Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip

Julia Bavendiek; Philip Maurer; Steffen Gräber; Andreas Pasch; Werner Karl Schomburg; Willi Jahnen-Dechent

doi:10.1371/journal.pone.0230493

Peer Review History

Original SubmissionAugust 8, 2019
8 Aug 2019 Author Response Attachments Attachment Submitted filename: Response to Reviewers- 19-22409.docx https://doi.org/10.1371/journal.pone.0230493.r001
8 Nov 2019 Decision Letter - Marc W. Merx, Editor PONE-D-19-22409 Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip PLOS ONE Dear Prof Jahnen-Dechent, 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 thoroughly addresses all points raised during the review process and quoted below. We would appreciate receiving your revised manuscript by Dec 23 2019 11:59PM. When you are 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. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. 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Thank you for including your ethics statement: RWTH Aachen University Clinics Ethical Board (www.medizin.rwth-aachen.de/EK; EK 300/14), data were analyzed anonymously Please add your ethics statement to the Methods section of the manuscript and ensure you specify that the named ethics committee specifically approved your study For additional information about PLOS ONE ethical requirements for human subjects research, please refer to http://journals.plos.org/plosone/s/submission-guidelines#loc-human-subjects-research. 3. Please provide additional details about the human serum samples used in this study in the Methods section of your manuscript. If the serum samples were obtained from a biobank or blood bank, please state this in the Methods. 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Competing interests can arise in relationship to an organization or another person. Please follow this link to our website for more details on competing interests: http://journals.plos.org/plosone/s/competing-interests [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 ******** 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: No ****** 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: In this manuscript, Bavendiek et al. present a newly designed microfluidic polymer chip which reduces times for measuring calcification propensity and may form a basis for future point of care tests. The most important part of this test is that measurement of calcification propensity can be achieved at much higher temperatures than the previous test thereby reducing measurement time from 600 to just a few minutes. It is a engineering-orientated paper with detailed reports on the fabrication and optimization of the microchip. Main comments: - What would be the benefit of a possible bedside test? Would there be changes in conducting dialysis or is calcification propensity rather a long-term parameter? - The introduction does not match the actual paper. Introduction handles kidney disease, hyperphosphatemia, morbidity, dialysis, clinical outcome etc, but the paper is an engineering-based approach. This should be adjusted to either more data on clinical use (e.g. adding a small clinical trial) or going all the way to an engineering paper. - The sera of 5 patients was used for validation of the test. But there is no data on the used test sera (Figure 7). Did the patients suffer from clinically relevant calcification? CAD? - Figure 1 seems to be redundant. Reviewer #2: Bavendiek et al demonstrated an improved temperature controlled T50 test format which reports Overall calcification propensity in serum. While this is an important topic, there are some major limitations of this manuscript which might it challenging to follow. The authors should more emphasize the aim of the study, shorten the introduction and extend the discussion. ****** 6. 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: 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 to be viewed.] 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 us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0230493.r002
Revision 1
20 Dec 2019 Author Response Response To Reviewers Our answer: We would like to thank all reviewers for their time and effort to improve the quality of our manuscript. We respond to their comments point-by-point. 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? 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: No 5. Review Comments to the Author Reviewer #1: In this manuscript, Bavendiek et al. present a newly designed microfluidic polymer chip which reduces times for measuring calcification propensity and may form a basis for future point of care tests. The most important part of this test is that measurement of calcification propensity can be achieved at much higher temperatures than the previous test thereby reducing measurement time from 600 to just a few minutes. It is a engineering-orientated paper with detailed reports on the fabrication and optimization of the microchip. Our answer: We thank reviewer 1 for their appreciation of our work and the suggestions made in the comments. Main comments: - What would be the benefit of a possible bedside test? Would there be changes in conducting dialysis or is calcification propensity rather a long-term parameter? Our answer: The benefit of a bedside, or better still, an in-line test could be patient individualized dialysis. An example may be taken from work cited in ref. 30 showing the “The Effect of Increasing Dialysate Magnesium on Serum Calcification Propensity in Subjects with End Stage Kidney Disease”. While such patient-individualized dialysis may still seem a long way off, an improved T50 test could be an important first step, because it better reflects the phosphorous woes (see ref 17) comprising phosphate, serum protein concentration, magnesium, pyrophosphate etc. than does serum phosphate alone. - The introduction does not match the actual paper. Introduction handles kidney disease, hyperphosphatemia, morbidity, dialysis, clinical outcome etc, but the paper is an engineering-based approach. This should be adjusted to either more data on clinical use (e.g. adding a small clinical trial) or going all the way to an engineering paper. Our answer: We thank the reviewer for this suggestion. In the introduction we toned done on clinical findings and focused on the task at hand, technical improvement of the T50 assay. - The sera of 5 patients was used for validation of the test. But there is no data on the used test sera (Figure 7). Did the patients suffer from clinically relevant calcification? CAD? Our answer: We have no information on the health status of the donors, because the samples were anonymized. To perform the repeat measurements shown in Fig 6 (was Fig 7) anonymized serum samples were blended to cover the range of published T50 studies, and to obtain sufficient amount of serum for repeat measurements. For clinical information we refer reviewer to the clinical studies employing the original T50 assay, some of which are cited. - Figure 1 seems to be redundant. Our answer: we deleted Fig 1. Reviewer #2: Bavendiek et al demonstrated an improved temperature controlled T50 test format which reports Overall calcification propensity in serum. While this is an important topic, there are some major limitations of this manuscript which might it challenging to follow. The authors should more emphasize the aim of the study, shorten the introduction and extend the discussion. Our answer: We thank the reviewer for this suggestion. In the introduction we toned done on clinical findings and focused on the task at hand, technical improvement of the T50 assay. At this stage we would like to refrain from further discussion of this proof-of-principle study until we have completed a first clinical study using the novel chip-based T50 assay. We hope the reviewers can agree with this. Attachments Attachment Submitted filename: Response To Reviewers.docx https://doi.org/10.1371/journal.pone.0230493.r003
27 Jan 2020 Decision Letter - Marc W. Merx, Editor PONE-D-19-22409R1 Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip PLOS ONE Dear Prof Jahnen-Dechent, 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 and quoted below. We would appreciate receiving your revised manuscript by Mar 12 2020 11:59PM. When you are 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. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. We look forward to receiving your revised manuscript. Kind regards, Marc W. Merx, MD Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] 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: (No Response) Reviewer #2: (No Response) ******** 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 present their revised manuscript on a newly designed microfluidic polymer chip which reduces times for measuring calcification propensity and may form a basis for future in-line tests. The most important part of this test is that measurement of calcification propensity can be achieved at much higher temperatures than the previous test thereby reducing measurement time from 600 to just a few minutes. It is an engineering-orientated paper with detailed reports on the fabrication and optimization of the microchip. The manuscript has significantly improved due to the changes made in the introduction part. Main comments: - We suggest to include one sentence/paragraph on the impact of reduced CPP measurements on dialysis quality in discussion. Reviewer #2: In the revised manuscript the authors have adequately addressed the reviewers' comments. I would recommend to add one phrase about the expected clinical benefit/ relevance into the abstract. ****** 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: 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 to be viewed.] 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 us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0230493.r004
Revision 2
28 Jan 2020 Author Response We thank both reviewers for their continued help and favorable judgement including their suggestion to comment on the potential clinical impact of this chip. We are happy to oblige. As before, we comment point-by-point quoting the original text. We hope the manuscript now meets the publication criteria. Kind regards Willi Jahnen-Dechent, on behalf of the authors Comments to the Author 6. Review Comments to the Author Reviewer #1: The authors present their revised manuscript on a newly designed microfluidic polymer chip which reduces times for measuring calcification propensity and may form a basis for future in-line tests. The most important part of this test is that measurement of calcification propensity can be achieved at much higher temperatures than the previous test thereby reducing measurement time from 600 to just a few minutes. It is an engineering-orientated paper with detailed reports on the fabrication and optimization of the microchip. The manuscript has significantly improved due to the changes made in the introduction part. Main comments: - We suggest to include one sentence/paragraph on the impact of reduced CPP measurements on dialysis quality in discussion. Our comment: Following reviewer’s suggestion we added a short paragraph to the discussion “Short analysis times are advantageous in clinical settings when T50 measurements inform about the completeness of dialysis or about the influence of varying dialysis protocols or fluids. Importantly, the T50 times reflect the combined calcification propensity including solutes, activators and inhibitors as well as CPP, which might serve as mineralization nuclei that are poorly reflected by conventional phosphate monitoring. Recently, CPP and their precursors were directly end elegantly measured by two independent methods [6, 34]. These and similar methods may further refine the assessment of calcification propensity if miniaturized and accelerated like the chip-based T50 assay presented here.” Reviewer #2: In the revised manuscript the authors have adequately addressed the reviewers' comments. I would recommend to add one phrase about the expected clinical benefit/ relevance into the abstract. Our comment: We added a sentence to the abstract “The speed and reproducibility of the T50 chip-based assay run at 75°C suggest that it may be suitable for rapid measurements, preferably in-line in a dialyser or in a portable microfluidic analytic device with the chip inserted as a disposable cartridge.” Attachments Attachment Submitted filename: Response To Reviewers R2.docx https://doi.org/10.1371/journal.pone.0230493.r005
3 Mar 2020 Decision Letter - Marc W. Merx, Editor Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip PONE-D-19-22409R2 Dear Dr. Jahnen-Dechent, We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, 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 enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and 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. With kind regards, Marc W. Merx, MD 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 ******** 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: (No Response) Reviewer #2: All reviewers' comments are adequately adressed. I would suggest to accept this manuscript for publication. ****** 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: No https://doi.org/10.1371/journal.pone.0230493.r006
Formally Accepted
5 Mar 2020 Acceptance Letter - Marc W. Merx, Editor PONE-D-19-22409R2 Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip Dear Dr. Jahnen-Dechent: I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, 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. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Prof. Dr. Marc W. Merx Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0230493.r007

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