Dear Dr. Rauf,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Academic Editor

PLOS ONE

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Additional Editor Comments :

We note that one or more reviewers have recommended that you cite specific previously published works. As always, we recommend that you please review and evaluate the requested works to determine whether they are relevant and should be cited. It is not a requirement to cite these works.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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2. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: N/A

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3. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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Reviewer #1: Comments:

The article titled "Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential" presents valuable findings, but it has following shortcomings that must be addressed before the publication of the articel

Recommendation: Major Revision

1. Provide overlaid FTIR spectra of extract and NPs and highlight areas of change of wavenumber.

2. EDX spectra show peaks for various elements e.g., Cl. What is the reason behind this peak?

3. Stability of the NPs is an important parameter while studying the biomedical applications. Provide Zeta potential of the particles at different pH, Temperature, and after different time intervals.

4. While the article discusses in vitro biological activities and some in vivo evaluations, the extent of in vivo studies appears limited. More comprehensive in vivo studies could provide a better understanding of the pharmacological effects and safety of the synthesized silver nanoparticles (Ag NPs).

5. The findings are based on specific conditions and concentrations used in the assays. The applicability of these results to broader biological contexts or different formulations may not be fully addressed, which could limit the generalizability of the conclusions drawn.

6. The article mentions that the Ag NPs are stabilized by the Ficus benghalensis extract, it does not provide long-term stability data. This information is crucial for assessing the practical applications of the synthesized nanoparticles in biomedical fields.

7. The introduction discusses the importance of metallic nanoparticles (NPs) and their applications, it does not clearly define the specific research gap that this study aims to fill. A more explicit statement regarding what is lacking in current literature would strengthen the rationale for the study.

8. While discussing the biological applications, update the introduction section with these recent reports, i) doi: https://doi.org/10.3390/ph17081053, ii) https://doi.org/10.1016/j.jep.2023.116503, iii) https://doi.org/10.3390/genes16010016 iv) https://doi.org/10.1016/j.optlaseng.2024.108688 v) https://doi.org/10.1016/j.bioorg.2024.107415 vi) https://doi.org/10.1016/j.apsb.2024.02.005

9. Although the introduction mentions the medicinal properties of Ficus benghalensis, it could benefit from a more detailed background on why this particular plant was chosen for the synthesis of silver nanoparticles. Providing more context on its unique properties or previous studies would enhance the reader's understanding of its relevance.

10. The introduction highlights the eco-friendly nature of green synthesis but does not delve into the environmental implications of using Ficus benghalensis specifically. Discussing the sustainability aspect in more detail could provide a stronger justification for the choice of plant extract.

11. Cite some recent reports while discussing green synthesis of metal nps https://doi.org/10.1016/j.jece.2024.112576, https://doi.org/10.1016/j.molliq.2023.123622. https://doi.org/10.1016/j.jece.2024.113350, https://doi.org/10.3390/antiox12061201

12. The introduction mentions various methods of synthesizing Ag NPs but lacks a comparative analysis of the advantages and disadvantages of green synthesis versus conventional methods. This could help emphasize the significance of the chosen approach in the study.

13. While the introduction states that NPs have extensive applications in various fields, it could be more focused on the specific applications relevant to the synthesized Ag NPs. Highlighting particular areas where these nanoparticles could be beneficial would provide a clearer direction for the research.

14. The introduction does not address any potential limitations or challenges associated with the synthesis and application of Ag NPs. Acknowledging these aspects could provide a more balanced view and prepare the reader for the discussion of results later in the article.

15. Compare the results of biological applications with following nanomaterials

i) https://doi.org/10.1039/D3RA05070J, ii) https://doi.org/10.1016/j.ijbiomac.2023.128009, iii) https://doi.org/10.1080/14786419.2023.2295936, iv) https://doi.org/10.3389/fchem.2023.1202252, v) https://doi.org/10.1016/j.enmm.2022.100735, vi) https://doi.org/10.3390/molecules27113363, vii) https://doi.org/10.1016/j.surfin.2024.104556

Reviewer #2: The manuscript is good, but it needs major revision before publishing.

Abstract:

• The abstract is very long, it should be made more concise.

Introduction:

• The introduction is lengthy. Condense it by focusing on the most relevant background information.

• "Various characteristics of NPs including particle size, shape, morphology, and increased surface area, set NPs apart from their bulk counterparts" is a long and awkward sentence. Revise for clarity.

• "Fe NPs have been successfully synthesized using extracts from..." This paragraph feels like a list. Integrate these examples more smoothly into the narrative and indicated each one to its respective citation.

• Add more citations, many statements are not backed up by research.

• The last paragraph of the introduction is very long. Divide it into smaller more focused paragraphs.

Materials and Methods:

• "Ethics" section: "killed with cervical dislocation which is an approved disposal method of animal ethics" is awkwardly phrased. Please revise the sentence for clarity and include the specific year of issue.

• "Plant collection and extraction": Specify the exact duration of drying and the temperature used.

• "Nanoparticles synthesis": Use standard font.

• "Instrumentations": "NCEG-UOP" needs to be defined.

• "Computational studies": "221 layers of Ag" is confusing. Did you mean a 2x2x1 supercell? Clarify.

• "Kollman chargres" should be "Kollman charges."

• "In vivo activities": it must be capitalized, and then revise all throughout the manuscript.

Results:

• "221 layers of Ag" still needs clarification.

• There are many instances of inconsistent units and formatting.

Discussion:

• "Ag NPs are synthesized by the green synthesis method. The synthesized Ag NPs are chemically and physically stable in extract use for it as a capping agent and out of degradation." This sentence is very poorly written. Please rewrite.

• The discussion section is very short, it needs to be expanded.

• The discussion section needs more citations.

• The discussion does not do a good job of comparing the results to other research.

References:

• Consistency: The reference formatting is inconsistent. Ensure all references adhere to a single style guide (e.g., APA, MLA, or the journal's specific guidelines). Pay attention to italics, capitalization, and punctuation.

• Accessibility: Some references lack DOIs or URLs, making it difficult for readers to access the source material. Provide complete and accessible information for each reference.

• Accuracy: Double-check all references for accuracy in author names, titles, publication years, and other details.

General Recommendations:

• Proofread the entire manuscript for grammatical errors and typos.

• Improve the clarity and flow of the writing.

• Expand the discussion section to provide a more thorough analysis of the results.

• Add more citations.

• Consider having a native English speaker review the manuscript.

By addressing these points, the authors can significantly improve the quality and clarity of the manuscript.

Reviewer #3: The manuscript Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential, the section wise comments are

Title: title is attractive, self explanatory and can gain readers attention hence no change required

Abstract: This section is poorly presented though the work is nice so I suggest authors to rewrite this section by adding some results to make this section more clear

Introduction: Some syntax/typo errors which should be checked carefully and corrected. There are some less supportive references in this section which should be replaced with interesting work, few suggestion are made in the references section.

Result and discussion: well written and managed, however the figures presented are of low quality, I suggest authors to provide high quality figures as per requirement of the journal

Experimental section is very well written and the procedures adopted are as per standard procedures adopted in the field

References: Some less supportive references should be replaced with the following interesting work

replace reference number 11 and 12 with

https://doi.org/10.1007/s10904-020-01763-8

https://doi.org/10.1155/2021/3475036

The inclusion of the mentioned work can increase the quality of the manuscript, with the above mandatory points I recommend this manuscript for publication

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what does this mean? ). If published, this will include your full peer review and any attached files.

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Reviewer #1: Yes:  Dr Azhar Abbas

Reviewer #2: Yes:  Yahya Al-Awthan

Reviewer #3: No

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Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential

PONE-D-25-01658R1

Dear Dr. Rauf,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Rajesh Kumar Singh, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

This manuscript entitled "Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential" Manuscript Id: PONE-D-25-01658R1 has been revised as per the reviewers comments but still few errors are existing.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #1: All comments have been addressed

Reviewer #3: All comments have been addressed

Reviewer #4: All comments have been addressed

Reviewer #5: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions??>

Reviewer #1: Yes

Reviewer #3: Yes

Reviewer #4: Yes

Reviewer #5: Yes

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3. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #1: Yes

Reviewer #3: Yes

Reviewer #4: N/A

Reviewer #5: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: Yes

Reviewer #3: Yes

Reviewer #4: Yes

Reviewer #5: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #3: Yes

Reviewer #4: Yes

Reviewer #5: Yes

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Reviewer #1: The author has addressed all the queries raised so i will recommend publication of the article in current from.

Reviewer #3: The authors have incorporated all the points so I feel pleasure to recommend this manuscript in its present form

Reviewer #4: The authors of the manuscript entitled “Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential” have addressed almost all comments, although the manuscript needs some grammatical corrections before it will be accepted.

Reviewer #5: The authors of the manuscript entitled “Ficus benghalensis extract mediated green synthesis of silver nanoparticles, its Optimization, Characterization, Computational studies and its in vitro and in vivo Biological potential” have addressed almost all comments, although the manuscript needs some grammatical corrections before it will be accepted.

Additionally, not necessary but if possible then add some key features or link of biological effects which shown in results with different diseases for further use or treatment.

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what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy

Reviewer #1: No

Reviewer #3: No

Reviewer #4: Yes:  Dr. Adwitiya Banerjee

Reviewer #5: No

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