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DOI: 10.1055/a-2418-2200
Synthesis and Characterization of Acacia-Stabilized Doxorubicin-Loaded Gold Nanoparticles for Breast Cancer Therapy
- Abstract
- Introduction
- Materials and methods
- Results
- Discussion
- Conclusion
- References
Abstract
The targeted delivery of drugs is vital in breast cancer treatment due to its ability to produce long-lasting therapeutic effects with minimal side effects. This study reports the successful development of doxorubicin hydrochloride (DOX)-loaded colloidal gold nanoparticles stabilized with acacia gum (AG). Optimization studies varied AG concentrations (0.25% to 3% w/v) to determine optimal conditions for nanoparticle synthesis. The resulting acacia stabilized gold nanoparticles (AGNPs) were characterized using various techniques including high-resolution transmission electron microscopy (HR-TEM), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FE-SEM), and selected area electron diffraction (SAED). In vitro drug release studies demonstrated a higher release rate of DOX in sodium acetate buffer (pH 5.0) compared to phosphate buffer saline (pH 7.4), suggesting an enhanced therapeutic efficacy in acidic tumor environments. Cytotoxicity of DOX-AGNPs and free DOX was assessed in human breast cancer cells (MDA-MB-231). The DOX-AGNPs exhibited significantly greater cytotoxicity, indicating enhanced efficacy in targeting cancer cells. This enhancement suggests that adsorbing DOX on the surface of gold nanoparticles can improve drug delivery and effectiveness, potentially reducing side effects compared to pure DOX and traditional delivery methods. Stability tests conducted over six months at 25±1°C showed significant changes in particle size and PDI, suggesting limited stability under these conditions. Overall, the acacia-stabilized gold nanoparticles synthesized in this study exhibit promising characteristics for drug delivery applications, particularly in cancer therapy, with effective drug loading, controlled release, and favorable physicochemical properties.
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Keywords
green synthesis - gold nanoparticles - breast cancer - acacia gum - doxorubicin hydrochlorideIntroduction
Breast cancer remains one of the most prevalent and deadly cancers affecting women worldwide [1]. Despite advancements in treatment, challenges such as drug resistance and systemic toxicity of chemotherapy drugs like doxorubicin (DOX) persist. Targeted drug delivery systems are increasingly being explored to enhance therapeutic efficacy and minimize side effects [2]. Nanotechnology offers a promising solution by enabling the development of nanoparticles that can deliver drugs directly to cancer cells, improving drug stability and bioavailability [3] [4].
Nanomaterials have gained significant attention in recent years for their potential applications across a range of fields, including chemotherapy, drug delivery, bioimaging, and sensing [5]. Nanotechnology provides a versatile platform for a wide variety of inorganic and organic compounds, such as gold, silver, platinum, palladium, and iron, each with unique characteristics [6]. Gold nanoparticles (AuNPs) have emerged as one of the most widely studied nanomaterials, largely due to their easy synthesis, low cytotoxicity, flexibility for surface modification, and unique physical, chemical, and optical-electronic properties [7]. These attributes make AuNPs particularly attractive for the development of advanced drug delivery systems, offering the potential for enhanced targeting, controlled release, and reduced systemic toxicity in therapeutic applications [8].
Doxorubicin hydrochloride (DOX), an anthracycline antibiotic, is a commonly used chemotherapeutic agent for treating various cancers, including breast cancer [9]. However, its use is limited by severe side effects and the development of drug resistance [10] [11]. To address these limitations, this study explores the synthesis and stabilization of colloidal gold nanoparticles (AuNPs) using acacia gum (AG) [12]. Acacia gum, also known as gum arabic, is a natural biopolymer derived from the exudates of Acacia trees. It is composed of complex polysaccharides with a variety of functional groups (e. g., hydroxyl, carboxyl), which interact effectively with nanoparticle surfaces to prevent aggregation. The structure of acacia gum allows it to form a dense, protective layer around nanoparticles, ensuring long-term stability, particularly in aqueous solutions [13] [14]. Its excellent water solubility enhances its ability to disperse nanoparticles uniformly in these systems, reducing the risk of aggregation or sedimentation. This is a notable advantage over other biopolymers like chitosan, which often require acid treatment for solubility [12]. Acacia gum also possesses inherent emulsifying and stabilizing properties due to its surface-active components, making it an ideal choice for stabilizing gold nanoparticles in various liquid mediums without the need for additional surfactants or emulsifiers [14] [15]. The molecular structure of acacia gum also allows for easy functionalization, which is beneficial in applications like drug delivery, where controlled release is important. It can be modified to improve interactions with nanoparticles, enabling better control over the release of encapsulated substances [12] [13]. The environmental benefits of acacia gum, especially within the context of green synthesis, are well-documented. As a biopolymer obtained from the sap of Acacia trees, acacia gum is both renewable and sustainable. Its production has a lower environmental impact compared to synthetic stabilizers or polymers derived from non-renewable resources [14]. Moreover, acacia gum is biodegradable, breaking down naturally in the environment without generating harmful byproducts, thereby minimizing its ecological footprint. Its non-toxic nature further enhances its appeal, making it safe for humans, animals, and the environment. The widespread use of acacia gum in the food and pharmaceutical industries underscores its safety profile and versatility [15] [16] [17].
The goal of this research is to develop and characterize acacia-stabilized colloidal gold nanoparticles loaded with doxorubicin (DOX-AGNPs) to enhance the delivery of this potent anticancer drug to breast tumors. By encapsulating DOX within acacia-stabilized AuNPs, we aim to address the drugʼs limitations, such as its short half-life and rapid degradation, thereby increasing its bioavailability and reducing off-target effects. This approach also has the potential to facilitate more efficient drug transport across biological barriers, potentially improving DOXʼs therapeutic efficacy while minimizing its adverse side effects.
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Materials and methods
Materials
Doxorubicin hydrochloride and hydrochloroauric acid (HAuCl4) were obtained from Sigma-Aldrich (St. Louis, Missouri, USA). Gum acacia was sourced from Loba Chemie Laboratory in Mumbai, Maharashtra. Reagents & Fine Chemicals Pvt. Ltd. (Mumbai, India) supplied additional reagents, and the buffer preparation involved chemicals acquired from CDH in Delhi, India.
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Preparation of Acacia gum solution
The 1% w/v gum stock solution was made by dissolving 1 g of the AG in 100 mL of MilliQ water. The mixture was stirred at room temperature for 30 min and then centrifuged (Heraeus Biofuge Stratos, Thermo Fisher Scientific, Germany) at 5000 rpm for 30 min. The resulting supernatant phase was collected and used for experiment [16] [18].
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Fabrication and optimization of gold nanoparticles
Gold nanoparticles were fabricated using established techniques with minor adjustments [16] [19] [20]. To begin, 100 µL of HAuCl4 solution (10 mM) was added to 5 mL of acacia gum solution (1% w/v), and the mixture was agitated at 80°±1°C. Within 15 to 30 minutes, the colour of the solution changed from colourless to ruby red. The process for creating acacia gum-stabilized gold nanoparticles (AGNP) was optimized for various concentrations of gum: 0.25%, 0.5%, 0.75%, 1%, 2%, and 3% (as discussed in previous study) [16]. After cooling to room temperature, the colloidal solution was refrigerated and stored in amber-coloured vials. Visualization of optimized Acacia gold nanoparticles (AGNPs) under natural light in transparent glass vials are presented in [Fig. 1].
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Drug capping
The 5 ml AGNP solution was loaded with 10 mg of DOX and agitated overnight at room temperature. To recover the DOX-loaded acacia gum stabilized gold nanoparticles (AGNP-DOX), the mixture was centrifuged at 5,000 rpm for 30 minutes [20]. The percent drug concentration of these samples was determined using a UV/Vis spectrophotometer (UV-1800, Shimadzu, Japan) set to 268 nm, according to the following formula:
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Characterization of doxorubicin loaded gold nanoparticles
Determination of hydrodynamic diameter and surface charges
Dynamic light scattering (DLS) is the most precise method for measuring the size and surface charge of colloidal nanoparticles. The nanoparticles were dispersed in deionized water, and their hydrodynamic diameter, polydispersity index, and zeta potential were determined using a particle size analyzer (ZEN 3690, Malvern Zetasizer, UK) [21].
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UV–Vis Spectrometry (UV)
A UV/Vis spectrophotometer (UV-1800, Shimadzu, Japan) was used to obtain UV-visible spectra for DOX, AGNP, and AGNP-DOX in the 200–800 nm range, confirming the synthetic blank AGNPs by their characteristic ruby red color [21].
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Fourier Transform Infrared Spectroscopy (FTIR)
Each sample (DOX, AGNP, and AGNP-DOX) was combined with 100 mg of potassium bromide using a graphite mortar and pestle, then compressed into a pellet using a hydraulic press. The materials were analyzed with an FTIR spectrophotometer (Spectrum BX, Perkin Elmer, UK) operating in the range of 4000 to 400 cm−1 [21].
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Powder X-ray Diffraction(P-XRD)
Measurements of powder X-ray diffraction were performed using an X-ray diffractometer (Smart Lab 3 kW, Rigaku Corporation, Japan) set at 200 V AC±10%, 50/60 Hz, 3\30 A, to measure diffraction angles between 1° and 80° [22].
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Nuclear Magnetic Resonance Spectroscopy (NMR)
DOX, AGNP, and AGNP-DOX were dissolved in deuterium oxide (D2O), and 1H NMR spectra were obtained using NMR spectrometers (AVANCE II 400, Bruker, Switzerland) at a frequency of 400 MHz [20].
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Differential Scanning Calorimetry (DSC)
Melting temperatures of DOX and AGNP-DOX were determined by heating (~5 mg) samples in covered aluminium pans from 200 to 400°C for 10 minutes each, using a differential scanning calorimeter (DSC - 7020 Hitachi, Japan) in an inert nitrogen environment. The heat flow graph was plotted as a function of temperature [21].
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Field Emission Scanning Electron Microscopy (FE-SEM)
The morphology of AGNP-DOX was examined using a Field Emission Scanning Electron Microscope (FE-SEM) (Supra 55, Carl Zeiss, Germany) with a tungsten filament. A transmission electron microscope (TEM) (JEM-2100Plus, Jeol, Japan) was employed to measure the size of the colloidal nanoparticles. The sample was drop-cast onto a copper grid covered with carbon, allowed to air dry at room temperature, and stained with 2% uranyl acetate [22].
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High-Resolution Transmission Electron Microscopy (HR-TEM)
A drop of the sample was placed on a copper grid coated with carbon, air-dried at room temperature, and stained with 2% uranyl acetate. High-resolution transmission electron microscopy (JEM-2100Plus, Jeol, Japan) was used to determine the nanoparticle size by averaging the sizes of 10 nanoparticles [21].
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In-Vitro Drug Release Studies
During the in-vitro drug release studies, dialysis membranes with a molecular weight cut-off (MWCO) between 12,000 and 14,000 Da were used. A sealed dialysis bag containing 5 mg of AGNP-DOX was immersed in 250 ml of release medium, which included pH 5.0 sodium acetate buffer and pH 7.4 phosphate buffer saline, at 37±1°C. Samples of 5 ml were taken at various intervals for up to 72 hours, and an equal amount of fresh medium was added to maintain volume. The drug content in the samples was determined using a UV/Vis spectrophotometer (UV-1800, Shimadzu, Japan) at 268.00 nm [20].
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In- Vitro Cytotoxicity Assay of Drug Loaded Nanoparticles
The in vitro cytotoxicity of DOX-AGNPs against the human breast cancer MDA-MB-231 cell line was assessed using the MTT assay, as previously described [21]. Cells were treated with DOX and DOX-AGNPs at concentrations ranging from 1 to 50 µg/mL. The results were compared with the cytotoxicity of pure DOX to evaluate the enhanced efficacy of DOX-AGNPs.
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Cellular uptake study
The AGNP was fluorescently labeled with FITC by replacing DOX and adding FITC. For qualitative cellular uptake studies, MCF-7 and MDA-MB-231 cells were seeded in 6-well plates. Cells were incubated with AUGNP-FITC nanoparticles for 6 hours, at an equivalent FITC concentration (0.1 mg/mL). Finally, cellular uptake was observed with a fluorescent microscope (Nikon, Japan).
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Stability Study
The stability of AGNP-DOX was evaluated over six months at room temperature (25±1°C). The physical stability was visually inspected, and the polydispersity index and mean particle size were measured [21].
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Results
Preparation and Optimization of Colloidal Gold Nanoparticles
Acacia gum served as both a stabilizing and reducing agent during the chemical reduction process used to produce colloidal gold nanoparticles. The hydrodynamic particle size was optimized at a specific ruby-red color with a 1% w/v gum concentration, as shown in [Table 1]. The average hydrodynamic colloidal particle size of AGNP-DOX was found to be 86.63 nm. The nanoparticles exhibited considerable polydispersity, with a value of 0.442±0.37, indicating a wide range of particle sizes. The high surface potential of − 15.4±1.88 mV suggested the presence of gum molecules on the surface of the nanoparticles and their excellent stability. This negative charge is due to polysaccharides and glycoproteins associated with arabinose and ribose. The increased particle size, polydispersity, and surface charge to 86.63 nm, 0.442±0.37, and − 15.4±1.88 mV upon DOX loading indicate the interaction between the positively charged amine group of DOX and the negatively charged acidic gum group ([Table 2] and [Fig. 2]).
|
S. No |
Gum concentration (% w/v) |
Particle Size (nm) |
Polydispersity Index |
Zeta Potential(mV) |
|---|---|---|---|---|
|
1. |
0.25% w/v |
227.5 nm±15.24 |
0.275±0.98 |
− 12.5±2.11 mV |
|
2. |
0.50% w/v |
69.14 nm±9.57 |
0.459±0.74 |
− 9.38±2.35 mV |
|
3. |
0.75% w//v |
89.57 nm±5.29 |
0.452±0.56 |
− 15.9±1.47 mV |
|
4. |
1% w/v |
78.15 nm±2.12 |
0.325±0.19 |
− 23.6±1.21 mV |
|
5. |
2%w/v |
60.38 nm±3.56 |
0.505±0.24 |
− 13.3±4.29 mV |
|
6. |
3%w/v |
63.20 nm±2.29 |
0.540±0.37 |
− 6.91±3.56 mV |
*Results are expressed as Mean±SD, n=3.
|
Sample Code |
Gum concentration (% w/v) |
Particle Size (nm) |
Polydispersity Index |
Zeta Potential (mV) |
|---|---|---|---|---|
|
AGNP |
1% w/v |
79.91±2.14 |
0.438±0.19 |
− 20.6±1.21 mV |
|
AGNP-DOX |
1% w/v |
86.63±4.28 |
0.442±0.37 |
15.4±1.88 mV |
*Results are expressed as Mean±SD, n=3.
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Drug Loading
The drug content of the samples was analyzed using a UV/Vis spectrophotometer (UV-1800, Shimadzu, Japan) set to measure the capped DOX on the surface of AuNPs at a wavelength of 268 nm. The analysis revealed that the AGNP surface contained 2.567±0.19 mg/mL of DOX. A photographic image of the doxorubicin-loaded acacia gold nanoparticles (AGNP-DOX) under natural light is shown in [Fig. 3].
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Characterization of Acacia Stabilized Colloidal Gold Nanoparticles
UV/Vis absorption spectroscopy, FTIR, PXRD analysis, DSC analysis, and NMR spectroscopy were used to assess DOX, AGNPs, and AGNP-DOX.
UV–Vis Spectrometry (UV)
The maximum absorption wavelength (λmax) of the Au solution in the UV spectrum is 290.5 nm. DOX exhibited a peak at 479.00 nm, while AGNP-DOX showed its highest absorbance at 487 nm. As shown in [Fig. 4], DOX demonstrated λmax at 479.00 nm and 487 nm, indicating the adsorption of DOX onto the surface of acacia stabilized gold nanoparticles.
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Fourier Transforms Infrared Spectroscopy (FTIR)
The FT-IR spectra of pure DOX were shown in [Fig. 5], where the hydroxyl group broad band vibration at 3387.62 cm−1, (C-H) stretching at 2934.34 cm-1, and NH Bending vibration at 1615.91 cm-1, 1210.04 cm-1, 1033.92 cm-1were visible. The same suggestive DOX peaks, however, were shown by AGNP-DOX, indicating successful drug adsorption at the AGNP surface.
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Powder X – Ray Diffraction (P-XRD)
P-XRD patterns were utilized to confirm the crystalline nature of AGNP-DOX and native DOX. The distinct crystalline character of native DOX was evidenced by sharp peaks in the 10° to 40° range. The adsorption of DOX on the surface of AuNPs was demonstrated by a reduction in the intensity of these crystalline peaks in DOX ([Fig. 6]).
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Nuclear Magnetic Resonance Spectroscopy (NMR)
The most reliable indicator of the spatial proximity is provided by NMR spectroscopy, which looks at the intermolecular dipolar cross interactions between the host and guest molecules. The 1H NMR spectra of AG, AGNP, DOX, and AGNP-DOX—all of which were acquired in D2O—are shown in [Fig. 7]. In the spectrum ([Fig. 7a]), AG signals one peak (1H, tetra-CH2) at (2.128 ppm), (3.490 ppm), and (1.236, 1.214, 1.167, 1.147 ppm), as well as two peaks (1H, singlet-OH, CH2). Two peaks (H-1, singlet-OH, CH2) and one peak (H-2, doublet-CH2) formed signals in the AGNP (Blank) spectra at 2.222 ppm, 3.482 ppm, and 1.260, 1.240 ppm. The AG tetra peaks combined to form a doublet in the metal ion complex, and the upper field moved accordingly. Since the protons (H-1 and H-2) of AGNP (Blank) lie beyond the structural cavity and the Au moleculeʼs exterior, they are thought to be the most often occurring binding sites in the metal ion complex. The metal ion complexʼs spectra also showed an interesting fluctuation of the H-1 and H-2 protons. According to the 1H NMR studies, AG was present on the Au outside cavity, and the proton pairs H-1 and H-2 most likely function as a binding site to stabilize the metal ion complex. Thus, these findings validate that the AG molecule is loaded into the external cavity of Au. (H-1, Tetra – OH), (H-1, Singlet-NH2), (H-1, Doublet-CH2), and (H-1, Singlet-CH3) appeared at (3.761, 3.712, 3.650, 3.622 ppm), (5.336 ppm), (2.130, 2.119 ppm), and (1.105 ppm) in the DOX spectrum ([Fig. 7b]). The AGNP-DOX signal of peaks (H-1, Tetra – OH), (H-1, Singlet-NH2), (H-1, Singlet-CH2), (H-1, Singlet-CH3), (5.039 ppm), (2.132 ppm), and (3.931, 3.666, 3.795, 3.647 ppm) may be found in the spectrum. DOX CH2-double peaks in AGNP nanocarriers are drug loaded and then shift into a single peak. It is clearly visible since the AG peaks cap the DOX CH2 peaks. In the AGNP-DOX formulation, the OH, NH2, CH2, and CH3 peaks of DOX are found to be the most totally binding sides with the AG since they are outside of the structural cavity. Due to the presence of AG-capped DOX peaks on the surface of the Au molecule, there is a slight shift in the proton signals, both upwards and downwards in ppm. Our data thus confirm the loading of DOX protons onto the external cavity of the AGNPs nanocarriers.
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Differential Scanning Calorimetric Analysis (DSC)
The DSC technique is an excellent technique for investigating temperature stability and phase transitions. The DSC thermograms of pure DOX and AGNP-DOX revealed endothermic transitions at 216°C and 108.718°C, respectively. The endothermic peak for DOX, typically found near its melting point of 195°C, was absent in the AGNP-DOX spectra, as shown in [Fig. 8a, b]. This absence indicates that DOX was present as a molecular or amorphous dispersion rather than precipitating on the nanoparticleʼs surface.
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Surface Morphology
FE-SEM analysis confirmed the surface morphology of AGNP-DOX, as depicted in [Fig. 9a], revealing that AGNP-DOX particles are spherical. HR-TEM, shown in [Fig. 9b], further assessed the surface morphology and actual particle size, indicating that the colloidal nanoparticles are spherical and approximately 20 nm in size. The corresponding selected area electron diffraction (SAED) pattern, presented in [Fig. 9c], displays clear diffraction spots. This pattern suggests an electrostatic interaction between DOX and AGNP-DOX, confirming the presence of DOX on AGNP-DOX.
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In Vitro Drug Release Studies
The In vitro drug release experiments conducted in phosphate buffer saline (pH 7.4) and sodium acetate buffer (pH 5.0) are illustrated in [Fig. 10]. The drug was completely released from the colloidal Au NPs within 72 hours. AGNP-DOX release was higher in sodium acetate buffer (96.23%) compared to phosphate buffer saline (92.21%). The pH 5.0 sodium acetate buffer is designed to mimic the internal environment of cancer cells, while the pH 7.4 phosphate buffer simulates blood plasma conditions.
The faster release at acidic pH is indicative of the potential for targeted drug release within the acidic tumor microenvironment, which could reduce systemic toxicity and improve the therapeutic index. However, the release of drugs from gold nanoparticles (AuNPs) was faster in acidic pH than in neutral because in an acidic environment the amine group in AuNPs is protonated causing cleavage the bonds between the DOX and AuNPs. Thus, the results show that DOX-AuNPs were able to particularly stimulate the delivery of drugs to cancer cells. Previous studies also reported rapid release of chemotherapeutic drug loaded on AuNPs in acidic conditions when compared to neutral conditions.
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In Vitro Cytotoxicity of DOX and DOX-AGNP
[Fig. 11] presents the results of the MTT assay conducted on MDA-MB-231 human breast cancer cells treated with varying concentrations (1–50 µg/mL) of DOX and DOX-AGNP. The data show that cytotoxicity increased with concentration for both formulations. At the highest concentration (50 µg/mL), cytotoxicity reached approximately 90% for both DOX and DOX-AGNP. Notably, DOX-AGNP exhibited slightly higher cytotoxicity than DOX alone at lower concentrations, indicating a potentially enhanced therapeutic effect. This improved efficacy of DOX-AGNP could be attributed to the increased cellular delivery of DOX facilitated by the gold nanoparticles.
Conversely, the cell viability assay displayed an inverse relationship to cytotoxicity. As the concentration increased, the viability of MDA-MB-231 cells decreased for both DOX and DOX-AGNP formulations. At the lowest concentration (1 µg/mL), cell viability was around 80% for both, but it dropped to below 10% at 50 µg/mL. The DOX-AGNP formulation consistently exhibited slightly lower cell viability compared to DOX alone, particularly at lower concentrations, suggesting a more potent cytotoxic effect when combined with acacia-stabilized gold nanoparticles.
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Cellular uptake study
In vitro cellular uptake of AGNPs loaded with FITC was investigated in MDA-MB-231 human breast cancer cell lines. Fluorescence microscopy revealed green fluorescence, indicating the successful internalization of AGNP-FITC in the cells at both 1 hour and 6 hours, as shown in [Fig. 12].
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Stability Studies
The colloidal stability of AGNP-DOX was assessed over a period of six months at 25±1°C. Initial measurements on day 1 indicated that AGNP-DOX particles had an average particle size of 86.63±1.13 nm and a PDI of 0.442±0.23. After six months, the particle size increased to 437.0±1.11 nm, and the PDI rose to 0.761. These changes in particle size and PDI suggest that the colloidal AGNP-DOX is unstable at 25±1°C showed in [Table 3].
|
Sample |
Particle size (nm) |
Polydispersity Index (PDI) |
||
|---|---|---|---|---|
|
Day 1 |
After 6th months |
Day 1 |
After 6th months |
|
|
AGNP-DOX |
86.63 nm |
437.0 nm |
0.442 |
0.761 |
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Discussion
Traditional chemotherapy treatments, such as those utilizing doxorubicin (DOX), often face significant challenges including systemic toxicity and the development of drug resistance [11]. Doxorubicin, an anthracycline drug, is widely used for treating various cancers including breast cancer, but its efficacy is limited by these issues [9]. DOX works by intercalating DNA and inhibiting topoisomerase II, leading to chromatin damage and cell cycle arrest [10]. However, its non-specific distribution results in severe side effects and limits the maximum tolerated dose [9].
Nanotechnology offers potential solutions to the limitations of traditional chemotherapy by enabling targeted drug delivery. Various nanoparticles, including those made from gold, silver, platinum, and other materials, have been explored for their ability to improve drug delivery, imaging, and therapeutic outcomes [5]. Gold nanoparticles (AuNPs) are particularly favoured due to their unique optical-electronic properties, ease of synthesis, and biocompatibility. They can be functionalized with various molecules to enhance their stability and targeting capabilities [6] [7]. The environmental impact and sustainability of acacia gum is well known, especially within the context of green synthesis. Acacia gum, also known as gum arabic, is a natural biopolymer derived from the exudates of Acacia trees. Its use in nanoparticle synthesis is considered environmentally friendly due to several reasons. Acacia gum is a naturally occurring material and is renewable, derived from trees that do not require intensive chemical processes for extraction. Its production is often sustainable, particularly when sourced from regions practicing sustainable harvesting. One of the most significant advantages of using acacia gum is its biodegradability. Unlike synthetic stabilizers, it naturally breaks down in the environment without producing harmful byproducts. This minimizes the ecological footprint associated with its use. Acacia gum is non-toxic and safe for humans, animals, and the environment. It is even used in the food and pharmaceutical industries, adding to its safety profile.
The use of natural substances like proteins, polysaccharides, and glycoproteins as reducing and stabilizing agents in the synthesis of nanoparticles is gaining traction due to their biocompatibility and low toxicity [7]. Acacia gum (AG), a complex polysaccharide, is one such material that has been recognized as safe by the FDA and is widely used in the food and pharmaceutical industries as an emulsifier and stabilizer. Its use in nanoparticle synthesis leverages its biocompatibility and cost-effectiveness [13].
Previous studies have demonstrated various approaches to loading DOX onto different nanocarrier platforms. These include liposomes, polymeric nanoparticles, and other inorganic nanoparticles like iron oxide and carbon nanotubes. These carriers have shown improved drug delivery efficiency and reduced toxicity compared to free DOX. However, issues such as complex synthesis processes, potential toxicity of the carriers, and stability concerns remain [9] [17]. The green synthesis approach using Acacia gum (AG) presents a novel and eco-friendly method for producing stabilized gold nanoparticles (AGNPs). The synthesis involves a simple and rapid process where AG acts as both the reducing and stabilizing agent. The use of AG ensures biocompatibility and reduces the environmental impact compared to traditional chemical synthesis methods [16] [18].
The development of AGNP-DOX combines the advantages of green-synthesized gold nanoparticles with the therapeutic efficacy of doxorubicin. This approach leverages the stabilizing properties of AG to produce a biocompatible and stable nanocarrier. The optimized concentration of AG (1% w/v) and the synthesis conditions (80±1°C for 30 minutes) result in nanoparticles with a hydrodynamic diameter of approximately 79.91 nm and a zeta potential of -20.6 mV, indicating good stability.
The cytotoxicity assays further emphasize the potential advantages of DOX-AGNPs. Compared to pure DOX, the nanoparticle formulation exhibited enhanced anti-proliferative effects against human breast cancer MDA-MB-231 cells. This improvement is likely due to the increased cellular uptake and retention of DOX within the target cells, facilitated by the nanoparticle delivery system.
Colloidal stability studies revealed that DOX-AGNPs exhibit good stability at 4±1°C, with minimal changes in particle size and surface charge over a period of 6 months. However, stability decreases at higher temperatures, indicating the need for controlled storage conditions to maintain the integrity of the nanoparticles. These findings provide essential insights into the handling and storage of DOX-AGNPs for future applications.
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Conclusion
The current study successfully produced and characterized AGNPs for DOX. Colloidal AGNP-DOX improved DOXʼs water solubility and drug release rate; this effect may have been caused by DOX being delivered on gold carrier. Enhanced drug release profile, enhanced water solubility, and a notable increase in particle size all contributed to an improvement in DOX targeting efficiency. When combined, the studyʼs findings may be useful in the creation of pharmaceutical and biopharmaceutical formulations.
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Acknowledgement
The authors are grateful to the Integral University Faculty of Pharmacy for providing all the necessary facilities required for the present work (Manuscript Communication Number: IU/R &D/2024-MCN0002794).
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- 20 Ahmad N, Bhatnagar S, Saxena R. et al. Biosynthesis and characterization of gold nanoparticles: Kinetics, in vitro and in vivo study. Mater Sci Eng C Mater Biol Appl 2017; 78: 553-564
- 21 López-Miranda JL, Molina GA, Esparza R. et al. Green Synthesis of Homogeneous Gold Nanoparticles Using Sargassum spp. Extracts and Their Enhanced Catalytic Activity for Organic Dyes. Toxics 2021; 9: 280
- 22 Soliman MKY, Salem SS, Abu-Elghait M. et al. Biosynthesis of Silver and Gold Nanoparticles and Their Efficacy Towards Antibacterial, Antibiofilm, Cytotoxicity, and Antioxidant Activities. Appl Biochem Biotechnol 2023; 195: 1158-1183
Correspondence
Publication History
Received: 29 July 2024
Accepted: 15 September 2024
Article published online:
08 October 2024
© 2024. Thieme. All rights reserved.
Georg Thieme Verlag KG
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