Human umbilical cord mesenchymal stem cell-derived exosomes mitigate acute radiation-induced intestinal oxidative damage via the Nrf2/HO-1/NQO1 signaling pathway

Hongyu Wang; Jinbao Wang; Gaosheng Yang; Yanjie Li; Weikai Chen; Jianping Yu; Xiaopeng Han

doi:10.1371/journal.pone.0324238

Peer Review History

Original SubmissionMarch 25, 2025
25 Mar 2025 Author Response https://doi.org/10.1371/journal.pone.0324238.r001
14 Apr 2025 Decision Letter - Sabata Martino, Editor Dear Dr. Han, 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. Please submit your revised manuscript by May 29 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. 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If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments : no comments [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? Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 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 ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: Weak points Minor weak points There are simply typos and minor adjustments to be made: • Line 39; 40; 68; 70; 87; 132; 181; 200; 203; 207; 323; 326; 337; 340; 358; 364; 388; 459; 471; 473 there is no space after the dot; • Line 168; 211 the number of the cells is not written with exponential, as previously done; • Line 327; 379; 381; 404; 418; 424; 425; 426; 427; 459; 462 the citation are written at the same level of the text and not as a superscript, as previously done; • Line 405 Ki67+ have the first letter written in lowercase and not in uppercase, as done before • The paragraph of the Flow cytometric analysis is written with a different style respect to the other, like a laboratory protocol, I consider to rewrite this paragraph with the same style of the other; • Line 200 what method was used to determine the protein concentration? Was a standard curve constructed first? If so, how? • Line 202-203 what instrument was used to measure the absorbance? • Figure 1A the scale bar is not readable and in Figure 1G and 1H the scale bar is not present; • Figure 1K which normalizing protein was used? I would insert its image; • Figure 4C the scale bar are not present Major weak points There are no major weak points Reviewer #2: The text is flowing in reading, there are no inconsistencies but the form of the text should be improved. Also, space between one paragraph and the next should be added. - Line 158, are present 2 dots - Line 126, 1x10^5 must be written 1x105 - Line 168, 1x106 must be written 1x106 - Line 185, which type of microscope? - Line 195, which type of microscope - Line 203, how did you measure the absorbance? - Line 211, 1x10^5 must be written 1x105 - Line 219, which instrument is it? - The style of the references is different in the conclusion than in the rest of the article. Check that all bibliographic references are uniform with each other and adhere to the style that the journal dictates. - Line 426, commas and References - Images 1, 2, 3 e 4 pay attention to the scale bar. Is not visible - Image 3C the words Dapi, Ros and Merge seem to refer to the image above; so bring it closer to the corresponding image. - “CON” to substitute with “CTRL” - Unit measures in image 4, graph B ****** 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: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org Attachments Attachment Submitted filename: External revision_PONE-D-25-12676.docx Attachment Submitted filename: External revision_.pdf https://doi.org/10.1371/journal.pone.0324238.r002
Revision 1
17 Apr 2025 Author Response Dear reviewer� Thank you for taking time to review our manuscript. We studied your comments and revised our draft accordingly. We also went through our manuscript and made corrections on the texts. Minor changes to the manuscript are highlighted. Hope that our revised draft will meet with your approval. Furthermore� we would like to show the details as follow� Reviewer #1 1.Line 39; 40; 68; 70; 87; 132; 181; 200; 203; 207; 323; 326; 337; 340; 358; 364; 388; 459; 471; 473 there is no space after the dot. The author's answer:We have corrected these errors in the article. 2.Line 168; 211 the number of the cells is not written with exponential, as previously done. The author's answer:We have corrected these errors in the article. 3.Line 327; 379; 381; 404; 418; 424; 425; 426; 427; 459; 462 the citation are written at the same level of the text and not as a superscript, as previously done The author's answer:We have corrected these errors in the article. 4.Line 405 Ki67+ have the first letter written in lowercase and not in uppercase, as done before The author's answer:We have corrected these errors in the article. 5.The paragraph of the Flow cytometric analysis is written with a different style respect to the other, like a laboratory protocol, I consider to rewrite this paragraph with the same style of the other. The author's answer:We have revised the sentence in the article. The details as follows: In this study�hucMSCs were analyzed for surface markers using flow cytometry. Single-cell suspensions were centrifuged, washed with PBS, and stained with PE-conjugated antibodies (CD73/CD90/CD105 for positivity, CD34/CD45 as negative controls) alongside isotype-matched controls. Cells were incubated at 4℃ for 30 min in darkness, washed thrice, and resuspended in 1% BSA-PBS. Samples were filtered (40 μm) and analyzed by flow cytometry, gating viable cells via FSC/SSC. A minimum of 10,000 events per sample were recorded, with data processed using FlowJo V10. 6.Line 200 what method was used to determine the protein concentration? Was a standard curve constructed first? If so, how? The author's answer:We have revised the sentence in the article. The details as follows: Protein concentrations were quantified using a bicinchoninic acid (BCA) assay. Briefly, serially diluted bovine serum albumin (BSA) standards and test samples were incubated with BCA working reagent at 37°C for 30 min. Absorbance at 562 nm was measured using a Microplate Reader (Multiskan FC, thermofisher, USA), with sample concentrations calculated through linear regression analysis of the standard curve. 7.Line 202-203 what instrument was used to measure the absorbance? The author's answer:We have revised the sentence in the article. The details as follows: Absorbance was measured at 450 nm using a Microplate Reader. 8.Figure 1A the scale bar is not readable and in Figure 1G and 1H the scale bar is not present The author's answer:We have revised the Figure 1A,G,H. 9.Figure 1K which normalizing protein was used? I would insert its image The author's answer:Thank you for your question.According to the protein marker identification criteria established by the International Society for Extracellular Vesicles (ISEV), the characterization of extracellular vesicles (EVs) should include detection of at least two transmembrane proteins and one cytosolic protein. Therefore, the standard practice involves analysis of three positive markers (mandatorily including one transmembrane protein and one cytosolic protein) alongside one negative marker. For exosomal positive protein markers, transmembrane proteins such as CD9, CD63, and CD81 can be selected, while cytosolic proteins may include TSG101, Alix, or HSP70. Appropriate negative protein markers for exosome identification comprise endoplasmic reticulum-associated proteins (e.g., Calnexin), nuclear proteins (e.g., Histone 3), or Golgi apparatus proteins (e.g., GM130). Given the qualitative nature of this analysis, detection of loading control proteins is generally omitted in most experimental protocols. 10.Figure 4C the scale bar are not present The author's answer:We have revised the Figure 4C Reviewer #2 1.Line 158, are present 2 dots The author's answer:We have corrected these errors in the article. 2.Line 126, 1x10^5 must be written 1x105 The author's answer:We have corrected these errors in the article. 3.Line 168, 1x106 must be written 1x106 The author's answer:We have corrected these errors in the article. 4.Line 185, which type of microscope? The author's answer:Goblet cells are typically observed under an optical microscope(Olympus, Japan) following PAS staining. 5.Line 195, which type of microscope The author's answer:Ki67+ immunohistochemical (IHC) staining sections are observed under an optical microscope (Olympus, Japan). 6.Line 203, how did you measure the absorbance? The author's answer:We have revised the sentence in the article. The details as follows: Absorbance was measured at 450 nm using a Microplate Reader(Multiskan FC, thermofisher, USA). 7.Line 211, 1x10^5 must be written 1x105 The author's answer:We have corrected these errors in the article. 8.Line 219, which instrument is it? The author's answer:It is the Microplate Reader 9.The style of the references is different in the conclusion than in the rest of the article. Check that all bibliographic references are uniform with each other and adhere to the style that the journal dictates. The author's answer:We have corrected these errors in the article. 10.Line 426, commas and References The author's answer:We have corrected these errors in the article. 11.Images 1, 2, 3 e 4 pay attention to the scale bar. Is not visible The author's answer:We have corrected these Figures in the article. 12.Image 3C the words Dapi, Ros and Merge seem to refer to the image above; so bring it closer to the corresponding image. The author's answer:We have corrected the Figure 3C in the article. 13.''CON'' to substitute with ''CTRL'' The author's answer:We have replaced ''CON'' with ''CTRL'' in the article. 14.Unit measures in image 4, graph B The author's answer:We have corrected the Figure 4B in the article. Thank you very much for your attention and time. Look forward to hearing from you. Best wishes, Xiaopeng Han 17 April , 2025 Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0324238.r003
23 Apr 2025 Decision Letter - Sabata Martino, Editor Human umbilical cord mesenchymal stem cell-derived exosomes mitigate acute radiation-induced intestinal oxidative damage via the Nrf2/HO-1/NQO1 signaling pathway PONE-D-25-12676R1 Dear Dr. Han, 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. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Sabata Martino, Ph.D Academic Editor PLOS ONE Additional Editor Comments (optional): no comments Reviewers' comments: https://doi.org/10.1371/journal.pone.0324238.r004
Formally Accepted
Acceptance Letter - Sabata Martino, Editor PONE-D-25-12676R1 PLOS ONE Dear Dr. Han, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Prof. Sabata Martino Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0324238.r005

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