INTRODUCTION:

At present, nanobiotechnology is a highly dynamic field of study within modern material science, wherein various plant products and plants themselves are being utilized extensively for nanoparticles (NPs) production. Particles measuring less than 100 nanometers (nm) in diameter are commonly denoted as NPs.

These particles exhibit entirely new and improved properties, including size, distribution, and morphology, when compared to the mass material¹. Silver nanoparticles (AgNPs) possess several notable characteristics, including a confined plasmon resonance, unique physicochemical features, a high surface-to-volume ratio, and a diverse array of applications in the medical field, and microelectronics^2,3. AgNPs have attracted significant attention in comparison to other metal NPs because of their extensive utilization in several pharmacologically important and commercial goods⁴. In the context of synthesis, the conventional procedures, including physical, thermal, hydrothermal, and chemical synthesis modes, are characterized by high costs, significant hazards, and the utilization of poisonous compounds. For this reason, an eco-friendly synthesis approach that utilizes biological resources is emphasized to formulate NPs efficiently⁵.

NPs have been successfully produced through green synthesis techniques, which involve the utilization of various biomolecules, including yeasts, algae, vitamins, enzymes, biodegradable polymers, and microorganisms, as well as plant sections including leaves, flowers, stems, roots, buds, gums, fruits, barks, and shells^6,7. Nanobiotechnology and its related products have distinctive characteristics, encompassing treatment methods, particle size, physical, chemical, and biological qualities, along with a wide array of applications⁸. The concept of nanobiotechnology is now in its early stages of development, mostly due to the limited adoption of revolutionary approaches on a big industrial scale⁹. In contrast, further progress is necessary to enhance the sector with modern technologies. Therefore, it is imperative to develop an economically feasible and sustainable practice for synthesizing AgNPs to satisfy the growing demand for this material in many industries. AgNPs have a considerable surface area, small dimensions, and desirable dispersibility. These particles, which are increasingly used in the fight against infections, are widely utilized in the medical industry¹⁰. The most crucial features of AgNPs include non-toxicity, high stability, hydrophilicity, thermal resistance, and increased resistance to microorganisms¹¹. Metal nanoparticles show antimicrobial properties due to their large surface area/volume ratio¹².

Currently, there is a limited amount of research on the sustainable production of NPs and their effectiveness in managing Leishmania parasites. Plant extracts are recommended among the many biosynthetic processes because they are easily accessible, cost-effective, and safe. Using plants as biomaterials for synthesizing NPs has shown promise for treating leishmaniasis. A study devised an environmentally friendly procedure to synthesise biogenic AgNPs and amphotericin B-bound biogenic silver nanoparticles (AmB-AgNPs) utilising Isatis tinctoria which led to improved outcomes¹³. The same promising outcome was demonstrated in another work by employing a natural sweetener (Stevia) extract for zinc oxide nanostructure production¹⁴. Aiming to improve drug delivery in cutaneous leishmaniasis (CL), nanogold and nanosilver have been used¹⁵.

Leishmaniasis is an infectious disease caused by the Leishmania protozoan, an obligate intracellular parasite of mammalian macrophages¹⁶. The most severely impacted Indian states by visceral leishmaniasis (VL) include Bihar, Assam, West Bengal, and eastern Uttar Pradesh, where anti-leishmanial treatment resistance and relapse are on the rise. A more recent investigation found an alarming 1,000,000 cases, 10,000 of which were resistant to antimonials, pentamidine, and amphotericin B¹⁷. Bihar is the most affected region in India, with more than 90% of all cases. The anti-leishmanial pharmacological arsenal still needs to be improved, despite the clinical usage of miltefosine and amphotericin B¹⁸.

More recently we have reported the antioxidant and anti-inflammatory activity of the silver nanoparticles synthesized from Phyllanthus emblica leaf extract further we observed antileishmanial activity of the synthesized AgNPs¹⁹.

This study has developed an innovative approach for the efficient and environmentally sustainable production of AgNPs from 1mM silver nitrate (AgNO₃) using aqueous extracts of an Indian medicinal plant, namely Carica papaya (papaya) leaf. The fruit and leaf of C. papaya are being utilised for controlling dengue fever²⁰ to increase RBC and platelet counts^21,22. In this study, the plant-mediated AgNPs were characterized using UV-Vis spectroscopy, fourier transform infrared spectrometer (FTIR), dynamic light scattering (DLS), zeta potential, and scanning electron microscopy (SEM) and investigation of their biomedical potential based on the antibacterial and anti-leishmanial activities.

MATERIAL AND METHODS:

Preparation of the leaf extract:

Fresh leaves of C. papaya were obtained from Bundelkhand University Campus, Jhansi, Uttar Pradesh, India and were washed with tap water and distilled water. The aqueous extract was made by placing 5g of fresh leaves in a 100mL flask along with 50mL of Milli Q water. The leaf mixture was boiled at 70°C for 15 min. The extract was filtered and thereafter stored at 4°C for the further synthesis of NPs.

Silver nanoparticle (AgNPs) synthesis:

The biosynthesis of AgNPs was carried out with a 1:4 ratio of the leaf extract and 1mM AgNO₃ (Sigma-Aldrich) solution. For the fabrication, this reaction mixture was heated in a water bath at 70ºC. Then it was periodically (hourly) observed for the change in colour (from light lime green to dark brown) (Fig. 1) and analyzed by a UV-Vis spectrophotometer at 300 to 700 nm range.

Figure 1. The extract colour changed to dark brown due to the addition of 1 mM AgNO₃

Characterization of synthesized NPs:

UV-Vis spectra analysis:

The UV-Vis spectrophotometer (Perkin Elmer, Germany) at 300-700nm characterized the production of AgNPs with C. papaya leaves. A 350 to 500nm absorption peak²³ demonstrates the presence of silver ions (Ag⁺) and a reduction in the analyzed materials. UV-visible spectra were observed at each hour for 0-6 h.

FTIR analysis:

The functional groups of the plant extract that are potentially responsible for the production of AgNPs were identified using the FTIR (Perkin Elmer Spectrum Version 10.03.06). These functional groups might aid in the stabilization, capping, and reduction of Ag⁺. The 400–4000 cm-1 range was performed using the FTIR spectral array.

Zeta potential analysis:

The size distribution analysis (DLS) of the AgNPs, hydro-dynamic diameter, and polydispersity index (PdI) value was determined by size analyzer (ZEN3600 Malvern, Nano series, HT Laser, Malvern, UK). The stability of the formulation and its surface charge are determined using a zeta potential study. Measuring the nanoparticle velocity enables to assessment of the colloidal stability of AgNPs produced by green synthesis. The particle's velocity as it moves toward the electrodes is measured under the influence of the electric field²⁴.

Scanning Electron Microscopy (SEM):

The shape, size, and dispersion of synthesized samples were tested using SEM. The dried samples were put on a double conductive tape that was attached to a sample container. The dried samples were then covered with gold using a coater and analyzed at an 80 kV voltage. The pictures of AgNPs were obtained in an SEM (ZEISS EVO-MA 10, Oberkochen, Germany). The information pertaining to the applied voltage, magnification employed, and dimensions of the image contents were embedded inside the photos themselves.

Estimation of antibacterial activity:

This study examines the relative antibacterial efficacy of various concentrations of AgNPs synthesized from the C. papaya leaf and was effectively accessed against two Gram +ve (Staphylococcus aureus, Bacillus cereus) bacteria and two Gram -ve (Escherichia coli, Klebsiella pneumoniae) bacteria. Well diffusion method²⁵ was followed for testing Ag NPs containing solution. Synthesized AgNPs and standard antibiotics were added steadily, followed by incubation for 24h at 37°C. The plates with nutrient agar medium were created by swabbing them with the microbial cultures. The measurement was taken to determine the size of the zone of inhibition.

Parasite culture and analysis of cell viability:

L. donovani promastigotes were routinely grown at 25±1°C in media (RPMI 1640) supplemented with fetal calf serum, streptomycin (100g/mL), and penicillin (100 g/mL). Promastigotes in the logarithmic phase were seeded (1×10⁶cells/mL) on 96-well plates along with different AgNP concentrations, followed by incubation at 25±1°C for 24, 48, and 72h. Then each well was filled with 100µl of MTT solution (5mg/mL) and incubated at 25±1°C for 4h. To finally dissolve the formed formazan, DMSO (100µl) was used. Then absorbance was recorded at 570nm, with DMSO (0.5%) serving as the control and miltefosine serving as the reference.

The percentage of cell viability was calculated with the following formula:²⁶

OD of cells with AgNPs

% Cell viability = --------------------------------- ×100

OD of cells without AgNPs

RESULTS AND DISCUSSION:

UV-Visible analysis:

The utilisation of C. papaya leaf as a reducing and stabilising agent in the sustainable production of AgNPs was principally demonstrated by the alteration in colour from light yellow or transparent to brown. The confirmation of AgNP formation was established by the observed change in colour, which confirmed the initiation of SPR (surface plasmon resonance) vibrations within these particles²⁷. The SPR spectra exhibit sensitivity to particle morphology, dimensions, interaction with the surroundings, refractive index, and the degree of charge shift occurring between the NPs and the medium²⁸. The UV-visible spectra recorded after intervals of 1 to 7 h observed from the initiation of the process are shown in Fig. (2). Absorption spectra of AgNPs have absorption maxima at 460nm. The UV-Vis spectra revealed a wider absorption peak, suggesting the existence of particles exhibiting a broad size range. This outcome is consistent with the conclusions of a prior investigation²⁹.

Figure 2. UV/Visible Spectra of AgNPs obtained from C. papaya

Fourier transform infrared spectroscopy (FTIR):

The FTIR spectra corresponding to the production of AgNPs from C. papaya are shown in Fig. (3). FTIR bands of AgNPs identify the probable functional groups present in the suspension. In AgNPs solutions, prominent bands of absorbance peaks were viewed which illustrate the presence of specified functional groups (Table 1). The bands mentioned by Huang et al. ³⁰ represent stretching vibrational bands that are associated with chemicals such as flavonoids and terpenoids. These bands are likely to have a role in effectively capping and stabilising the AgNPs that are formed. Furthermore, the biomolecules have a pronounced affinity for metal binding, indicating the potential creation of a protective layer around metal NPs. This layer acts as a capping agent, effectively inhibiting agglomeration and ensuring stability within the surrounding media³¹.

Figure 3. FTIR Spectra of AgNPs obtained from C. papaya

Table. 1: FTIR peak value represents the functional groups present in AgNPs fabricated from C. papaya

Peak Value	Functional Group
3304	O-H / N-H
2123	N=C=N /N=C=S
1636	C=C / N-H
588	C-Cl
581	C-Cl
565	C-Cl
546	C-l
536	C-l
526	C-l
509	C-l

DLS measurements and zeta potential:

DLS (dynamic light scattering) is a highly versatile technique employed to assess the shape, size, and dispersion of AgNPs. Fig. (4) illustrates the DLS analysis of the mean size of AgNPs. The AgNPs generated by C. papaya had a mean size of around 68.40 nm, with a PdI value of 15.51. The PdI indicates the level of uniformity in the AgNPs. The DLS is employed to ascertain the hydrodynamic properties of NPs. This measurement is conducted in a liquid suspension that includes a metallic centre, ions, and biological macromolecules that are bound around the nanoparticles ³².

Zeta potential (ZP) tests can be used to assess the stability of the created NPs suspension. The ZP of the nanosuspension was determined at pH 7. C. papaya yielded a significantly low ZP value of -28.7mV. The ZP may have negative values due to the presence of negatively charged substances in the vicinity of the NPs' surface³³.

Figure 4. DLS measurement of the average size of phytosynthesized AgNPs obtained from C. papaya

SEM analysis:

Among other electron microscopy techniques, SEM can analyse the surface features of nanoparticles, such as their shape, size, and size distribution³⁴. Field emission SEM (FE-SEM) involves the emission of electrons that are accelerated using a powerful electric field. Fig. (5) displays the SEM image of the AgNPs. AgNPs are typically spherical, cubical, triangular, oval, pebble-like, and circular, and appear as single or aggregated particles ³⁵. The average diameter of the AgNPs obtained from C. papaya leaves was confirmed to be 15 nm less than the value obtained by DLS. The SEM results indicated the spherical and cuboidal shape of synthesized nanoparticles. This could be linked to the varying quantities and characteristics of capping agents found in the leaf extracts. Additionally, the shifts and variations in peak areas seen in the FTIR analysis further evidence for this assertion. The findings shown here are consistent with the research conducted by Mohamed et al. ³⁶ and Tippayawat et al. ³⁷, which documented the synthesis of spherical silver nanoparticles using Aloe vera extract, with sizes ranging from 70 to 190nm.

Figure 5. SEM image of silver nanoparticles. Formed by the reaction of 1 mM silver nitrate and extract of C. papaya leaves.

Antibacterial study:

The antibacterial activity of AgNPs was examined by growing Gram +ve bacteria (Staphylococcus aureus, Bacillus cereus) and Gram −ve bacteria (Escherichia coli, Klebsiella pneumonia) colonies on nutrient agar media applied with different concentrations (250, 500 and 1000 μg/mL) of AgNPs (Fig. 6). The inhibition zones indicate the maximum antibacterial property of the AgNPs. Results obtained in earlier studies^9,29 also validate the antibacterial efficiency of AgNPs. The synthesized AgNPs at 1000μg/mL concentration exhibited a significant antibacterial efficacy on E. coli more than antibiotics in contrast to the other bacterial strains. This efficacy of the synthesised AgNPs exhibited a dose-dependent pattern. The results can be inferred based on the observation that these particles exhibited the shortest diameter. Furthermore, when comparing antibiotics with AgNPs, it is evident that antibiotics do not exhibit significant antibacterial efficacy against B. cereus, E. coli, and K. pneumonia. The control did not exhibit any zone of inhibition. Previous studies have put forth several techniques to explain the antibacterial action of AgNPs³⁸.

The cell wall of Gram +ve bacteria is distinguished by a significant peptidoglycan layer, consisting of linear polysaccharide chains that are interconnected by short peptides. The inflexible arrangement of the cell wall creates an obstacle to the entry of AgNPs into the bacterial cell. On the other hand, Gram –ve bacteria possess a less substantial peptidoglycan layer within their cell wall³⁹.

Figure 6. The sizes of inhibition zones (mm) representing the antibacterial activity of AgNPs at different concentrations and antibiotic standards (blue columns): S. aureus, B. cereus, E. coli, K. pneumonia

Anti-leishmanial activity:

The impact of various quantities of AgNPs (20-100 µg/mL) after 24, 48, and 72h of exposure to L. donovani promastigotes was studied in vitro. The IC₅₀ content of AgNPs after 24, 48, and 72h on L. donovani was calculated to be 45.88, 36.86, and 24.81µg/mL, respectively. The highest anti-leishmanial effect (100%) was observed at 100µg/mL after 72 h of exposure. Table 2 and Fig. (7) present the per cent cell viability of Leishmania promastigotes being exposed to various concentrations of AgNPs after incubation of 24, 48, and 72h. The AgNPs exhibited strong anti-leishmanial activities but comparatively, had lower activities than the anti-leishmanial drug miltefosine (positive control) against Leishmanial promastigotes. Ghosh et al. ⁴⁰ evaluated the anti-parasitic activity of isolated compounds from the chloroform extract of C. papaya.

Figure 7. Anti-leishmanial activities of AgNPs against Leishmania donovani at 24, 48, and 72 h

CONCLUSION:

The current study holds importance within the area of nanotherapeutics based on its investigation into the phytosynthesis of AgNPs (silver nanoparticles) via extract derived from C. papaya leaves. This approach is environmentally benign, characterised by its cost-effectiveness, efficiency, simplicity, and absence of hazardous substances. The fabricated AgNPs have been characterized with UV-Vis, FTIR, DLS, and SEM. The AgNPs were spherical, with a mean diameter of 68.40 nm. Additionally, the AgNPs have shown substantial antibacterial effectiveness against different bacterial strains. All these results emphasize the high efficacy of AgNPs as new, safer and effective anti-leishmanial formulations. The findings of this research present novel opportunities for the utilisation of NPs derived from extracts of C. papaya leaves in several domains such as pharmaceuticals, cosmetics, biomedicine, and nanomedicine.

CONFLICT OF INTEREST:

The authors have no relevant financial or non-financial interests to disclose.

REFERENCES:

1. Van den Wildenberg W: Roadmap report on nanoparticles. W and W Espana sl, Barcelona, Spain; 2005.

2. Nouri A, Yaraki MT, Lajevardi A, Rezaei Z, Ghorbanpour M, Tanzifi M. Ultrasonic-assisted green synthesis of silver nanoparticles using Menthaaquatica leaf extract for enhanced antibacterial properties and catalytic activity. Colloids Interface Sci Commun. 2020; 35: 100252. https://doi.org/10.1016/j.colcom.2020.100252

3. Gul AR, Shaheen F, Rafique R. Grass-mediated biogenic synthesis of silver nanoparticles and their drug delivery evaluation: A biocompatible anti-cancer therapy. Chem Eng J. 2021; 407: 127202. https://doi.org/10.1016/j.cej.2020.127202

4. Atoussi Ouidad, Chetehouna Sara, Derouiche Samir. Biological properties and Acute Toxicity Study of Copper oxide nanoparticles prepared by aqueous leaves extract of Portulaca oleracea (L). Asian J. Pharm. Res. 2020; 10(2): 89-94. https://doi.org/10.5958/2231-5691.2020.00017.9

5. Saira Sehar, Amiza, I. H Khan. Role of ZnO Nanoparticles for improvement of Antibacterial Activity in Food Packaging. Asian Journal of Pharmaceutical Research. 2021; 11(2): 128-1. https://doi.org/10.52711/2231-5691.2021.00024

6. Mankodi HR. Studies on different type of sutures using aloe vera gel coating. Int J Text Fashion Technol. 2013; 4: 11–6

7. Saira Sehar, Mohsin Sher Ali Khan, Amiza, Touqir Hussain, M. Zahid, Moazina Mobeen, Imran Raza, Minnatullah. Synthesis of Zinc Oxide Nano particles from Moringa Tree leaves by Green Method and also check its Antibacterial activity against Gram Positive and Gram Negative Bacteria. Asian Journal of Pharmacy and Technology. 2022; 12(3): 213-7. https://doi.org/10.52711/2231-5713.2022.00035

8. Reshma Chauhan, Charmi Patel, Jitendriya Panigrahi. Greener approach for copper nanoparticles synthesis from Catharanthus roseus and Azadirachta indica leaf extract and their antibacterial and antioxidant activities. Asian J. Res. Pharm. Sci. 2018; 8(2): 81-90. https://doi.org/10.5958/2231-5659.2018.00016.4

9. Banerjee P, Satapathy M, Mukhopahayay A, Das P. Leaf extract mediated green synthesis of silver nanoparticles from widely available Indian plants: synthesis, characterization, antimicrobial property and toxicity analysis. Bioresour Bioprocess. 2014; 1: 1–10. https://doi.org/10.1186/s40643-014-0003-y

10. Franci G, Falanga A, Galdiero S, Palomba L, Rai M, Morelli G, et al. Silver nanoparticles as potential antibacterial agents. Molecules. 2015; 20(5): 8856–74. https://doi.org/10.3390/molecules20058856

11. Zhang XF, Liu ZG, Shen W, Gurunathan S. Silver nanoparticles: synthesis, characterization, properties, applications, and therapeutic approaches. Int J Mol Sci. 2016; 17(9): 1534. https://doi.org/10.3390/ijms17091534

12. M.C. Purohit, Anuj Kandwal, Reena Purohit, A.R. Semwal, Shama Parveen, Arun K. Khajuria. Antimicrobial Activity of Synthesized Zinc Oxide Nanoparticles using Ajuga bracteosa Leaf Extract. Asian Journal of Pharmaceutical Analysis. 2021; 11(4): 275-0. https://doi.org/10.52711/2231-5675.2021.00047

13. Ahmad A, Wei Y, Syed F. Isatis tinctoria mediated synthesis of amphotericin B-bound silver nanoparticles with enhanced photoinduced antileishmanial activity: a novel green approach. J Photochem Photobiol B: Biol. 2016; 161: 17–24. https://doi.org/10.1016/j.jphotobiol.2016.05.003

14. Khatami M, Alijani HQ, Heli H, Sharifi I. Rectangular shaped zinc oxide nanoparticles: green synthesis by Stevia and its biomedical efficiency. Ceram Int. 2018; 44: 15596–602. https://doi.org/10.1016/j.ceramint.2018.05.224

15. Basu M, Das PK. Role of Reactive Oxygen Species in Infection by the Intracellular Leishmania Parasites. In: Chakraborti, S., Chakraborti, T., Chattopadhyay, D., Shaha, C. (eds) Oxidative Stress in Microbial Diseases. Springer, Singapore. 2019 https://doi.org/10.1007/978-981-13-8763-0_16

16. Tyler KM, Myler PJ, Fasel N. Leishmania – After the Genome. Parasit Vectors. 2008; 1: 11. https://doi.org/10.1186/1756-3305-1-11

17. Sundar S. Drug resistance in Indian visceral leishmaniasis. Trop Med Int Health. 2001; 6: 849–54. https://doi.org/10.1046/j.1365-3156.2001.00778.x

18. Richard JV, Werbovetz KA. New antileishmanial candidates and lead compounds. Curr Opin Chem Biol. 2010; 14: 447–55. https://doi.org/10.1016/j.cbpa.2010.03.023

19. Sharma S, Kumar SK, Pai K, Kumar R. Synthesis of silver nanoparticles using Phyllanthus emblica leaf extract: Characterization, antioxidant, anti-inflammatory and antileishmanial activity against L. donovani. Nanomed Res J. 2024; Mar 1; 9(1): 9-19. https://doi.org/10.22034/nmrj.2024.01.002

20. Ahmad N, Fazal H, Ayaz M, Abbasi BH, Mohammad I, Fazal L. Dengue fever treatment with Carica papaya leaves extracts. Asian Pac J Trop Biomed. 2011; 1(4): 330–3. https://doi.org/10.1016/S2221-1691(11)60055-5

21. Dharmarathna SL, Wickramasinghe S, Waduge RN, Rajapakse RP, Kularatne SA. Does Carica papaya leaf-extract increase the platelet count? An experimental study in a murine model. Asian Pac J Trop Biomed. 2013; 3(9): 720–4. https://doi.org/10.1016/S2221-1691(13)60145-8

22. Anamika, Geetika Chandra. A Brief Review Article on Carica papaya and its Medical Advantages. Research Journal of Pharmacognosy and Phytochemistry. 2024; 16(3): 180-4. https://doi.org/10.52711/0975-4385.2024.00034

23. Prabhu N, Divya RT, Yamuna GK. Synthesis of silver phytonanoparticles and their antibacterial efficacy. Dig J Nanomater Biostructures. 2010; 5: 185–9

24. Silva LP, Pereira TM, Bonatto CC. Frontiers and perspectives in the green synthesis of silver nanoparticles. In: Green synthesis, characterization and applications of nanoparticles,. Elsevier; 2019. https://doi.org/10.1016/B978-0-08-102579-6.00007-1

25. Vidhu VK, Aromal SA, Philip D. Green synthesis of silver nanoparticles using Macrotyloma uniflorum. Spectrochim Acta A Mol Biomol Spectros. 2011; 83: 392–7. https://doi.org/10.1016/j.saa.2011.08.051

26. Sharma U, Singh D, Kumar P. Antiparasitic activity of plumericin and isoplumericin isolated from Plumeria bicolor against Leishmania donovani. Indian J Med Res. 2011; 134: 709–16. https://doi.org/10.4103/0971-5916.91005

27. Boken J, Khurana P, Thatai S. Plasmonic nanoparticles and their analytical applications: A review. Appl Spectrosc Rev. 2017; 52: 774–820. https://doi.org/10.1080/05704928.2017.1312427

28. Rajkumar T, Sapi A, Das G, Debnath T, Ansari A, Patra JK. Biosynthesis of silver nanoparticle using extract of Zea mays (corn flour) and investigation of its cytotoxicity effect and radical scavenging potential. J Photochem Photobiol B: Biol. 2019; 193: 1–7. https://doi.org/10.1016/j.jphotobiol.2019.01.008

29. Alwahibi MS, Soliman DA, Alonaizan A. Green biosynthesis of silver nanoparticle using Commiphora myrrh extract and evaluation of their antimicrobial activity and colon cancer cells viability. J King Saud Univ Sci. 2020; 32: 3372–9. https://doi.org/10.1016/j.jksus.2020.09.024

30. Huang J, Li Q, Sun D. Biosynthesis of silver and gold nanoparticles by novel sundried Cinnamomumcamphora leaf. Nanotechnology. 2007; 18: 105104. https://doi.org/10.1088/0957-4484/18/10/105104

31. Aslany S, Tafvizi F, Naseh V. Characterization and evaluation of cytotoxic and apoptotic effects of green synthesis of silver nanoparticles using Artemisia Ciniformis on human gastric adenocarcinoma. Mater Today Commun. 2020; 24: 101011. https://doi.org/10.1016/j.mtcomm.2020.101011

32. Moteriya P, Chanda S. Green synthesis of silver nanoparticles from Caesalpinia pulcherrima leaf extract and evaluation of their antimicrobial, cytotoxic and genotoxic potential. J Inorg Organomet Polym Mater. 2020; 30: 3920–32. https://doi.org/10.1007/s10904-020-01532-7

33. Baláž M, Bedlovičová Z, Kováčová M, Salayová A, Balážová Ľ. Green and bio-mechanochemical approach to silver nanoparticles synthesis, characterization and antibacterial potential. In: Prasad, R., Siddhardha, B., Dyavaiah, M. (eds) Nanostructures for Antimicrobial and Antibiofilm Applications. Nanotechnology in the Life Science. 2020; Springer: Cham pp. 145-83. https://doi.org/10.1007/978-3-030-40337-9_72020:

34. Sadeghi B, Gholamhoseinpoor F. A study on the stability and green synthesis of silver nanoparticles using Ziziphora tenuior (Zt) extract at room temperature. Spectrochim Acta A Mol Biomol Spectrosc. 2015; 134: 310–5. https://doi.org/10.1016/j.saa.2014.06.046

35. Ghabban H, Alnomasy SF, Almohammed H, Al Idriss OM, Rabea S, Eltahir Y. Antibacterial, cytotoxic, and cellular mechanisms of green synthesized silver nanoparticles against some cariogenic bacteria (Streptococcus mutans and Actinomyces viscosus. J Nanomater. 2022; 1–8. https://doi.org/10.1155/2022/9721736

36. Mohamed N, Elmasry H. Aloe Vera gel extract and sunlight mediated synthesis of silver nanoparticles with highly effective antibacterial and anticancer activity. Journal of Nanoanalysis. 2020: 7(1): 73-82

37. Tippayawat P, Phromviyo N, Boueroy P, Chompoosor A. Green synthesis of silver nanoparticles in aloe vera plant extract prepared by a hydrothermal method and their synergistic antibacterial activity. PeerJ. 2016; 4: e2589. https://doi.org/10.7717/peerj.2589

38. Panda MK, Singh YD, Behera RK, Dhal NK. Biosynthesis of nanoparticles and their potential application in food and agricultural sector. Green Nanoparticles. Springer, Cham; 2020. https://doi.org/10.1007/978-3-030-39246-8_10

39. G. Alagumuthu, R. Kirubha. Biogenic Synthesis of Silver Nanoparticles using Aegle marmelos fruit extract and their Antibacterial potential. Asian J. Research Chem. 2013; 6(9): 839-844.

40. Ghosh S, Saha M, Bandyopadhyay PK, Jana M. Extraction, isolation and characterization of bioactive compounds from chloroform extract of Carica papaya seed and it’s in vivo antibacterial potentiality in Channa punctatus against Klebsiella PKBSG14. Microb Pathog. 2017; 111: 508–18. https://doi.org/10.1016/j.micpath.2017.08.033

Received on 14.10.2024 Revised on 11.02.2025

Accepted on 25.04.2025 Published on 01.12.2025

Available online from December 06, 2025

Research J. Pharmacy and Technology. 2025;18(12):5964-5970.

DOI: 10.52711/0974-360X.2025.00862

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.