Delayed delivery of antibiotics by ultrasound-mediated rupture of polylactic acid pockets: In vitro and in vivo studies

Selin Isguven Billmyer; Priscilla Machado; Ryan E. Tomlinson; Lauren J. Delaney; Ji-Bin Liu; Alexander H. Harris; Eric McLaughlin; Noreen J. Hickok; Flemming Forsberg

doi:10.1371/journal.pone.0337717

Peer Review History

Original SubmissionMarch 18, 2025
28 Apr 2025 Decision Letter - Dola Sundeep, Editor Dear Dr. Forsberg, 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 Jun 12 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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We look forward to receiving your revised manuscript. Kind regards, Dola Sundeep Academic Editor PLOS ONE Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. To comply with PLOS ONE submissions requirements, in your Methods section, please provide additional information regarding the experiments involving animals and ensure you have included details on (1) methods of sacrifice, (2) methods of anesthesia and/or analgesia, and (3) efforts to alleviate suffering. 3. Thank you for stating the following financial disclosure: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University” Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 4. Thank you for stating the following in the Acknowledgments Section of your manuscript: “We thank Hebah Falatah for Definity droplet preparation and Catherine Gurr for help with data presentation. Research reported in this manuscript was supported by the National Institute of Arthritis and Musculoskeletal Diseases of the National Institutes of Health under award number R01AR069119 (Hickok, Forsberg, Isguven) and by the Mullin Fund for Spinal Research at Thomas Jefferson University. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. “ We note that you have provided funding information that is currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 5. Thank you for stating the following in the Competing Interests section: “I have read the journal's policy and the authors of this manuscript have the following competing interests: Selin Isguven Billmyer, Noreen Hickok, Flemming Forsberg reports financial support was provided by National Institutes of Health. Flemming Forsberg reports a relationship with GE HealthCare that includes: consulting or advisory. Flemming Forsberg reports a relationship with Lantheus Medical Imaging Inc that includes: consulting or advisory. Selin Isguven Billmyert, Noreen Hickok, Flemming Forsberg has patent PCT Application No. PCT/US2023/077854 pending to Thomas Jefferson University. The other authors, declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.” Please confirm that this does not alter your adherence to all PLOS ONE policies on sharing data and materials, by including the following statement: "This does not alter our adherence to PLOS ONE policies on sharing data and materials.” (as detailed online in our guide for authors http://journals.plos.org/plosone/s/competing-interests). If there are restrictions on sharing of data and/or materials, please state these. Please note that we cannot proceed with consideration of your article until this information has been declared. Please include your updated Competing Interests statement in your cover letter; we will change the online submission form on your behalf. 6. In this instance it seems there may be acceptable restrictions in place that prevent the public sharing of your minimal data. However, in line with our goal of ensuring long-term data availability to all interested researchers, PLOS’ Data Policy states that authors cannot be the sole named individuals responsible for ensuring data access (http://journals.plos.org/plosone/s/data-availability#loc-acceptable-data-sharing-methods). Data requests to a non-author institutional point of contact, such as a data access or ethics committee, helps guarantee long term stability and availability of data. Providing interested researchers with a durable point of contact ensures data will be accessible even if an author changes email addresses, institutions, or becomes unavailable to answer requests. Before we proceed with your manuscript, please also provide non-author contact information (phone/email/hyperlink) for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If no institutional body is available to respond to requests for your minimal data, please consider if there any institutional representatives who did not collaborate in the study, and are not listed as authors on the manuscript, who would be able to hold the data and respond to external requests for data access? If so, please provide their contact information (i.e., email address). Please also provide details on how you will ensure persistent or long-term data storage and availability. 7. Please review your reference list to ensure that it is complete and correct. 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: Subject: Decision on Your Manuscript PONE-D-25-14645 Dear Dr. Forsberg, We have now received the required number of reviewer reports for your manuscript titled "Delayed delivery of antibiotics by ultrasound-mediated rupture of polylactic acid pockets: in vitro and in vivo studies" (Manuscript Number: PONE-D-25-14645). Based on the reviewers' comments, the decision on your manuscript is Minor Revision. We look forward to receiving your revised submission. Thank you for your continued interest in our journal. Best regards, Dr. Dola Sundeep 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: This study introduces a promising and innovative ultrasound-triggered antibiotic delivery system for spinal surgery infections. The approach is highly relevant and potentially impactful, offering a non-invasive method for localized infection control. However, to strengthen the manuscript, several areas need improvement: Statistical Analysis – Key outcomes like rupture rates and antibiotic release should include significance testing. Long-Term Efficacy – Additional data on stability and prolonged effectiveness are needed. Comparative Controls – Comparing with other delivery systems (e.g., beads, hydrogels) would add context. Tissue Compatibility – More information on inflammatory responses or biocompatibility is recommended. Ultrasound Parameters – Clearer details (e.g., frequency, intensity, duration) would aid reproducibility. Limitations & Future Work – The authors should discuss the small sample size and outline next steps. Overall, this is a well-designed, novel study with strong potential, but a few enhancements would improve its clarity, reproducibility, and clinical applicability. Reviewer #2: PONE-D-25-14645 Review I would recommend publishing however, authors should explain the points below and consider the following revisions. 1) Authors should explain why the cone shaped geometry was selected for this application. 2) Authors should consider and report MeB and VAN release data as % release / surface area of the pocket 3) How many pockets will be needed to deliver the typical bolus dose of VAN post-surgery ? 4) Have the authors altering the hydrophilicity of PLA , by replacing it with hydrophobic PLGA in the thin film ? 5) Detailed surface characterization using AFM and visualization using SEM, might help explain the difference between invitro and invivo disconnect that the authors observed. ****** 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: Yes: Nour H. Aboalhaija 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 https://doi.org/10.1371/journal.pone.0337717.r001
Revision 1
13 Oct 2025 Author Response August 12, 2025 Dola Sundeep Academic Editor PLOS ONE Re: Delayed delivery of antibiotics by ultrasound-mediated rupture of polylactic acid pockets: in vitro and in vivo studies; Manuscript no: PONE-D-25-14645 Dear Dr. Sundeep Thank you for your email of April 28th, 2025 and for the reviewer’s insightful comments on our manuscript referenced above. Our responses to the questions raised can be found below. All line references refer to the revised and marked version of the manuscript. Finally, we discovered a few trivial typographical and linguistic errors in the original manuscript and those have also been corrected. Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf We have carefully gone through the entire manuscript and made sure it conforms to the journal guidelines. 2. To comply with PLOS ONE submissions requirements, in your Methods section, please provide additional information regarding the experiments involving animals and ensure you have included details on (1) methods of sacrifice, (2) methods of anesthesia and/or analgesia, and (3) efforts to alleviate suffering. As requested we have expanded the information regarding our animal experiments, which now reads (in part): “Animal experiments were performed according to a protocol (Protocol Number: 22-11-606) approved by the Institutional Animal Care & Use Committee (IACUC) of Thomas Jefferson University and in accordance with the National Research Council's “Guide for the Care and Use of Laboratory Animals.” All surgery was performed under 1-4% isoflurane anesthesia, and all efforts were made to minimize suffering. Rabbits were pre-medicated with ketamine 30-40 mg/kg, xylazine 2-5 mg/kg, and acepromazine 0.25-1.00 mg/kg. Anesthesia was induced with 4-5% isoflurane and maintained with 1-4% isoflurane during the entire procedure. ... Following implantation, the rabbits were allowed unrestricted ambulation and observed for activity and recovery. Wounds were inspected daily for drainage, erythema, warmth, and swelling, as well as MeB release … for a total of 6 days, at which time all animals were euthanized with a barbiturate overdose in accordance with AVMA recommendations.” (lines 247 - 268) 3. Thank you for stating the following financial disclosure: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University” Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. Thank you for your guidance. The cover letter now includes the statement: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” 4. Thank you for stating the following in the Acknowledgments Section of your manuscript: “We thank Hebah Falatah for Definity droplet preparation and Catherine Gurr for help with data presentation. Research reported in this manuscript was supported by the National Institute of Arthritis and Musculoskeletal Diseases of the National Institutes of Health under award number R01AR069119 (Hickok, Forsberg, Isguven) and by the Mullin Fund for Spinal Research at Thomas Jefferson University. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. “ We note that you have provided funding information that is currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. Thank you for your guidance. The cover letter now includes the statement: “NIH R01 AR069119 The Mullin Fund for Spinal Research at Thomas Jefferson University The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” Moreover, we have removed the funding information from the Acknowledgement section, which now reads” “We thank Hebah Falatah for Definity droplet preparation and Catherine Gurr for help with data presentation.” (lines 561 - 562) 5. Thank you for stating the following in the Competing Interests section: “I have read the journal's policy and the authors of this manuscript have the following competing interests: Selin Isguven Billmyer, Noreen Hickok, Flemming Forsberg reports financial support was provided by National Institutes of Health. Flemming Forsberg reports a relationship with GE HealthCare that includes: consulting or advisory. Flemming Forsberg reports a relationship with Lantheus Medical Imaging Inc that includes: consulting or advisory. Selin Isguven Billmyert, Noreen Hickok, Flemming Forsberg has patent PCT Application No. PCT/US2023/077854 pending to Thomas Jefferson University. The other authors, declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.” Please confirm that this does not alter your adherence to all PLOS ONE policies on sharing data and materials, by including the following statement: "This does not alter our adherence to PLOSONE policies on sharing data and materials.” (as detailed online in our guide for authors http://journals.plos.org/plosone/s/competing-interests). If there are restrictions on sharing of data and/or materials, please state these. Please note that we cannot proceed with consideration of your article until this information has been declared. Please include your updated Competing Interests statement in your cover letter; we will change the online submission form on your behalf. Thank you for your guidance. The cover letter now includes the statement: “I have read the journal's policy and the authors of this manuscript have the following competing interests: Selin Isguven Billmyer, Noreen Hickok, Flemming Forsberg report financial support was provided by National Institutes of Health. Flemming Forsberg reports a relationship with GE HealthCare that includes: consulting or advisory. Flemming Forsberg reports a relationship with Lantheus Medical Imaging Inc that includes: consulting or advisory. Selin Isguven Billmyer, Noreen Hickok, Flemming Forsberg have patent PCT Application No. PCT/US2023/077854 pending to Thomas Jefferson University. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. This does not alter our adherence to PLOS One policies on sharing data and materials.” 6. In this instance it seems there may be acceptable restrictions in place that prevent the public sharing of your minimal data. However, in line with our goal of ensuring long-term data availability to all interested researchers, PLOS’ Data Policy states that authors cannot be the sole named individuals responsible for ensuring data access (http://journals.plos.org/plosone/s/data-availability#loc-acceptable-data-sharing-methods). Data requests to a non-author institutional point of contact, such as a data access or ethics committee, helps guarantee long term stability and availability of data. Providing interested researchers with a durable point of contact ensures data will be accessible even if an author changes email addresses, institutions, or becomes unavailable to answer requests. Before we proceed with your manuscript, please also provide non-author contact information (phone/email/hyperlink) for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If no institutional body is available to respond to requests for your minimal data, please consider if there any institutional representatives who did not collaborate in the study, and are not listed as authors on the manuscript, who would be able to hold the data and respond to external requests for data access? If so, please provide their contact information (i.e., email address). Please also provide details on how you will ensure persistent or long-term data storage and availability. Our institution does not have a data access committee or anything similar. We will provide a complete copy of all the data to Dr. John Eisenbrey (Professor of Radiology) who will maintain long term availability of the data from this study and who will handle any requests regarding access. His email is: john.eisenbrey@jefferson.edu 7. Please review your reference list to ensure that it is complete and correct. 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. We are not aware of any retractions among our reference list have updated the list to reflect the PLOS One style, as requested. Moreover, we replaced the abstract cited as (26) with the peer-reviewed paper of this study. Finally, three new references were added to comply with the expanded discussion requested by the reviewers (32, 33, 50). Additional Editor Comments: Subject: Decision on Your Manuscript PONE-D-25-14645 Dear Dr. Forsberg, We have now received the required number of reviewer reports for your manuscript titled "Delayed delivery of antibiotics by ultrasound-mediated rupture of polylactic acid pockets: in vitro and in vivo studies" (Manuscript Number: PONE-D-25-14645). Based on the reviewers' comments, the decision on your manuscript is Minor Revision. We look forward to receiving your revised submission. Thank you for your continued interest in our journal. Best regards, Dr. Dola Sundeep 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 ________________________________________ 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 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 ________________________________________ 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: Yes Reviewer #2: Yes ________________________________________ 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: This study introduces a promising and innovative ultrasound-triggered antibiotic delivery system for spinal surgery infections. The approach is highly relevant and potentially impactful, offering a non-invasive method for localized infection control. However, to strengthen the manuscript, several areas need improvement: Statistical Analysis – Key outcomes like rupture rates and antibiotic release should include significance testing. We tested the rupture rates for neat PLA and PLA-VAN pockets and found no statistically significant difference (p > 0.99). We also clarified the issue with the one excluded pocket and the text now reads: “Importantly, all 5 of the intact pockets (3 neat PLA and 2 PLA-VAN) that were allocated for insonation ruptured following US (100%; p > 0.99). For the neat PLA pockets, 5 out of 9 were intact on day 3, while 4 out of 6 PLA-VAN pockets survived until day 3 (p > 0.99). An additional pocket from Rabbit #5 was excluded from analysis, due to the emptying of the pocket contents occurring without staining, making it impossible to determine the timeline of leakage.” (lines 433 - 438). The antibiotic release rates were converted to % (as requested by Reviewer 2) and testing showed no increase by day 15 (p = 0.19) but after 27 days the release was statistically significantly higher (p = 0.0436). Both p-values have now been added to the text, which was rewritten to: ” For a first trial, the percent VAN release from the film in the first day and up to 15 days (Fig. 11-A) are shown. Total release values are close to 2.5 mg or 2,500 µg (n = 3; p = 0.19). Despite the variability in each sample, superficial VAN dissolved in the first couple of hours, in keeping with many passive elution systems. We next investigated if VAN release was still present at 27 days. Again, we measured immediate VAN release expressed as percent of the total loading (n = 3) on the first day and up to 27 days, which was statistically significantly higher (p = 0.0436; Fig. 11-B). Most of the VAN dissolved early in the incubation period, although some additional release was measured after the first day, which we speculate is due to some PLA degradation which releases additional VAN.“ (lines 403 - 416). Long-Term Efficacy – Additional data on stability and prolonged effectiveness are needed. The premise of our local drug delivery system is that all pockets must be activated within 6 days to reduce the possibility of bacterial infections taking hold (i.e., for prophylaxis). There is therefore no need for long-term efficacy studies, and we have not made any changes to the manuscript in this regard. Comparative Controls – Comparing with other delivery systems (e.g., beads, hydrogels) would add context. While we do agree with the reviewer, we feel that such experiments are outside the scope of this manuscript. We do ho Attachments Attachment Submitted filename: Response to reviewers.docx https://doi.org/10.1371/journal.pone.0337717.r002
12 Nov 2025 Decision Letter - Dola Sundeep, Editor Delayed delivery of antibiotics by ultrasound-mediated rupture of polylactic acid pockets: in vitro and in vivo studies PONE-D-25-14645R1 Dear Dr. Flemming Forsberg, 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. For questions related to billing, please contact billing support . 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, Dola Sundeep Academic Editor PLOS ONE Additional Editor Comments (optional): Dear Dr. Flemming Forsberg, Thank you for submitting your manuscript entitled “Delayed Delivery of Antibiotics by Ultrasound-Mediated Rupture of Polylactic Acid Pockets: In Vitro and In Vivo Studies” (Manuscript ID: PONE-D-25-14645R1) to PLOS ONE. Based on the reviewers’ comments and the satisfactory revisions made in response, I am pleased to inform you that the final editorial decision is to Accept your manuscript for publication in PLOS ONE. We appreciate your careful attention to the reviewers’ suggestions and the improvements made to enhance the quality and clarity of your work. Congratulations on this achievement, and thank you for choosing PLOS ONE as the venue for your research. Kind regards, Dr. Dola Sundeep Academic Editor PLOS ONE Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #2: Yes ****** Reviewer #2: (No Response) ****** 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 #2: No ******** https://doi.org/10.1371/journal.pone.0337717.r003
Formally Accepted
Acceptance Letter - Dola Sundeep, Editor PONE-D-25-14645R1 PLOS ONE Dear Dr. Forsberg, 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. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. 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 Dr. Dola Sundeep Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0337717.r004

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