Visible light photocatalytic degradation and biological activities of Aegle marmelos-mediated cerium oxide nanoparticles

Santhoshkumar Jayakodi; Mohana Sriramulu; Mani Govindasamy; Elumalai Pandian; Rajeshkumar Shanmugam; Fuad Ameen

doi:10.1515/gps-2024-0060

Article Open Access

Visible light photocatalytic degradation and biological activities of Aegle marmelos-mediated cerium oxide nanoparticles

Santhoshkumar Jayakodi , Mohana Sriramulu , Mani Govindasamy , Elumalai Pandian , Rajeshkumar Shanmugam and Fuad Ameen

Published/Copyright: November 26, 2024

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From the journal Green Processing and Synthesis Volume 13 Issue 1

Abstract

This study investigated the antibacterial and photocatalytic properties of cerium oxide nanoparticles (CeO₂ NPs), which were synthesized using Aegle marmelos extract. CeO₂ NPs were characterized by different spectroscopy methods such as ultraviolet-visible spectroscopy, powder X-ray diffraction, and Fourier transform-infrared spectroscopy. The shape and elemental analysis of synthesized CeO₂ NPs were evaluated using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. Further, the size and stability of the synthesized CeO₂ NPs were characterized by ZETA potential and dynamic light scattering analysis. Further, the photocatalytic efficiency of synthesized NPs was verified by 98% of methylene blue dye degradation exhibited within 180 min at the visible region and the first-order kinetic value was 0.2038 min⁻¹. The green synthesized CeO₂ NPs were found to have significant antibacterial activity against bacterial organisms. The effective antibacterial activity of E. coli (30 ± 0.02), P. aeruginosa (30 ± 0.07), S. aureus (24 ± 0.04), and S. epidermis (29 ± 0.04) at 100 µg·mL⁻¹. Therefore, Aegle marmelos CeO₂ NPs were used as a photocatalytic and antimicrobial therapeutic agent. The synthesized CeO₂ NPs also displayed enhanced anticancer efficacy with higher concentrations against MCF-7 cells. Thus, CeO₂ NPs have antibacterial, anticancer, and photocatalytic properties, according to this study.

Keywords: Aegle marmelos extract; DLS analysis; methylene blue; visible light; antibacterial activity

1 Introduction

Cerium oxide nanoparticles (CeO₂ NPs) have a widespread diversity of applications in fuel cells [1], photocatalyst [2], sunscreens [3], phosphorescent materials [4], solar cells [5], and sensor [6]. The oxidation states of cerium, a rare earth element, range from Ce(iii) to Ce(iv). CeO₂ NPs enable them to function as regenerative catalysts by converting between the Ce(iii) and Ce(iv) oxygen states [7]. Dyes from textiles and paper are the main examples of water pollution. Recently, organic dyes discharged from industrial wastewater have been determined to cause severe health issues [8]. Methylene blue is extensively used as colorant in water industries. Methylene blue has been described as a cancer-causing agent [9], Which produces various environmental issues through discharging disease-causing toxic substances into water. Therefore, there is an urgent requirement for an effective method for the degradation of methylene blue dye [10]. Numerous techniques are available for the polluted water treatment used for toxic dye degradation such as physical techniques, electrochemical techniques, photocatalytic degradation, and biodegradation. Among these techniques, dye degradation through photocatalytic activity is one of the most noticeable techniques [11]. Various metal catalysts like CuO, ZnO, TiO₂, ZrO₂, and CeO₂ NPs are mainly used for methylene blue dye degradation [12,13,14,15]. Among these, CeO₂ NPs play a major role as environmental photocatalysts due to their excellent redox capacity, low cost, nontoxicity, and long-term stability [16].

CeO₂ NPs have a significant physico-chemical characterization and antibacterial activity. The release of cerium ions and stability are the key features of their activity [17]. Various literature reporting the study of antibacterial activity and antimicrobial activity of CeO₂ NPs is available[18,19]. Several studies have reported the synthesis of CeO₂ NPs using various sources, stabilizing agents, solvents, and reactants. The added organic solvents directly react with the stabilizing agent in the synthesis process of CeO₂ NPs [20]. We report Aegle marmelos-mediated CeO₂ nanoparticles (NPs), which have been verified to exhibit proficient photocatalytic, antibacterial, and anticancer properties due to their non-toxicity, stability, physicochemical properties, and optimal size.

Aegle marmelos leaves are one of the sources of natural products used in medicinal and ayurvedic products. These leaves are used to treat neurological diseases, dyspepsia, dysentery, oedema, and rheumatism [21]. Also, Leaves of the Aegle marmelos contains alkaloids, phenylethyl cinnamates, rutin, mermesinin, aegelinosides, anhydromarmeline, and sterols [22]. The phytochemical compounds of these leaves are used to treat diarrhoea, peptic ulcers, dysentery, and laxative for causticity [23]. In this study, Aegle marmelos leaves extracts with medicinal values were used for the synthesis of NPs. The synthesized NPs potential was determined using dynamic light scattering (DLS) analysis. A study was also conducted on the antibiotic activity of microorganisms, the anticancer activity of bacteria, and the photocatalytic activity of methylene blue dye degradation.

2 Experimental methods

2.1 Synthesis of NPs

CeO₂ NPs were synthesized using Aegle marmelos leaves. These leaves were boiled in distilled water and stirred for 20 min to get brownish Aegle marmelos leaf extract. 10 mL of Aegle Marmelos leaf extract were combined with 2 g cerium nitrate, which was weighed and stirred for about 30 min at 60℃.

2.2 Characterization

The obtained CeO₂ NPs was established by using ultra violet visible spectroscopy (Jasco V670 Spectrophotometer). The crystalline structure of the CeO₂ NPs was examined using powder x-ray diffraction (XRD; D8 Advance BRUKER). The functional molecules of CeO₂ NPs were studied using a Fourier transform-infrared (FTIR) spectrophotometer (Shimadzu IR affinity 1). Scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy (SEM-EDAX; ZEISS EVO18) was used to examine the NPs’ morphology and elemental makeup. By using DLS, the stability and particle size of the CeO₂ NPs were assessed (Horiba Scientific SZ-100).

2.3 Antimicrobial activity

Gram-negative (E. coli and P. aeruginosa) and gram-positive (S. aureus and S. epidermis) bacteria were separated, sub cultured on nutrient broth, and then incubated at 37°C for 24 h. Using a sterile L rod, a bacterial cell from the nutrient broth was injected into a nutrient agar plate. Nutrient agar was mixed with 30 µg·mL⁻¹ of NPs, antibiotic, and negative control (water), and incubated for 24 h at 37°C. Bacterial inhibition was measured after a 24 h period in order to investigate the antibacterial activity.

2.4 Photocatalytic experiment

An electric tungsten lamp of 500 W was used to evaluate photocatalytic activity in visible light. CeO₂ NPs were mixed with 50 mL of methylene blue solution (10 ppm). The NPs and dye-containing solution were stirred for 1 h, finally, that solution reached the equilibrium between methylene blue dye and the suspension. The photodegradation of methylene blue was performed in a photoreactor under visible light. A volume of 3 mL was extracted for the ultraviolet examination at regular intervals. Ultraviolet-visible (UV) spectrophotometer was used to analyse the dye solution. In order to determine the degradation of methylene blue dye, the following formula was used:

(1) Photocatalytic activity = C 0 − C t / C 0 × 100

where C ₀ is the initial concentration of methylene blue dye and C _t is the methylene blue dye degradation at various time t.

2.5 Anti-cancer activity of CeO₂ NPs

MTT test was used to assess the cytotoxic activity of CeO₂ NPs mediated by Aegle marmelos. First, 96 wells were established with around 5 × 10⁻⁵, and various concentrations of CeO₂ NPs (50–500 μg·mL⁻¹) were added; in the meantime, the control wells received no supplementation. Twenty microlitres of MTT stain were combined following a 24 h incubation period. After 4 h of incubation at 37°C, the microtiter plate was exposed to 150 μL of DMSO, and the absorbance was measured calorimetrically using 570 nm as the standard wavelength. By calculating the 50% inhibitory concentration, these values were additionally utilized to assess the percentage of cell viability.

3 Result

3.1 UV-visible spectroscopy

The optical band gap of CeO₂ NPs were estimated using Tauc’s relation in Eq. 2

(2) α h γ = A ( h γ − E g )

where α is the absorption coefficient, h is the Plank’s constant, γ is the photon frequency, A is a constant, and E _g is the optical bandgap. The bandgap of Aegle marmelos-mediated CeO₂ NPs is 2.52 eV. The band gap energy of commercially available CeO₂ nanoparticles was estimated to be 2.7 eV shown in Figure 1. The reduced band gap energy value represents the oxygen values present in the green synthesised NPs.

Figure 1

UV-DRS analysis of Aegle marmelos analysis of CeO₂ NPs.

3.2 Powder XRD analysis

The XRD pattern of CeO₂ NPs displays four major peaks at around 28.78, 33.26, 47.59, and 58.02^o corresponding to (111), (200), (220), and (311) orientation planes. The observed XRD diffraction pattern of CeO₂ NPs is reliable with JCPDS file No#89-8436. No extra peaks appear from the XRD pattern on behalf of the high purity of the CeO₂ NPs. The broad XRD diffraction peaks are shown in Figure 2 depicting that the Aegle marmelos synthesized CeO₂ NPs are nanosized.

Figure 2

Powder XRD analysis of Aegle marmelos analysis of CeO₂ NPs.

3.3 FTIR analysis

The FTIR spectra of CeO₂ NPs revealed absorption peaks at 3,421, 1,562, 1,369, 1,150, and 838 cm⁻¹ shown in Figure 3. Stretching vibrations of Ce–O–Ce are accountable for the peak at 800–400 cm⁻¹. The C–N is responsible for the peaks at 1,150 cm⁻¹. The stretching vibration of C–O is responsible for the absorbance peak at 1,369 cm⁻¹ [29]. In this case, the C–H bond corresponds to the peak at 1,562 cm⁻¹. Stretching vibration at 3,421 cm⁻¹ confirmed the OH group, which is in good agreement with the earlier published literature [21].

Figure 3

FTIR analysis of Aegle marmelos analysis of CeO₂ NPs.

3.4 SEM and EDAX analysis

Figure 4b shows the EDAX spectrum of CeO₂ NPs. The elemental composition and morphological characteristics of CeO₂ NPs were investigated using SEM with EDAX analysis. The SEM image of the CeO₂ nanoparticles demonstrated that the particles were agglomerated and exhibited a relatively uniform distribution across the surface. Additionally, the nanoparticles displayed irregular shapes, often forming clusters due to aggregation (Figure 4a). EDAX analysis confirmed the presence and elemental composition of cerium (Ce) and oxygen (O) in the synthesized CeO₂ nanoparticles, verifying successful synthesis and purity.

Figure 4

(a) SEM analysis and (b) EDAX analysis of Aegle marmelos synthesized CeO₂ NPs.

3.5 Transmission electron microscopy (TEM) analysis

TEM was used to examine the shape and structure of the CeO₂ NPs (Figure 5a and b). The produced CeO₂ NPs exhibited a variety of morphologies, including cubic and spherical shapes, with diameters ranging from 15 to 30 nm and averaging approximately 25 nm.

Figure 5

TEM images analysis of (a) 50 nm (b) 100 nm of CeO₂ NPs.

3.6 Zeta potential and DLS analysis

Dynamic light scattering was employed to analyse the particle size of the synthesized CeO₂ NPs depicted in the EDAX spectrum. The size of CeO₂ NPs spans from 2 to 20 nm, as illustrated in Figure 6b. The zeta potential value of the Aegle marmelos-mediated CeO₂ nanoparticles was measured at –3 mV, which generally indicates that the synthesized nanoparticles are stable (Figure 6a).

Figure 6

(a) Zeta potential and (b) DLS analysis of Aegle marmelos synthesized CeO₂ NPs.

3.7 Antibacterial activity

The microbial ZOI assay was used to examine the antibacterial properties of CeO₂ NPs against E. Coli, P. aeruginosa, S. aureus, and S. epidermis. When 100 µg·mL⁻¹ of gram-negative pathogens (E. Coli and P. aeruginosa) were present, a ZOI measuring approximately 30 ± 0.02 mm and 30 ± 0.07 mm was found. At concentrations of 25 and 50 µg·mL⁻¹, the nanoparticles exhibited zones of inhibition (ZOI) of 24 ± 0.14 mm and 26 ± 0.01 mm against E. coli, and 14 ± 0.05 mm and 18 ± 0.01 mm against P. aeruginosa, respectively (Figure 7). The data from Table 1 clearly displays that the increase in the concentration of CeO₂ NPs simultaneously increased the ZOI values gram-negative microorganisms. In comparison to other gram-positive bacteria, the inhibition values of both types were significantly higher. They show greater inhibitory action in gram-positive bacteria including S. aureus and S. epidermis. The CeO₂ NPs at 100 µg·mL⁻¹ concentration of S. aureus (24 ± 0.04) revealed the ZOI. Further, S. epidermis exhibited the ZOI at (29 ± 0.04). The 25 and 50 µg·mL⁻¹ concentration of CeO₂ NPs displayed the lowest ZOI when compared to 100 µg·mL⁻¹ of CeO₂ NPs. The chloramphenicol antibiotic values are displayed in Table 1. This experiment demonstrated unequivocally how the concentration of CeO₂ NPs affects the ZOI.

Figure 7

Antibacterial activity of (a) Aegle marmelos synthesized of CeO₂ NPs and (b) standard.

Table 1

Antibacterial activity of synthesized CeO₂ NPs

S. no.	Organisms	ZOI
S. no.	Organisms	CeO₂ NPs (5 mg·mL⁻¹)			Chlormaphenicol (5 mg·mL⁻¹)
1	E. coli	25 µg·mL⁻¹	50 µg·mL⁻¹	100 µg·mL⁻¹	25 µg·mL⁻¹	50 µg·mL⁻¹	100 µg·mL⁻¹
1	E. coli	24 ± 0.05	26 ± 0.01	30 ± 0.02	28 ± 0.05	30 ± 0.02	32 ± 0.02
2	P. aerogenosa	14 ± 0.05	18 ± 0.01	30 ± 0.07	32 ± 0.07	34 ± 0.01	38 ± 0.02
3	S. aureus	12 ± 0.01	20 ± 0.02	24 ± 0.04	24 ± 0.02	28 ± 0.03	30 ± 0.05
4	S. epidermis	22 ± 0.02	25 ± 0.02	29 ± 0.04	27 ± 0.05	29 ± 0.04	32 ± 0.01

3.8 Photocatalytic activity

The synthesized CeO₂ NPs were evaluated for their photocatalytic performance under visible light conditions, as depicted in Figure 8a. A photocatalytic degradation experiment of methylene blue dye using CeO₂ NPs was carried out, and the process was completed within 180 min. Further, first-order kinetics (0.2038 min⁻¹) were studied for methylene blue dye degradation using CeO₂ NPs and the rate of constant is shown in Figure 8b.

Figure 8

Photocatalytic activity of Aegle marmelos synthesized CeO₂ NPs. (a) UV-Visible analysis of photocatalytic activity and (b) Kinetics studies of CeO₂ NPs.

3.9 Anticancer activity

The in vitro cytotoxic activity of CeO₂ NPs synthesized by Aegle marmelos was evaluated using MCF-7 cell lines. According to the interpretation of the results, the proliferation of the cells decreased as the concentration of NPs increased. The cell viability shown by synthesized CeO₂ NPs was 78% at lower concentrations and 39% at higher concentrations (Figure 9a). The morphological examination of MCF-7 cells lines subjected to both treated and untreated conditions with CeO₂ NPs is depicted in Figure 9b. Within this figure, MCF-7 cell lines treated with NPs displayed irregular shapes, distortion, and shrinkage compared to their untreated counterparts. Conversely, no discernible changes were evident in the untreated MCF-7 cell lines. As a result, CeO₂ NPs mediated by Aegle marmelos had a potent growth suppression effect that was comparable to that of a typical drug.

Figure 9

(a) MTT Assay and (b) Control and test MCF-7 cell lines.

4 Discussion

Ravishankar et al. [24] reported CeO₂ NPs using solution combustion and band gap energy (4.1\eV). Arumugam et al. [25] reported that the particle size of the CeO₂ NPs controlled by the charge transition of cerium ions may play the main role in the decrease and increase in the band gap energy of synthesised CeO₂ NPs. So, the band gap energy values depend on the synthesis method. Previous researchers suggested reduction in band gap energy significantly increases the photocatalyst activity. Accordingly, the Aeges marmelos synthesized CeO₂ NPs were used for the significant photocatalytic activity. The obtained XRD peaks of CeO₂ NPs were in good agreement with that reported by Tamizhdurai et al. [26]. The crystalline size was calculated by using Scherrer equation [27,28] and the crystalline size of the CeO₂ NPs is 25 nm. The functional groups from the A. marmelos employed for the manufacture of CeO₂ NPs are confirmed by the FTIR analysis. The exposed crystal planes on CeO₂ NPs surface were visible in high-resolution TEM pictures, indicating the ceria NPs’ highly crystalline structure. The corresponding spherical diameter of a sphere with a volume equal to the particle size was used to characterize the particle size. The average particle size of the verticillium-mediated CeO₂ NPs, as reported by Mukherjee et al. [30], was 25 nm. Additionally, the average particle size of CeO₂ NPs synthesized using Penicillium sp. was found to be 60 nm [31]. According to Gopinath et al. [32], Aspergillus niger-mediated CeO₂ NPs have a particle size ranging from 3 to 30 nm. Next, spherical-shaped A. terreus-mediated CeO₂ nanoparticles were studied, exhibiting sizes ranging from 1 to 20 nm. [33]. CeO₂ NPs antibacterial activity was mediated by their electrostatic contact through the bacterial cell wall [34]. This interaction controls the surface of the bacterial cell wall and generates reactive oxygen species (ROS) species which leads to bacterial cell death [32]. CeO₂ NPs demonstrated nearly comparable zones of inhibition against gram-positive and gram-negative bacteria in this study. Similar to this, Alpaslan et al. [34] looked into the suppression of gram-positive and gram-negative pathogens by pH-controlled CeO₂ NPs. This study demonstrated that gram-negative bacteria had significantly more antibacterial activity than gram-positive bacteria. These findings verified that the ZOI value increased together with the concentration of Aegle marmelos-mediated CeO₂ NPs. Sabouri et al. [35] studied Rheum turkestanicum extract-mediated CeO₂ NPs and its methylene blue dye degradation activity. 33% of dye degradation was achieved at 180 min in the existence of UV light. Meanwhile Sultan Irshad et al. [36] reported that effective methylene blue dye degradation by orange peel extract-mediated CeO₂ NPs was confirmed at 30 mins exposure to sunlight. The primary purpose of the photocatalytic activity of green-synthesized nanoparticles is attributed to their preparation through a green method. The enhanced discoloration rate of the photocatalyzed reaction was due to the introduction of hydroxyl ions into the nanoparticles [37]. To enhance the degradation efficiency, hydroxyl radicals were generated to regulate the photocatalyst surface as studied by Gnanaprakasam et al. [37]. Sepahvand et al. reported that the photocatalytic activity was an efficient and effective catalyst for dye degradation from the aqueous solution, because of the high degradation efficiency, easy recoverability, and good mineralization [38]. The biomolecules included in the sample extract were used to produce CeO₂ NPs, which are effective against MCF-7 cells [39]. CeO₂ NPs induced by extract demonstrated antitumor efficacy against A549 cells [40]. Apoptosis is triggered by the generation of ROS, activation of apoptosis pathways, and disruption of cellular haemolysis, mitochondrial dysfunction, cell cycle arrest, and DNA fragmentation are all suggested as possible causes of the CeO₂ NPs’ anticancer effect [41].

5 Conclusion

In this study, the leaves of Aegle marmelos were shown to be a good capping and reducing agent in the synthesis of CeO₂ NPs. The analysis of the synthesized nanoparticles revealed that CeO₂ NPs were formed and matched well with JCPDS #89-8436. The synthesized NPs showed a gap energy of 2.52 eV. The stability of the synthesized NPs was confirmed based on zeta potential measurements (−3 mV), while DLS analysis revealed a NP size of 12 nm. Remarkably, significant antibacterial activity of CeO₂ NPs against both gram-positive and gram-negative bacteria was observed. Additionally, significant photocatalytic activity of CeO₂ NPs was demonstrated after exposure to visible light following methylene blue dye treatment. The anticancer activity of CeO₂ NPs exhibited significant cell viability at higher concentrations. The synthesized CeO₂ NPs exhibited strong antibacterial and photocatalytic activities. We may conclude from the current research that the synthesized CeO₂ NPs mediated by Aeges marmoles have strong antibacterial, anti-cancer, and photocatalytic activities.

Acknowledgements

This research was funded by Researchers Supporting Project number (RSP2024R364), King Saud University, Riyadh, Saudi Arabia. Thank you to Department of Biotechnology, Saveetha School of Engineering, Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, India for providing the research facilities for this study.

Funding information: This research was funded by Researchers Supporting Project number (RSP2024R364), King Saud University, Riyadh, Saudi Arabia. The authors thank the Department of Biotechnology, Saveetha School of Engineering, Saveetha Institute of Medical and Technical Sciences, Chennai, Tamil Nadu, India for providing the research facilities for this study.
Author contributions: Santhoshkumar Jayakodi: formal analysis, methodology, investigation, writing – original draft, and supervision. Mohana Sriramulu: methodology, investigation, and writing – original draft. Mani Govindasamy: conceptualization and methodology. Elumalai Pandian: methodology, review, and formal correction. Rajeshkumar Shanmugam: review and editing. Fuad Ameen: review, funding, and formal correction.
Conflict of interest: Authors state no conflict of interest.
Data availability statement: The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Received: 2024-03-20

Accepted: 2024-10-12

Published Online: 2024-11-26

This work is licensed under the Creative Commons Attribution 4.0 International License.

Articles in the same Issue

https://doi.org/10.1515/gps-2024-0060

Keywords for this article

Aegle marmelos extract; DLS analysis; methylene blue; visible light; antibacterial activity

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BY 4.0

Visible light photocatalytic degradation and biological activities of Aegle marmelos-mediated cerium oxide nanoparticles

Article

Abstract

1 Introduction

2 Experimental methods

2.1 Synthesis of NPs

2.2 Characterization

2.3 Antimicrobial activity

2.4 Photocatalytic experiment

2.5 Anti-cancer activity of CeO2 NPs

3 Result

3.1 UV-visible spectroscopy

3.2 Powder XRD analysis

3.3 FTIR analysis

3.4 SEM and EDAX analysis

3.5 Transmission electron microscopy (TEM) analysis

3.6 Zeta potential and DLS analysis

3.7 Antibacterial activity

3.8 Photocatalytic activity

3.9 Anticancer activity

4 Discussion

5 Conclusion

Acknowledgements

References

Articles in the same Issue

Articles in the same Issue

Articles in the same Issue

2.5 Anti-cancer activity of CeO₂ NPs