-->PONE-D-24-32691-->-->EVs Therapy for Cisplatin-Induced Testicular Tissue Toxicity-->-->PLOS ONE

Dear Dr. TOZAK YILDIZ,

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Roland Eghoghosoa Akhigbe

Academic Editor

PLOS ONE

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Additional Editor Comments:

The study by Halime et al. is interesting and demonstrates the therapeutic value of EVs in attenuating cisplatin-induced testicular toxicity. However, just like both reviewers have suggested a major revision, the manuscript needs a thorough revision to meet the standard of this journal. Authors should also consider editing the manuscript for grammar and syntax. In addition to the reviewers' comments, these comments would be useful:

Abstract

1. What are the metabolites of cisplatin that exert its effect

2. “MDA activity”. Is MDA an enzyme?

3. Include recommendations

Introduction

1. Paragraph 1: The effects should precede the mechanisms

2. Paragraph 3: The note on autophagy is hanging. It is not linked with cisplatin nor EV

Methods/Discussion

1. The immunolocalization of the assayed molecules should be discussed rather than the expression only

2. Since cisplatin also suppresses hypothalamic-pituitary-testicular axis, it is important to evaluate gonadotropins and GnRH

3. How long did the study last?

4. Was spermatogenic cycle studied?

5. Why is the histology of the epididymis not presented?

6. It is not sufficient to report sperm count and motility and ignore other sperm physiological functions. Are others not relevant?

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

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

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

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

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: No

Reviewer #2: No

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-->5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: Although this study is interesting, there are many problems with this manuscript, mainly due to missing information in the text, improperly explained facts, and missing references. For all the details, I attach a PDF with all reviewer comments.

Reviewer #2: Hi Dr Yildiz

I have gone through your work and want to commend you for the effort which you have made to put this together, especially seeing that this project is likely self-funded. However, I have the following comments to make this better for the entire scientific community;

1. The use of English in this draft does not represent the best scientific expertise expected for a paper like this.

2. From the opening sentence of your abstract , you failed to ignite my enthusiasm about this project. The significance of cisplatin...why cisplatin? Justification for the study? The purpose section in your abstract must be rewritten. This manuscript have great potential if you can do that. Then, the methods... Rewrite your methods so we understand your methodology well...length of experiment, animals? How many? You left me clueless about this experiment here! Then, how did you get your data? Laboratory analysis? Statistical tests? You said testicular morphology was examined using H&E?? Really???? Histoarchitecture or morphology ?? Please add the methods you used for the measurement of hormones, sperm parameters. Your methodology is very poor in this abstract in that... You omitted so many details....

3. The results subsection of your abstract is also faultable. I expect that the results will be presented in a logical sense that follows a thorough discussion of effect and mechanism.

For example; first state the effects on structure, Hormone (function), sperm parameters (function) then the mechanisms of these negative or positive changes (oxidative stress, autophagy)

4. The conclusion of the abstract does not show comprehensive snapshot of your results. I guess you were have a wordcount limit for your abstract but you have to do these corrections to put this manuscript in top shape.

5. Your introduction is needs to be overhauled. From the first sentence with reference (Cisplatin I'd a potent....) to " it has been reported" when you actually didn't add those studies that reported. The third sentence on the genotoxic effects of Cis also lacks merit? Formation of primordial follicle is a genotoxic effects?? Are you sure? You indicated that Cis reduces spermatogenesis? How? What sperm parameters? What stage of spermatogenesis?

6. Then you described EVs as if you didn't get the description somewhere, why? This dampens my enthusiasm, honestly! Each factual statement you make should be backed by a reference.

7. The next paragraph is same. So many missing references. Each factual statement should end with a reference.

8. Replace "long-lived" with a more appropriate adjective.

9. I see no need for the "much research has been focused on starvation-induced autophagy..." What are you trying to say? A lot of missing connections in the paragraph.

10. These paragraphs in the introduction do not connect.

11. I was expecting your introduction will shadow this line of thought

Few sentences on general info about cisplatin. Then it's toxicity on general physiology in humans and animal models. Then, the reprotoxic effects. This should be detailed and more of males. After stating the effects, you discuss the mechanisms. Then, the following paragraph will tow...

"On the other hand, EVs are bla bla..." After the general info about EVs you should talk about effects in several reprotoxicity models , preferably cisplatin or anticancer drugs (if available). You should discuss the mechanisms of its effects exhaustively.

11. Be sure to cite similar works https://doi.org/10.1007/s43188-024-00250-3 in your methodology especially for spermatogenesis, hormones, oxidative stress.

12. Your discussion should tow the same line for the presentation of result. Always be sure to first summarize your findings in the first paragraph of discussion, then discuss histology and morphological parameters in the next paragraph. The following paragraph should discuss findings on hormones...

13. Next, spermatogenesis and sperm parameters, then go to the mechanisms in the subsequent paragraph.

14. Your discussion should include strengths and weaknesses of this study. Your methodology is unique and I am sure there are good things and limitations to it.

15. If possible include graphical abstract. This is not compulsory though.

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Reviewer #1: No

Reviewer #2: Yes:  Oyedokun PRECIOUS ADEOYE

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Attachments

Attachment

Submitted filename: PONE-D-24-32691_reviewer.pdf

Attachment

Submitted filename: Cis, EVs, Testicular toxicity Yildiz.docx

Extracellular Vesicles Therapy Alleviates Cisplatin-Induced Testicular Tissue Toxicity in a Rat Model

PONE-D-24-32691R1

Dear Dr. TOZAK YILDIZ,

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,

Roland Eghoghosoa Akhigbe

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.-->

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: The authors have basically reworked their submission and thereby have improved it significantly. Except for a few minor editorial corrections that should be done, I am satisfied. I attach a PDF in which I indicated these issues.

Reviewer #2: The authors have addressed all comments previously raised. I am satisfied with the current version of the paper.

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-->7. PLOS authors have the option to publish the peer review history of their article (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 #2: Yes:  Oyedokun Precious Adeoye

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