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Review Article | DOI: https://doi.org/10.31579/2690-8816/163
1Professor, Department of Pharmaceutics, Sri Ramakrishna Institute of Paramedical Sciences, College of Pharmacy, Tamil Nadu, Coimbatore – 44, India.
2B. Pharm, Sri Ramakrishna Institute of Paramedical Sciences, College of Pharmacy, Tamil Nadu, Coimbatore – 44, India.
*Corresponding Author: J Bagya lakshmi. Professor, Department of Pharmaceutics, Sri Ramakrishna Institute of Paramedical Sciences, College of Pharmacy, Tamil Nadu, Coimbatore – 44, India.
Citation: J Bagya lakshmi, K Vibisha, M Vignesh, N Vijay, R Vijayalakshmi, et al, (2025), Green Synthesized Silver Nanoparticles of Artocarpus Heterophyllus Peel Extract and Its Evaluation for Anti-Inflammatory Activity, J Clinical Research Notes, 6(2); DOI:10.31579/2690-8816/163
Copyright: © 2025, J Bagya Lakshmi. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received: 18 March 2025 | Accepted: 26 March 2025 | Published: 01 April 2025
Keywords: artocarpus heterophyllus; silver nanoparticles;green synthesis; anti-inflammatory
The aim of the study was to formulate Artocarpus heterophyllus peel extract into silver nanoparticles using green synthesis method and evaluating its Anti-inflammatory activity. It has various pharmacological benefits like anti-inflammatory, carcinogenic, anti-tubercular, anti-diabetic, anti-infective, anti-fungal, anti-viral, anti-oxidant. Artocarpus heterophyllus peel was extracted using methanol by maceration method and synthesized into silver nanoparticles using 1Mm silver nitrate by green synthesis. Then the nanoparticle was evaluated using particle size, zeta potential, UV-visible, and visual examination. The in vitro anti-inflammatory protein denaturation method using egg albumin in green synthesized Artocarpus heterophyllus peel extract silver nanoparticle was eco-friendly, non- toxic when compared to synthetic anti- inflammatory drugs.
Silver Nanoparticles
The investigation of the therapeutic properties of AgNPs has allowed studies to be carried out on the mechanism for modulating cytokines. This research could allow the development of new treatments, define the effective therapeutic concentrations and indications for use as well as evaluate the cytotoxic, genotoxic and carcinogenic potential and associated environmental impact. AgNPs show a reduction in pro-inflammatory proteins and in the selective inhibition of the COX-2 pathway. It was concluded that AgNPs presents an anti-inflammatory action in vivo through mechanisms involving the increase of anti -inflammatory molecules.
Green Synthesis
Green synthesis is an emerging area in the field of science and provides both economic and environmental benefits. In this study, non- toxic reagents that are eco- friendly and biosafe are used. The use of herbal extracts in the green synthesis of metal oxide nanoparticles is an increasing approach.
Inflammation
Inflammation that can be triggered by a variety of factors including pathogens, damaged cells and toxic compounds. These factors may induce acute and chronic inflammatory responses in the heart, pancreas, lung, brain, intestinal tract and reproductive system potentially leading to tissue damage or disease. Both infectious and non -infectious agents and cell damage activate inflammatory cells and trigger inflammatory signaling pathways, most commonly the NK-B, MAPK and JAK STAT pathways. These signals activate LEUKOCYTE CHEMOTAXIS from the general circulation to sites of damage. These activated leukocytes produce cytokines that induce inflammatory responses.
Anti Inflammatory Activity
Anti-inflammatory drugs that inhibit H1 Histamine, COX enzyme and reduce the synthesis of prostanoids. Corticosteroids prevent the formation of both PGs and LTs. The inhibition of phospholipase A2 reduces arachidonic acid release and suppresses the inflammation.
Materials
Artocarpus heterophyllus peel powder
Chemicals Reagents
Silver nitrate (AgNO3), distilled water.
Equipment Required
Magnetic stirrer, UV-Visible spectrophotometer, Homogenizer, Malvern Zeta size analyzer
Procedure:
Sample Collection
Artocarpus heterophyllus peels were collected from Coimbatore district.
Authentication
The plant specimen was identified and authenticated from Tamil Nadu Agriculture university.
Drying and Pulverizing
The peels were collected and shade dried. It was chopped into small pieces and kept for extraction.
Preparation of extract of peel of Artocarpus heterophyllus
Artocarpus heterophyllus fruits in total were collected, and each fruit's surface was cleaned with water and allowed to air dry. After cutting each fruit, the peel and pulp were separated. Carefully, the rinds, skin, and spines were separated. All of the fruits' spine, and skin samples were taken, chopped into tiny bits, and then blended. Each smashed piece was weighed separately. For 72 hours, 300 g of each were soaked in a 1:5 ratio (100 gm in 500 ml) of seven distinct solvents (polar-ethanol, methanol, ethyl acetate, non-polar chloroform, n-hexane, dimethyl sulfoxide, and petroleum ether) at room temperature. Filtration was used to separate the supernatant fluid, and a hot water bath was used to dry the residue. It was taken for analysis since the ethanolic extraction yield from all three sections was determined to be higher.
Synthesis of Silver Nanoparticles
In a typical reaction procedure, the 1 mM (mM) of AgNO3 aqueous solution was prepared in 100 ml Distilled water in 250 ml beaker and was used to produce silver Nanoparticles. The 40 ml of 1 mM AgNO3 solution was taken and treated with 10 ml of peel extract filtrate and heated the mixture for 1 hour at 40-50°C for reduction into Ag ions. For 1 mM silver nitrate solution the leaf extract acts as a reducing and stabilizing agent. The resulting solution becomes grey-black in colour after 48 hours, indicating the formation of nanoparticles. The concentrations of peel extract were also varied from 4 ml to 10 ml leaves extract as well as temperature and time duration are also varied.
Preformulation Studies
Solubility
Solubility test of Artocarpus heterophyllus peel were tested using different solvents such as ethanol, methanol, water and the results were given below.
| SOLVENTS | SOLUBILITY |
| Water | Insoluble |
| Ethanol | Completely soluble |
| Methanol | Completely soluble |
Table 1: Solubility of Artocarpus heterophyllus peel extract
Characterization of Synthesized Artocarpus heterophyllus Silver Nanoparticle
Characterization of Artocarpus heterophyllus silver nanoparticles is done to evaluate the functional properties. Various analytical techniques are utilized to confirm their formation and evaluate their characteristics. These techniques include UV-Visible spectroscopy, particle size analysis and zeta potential measurement.
Visual Examination
The primary confirmation of the synthesized Artocarpus heterophyllus silver nanoparticles is done by visual basis. The colour change of reaction mixture (silver nitrate solution and peel extract) with respect to time is observed.
UV-VIS spectroscopy
UV-VIS spectroscopy is a valuable technique to characterize the transmission, absorption and reflectivity of several compounds and for scientifically important material, such as coating and pigments etc. The ultraviolet and visible spectrum have wide features that are of use for sample identification but are extremely helpful for quantitative measurements.
Determination of particle size
The dried powders of Artocarpus heterophyllus Silver Nanoparticles dispersed in water to obtain proper scattering intensity of Artocarpus heterophyllus silver nanoparticles. The particle size was determined by the Malvern zeta size analyser.
Determination of zeta potential
The zeta potential was measured by using Zeta Sizer (Malvern Instruments) having zeta cells,
polycarbonate cell with gold-plated electrodes and using water as medium for sample preparation. Zeta potential determines the surface potential of silver nanoparticles and it is essential for the characterization of stability of nanoparticles.
In-Vitro Methods
Anti-arthritic studies by protein denaturation method
In Vitro anti-arthritic assay was conducted by protein denaturation method given by Mizushima and Kobayashi (1968). The reaction mixture (5 ml) consisted of 0.2 ml of egg albumin, 2.8 ml of phosphate-buffered saline (PBS with pH 6.4), and 2 ml of varying concentrations (10, 20,50, 10 ug/L) of plant extract. A similar volume of double distilled water is used as control. Then the mixture was incubated at 37 °C in a BOD incubator for 15 min and then heated at 70 °C for 5 min. After cooling, their absorbance was measured at 660 m by using the vehicle as blank.
The percentage of inhibition of protein denaturation was calculated by using the following formula;
Inhibition = AC660-AT660 x 100 AC600
AC = Absorbance of control solution, AT = Absorbance of the test sample.
Egg Albumin Assay
The anti-inflammatory activity of unknown crude extracts can be determined in vitro for inhibition of the denaturation of egg albumin (protein).
* 0.2 mL of 1-2% egg albumin solution (from fresh hen's egg or commercially available egg albumin powder), 2 mL of sample extract or standard (methotrexate) at varying concentrations, and 2.8 mL of phosphate buffer saline (pH 7.4) were mixed to form a reaction mixture of a total volume of 5 ml.
* A total volume of 5 mL of the control was created by combining 2 mL of triple-distilled water, 0.2 mL of 1-2% egg albumin solution, and 2.8 mL of phosphate-buffered saline.
* The reaction mixtures were then incubated at 37±2°C for 30 min and will be heated in a water bath at 70±2°C for 15 min.
* After cooling, the absorbance was measured at 280 nm by a suitable UV/Vis spectrophotometer using triple distilled water as the blank.
* The following equation was used to determine the percentage inhibition of protein denaturation:
Percentage Inhibition = (Absorbance of control - Absorbance of test sample) / Absorbance of control * 100
* Then plant extract/positive control concentration for 50% inhibition (IC50) was determined by plotting percentage inhibition concerning control against concentration. All determinations were carried out in triplicate.
Characterization of green synthesized artocarpus heterophyllus silver nanoparticles
Visual Examination
The colour of Artocarpus heterophyllus extract in the initial stage was yellow, after the addition of silver nitrate solution it turned into brown colour. There is no colour change after 90minutes, indicating that the reaction is finished. After 48 hrs the colour change from yellow to brown indicates the formation of silver nanoparticles. The image of Artocarpus heterophyllus extract is depicted in figure 6: The image of Artocarpus heterophyllus extract is depicted in figure 7: which denotes the formation of silver nanoparticles after 48hrs.
UV VISIBLE Spectral Analysis OF Artocarpus heterophyllus Silver Nanoparticles
Silver nanoparticles production and completion are characterized by UV-Visible spectral analysis, which makes use of a UV-Visible spectrophotometer.
The production of silver ions to silver nanoparticles was indicated by colour change from yellow to brown colour and it was reflected in spectral data obtained by using UV-Visible spectrophotometer. The silver nanoparticles Surface Plasmon Resonance (SPR) is characterized by the coloration. It shows an absorption peak at 425nm, confirming the formations of Artocarpus heterophyllus silver nanoparticles.
Particle size measurement:
The particle size is one of the most important parameters for Characterization nanoparticles. The mean particle size(z -average), polydispersity index (PI) of Artocarpus heterophyllus silver nanoparticles were determined by using a zeta size analyser (Malvern Instrument). The average particle size of Artocarpus heterophyllus was found to be 60.45nm. Particle size analysis showed the presence of nanoparticles with polydispersity indices PDI value 0.356 with intercept 0.845.
Zeta Potential
Zeta potential was used to determine the stability of prepared nanoparticles. Zeta potential was determined using a zeta-sizer instrument. The surface charge of the particles and stability of the solution was characterized by zeta potential. For Artocarpus heterophyllus silver nanoparticles, zeta potential was found to be 0.304mV with peak area 46.9 intensity. These values indicate that Artocarpus heterophyllus silver nanoparticles are stable.
Invitro Anti- Inflammatory Activity
Anti- inflammatory activity of standard drug aspirin and the Artocarpus heterophyllus silver nanoparticles were determined using protein denaturation assay. The anti- inflammatory activity of unknown crude extracts can be determined by using in vitro for inhibition of the denaturation of egg albumin. Different concentrations of drug sample and egg albumin solution are taken and mixed with phosphate buffer on PH 7.4 and incubated for 30min. Reaction mixture was heated at the water bath for 70°C for 15minutes. After cooling, the absorbance was measured at 280 nm by a suitable UV/Vis spectrophotometer using triple distilled water as the blank. Aspirin is used as standard and the drug sample was compared with standard.
From the above standard drug is compared with our plant extract and the inhibition concentration value for inflammatory prepared nanoparticles of Artocarpus heterophyllus was slightly higher when compared to the standard drug aspirin.
It is believed that protein denaturation is the factor that promotes inflammation. Inhibition activity of Artocarpus heterophyllus silver nanoparticles against Rheumatoid were observed, hence it can be used as a potential alternative to synthetic drugs as a non-toxic safe way of green synthesis.
Figure 1: UV Visible spectrum of Formulation of Artocarpus heterophyllus Nanoparticles
Figure 2: Determination of Particle Size of Artocarpus heterophyllus Silver Nanoparticle
Figure 3: Determination of Zeta Potential of Artocarpus heterophyllus silver nanoparticles
Concentration (µg/ml) | Absorbance | % Inhibition | Average | ||||
| T1 | T2 | T3 | T1 | T2 | T3 | ||
| 10 | 0.0665 | 0.0652 | 0.0646 | 67.08 | 63.81 | 62.31 | 64.40 |
| 20 | 0.0618 | 0.0630 | 0.0627 | 58.27 | 58.29 | 57.03 | 57.03 |
| 30 | 0.0491 | 0.0496 | 0.0506 | 23.37 | 24.62 | 27.14 | 25.04 |
| 40 | 0.0659 | 0.0672 | 0.0661 | 65.57 | 68.84 | 66.08 | 66.82 |
| 50 | 0.0558 | 0.0573 | 0.0580 | 40.20 | 43.97 | 45.73 | 43.30 |
Table 2: Protein Inhibitory Data of Control Aspirin
Concentration (µg/ml) | Absorbance | % Inhibition | Average | ||||
| T1 | T2 | T3 | T1 | T2 | T3 | ||
| 10 | 0.0445 | 0.0450 | 0.0456 | 11.30 | 13.06 | 14.57 | 13.14 |
| 20 | 0.0623 | 0.0633 | 0.0634 | 56.53 | 59.05 | 59.55 | 58.37 |
| 30 | 0.0712 | 0.0725 | 0.0732 | 78.89 | 82.16 | 83.91 | 81.65 |
| 40 | 0.0841 | 0.0832 | 0.0830 | 111.30 | 109.04 | 108.54 | 109.62 |
| 50 | 0.1092 | 0.1097 | 0.1101 | 174.37 | 175.62 | 176.63 | 175.54 |
Table 3: Protein Inhibitory Data of Artocarpus heterophyllus Silver Nanoparticles
The study shows green synthesized silver nanoparticles of Artocarpus heterophyllyus has anti-inflammatory activity. These nanoparticles were characterized and evaluated by various test such as visual examination, UV-VIS Spectral analysis, zeta potential. Invitro anti-inflammatory activity was studied. Perform invivo methods for further confirmation of above mentioned property.
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To Dear Erin Aust, I would like to express my heartfelt appreciation for the opportunity to have my work published in this esteemed journal. The entire publication process was smooth and well-organized, and I am extremely satisfied with the final result. The Editorial Team demonstrated the utmost professionalism, providing prompt and insightful feedback throughout the review process. Their clear communication and constructive suggestions were invaluable in enhancing my manuscript, and their meticulous attention to detail and dedication to quality are truly commendable. Additionally, the support from the Editorial Office was exceptional. From the initial submission to the final publication, I was guided through every step of the process with great care and professionalism. The team's responsiveness and assistance made the entire experience both easy and stress-free. I am also deeply impressed by the quality and reputation of the journal. It is an honor to have my research featured in such a respected publication, and I am confident that it will make a meaningful contribution to the field.
"I am grateful for the opportunity of contributing to [International Journal of Clinical Case Reports and Reviews] and for the rigorous review process that enhances the quality of research published in your esteemed journal. I sincerely appreciate the time and effort of your team who have dedicatedly helped me in improvising changes and modifying my manuscript. The insightful comments and constructive feedback provided have been invaluable in refining and strengthening my work".
I thank the ‘Journal of Clinical Research and Reports’ for accepting this article for publication. This is a rigorously peer reviewed journal which is on all major global scientific data bases. I note the review process was prompt, thorough and professionally critical. It gave us an insight into a number of important scientific/statistical issues. The review prompted us to review the relevant literature again and look at the limitations of the study. The peer reviewers were open, clear in the instructions and the editorial team was very prompt in their communication. This journal certainly publishes quality research articles. I would recommend the journal for any future publications.
Dear Jessica Magne, with gratitude for the joint work. Fast process of receiving and processing the submitted scientific materials in “Clinical Cardiology and Cardiovascular Interventions”. High level of competence of the editors with clear and correct recommendations and ideas for enriching the article.
We found the peer review process quick and positive in its input. The support from the editorial officer has been very agile, always with the intention of improving the article and taking into account our subsequent corrections.
My article, titled 'No Way Out of the Smartphone Epidemic Without Considering the Insights of Brain Research,' has been republished in the International Journal of Clinical Case Reports and Reviews. The review process was seamless and professional, with the editors being both friendly and supportive. I am deeply grateful for their efforts.
To Dear Erin Aust – Editorial Coordinator of Journal of General Medicine and Clinical Practice! I declare that I am absolutely satisfied with your work carried out with great competence in following the manuscript during the various stages from its receipt, during the revision process to the final acceptance for publication. Thank Prof. Elvira Farina
