, and potentially even reversing preformed aggregates [ 46 ]. For instance, modifying iron oxide NPs surface with leucine made them more hydrophobic and significantly blocked mTTR’s aggregation and fibrillation pathways [ 47 ]. Additionally, these modifications have the potential to sequester misfolded states or even correct their conformation. GNPs have been extensively studied for their interactions with amyloidogenic proteins, which are associated with neurodegenerative diseases [ 46 ]. Specifically, GNPs-BA may interact with α-Syn and lead to inhibiting its aggregation and fibrillation processes. The presence of C-G-BA & U-G-BA NPs alters the aggregation kinetics of α-Syn The kinetics of α-Syn fibril formation were studied in the absence and presence of increasing concentrations of bare GNPs, GNP-BA, and pure BA. This process was monitored by tracking the characteristic increase in Thioflavin T (ThT) fluorescence intensity [ 53 – 54 ]. Typically, amyloid aggregation kinetics are characterized by a sigmoidal curve, which includes a lag phase, a growth phase, and a final equilibrium phase[ 50 ]. The impact of GNP-BA on both the nucleation (lag time) and elongation (exponential phase) processes was quantified using kinetic parameters derived from data fitting. Regardless of the specific amyloid protein being studied, experimental data are generally fitted to this sigmoidal model [ 51 ] (Eq. 1 ): $$\:F=\:\frac{{F}_{final}-\:{F}_{0}}{1+exp(-{k}_{app}(t-\:{t}_{1/2}\left)\right)}$$ 1 In this equation, F represents the fluorescence intensity at time t , F final denotes the maximum fluorescence intensity, \(\:{t}_{\raisebox{1ex}{$1$}\!\left/\:\!\raisebox{-1ex}{$2$}\right.}\) is the time required to reach half of the maximum fluorescence intensity (corresponding to the midpoint between nuclei formation and fibril growth), and k app stands for the apparent first-order aggregation constant. The lag time is defined as the point where the tangent at the maximum fibrillation rate intersects the abscissa, as given by [ 52 ] (Eq. 2 ): $$\:{t}_{lag}=\:{t}_{1/2}-\:{2k}^{-1}$$ 2 In several studies, BA has been utilized to treat neurodegenerative diseases at concentrations ranging from 1 to 100 µM [ 53 ], [ 54 ]. In our experiments, we investigated concentrations from 1.25 to 95 µM. Consistent with previous reports, we found that concentrations up to 20 µM inhibit aggregation (Table 2 and Fig. 3 ). However, at higher concentrations, BA appears to promote fibril formation by decreasing the lag time (Fig. 3 a). We observed that negatively charged C-G (citrate-capped), slightly accelerated the aggregation of α-Syn at concentrations ranging from 1.25 to 5 µM, as evidenced by a reduction in the lag time (Table 2 and Fig. 3 b). These findings are consistent with previous research, which also reported that NP surface charge and concentration can modulate protein aggregation pathways. This underscores the dual role of NP in either inhibiting or promoting fibril formation, depending on their physicochemical properties and the environment [ 55 ]. α-Syn consists of three structural domains: the N-terminal domain (residues 1–60), the central non-amyloid component (NAC) domain (residues 61–95), and the C-terminal domain (residues 96–140). The N-terminal region is predominantly positively charged due to the presence of 11 lysine residues, whereas the C-terminal domain is negatively charged [ 56 ]. C-G appears to interact with the N-terminal region of α-Syn, potentially exposing the NAC region. This exposure may result in higher local concentrations of the NAC domain, thereby facilitating aggregation [ 55 ]. When studying C-G-BA at concentrations ranging from 1.25 to 5 µM, fibril formation declined compared to C-G alone. The rate of fibril formation also decreased with increasing concentrations of C-G-BA (Table 2 and Fig. 3 c). This suggests that the presence of BA slows down the growth kinetics of fibril formation. We observed that positively charged U-G Triton (X-100 capped), at concentrations ranging from 4 to 95 µM, decreased the aggregation of α-Syn, as indicated by an increase in the lag time (Table 2 and Fig. 4 a). At higher concentrations, U-G further inhibited fibril formation. Previous studies have shown that positively charged NPs can bind to the C-terminus of α-Syn. The positively charged U-G are expected to adsorb/capture a large number of α-syn monomers, making them promising candidates to prevent or delay the fibrillation process.[ 57 ]. The U-G-BAs, studied at concentrations ranging from 4 to 95 µM, significantly extended the lag phase and impacted the elongation phase (Table 2 and Fig. 4 b), tackling the aggregation of α-Syn. This inhibitory effect of U-G-BA was concentration-dependent, with the most pronounced reduction in the lag phase and half-time occurring at the highest concentration of 95 µM. These effects correlated with the surface properties of the synthesized U-G and its non-covalent interactions with BA. The interactions between the U-G-BA surface and α-Syn monomers and/or oligomers might create unfavorable conditions for fibril growth by obstructing binding sites for the addition of new monomers [ 58 ]. Moreover, the behavior of U-G-BA in inhibiting fibril formation is reminiscent of chaperone-like activity. Chaperone proteins assist in the proper folding of other proteins and prevent misfolding and aggregation. U-G-BA, through its multivalent interactions and enhanced solubility, mimics this chaperone-like function by stabilizing α-Syn in its non-fibrillar form and preventing the progression of aggregation [ 59 ]. Table 2 Relative half-time value ( \(\:{\varvec{t}}_{1/2}\) ), relative growth rate ( \(\:{\varvec{k}}_{\varvec{a}\varvec{p}\varvec{p}}\) ), and relative lag time ( \(\:{\varvec{t}}_{\varvec{l}\varvec{a}\varvec{g}}\) ) of fibril formation versus GNPs and GNP-BA concentration for α-Syn fibril formation. k app (h − 1 ) t 1/2 (h) t lag (h) BA (0.05) 0.10444 \(\:\pm\:\) 0.015 32.86 \(\:\pm\:\) 1.088 13.71025 \(\:\pm\:\) 2.706 BA (0.2) 0.10782 \(\:\pm\:\) 0.020 31.75 \(\:\pm\:\) 0.991 13.20057 \(\:\pm\:\) 2.463 BA (1) 0.10888 \(\:\pm\:\) 0.026 31.07 \(\:\pm\:\) 1.219 12.70115 \(\:\pm\:\) 2.391 BA (4) 0.10894 \(\:\pm\:\) 0.015 31.11 \(\:\pm\:\) 1.392 12.75127 \(\:\pm\:\) 2.387 BA (20) 0.09668 \(\:\pm\:\) 0.020 30.08 \(\:\pm\:\) 1.219 9.393198 \(\:\pm\:\) 3.370 BA (95) 0.11582 \(\:\pm\:\) 0.038 27.32 \(\:\pm\:\) 0.993 10.05183 \(\:\pm\:\) 1.973 C-G (0.05) 0.11566 \(\:\pm\:\) 0.026 34.06 \(\:\pm\:\) 1.088 16.76794 \(\:\pm\:\) 1.973 C-G (0.2) 0.10572 \(\:\pm\:\) 0.154 33.28 \(\:\pm\:\) 0.991 14.3621 \(\:\pm\:\) 1.982 C-G (1) 0.10384 \(\:\pm\:\) 0.020 32.53 \(\:\pm\:\) 1.219 13.2696 \(\:\pm\:\) 2.611 C-G-BA (0.05) 0.0903 \(\:\pm\:\) 0.020 33.52 \(\:\pm\:\) 1.088 11.37161 \(\:\pm\:\) 2.752 C-G-BA (0.2) 0.10116 \(\:\pm\:\) 0.026 34.03 \(\:\pm\:\) 0.991 14.25934 \(\:\pm\:\) 4.058 C-G-BA (1) 0.12282 + 0.038 34.03 \(\:\pm\:\) 1.219 17.74601 \(\:\pm\:\) 2.967 U-G (4) 0.10422 \(\:\pm\:\) 0.023 33.05 \(\:\pm\:\) 1.392 13.85983 \(\:\pm\:\) 1.623 U-G (20) 0.12736 \(\:\pm\:\) 0.022 33.52 \(\:\pm\:\) 1.013 17.81648 \(\:\pm\:\) 2.723 U-G (95) 0.1143 \(\:\pm\:\) 0.022 29.4 \(\:\pm\:\) 0.988 11.90219 \(\:\pm\:\) 1.428 U-G-BA (4) 0.11272 \(\:\pm\:\) 0.021 30.65 \(\:\pm\:\) 1.032 12.90692 \(\:\pm\:\) 2.150 U-G-BA (20) 0.10644 \(\:\pm\:\) 0.017 32.18 \(\:\pm\:\) 1.234 13.39007 \(\:\pm\:\) 2.559 U-G-BA (95) 0.09456 \(\:\pm\:\) 0.013 36.28 \(\:\pm\:\) 1.876 15.12941 \(\:\pm\:\) 3.582 To address this issue, ThT kinetics data were confirmed by AFM and far-UV circular dichroism (CD) analyses, which provide valuable information on the morphology and extent of α-syn fibrils. The Far-UV CD spectra of α-Syn (Fig. 5 ) resemble the classical spectrum of a disordered protein, while the β-sheet structures of fibrils are characterized by a negative minimum around 218 nm and a positive peak at 202 nm respectively [ 60 ]. The Far-UV CD signature became more complex in the presence of GNP-BA, suggesting alterations in the α-syn's secondary structure. Conversely, C-G-BA exhibited a weak band at 218 nm, indicative of reduced fibril formation and the potential emergence of alternative fibril morphologies. U-G-BA effectively preserved α-Syn’s native secondary structure and inhibited fibril formation, as reflected by the absence of notable spectral changes. Additionally, the Far-UV CD spectrum of α-Syn in the presence of BA at 5 µM suggested the protein predominantly remained monomeric, whereas, at 95 µM, a transition to aggregated states was observed, characterized by a negative minimum at 216 nm. AFM has been utilized as a suitable tool to determine possible rearrangements between the protofibrils producing a fibrillar polymorphism in the mature fibril. AFM in the control sample, which was untreated, the fibrils exhibited an elongated and intertwined morphology with heights reaching up to 15.40 nm and average lengths of 1.2 µm. These fibrils were uniformly structured, indicating a stable morphology with minimal height variation (Fig. 6a). Treatment with C-G-BA led to more complex fibril structures, with heights extending up to 21.13 nm and average lengths of 0.54 µm. The AFM images revealed a heterogeneous arrangement of fibrils, including straight and curved forms with varying densities among treated samples. This suggests that C-G-BA treatment induced significant structural changes, resulting in a more pronounced and varied fibril morphology compared to the control. The increased height range and diverse structural features indicated a more extensive aggregation pattern (Fig. 6b). In contrast, U-G-BA treatment resulted in fibrils with heights up to 12.55 nm and average lengths of 0.39 µm. The AFM images showed elongated, thread-like structures with less pronounced height variation and a more dispersed distribution. This pattern suggests that U-G-BA treatment is associated with reduced aggregation compared to C-G-BA, as indicated by the smaller and fewer aggregates. The less pronounced height variation in the U-G-BA-treated fibrils further supports the notion that this treatment leads to a less structured fibril network (Fig. 6c) [ 61 ]. U-G-BA largely determines the polymorphism of fibrils [ 62 ]. CONCLUSION PD is characterized by the toxic oligomeric and fibrillar phases formed by monomeric α-Syn. Certain NPs have been shown to promote protein aggregation, while others have been found to prevent this process. In the current study, we investigated the effects of these opposing behaviors using GNPs conjugated with BA under two different synthesis methods: citrate-capped gold nanoparticles (C-G) and their covalently conjugated form with BA (C-G-BA), as well as Triton X-100 capped gold nanoparticles (U-G) with non-covalent interaction with BA (U-G-BA). The noncovalent GNP-BA conjugates, particularly those synthesized via a photochemical method, showed significant inhibitory effects on the kinetics of α-Syn aggregation. These findings demonstrate the importance of synthesis and conjugation methods and open new avenues for developing nanoparticle-based treatments for neurodegenerative diseases, overcoming the pharmacokinetic limitations of natural compounds like Boswellic acid. Further, in vivo studies are needed to assess the therapeutic potential of GNP-BA conjugates in PD models. Declarations Author Contribution Masoumeh Gharb: Data collection and initial manuscript drafting.Farima Mozafari: Manuscript writing and preparation of figures.Payam Arghavani: Final revisions and validation.Ali Akbar Saboury: Methodology design and revisions.Gholamhossein Riazi: Corresponding author, methodology, and final revisions Data availability The datasets used and/or analyzed during the current study available from the corresponding author on reasonable request. References A. Kluge et al. , “α-Synuclein Pathology in PRKN-Linked Parkinson’s Disease: New Insights from a Blood-Based Seed Amplification Assay,” Ann. Neurol. , vol. 95, no. 6, pp. 1173–1177, 2024, doi: 10.1002/ana.26917. A. Recasens et al. , “Lewy Body extracts from Parkinson’s Disease Brains trigger α-Synuclein Pathology,” Ann. Neurol. , vol. 75, pp. 351–362, 2014. H. Mohammad-Beigi et al. , “Oleuropein derivatives from olive fruit extracts reduce - Synuclein fibrillation and oligomer toxicity,” J. Biol. 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Supplementary Files DLS.xlsx FTIRCG.pzfx UVvisabsorptionspectradistribution.pzfx zeta.pzfx SupplementaryMaterial.docx Cite Share Download PDF Status: Published Journal Publication published 17 Jul, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 03 Feb, 2025 Reviews received at journal 03 Jan, 2025 Reviewers agreed at journal 18 Dec, 2024 Reviews received at journal 15 Dec, 2024 Reviewers agreed at journal 04 Dec, 2024 Reviewers agreed at journal 03 Dec, 2024 Reviewers invited by journal 15 Nov, 2024 Editor assigned by journal 15 Nov, 2024 Editor invited by journal 07 Nov, 2024 Submission checks completed at journal 06 Nov, 2024 First submitted to journal 03 Nov, 2024 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. 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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-5383385","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":379461112,"identity":"d2544e8e-049d-4a28-8d06-35f796e49eb2","order_by":0,"name":"Masoumeh Gharb","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"prefix":"","firstName":"Masoumeh","middleName":"","lastName":"Gharb","suffix":""},{"id":379461113,"identity":"6316211f-177b-4b16-bc80-d371911e5a12","order_by":1,"name":"Farima Mozafari","email":"","orcid":"","institution":"University of Birmingham","correspondingAuthor":false,"prefix":"","firstName":"Farima","middleName":"","lastName":"Mozafari","suffix":""},{"id":379461114,"identity":"baa7ef75-d935-4053-acb8-559219dd8e7a","order_by":2,"name":"Payam Arghavani","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"prefix":"","firstName":"Payam","middleName":"","lastName":"Arghavani","suffix":""},{"id":379461115,"identity":"464b5c61-fbed-4da8-8038-26ded27b2e59","order_by":3,"name":"Ali Akbar Saboury","email":"","orcid":"","institution":"University of Tehran","correspondingAuthor":false,"prefix":"","firstName":"Ali","middleName":"Akbar","lastName":"Saboury","suffix":""},{"id":379461116,"identity":"28178a6d-212d-4638-8242-3bbdd8066b70","order_by":4,"name":"Gholamhossein Riazi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAuklEQVRIiWNgGAWjYDACCQaGA0BSDsElVosxAxspWkAgsYGNWHfJR/c+PPDjj0X6hvsNjB9+MFjkE9RieOe4wcEeHoncDccYmCV7GCQsGwhqmZHGcIBHAqyFQRroTgPCtgC1HPxjIJFuALTlN1Fa5CXSGA7zJEgkALWwEWeLAUiLzAEJw5nHEtssewyIsWVGGvPHN3/q5PkOHz5840dFHRG2HIAzGRuAXIIagLY0EKFoFIyCUTAKRjgAAJAdNDn27xOBAAAAAElFTkSuQmCC","orcid":"","institution":"University of Tehran","correspondingAuthor":true,"prefix":"","firstName":"Gholamhossein","middleName":"","lastName":"Riazi","suffix":""}],"badges":[],"createdAt":"2024-11-03 19:08:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5383385/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5383385/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-11107-6","type":"published","date":"2025-07-17T16:05:37+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":69659614,"identity":"ab97a554-465a-4dfc-97b9-6a024406baf2","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1098609,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCharacterization of C-G. (a) Schematic of the cross-linking GNP reactions with BA Functionalized by EDC/NHS in covalent conjugation (C-G-BA) (b) TEM image displaying C-G and its associated size distribution. (c) HR-TEM image of C-G. (d) Selected area electron diffraction (SAED) pattern. (e) EDX analysis of C-G via FE-SEM\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/ad143a16467871ccc252e0e4.png"},{"id":69659616,"identity":"6e472ca2-16fa-4e5b-a78e-3bf729600375","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1123098,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCharacterization of U-G. (a) Schematic of the noncovalent conjugation of GNPs with BA (U-G-BA). (b) TEM image showing U-G and its corresponding size distribution. (c) HR-TEM image of U-G. (d) Selected area electron diffraction (SAED) pattern. (e) EDS analysis of U-G via FE-SEM.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/02bac8839e1182af68e16e07.png"},{"id":69660188,"identity":"8c71f6be-6c47-4925-9d34-bc8d28b245f9","added_by":"auto","created_at":"2024-11-22 18:45:55","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":215026,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAggregation profile of α-Syn in the presence of chemically prepared GNPs, measured by ThT assay. Effect of the (a) BA concentration (1.25−95 μM), (b) C-G concentration (1.25−5 μM), and (c) C-G-BA concentration (1.25−5 μM).\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/48b6969b2227f43b482cd83d.png"},{"id":69659620,"identity":"d2ec4132-ce32-4963-bfa2-51138c17327b","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":210344,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAggregation profile of α-Syn in the presence of physically prepared GNPs, measured by ThT assay. Effect of the (a) U-G concentration (4-95 μM), and (b) U-G-BA concentration (4−95 μM).\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/b3b14455deffb762b0068f76.png"},{"id":69659617,"identity":"a6d6189a-2af7-455c-9982-f0fb2027a266","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":154097,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFar-UV CD spectra showing α-Syn monomers and fibrils, comparing samples formed in the absence (control) versus the presence of C-G-BA (5 μM) and U-G-BA (95 μM).\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/93db0fda203d043b97d2b4b0.png"},{"id":69660190,"identity":"4658d67a-546b-4eb9-acae-f14850ab613a","added_by":"auto","created_at":"2024-11-22 18:45:55","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":4113341,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAFM images of (a) α-Syn fibril, (b) α-Syn fibrils formed in the presence of 5 μM concentration of C-G-BA, and (c) α-Syn fibrils formed in the presence of 95 μM concentration of U-G-BA.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/38533d4fe05c4858c07369ff.png"},{"id":88507045,"identity":"966d4c3d-5dcf-4e8a-883d-fe6e5e5745dc","added_by":"auto","created_at":"2025-08-07 07:36:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":12227629,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/f8cd559f-f97f-4ca6-a7bf-3800d9cdb434.pdf"},{"id":69659615,"identity":"8676294f-3447-467a-9599-60f849b431b3","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":10758,"visible":true,"origin":"","legend":"","description":"","filename":"DLS.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/888f3f838406a49bf4b47b88.xlsx"},{"id":69660189,"identity":"43229446-31d4-4c64-b499-37a34bba97dd","added_by":"auto","created_at":"2024-11-22 18:45:55","extension":"pzfx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":233937,"visible":true,"origin":"","legend":"","description":"","filename":"FTIRCG.pzfx","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/4dbc09ca29b8d0c03e86d3e6.pzfx"},{"id":69660748,"identity":"89c16a40-c7bf-46b0-a5ff-3d9d918fe30f","added_by":"auto","created_at":"2024-11-22 19:01:55","extension":"pzfx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":110641,"visible":true,"origin":"","legend":"","description":"","filename":"UVvisabsorptionspectradistribution.pzfx","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/dda060687415f6cf94ee7400.pzfx"},{"id":69659623,"identity":"1bbb4b34-0af5-40bd-bdf0-89a6a38ff08e","added_by":"auto","created_at":"2024-11-22 18:37:55","extension":"pzfx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":398598,"visible":true,"origin":"","legend":"","description":"","filename":"zeta.pzfx","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/c5646e384e369b45d3526aca.pzfx"},{"id":69660491,"identity":"f18f8b70-c521-4af1-88fa-81972dd95b91","added_by":"auto","created_at":"2024-11-22 18:53:55","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":282520,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterial.docx","url":"https://assets-eu.researchsquare.com/files/rs-5383385/v1/fdde4f6cde4718e1a45cb508.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Anti-fibrillation Effect of Gold Nanoparticles Conjugated with Boswellic Acid on α-synuclein","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe role of α-synuclein (α-Syn) in Parkinson's disease (PD) has led to new understandings of the mechanisms behind this debilitating disorder [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. The major pathological hallmarks of PD are degeneration of dopamine neurons in the substantia nigra and abnormal deposition of α-Syn into fibrillar Lewy bodies [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Although the physiological function of α-Syn remains unresolved, neuron degeneration in the affected brain regions is associated with the formation of α-Syn oligomers during its fibrillation process [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. During this self-aggregation, α-Syn undergoes a structural transformation from intrinsically disordered monomers to form small cytotoxic soluble oligomers, which further assemble into protofibrils and eventually insoluble β-sheet rich fibrils [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. These intermediate oligomers have been identified as highly damaging to cellular function, and are considered the most neurotoxic forms of α-Syn [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. The toxicity of certain α-Syn oligomers is attributed to specific structural features, which lead to a range of detrimental effects including disrupting cell membranes especially mitochondrial, and synaptic impairment [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. As people age, certain small protein fragments and peptides become more prone to aggregation, which can lead to the formation of harmful species. This process is especially apparent in progressive amyloidosis disorders such as PD. The increasing incidence of such diseases in an aging population has led to significant societal costs [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Therefore, the identification of compounds capable of inhibiting or delaying the aggregation mechanisms associated with neurotoxicity could be a promising approach to preventing or treating PD.\u003c/p\u003e \u003cp\u003eNatural products, with their complex molecular frameworks, offer a diverse array of chemical species for medicinal chemists to explore in discovering chemical probes and drugs. For a long time, these natural compounds have been a valuable resource in drug development and treating various diseases. Recently, certain natural products, such as Boswellic acid, have gained significant attention for their potential in treating neurodegenerative complications [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. β-Boswellic acid (BA), a pentacyclic triterpene derived from frankincense resin obtained from the Boswellia serrata tree, has been studied extensively. BA has demonstrated potent efficacy in treating various neurological diseases [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, bioavailability has been a major hurdle in translating the preclinical potential of BA into therapeutic effects [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Various approaches have been employed to improve BA's bioavailability, including lecithin formulation, standardized meal administration, and oral co-administration, which have shown enhanced bioavailability and therapeutic effects [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Notably, novel drug delivery systems like nanoparticles (NPs) introducing chemically active surfaces and high surface-to-volume ratios have been proven effective in increasing the bioavailability and pharmacokinetic properties of BA [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eGold nanoparticles (GNPs) are considered one of the most biocompatible nanocarriers for drug delivery due to their physicochemical properties, ease of synthesis in various sizes and shapes, minimal toxicity, and excellent penetration into the blood-brain barrier. Due to their significant surface area, GNPs can carry high drug payloads and enhance drug efficacy in a controlled manner while minimizing the potential for adverse effects [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eConsequently, there has been growing interest in combining BA with GNPs, leveraging the unique physical, chemical, and biological properties of the resulting hybrid materials. In addition to the valuable pharmaceutical effects of BA, conjugation of GNPs with BA is a functionalization approach of NPs in nature, a promising strategy to achieve significant biocompatibility and improved colloidal stability as well as minimizing unwanted biological responses such as the challenge protein corona formation [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. The inhibition of α-Syn fibrillation using metal NPs has been studied broadly [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. suggesting that GNPs potentially may interact and disassemble amyloid fibrils. Therefore, in this investigation, we aim to examine the inhibitory effect of GNP-BA on the fibrillation of α-Syn in vitro.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e \u003cb\u003eMaterials.\u003c/b\u003e Tetrachloroauric acid (HAuCl4), 1-ethyl-3-(3dime-thylaminopropyl) carbodiimide (EDC), N-hydroxy succinic-mide (NHS), 2-(N-111Morpholino) ethane sulfonic acid (MES), trisodium citrate, dimethyl sulfoxide (DMSO), thioflavin T (ThT), isopropyl-D-1-thiogalactopyranoside (IPTG) from Sigma-Aldrich (Munich, Germany). PT7-7 α-Syn WT plasmid containing the α-Syn gene was obtained from Addgene. The HiTrap Q FF anion exchange chromatography column was from GE Healthcare. Tween 20, and Triton X-100 (TX-100) were purchased from Merck (Darmstadt, Germany). Boswellic acid was a gift from Kondor Pharma Inc. (Canada). Deionized water was used for making all solutions.\u003c/p\u003e \u003cp\u003e \u003cb\u003ePreparation of GNP-BAs Using the Chemical Method.\u003c/b\u003e GNPs were synthesized using the Turkevich method (C-G). Trisodium citrate (1%) was added to boiling chloroauric acid (0.8 mM), changing its color to red in 15 min. After the solution cooled, BA was conjugated to the GNPs using EDC/NHS chemistry. The GNPs were first mixed with Tween 20 and cysteamine in an MES buffer. EDC (2 µM) was then added to BA, followed by NHS (4 µM), and the mixture was added to the cysteamine-coated GNPs. This reaction was gently shaken for 48 h. The GNPs were then centrifuged at 10,000 × g for 30 minutes to remove excess reagents. The final GNP-BA conjugates (C-G-BA) were stored at 4°C [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003ePreparation of GNP-BAs Using the Physical Method.\u003c/b\u003e GNPs were synthesized by subjecting a solution of HAuCl₄ and TX-100 to UV irradiation for 10 minutes (U-G). For conjugation, the GNPs were mixed with BA (5 mM) in DMSO and incubated for 48 h. After incubation, the mixture was centrifuged to remove excess BA, and the GNP-BA conjugates (U-G-BA) were stored at 4°C [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003ePhysicochemical Properties of GNP and GNP-BA\u003c/b\u003e. The spectra of GNPs and GNP-BA were measured using a UV − visible spectrophotometer (Carry 100 Bio Varian) over a wavelength range of 200–800 nm. A Fourier transform infrared spectrometer (Irprestige-21, Shimadzu) was employed to assess the chemical interactions of different functional groups, with spectra recorded in the range of 4000 to 400 cm⁻¹ at a resolution of 4 cm⁻¹. The particle size, size distribution, and surface charge were analyzed using dynamic light scattering (DLS) and ζ potential measurements (Brookhaven ZetaPlus ζ Potential Analyzer). The surface morphology, size distribution, and crystallinity of the GNPs were examined using a high-resolution transmission electron microscope (FEI Tecnai G2 F20 SuperTwin) operating at an accelerating voltage of 200 kV. A drop of each sonicated and monodispersed GNP sample in deionized water was placed on a transparent carbon-coated copper grid. Elemental composition analysis was performed on a field emission scanning electron microscope (Zeiss 436 Sigma VP) with a carbon-coated copper tape grid. Images were analyzed using ImageJ software.\u003c/p\u003e \u003cp\u003e \u003cb\u003eExpression and purification of αSyn.\u003c/b\u003e α-Syn was expressed in \u003cem\u003eEscherichia coli\u003c/em\u003e BL21(DE3) cells transfected with pT7-7 α-Syn wild type (WT) plasmid, following the method of Hoyer et al. with some modifications [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Briefly, the transfected \u003cem\u003eE. coli\u003c/em\u003e cells were grown overnight in Luria broth (LB) containing 100 µg/mL ampicillin. The next day, the pre-culture was used to inoculate fresh LB medium. When the culture reached an OD\u003csub\u003e600\u003c/sub\u003e of 0.6, α-Syn expression was induced by adding 1 mM IPTG, then incubated at 37°C while shaking at 180 rpm for 4 h. Cells were then harvested by centrifugation at 6000 rpm for 5 minutes at 4°C to obtain the cell pellet. The pellet was resuspended in lysis buffer (20 mM Tris base pH 8.0, 1 mM EDTA, and 1 mM PMSF), and the cells were lysed by sonication (50 W, 10 seconds on and 10 seconds off). The lysed cells were then incubated in boiling water for 20 minutes, followed by centrifugation at 18,000\u003cem\u003eg\u003c/em\u003e for 30 minutes at 4°C. The supernatant was collected, and ammonium sulfate was slowly added to a final concentration of 0.36 g/mL. After stirring for 30 minutes at 4°C, the mixture was centrifuged at 18,000\u003cem\u003eg\u003c/em\u003e for 20 minutes at 4°C. The resulting pellet was resuspended in 20 mM Tris buffer (pH 8.0) and loaded onto a HiTrap Q FF anion exchange chromatography column. α-Syn was eluted using 300 mM NaCl, and its purity (≥ 95%) was confirmed by SDS-PAGE. The purified α-Syn was dialyzed against 20 mM Tris base buffer (pH 7.5), and its concentration was determined by measuring absorbance at 275 nm (ε\u003csub\u003e275\u003c/sub\u003e = 5600 M\u003csup\u003e− 1\u003c/sup\u003e.cm\u003csup\u003e− 1\u003c/sup\u003e). Notably, the purified α-Syn used in our studies didn’t contain any additional motifs, such as a His-tag, that could interact with the metal ions used in our experiments [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eAggregation of αSyn\u003c/b\u003e. To study the effect of BA, GNP, and GNP-BA on α-Syn fibril formation, 90 µM of α-Syn in 20 mM Tris buffer (pH 7.5) was incubated with each sample at 37°C while stirring at 1000 rpm for 60 h. As a control, α-Syn was incubated in Tris buffer alone. Kinetic studies were conducted during the incubation period, followed by characterization of the fibrils after the completion of fibril formation [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003eThioflavin (ThT) fluorescence assay.\u003c/b\u003e A steady-state ThT kinetic assay was conducted to study α-Syn aggregation, with excitation at 440 nm and emission at 482 nm, using a Cary Eclipse fluorescence spectrophotometer (Varian). Cuvette wells were filled with a final sample volume of 200 µL, containing 90 µM α-Syn and 20 µM ThT.\u003c/p\u003e \u003cp\u003e \u003cb\u003eCircular dichroism spectropolarimetry (CD).\u003c/b\u003e Far-UV circular dichroism (CD) spectra were recorded using an AVIV 215 spectropolarimeter to analyze the secondary structural changes of α-Syn at the end of fibril formation. The measurements were conducted at room temperature in the presence of C-G-BA and U-G-BA, covering the wavelength range of 190 − 260 nm.\u003c/p\u003e \u003cp\u003e \u003cb\u003eAtomic force microscopy (AFM).\u003c/b\u003e α-Syn fibrils formed in the presence and absence of C-G-BA and U-G-BA were analyzed by AFM. Images were obtained in semi-contact mode using an AFM (NTEGRA, NT-MDT, Russia) and processed with Nova software (version 1.26.0.1443).\u003c/p\u003e "},{"header":"RESULT AND DISCUSSION","content":"\u003cp\u003eIn this study, we have meticulously synthesized GNPs through two distinct methods. The physicochemical attributes of the synthesis process play a pivotal role in the size, morphology, and functional properties of these NPs [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e–\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Subsequently, we employed BA, known for its therapeutic potential in addressing neurodegenerative disorders, as a conjugation agent with GNPs. Our objective was to enhance potential GNP efficacy in neurodegenerative treatment alongside improving their stability, biocompatibility, and bioavailability. The results presented in this article explore the intricacies of this innovative strategy.\u003c/p\u003e\u003cp\u003eThe Turkevich method, the most common chemical method for the synthesis of GNPs in biological applications, involves the reduction of tetrachloroauric acid (HAuCl4) with trisodium citrate to produce GNPs (C-G) [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Over the past two decades, the development of covalent conjugation techniques has significantly advanced research in biomedicine, materials science, and nanotechnology [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. In this study, GNPs were conjugated with BA using cysteamine (CysA) as a linker and EDC/NHS as a cross-linker to establish covalent bonds (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea) [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAnother effective method for producing GNPs (U-G) is photochemical synthesis [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. While each method for synthesizing GNPs has its advantages and disadvantages, chemical methods are efficient but often involve toxic reducing agents, which pose biological risks. In contrast, photochemical synthesis is a non-toxic, eco-friendly alternative that is increasingly recognized for producing GNPs suitable for various biological applications. This method utilizes UV irradiation in the presence of Triton X-100 micelles, which play a crucial role in stabilizing the GNPs and influencing their morphology (U-G) [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eCovalent interactions lead to the creation of new molecules with distinct properties and are generally stronger than noncovalent interactions [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e–\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. However, for smaller drug molecules bound to NP surfaces via covalent bonds, these bonds must be broken for the drug release to function effectively. Therefore, for targeted therapy, using non-covalent interactions to create nanoconjugates may be more effective. Such nanoconjugates are stable enough to reach the intended site in the body after administration [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. This approach ensures both stability and functionality, thereby enhancing drug delivery efficacy. Accordingly, in this study, the GNPs were also coated with BA using noncovalent electrostatic interactions (U-G-BA) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea).\u003c/p\u003e\u003cp\u003e \u003cb\u003eFabrication and characteristics of C-G-BA and U-G-BA.\u003c/b\u003e The chemical and physical properties of the synthesized GNPs were characterized using a combination of UV-Vis spectroscopy, DLS, ζ potential, FT-IR, and HR-TEM. The formation of GNPs was further confirmed by EDX analysis via FE-SEM. Visual inspection was used to track the formation of C-G-BA and U-G-BA, with a change from light-yellow color to wine red confirming the synthesis. This color transformation corresponds to the surface plasmon resonance (SPR) feature of the produced GNPs [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. After three rounds of centrifugation and washing with deionized water, the GNP samples were examined with UV–Vis spectroscopy. SPR band of C-G was observed at 522 nm, and after conjugation with BA (C-G-BA), a redshift to 524 nm was noted. To optimize BA concentration for conjugation with C-G, various concentrations (1, 5, 10, 15, 20, and 25) were tested, with the results indicating that a BA concentration of 5 was the most effective. Similarly, the surface plasmon band of U-G-BA exhibited a slight redshift, shifting from 530 nm to 536 nm compared to U-G (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003ea). This redshift is due to a change in the dielectric environment of the GNPs, confirming the effective conjugation of BA to GNPs in both cases. DLS measurements were carried out to measure the hydrodynamic diameter of GNPs before and after they were conjugated with BA. The results showed an increase in particle size for GNPs-BA compared to naked GNPs (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003eb and Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This increase aligns with the expected rise in light scattering due to the adsorption of larger molecules on the particle surface [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eU-G-BA had a bigger diameter than C-G-BA, suggesting a higher number of BA equivalents. Nonetheless, the DLS data indicated a monodispersed system for all GNPs and GNP-BA samples (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003eb).\u003c/p\u003e\u003cp\u003eThe higher ξ potential value indicated that C-G and C-G-BA carry a negative charge, while the U-G exhibits a positive ξ potential. After interacting with BA, the ξ potential of U-G-BA shifts to a negative value (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). When particles in a suspension have a high negative or positive ξ potential, they repel one another, which enhances their stability by preventing aggregation. In contrast, if particles have a low ξ potential, there is insufficient repulsive force to prevent interaction, leading to flocculation [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The phase plots for C-G and C-G-BA exhibited a similar pattern, while those for U-G and U-G-BA were inverted due to the electrostatic interaction between BA and U-G. This suggests that U-G had a positive surface charge, and since BA is negatively charged, electrostatic interactions between them were favored. Consequently, the size and charge of the particles play a crucial role in effective conjugation ( Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003ec) [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eHR-TEM analyses were conducted to evaluate the size and morphology of the GNPs. The images depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb show spherical-shaped GNPs with an average diameter of 16.48 ± 2.13 and 31.23 ± 6.21 for C-G and U-G respectively. The selected area electron diffraction (SAED) pattern confirmed the crystalline nature of the GNPs, revealing bright circular rings (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ed). These rings corresponded to reflections from the standard Bragg planes (111), (200), (220), and (311), indicating that the GNPs had a cubic crystal structure (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec) [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Energy-dispersive X-ray spectroscopy (EDX) revealed strong peaks at 2.15 keV, typical of metallic gold nanocrystallites, due to surface plasmon resonance (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ee and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ee) [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"±\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSurface charge and size of synthesized GNPs.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNanoparticles\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eξ potential (mV)\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eparticle size (nm)\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDLS (nm)\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHR-TEM (nm)\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e \u003cp\u003e−25.59 ± 2.02\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e23.4\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.48 ± 2.13\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G-BA\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e \u003cp\u003e−15.74 ± 0.76\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e27.4\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e \u003cp\u003e+ 8.16 ± 0.86\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e48.2\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e31.23 ± 6.21\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G-BA\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\"±\" colname=\"c3\"\u003e \u003cp\u003e−22.56 ± 1.51\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e55.5\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003eThe FTIR spectrum of BA displayed characteristic absorption bands at 3673, 2925, 1697, 1453, 1380, and 1242 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e corresponding to the following: O–H stretching vibrations, C − H stretching vibrations, C = O stretching vibrations of aryl acid, C − H bending, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:-\\text{C}\\text{O}\\text{O}-\\)\u003c/span\u003e\u003c/span\u003e symmetric stretching vibrations of carboxylates and C − COC stretching vibrations of aryl ketone, respectively (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003ea) [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe synthesized C-G was analyzed by FTIR, revealing peaks at 3302 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e, 1635 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e, 1024 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e, and 871 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e ascribed to the presence of O–H, C = C, C–O, and C–C groups in the sample. The O–H peak is linked to water molecules and O–H stretching of citrate molecules, while the C = C, C–O, and C–C peaks confirmed the presence of citrate molecules along with the C-G (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003ea) [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe lack of characteristic peaks of S–H stretching peak around 2601 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e, combined with the presence of N–H bending vibrations at 1603 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\:\\)\u003c/span\u003e\u003c/span\u003eand 824 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e, suggests that Cy is linked to AuNPs via Au–S bonds (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003ea). The intense peak at 1641 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003ea) is associated with the carbonyl stretching vibration of the carboxyl group present in BA [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. In the case of U-G, the spectrum underwent an increase in the intensity of a band around 1099 \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\text{c}\\text{m}}^{-1}\\)\u003c/span\u003e\u003c/span\u003e which corresponds to C-O stretching in the TX-100 molecule. This increase in intensity is due to the coordination of AuNPs with the O-H group of TX-100, leading to a polarity change in the C-O bond and a boost in the intensity of the C = O stretching band (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eb) [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTX-100 contains hydroxyl and oxyethylene groups that may interact with the BA molecules through hydrogen bonding. This suggests that TX-100 (non-ionic surfactant) with a hydrophilic chain of O–H groups interacts with the N − H groups of BA molecules. The bands at 1404–1387 cm-1 are attributed to the C-H group vibrations found in phenyl rings of TX-100 [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eU-G-BA had changes associated with decreased C − O stretching vibration and increased C-O stretching vibration, respectively [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. These spectra confirmed that both types of GNPs have incorporated BA, forming GNPs-BA (Figure \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003eb).\u003c/p\u003e\u003cp\u003eNPs hydrophobicity, size, and surface charge are physicochemical properties that are known to affect protein aggregation [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. It is well documented that surface modifications of NPs can alter protein aggregation pathways, either slowing down or accelerating aggregation, and potentially even reversing preformed aggregates [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. For instance, modifying iron oxide NPs surface with leucine made them more hydrophobic and significantly blocked mTTR’s aggregation and fibrillation pathways [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]. Additionally, these modifications have the potential to sequester misfolded states or even correct their conformation. GNPs have been extensively studied for their interactions with amyloidogenic proteins, which are associated with neurodegenerative diseases [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Specifically, GNPs-BA may interact with α-Syn and lead to inhibiting its aggregation and fibrillation processes.\u003c/p\u003e\u003ch3\u003eThe presence of C-G-BA \u0026amp; U-G-BA NPs alters the aggregation kinetics of α-Syn\u003c/h3\u003e\u003cp\u003eThe kinetics of α-Syn fibril formation were studied in the absence and presence of increasing concentrations of bare GNPs, GNP-BA, and pure BA. This process was monitored by tracking the characteristic increase in Thioflavin T (ThT) fluorescence intensity [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e–\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Typically, amyloid aggregation kinetics are characterized by a sigmoidal curve, which includes a lag phase, a growth phase, and a final equilibrium phase[\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. The impact of GNP-BA on both the nucleation (lag time) and elongation (exponential phase) processes was quantified using kinetic parameters derived from data fitting. Regardless of the specific amyloid protein being studied, experimental data are generally fitted to this sigmoidal model [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e] (Eq.\u0026nbsp;\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e):\u003c/p\u003e\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e\n$$\\:F=\\:\\frac{{F}_{final}-\\:{F}_{0}}{1+exp(-{k}_{app}(t-\\:{t}_{1/2}\\left)\\right)}$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\u003c/div\u003e\u003cp\u003eIn this equation, F represents the fluorescence intensity at time \u003cem\u003et\u003c/em\u003e, \u003cem\u003eF\u003c/em\u003e\u003csub\u003e\u003cem\u003efinal\u003c/em\u003e\u003c/sub\u003e denotes the maximum fluorescence intensity, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{t}_{\\raisebox{1ex}{$1$}\\!\\left/\\:\\!\\raisebox{-1ex}{$2$}\\right.}\\)\u003c/span\u003e\u003c/span\u003e is the time required to reach half of the maximum fluorescence intensity (corresponding to the midpoint between nuclei formation and fibril growth), and \u003cem\u003ek\u003c/em\u003e\u003csub\u003e\u003cem\u003eapp\u003c/em\u003e\u003c/sub\u003e stands for the apparent first-order aggregation constant. The lag time is defined as the point where the tangent at the maximum fibrillation rate intersects the abscissa, as given by [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e] (Eq.\u0026nbsp;\u003cspan refid=\"Equ2\" class=\"InternalRef\"\u003e2\u003c/span\u003e):\u003c/p\u003e\u003cdiv id=\"Equ2\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ2\" name=\"EquationSource\"\u003e\n$$\\:{t}_{lag}=\\:{t}_{1/2}-\\:{2k}^{-1}$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e2\u003c/div\u003e\u003c/div\u003e\u003cp\u003eIn several studies, BA has been utilized to treat neurodegenerative diseases at concentrations ranging from 1 to 100 µM [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e], [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. In our experiments, we investigated concentrations from 1.25 to 95 µM. Consistent with previous reports, we found that concentrations up to 20 µM inhibit aggregation (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). However, at higher concentrations, BA appears to promote fibril formation by decreasing the lag time (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea).\u003c/p\u003e\u003cp\u003eWe observed that negatively charged C-G (citrate-capped), slightly accelerated the aggregation of α-Syn at concentrations ranging from 1.25 to 5 µM, as evidenced by a reduction in the lag time (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). These findings are consistent with previous research, which also reported that NP surface charge and concentration can modulate protein aggregation pathways. This underscores the dual role of NP in either inhibiting or promoting fibril formation, depending on their physicochemical properties and the environment [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eα-Syn consists of three structural domains: the N-terminal domain (residues 1–60), the central non-amyloid component (NAC) domain (residues 61–95), and the C-terminal domain (residues 96–140). The N-terminal region is predominantly positively charged due to the presence of 11 lysine residues, whereas the C-terminal domain is negatively charged [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e]. C-G appears to interact with the N-terminal region of α-Syn, potentially exposing the NAC region. This exposure may result in higher local concentrations of the NAC domain, thereby facilitating aggregation [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eWhen studying C-G-BA at concentrations ranging from 1.25 to 5 µM, fibril formation declined compared to C-G alone. The rate of fibril formation also decreased with increasing concentrations of C-G-BA (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). This suggests that the presence of BA slows down the growth kinetics of fibril formation.\u003c/p\u003e\u003cp\u003eWe observed that positively charged U-G Triton (X-100 capped), at concentrations ranging from 4 to 95 µM, decreased the aggregation of α-Syn, as indicated by an increase in the lag time (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea). At higher concentrations, U-G further inhibited fibril formation. Previous studies have shown that positively charged NPs can bind to the C-terminus of α-Syn. The positively charged U-G are expected to adsorb/capture a large number of α-syn monomers, making them promising candidates to prevent or delay the fibrillation process.[\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe U-G-BAs, studied at concentrations ranging from 4 to 95 µM, significantly extended the lag phase and impacted the elongation phase (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb), tackling the aggregation of α-Syn. This inhibitory effect of U-G-BA was concentration-dependent, with the most pronounced reduction in the lag phase and half-time occurring at the highest concentration of 95 µM. These effects correlated with the surface properties of the synthesized U-G and its non-covalent interactions with BA. The interactions between the U-G-BA surface and α-Syn monomers and/or oligomers might create unfavorable conditions for fibril growth by obstructing binding sites for the addition of new monomers [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMoreover, the behavior of U-G-BA in inhibiting fibril formation is reminiscent of chaperone-like activity. Chaperone proteins assist in the proper folding of other proteins and prevent misfolding and aggregation. U-G-BA, through its multivalent interactions and enhanced solubility, mimics this chaperone-like function by stabilizing α-Syn in its non-fibrillar form and preventing the progression of aggregation [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e].\u003c/p\u003e\u003cdiv class=\"gridtable\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eRelative half-time value (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\varvec{t}}_{1/2}\\)\u003c/span\u003e\u003c/span\u003e), relative growth rate (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\varvec{k}}_{\\varvec{a}\\varvec{p}\\varvec{p}}\\)\u003c/span\u003e\u003c/span\u003e), and relative lag time (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:{\\varvec{t}}_{\\varvec{l}\\varvec{a}\\varvec{g}}\\)\u003c/span\u003e\u003c/span\u003e) of fibril formation versus GNPs and GNP-BA concentration for α-Syn fibril formation.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003c/colgroup\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003ek\u003c/em\u003e\u003csub\u003e\u003cem\u003eapp\u003c/em\u003e\u003c/sub\u003e (h\u003csup\u003e− 1\u003c/sup\u003e)\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003et\u003c/em\u003e\u003csub\u003e\u003cem\u003e1/2\u003c/em\u003e\u003c/sub\u003e (h)\u003c/p\u003e \u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003et\u003c/em\u003e\u003csub\u003e\u003cem\u003elag\u003c/em\u003e\u003c/sub\u003e (h)\u003c/p\u003e \u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (0.05)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10444\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.015\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.86\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.088\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.71025\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.706\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (0.2)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10782\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.020\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.75\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.991\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.20057\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.463\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (1)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10888\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.026\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.07\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.219\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.70115\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.391\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (4)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10894\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.015\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e31.11\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.392\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.75127\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.387\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (20)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.09668\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.020\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.08\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.219\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e9.393198\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e3.370\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBA (95)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.11582\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.038\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e27.32\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.993\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10.05183\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.973\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G (0.05)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.11566\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.026\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.06\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.088\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e16.76794\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.973\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G (0.2)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10572\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.154\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.28\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.991\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.3621\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.982\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G (1)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10384\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.020\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.53\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.219\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.2696\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.611\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G-BA (0.05)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.0903\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.020\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.52\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.088\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.37161\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.752\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G-BA (0.2)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10116\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.026\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.03\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.991\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.25934\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e4.058\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC-G-BA (1)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.12282 + 0.038\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e34.03\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.219\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.74601\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.967\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G (4)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10422\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.023\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.05\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.392\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.85983\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.623\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G (20)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.12736\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.022\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.52\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.013\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17.81648\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.723\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G (95)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.1143\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.022\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e29.4\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.988\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.90219\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.428\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G-BA (4)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.11272\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.021\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.65\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.032\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.90692\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.150\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G-BA (20)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.10644\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.017\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e32.18\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.234\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13.39007\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e2.559\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eU-G-BA (95)\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.09456\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e0.013\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e36.28\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e1.876\u003c/p\u003e \u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.12941\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e3.582\u003c/p\u003e \u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/table\u003e\u003c/div\u003e\u003cp\u003eTo address this issue, ThT kinetics data were confirmed by AFM and far-UV circular dichroism (CD) analyses, which provide valuable information on the morphology and extent of α-syn fibrils. The Far-UV CD spectra of α-Syn (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e) resemble the classical spectrum of a disordered protein, while the β-sheet structures of fibrils are characterized by a negative minimum around 218 nm and a positive peak at 202 nm respectively [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. The Far-UV CD signature became more complex in the presence of GNP-BA, suggesting alterations in the α-syn's secondary structure. Conversely, C-G-BA exhibited a weak band at 218 nm, indicative of reduced fibril formation and the potential emergence of alternative fibril morphologies. U-G-BA effectively preserved α-Syn’s native secondary structure and inhibited fibril formation, as reflected by the absence of notable spectral changes. Additionally, the Far-UV CD spectrum of α-Syn in the presence of BA at 5 µM suggested the protein predominantly remained monomeric, whereas, at 95 µM, a transition to aggregated states was observed, characterized by a negative minimum at 216 nm.\u003c/p\u003e\u003cp\u003eAFM has been utilized as a suitable tool to determine possible rearrangements between the protofibrils producing a fibrillar polymorphism in the mature fibril. AFM in the control sample, which was untreated, the fibrils exhibited an elongated and intertwined morphology with heights reaching up to 15.40 nm and average lengths of 1.2 µm. These fibrils were uniformly structured, indicating a stable morphology with minimal height variation (Fig.\u0026nbsp;6a). Treatment with C-G-BA led to more complex fibril structures, with heights extending up to 21.13 nm and average lengths of 0.54 µm.\u003c/p\u003e\u003cp\u003eThe AFM images revealed a heterogeneous arrangement of fibrils, including straight and curved forms with varying densities among treated samples. This suggests that C-G-BA treatment induced significant structural changes, resulting in a more pronounced and varied fibril morphology compared to the control. The increased height range and diverse structural features indicated a more extensive aggregation pattern (Fig.\u0026nbsp;6b). In contrast, U-G-BA treatment resulted in fibrils with heights up to 12.55 nm and average lengths of 0.39 µm. The AFM images showed elongated, thread-like structures with less pronounced height variation and a more dispersed distribution. This pattern suggests that U-G-BA treatment is associated with reduced aggregation compared to C-G-BA, as indicated by the smaller and fewer aggregates. The less pronounced height variation in the U-G-BA-treated fibrils further supports the notion that this treatment leads to a less structured fibril network (Fig.\u0026nbsp;6c) [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. U-G-BA largely determines the polymorphism of fibrils [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e].\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003ePD is characterized by the toxic oligomeric and fibrillar phases formed by monomeric α-Syn. Certain NPs have been shown to promote protein aggregation, while others have been found to prevent this process. In the current study, we investigated the effects of these opposing behaviors using GNPs conjugated with BA under two different synthesis methods: citrate-capped gold nanoparticles (C-G) and their covalently conjugated form with BA (C-G-BA), as well as Triton X-100 capped gold nanoparticles (U-G) with non-covalent interaction with BA (U-G-BA).\u003c/p\u003e \u003cp\u003eThe noncovalent GNP-BA conjugates, particularly those synthesized via a photochemical method, showed significant inhibitory effects on the kinetics of α-Syn aggregation. These findings demonstrate the importance of synthesis and conjugation methods and open new avenues for developing nanoparticle-based treatments for neurodegenerative diseases, overcoming the pharmacokinetic limitations of natural compounds like Boswellic acid. Further, in vivo studies are needed to assess the therapeutic potential of GNP-BA conjugates in PD models.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eMasoumeh Gharb: Data collection and initial manuscript drafting.Farima Mozafari: Manuscript writing and preparation of figures.Payam Arghavani: Final revisions and validation.Ali Akbar Saboury: Methodology design and revisions.Gholamhossein Riazi: Corresponding author, methodology, and final revisions\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eA. 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Tech.\u003c/em\u003e, vol. 87, no. 6, pp. 1131\u0026ndash;1145, 2024.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"α-synuclein, Boswellic acids, gold nanoparticle, protein fibrillation","lastPublishedDoi":"10.21203/rs.3.rs-5383385/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5383385/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNeurodegenerative diseases such as Alzheimer\u0026rsquo;s and Parkinson\u0026rsquo;s are characterized by the death of neurons in specific brains. α-synuclein (α-Syn) is a key factor in Parkinson\u0026rsquo;s disease (PD), forming toxic fibrils when misfolded. Natural products, such as \u003cem\u003eBoswellia serrata\u003c/em\u003e, have shown promise in treating neurodegenerative diseases. However, the poor pharmacological performance of Boswellia acids (BAs) limits their effectiveness. Enhancing the bioavailability of BAs through nanocarriers could be a solution. This study explores the potential of β-Boswellic acid conjugated to gold nanoparticles (GNPs) as a novel PD treatment. Covalent and noncovalent conjugations of β-Boswellic acid to GNPs (GNP-BA) were developed to study their impact on α-Syn fibrillation \u003cem\u003ein vitro\u003c/em\u003e. The successful synthesis of spherical GNPs (\u0026lt;\u0026thinsp;32 nm) was confirmed using high-resolution transmission electron microscopy (HR-TEM) and field emission scanning electron microscopy (FESEM). UV-visible and Fourier-transform infrared (FTIR) spectroscopies confirmed the conjugation of BA to GNPs. Specific interactions between α-Syn and GNP-BA conjugates were observed, with GNPs noncovalently bound to BA effectively inhibiting fibril formation. Thioflavin T (ThT) assay and atomic force microscopy (AFM) further supported the inhibitory effect of designed GNPs on α-Syn fibrillation, suggesting a potential therapeutic approach for PD treatment.\u003c/p\u003e","manuscriptTitle":"Anti-fibrillation Effect of Gold Nanoparticles Conjugated with Boswellic Acid on α-synuclein","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-22 18:37:50","doi":"10.21203/rs.3.rs-5383385/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-02-04T04:43:25+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-01-03T05:34:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"5971514560427854155777184427346689592","date":"2024-12-19T03:12:43+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-12-15T21:39:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"77721334408792898263128123289315206362","date":"2024-12-04T15:58:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"211158445114340024046416690524460158438","date":"2024-12-03T15:52:55+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-11-15T08:28:28+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-11-15T08:27:42+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-11-07T08:48:36+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-11-07T03:09:19+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-11-03T19:06:58+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d0b06c58-7f5d-41ea-9825-23abbec8125d","owner":[],"postedDate":"November 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-08-07T07:29:05+00:00","versionOfRecord":{"articleIdentity":"rs-5383385","link":"https://doi.org/10.1038/s41598-025-11107-6","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-07-17 16:05:37","publishedOnDateReadable":"July 17th, 2025"},"versionCreatedAt":"2024-11-22 18:37:50","video":"","vorDoi":"10.1038/s41598-025-11107-6","vorDoiUrl":"https://doi.org/10.1038/s41598-025-11107-6","workflowStages":[]},"version":"v1","identity":"rs-5383385","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5383385","identity":"rs-5383385","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}