PONE-D-24-35346Efficacy of Silver Needle Thermal Conduction Therapy on Myofascial Pain Syndrome Rats: Based on Quantitative High-resolution Magnetic Resonance T2 Imaging, Shear Wave Elastrography and Muscle Mitochondrial Ultrastructure.PLOS ONE

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Guest Editor

PLOS ONE

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Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: I Don't Know

Reviewer #2: Yes

**********

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Reviewer #1: Yes

Reviewer #2: Yes

**********

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Reviewer #1: Yes

Reviewer #2: Yes

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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. 

Reviewer #1: Dear Authors,

Thank you very much for your article titled "Efficacy of Silver Needle Thermal Conduction Therapy on Myofascial Pain Syndrome Rats: Based on Quantitative High-resolution Magnetic Resonance T2 Imaging, Shear Wave Elastrography and Muscle Mitochondrial Ultrastructure". I found your research very interesting and relevant, and I believe it has the potential to make a significant contribution to the understanding of myofascial pain syndrome and its treatment.

However, I have a few observations and suggestions that I hope will help you improve the clarity and rigor of your manuscript.

Points for Clarification:

Inconsistent Rat Numbers: You mention using 24 rats, divided into three groups of 5. However, the HE staining and TEM analyses mention 6 rats per group. Please standardize the number of rats per group for all analyses.

MPS Model: The description of the MPS induction method should be more detailed. Please specify the frequency and duration of the impacts, as well as the type and duration of the physical training imposed on the rats.

Temperature Justification: You indicate that the needle temperature is 110°C, but that the skin surface temperature is 42°C. Explain the choice of these temperatures and how the skin temperature was controlled and maintained constant.

Suggested Improvements:

SWE Analysis: Please specify the size and shape of the ROIs used for the SWE analysis, as well as the method of selecting these ROIs.

SIRT3 Interpretation: The increase in SIRT3 expression alone is not enough to conclude mitochondrial repair. Additional analyses of mitochondrial function would be necessary to support this conclusion.

Limitations: It would be wise to include a section in the discussion to address the limitations of the study, including the small sample size and the lack of confirmation of mitochondrial repair.

Reviewer #2: This manuscript explores the effects of silver needle thermal conduction therapy on myofascial trigger points (TrPs) using advanced imaging modalities like T2 mapping and strain elastography (STE), complemented by mitochondrial evaluations. The study provides intriguing data and demonstrates high scientific value. However, several critical points need to be addressed to enhance its rigor and clarity.

Strengths

Sophisticated Design: The use of high-resolution magnetic resonance imaging (T2 mapping) and STE to quantitatively assess TrPs is commendable.

Innovative Approach: Employing silver needle thermal conduction therapy and evaluating its effects on mitochondrial structure and function is novel.

Comprehensive Analysis: Combining structural (imaging) and functional (mitochondrial) parameters adds depth to the study.

Major Concerns

Targeting Accuracy of TrPs:

The study does not clearly address whether TrPs were precisely targeted. Imaging-based guidance (e.g., ultrasound) should be detailed to verify accurate targeting before and after the therapy.

Referencing established protocols for TrP detection using ultrasound, such as those by Bubnov et al., would improve the methodological section [Bubnov R.V. Evidence-based pain management: is the concept of integrative medicine applicable?. EPMA Journal 3, 13 (2012). https://doi.org/10.1186/1878-5085-3-13

Bubnov RV: The use of trigger point dry needling under ultrasound guidance for the treatment of myofascial pain (technological innovation and literature review). Lik Sprava. 2010, 5–6: 56-64.]

US Imaging and Needling:

Ultrasound images should be included to showcase how TrPs were detected and evaluated pre- and post-treatment.

Dry needling, widely recognized as effective for TrPs, should be compared to silver needle thermal conduction therapy. This will establish the added value of the thermal conduction method.

Mechanism of Action:

The manuscript does not adequately explain why silver needles were chosen. What unique mechanisms (chemical, mechanical, or thermal) contribute to their effects?

The absence of discussion regarding local twitch responses evoked during treatment is a missed opportunity to connect EMG findings with imaging data.

Title and Focus:

The title should explicitly reflect needling for TrPs instead of the generalized term "Myofascial Pain Syndrome."

Consider shortening the title to improve clarity and focus.

Visualization:

The images do not sufficiently reflect TrPs as a target. Highlighting the precise region of TrPs in T2 mapping or STE images is essential.

Clear visualization would strengthen the study's impact and reproducibility.

Mitochondrial Analysis:

While the mitochondrial findings are compelling, their relevance to the clinical outcomes should be elaborated. Does repairing mitochondrial damage directly correlate with pain relief or muscle function restoration?

Recommendations

Methods: Include a detailed description of ultrasound imaging protocols for TrP detection with references, and explain pre- and post-evaluation methods.

Comparative Analysis: Provide a robust comparison of silver needle therapy with dry needling or other mechanical stimulation methods.

Mechanisms: Discuss the rationale for silver needles and clarify the dominant mechanism (thermal vs. mechanical).

Images: Add annotated imaging of TrPs for better visualization and reproducibility.

Title Revision: Refine the title to emphasize needling for TrPs and its therapeutic implications

**********

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PONE-D-24-35346R1Quantitative evaluation of therapeutic effect of silver needle thermal conduction therapy on myofascial trigger pointPLOS ONE

Dear Dr. Qin,

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 Apr 21 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.

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). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Jiajia Ye

Guest Editor

PLOS ONE

Journal Requirements:

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:

The manuscript has been significantly improved since the initial submission, with clearer explanations and refined methodology. However, a major conceptual issue remains unresolved: the justification for selecting silver needle thermal conduction therapy (SNTCT) over conventional dry needling (DN). This aspect must be explicitly addressed throughout the manuscript to enhance its scientific rigor and clinical relevance.

Major Issues:

1. Justification for Silver Needle Thermal Conduction Therapy (SNTCT)

The manuscript should clearly define why the authors chose this method over conventional DN. The following aspects require detailed discussion:

Why introduce silver and heat to the needle when standard fine needles are already effective for trigger point inactivation?

What specific physiological or biomechanical advantages does silver provide in this context? If its antimicrobial properties or conductivity are relevant, these should be supported with evidence.

How does thermal conduction enhance the therapeutic effect beyond what is achieved with precise DN?

Is there prior research supporting the addition of these variables in needling therapy? If not, a stronger theoretical basis should be provided.

If silver is only used as a thermal conductor, why not use another material?

2. Mechanism of Action & Physiological Effects

The study should clarify the hypothesized mechanisms through which silver and heat alter the effects of DN:

The standard effectiveness of DN is largely attributed to precise intervention and eliciting localized twitch responses (LTRs) in trigger points. How do heat and silver modify this response?

Could thermal conduction alter needle sensitivity, thereby influencing tactile feedback and the ability to localize trigger points?

If heat increases circulation or pain modulation, was this measured or supported by prior research?

Were adverse effects (e.g., risk of burns, altered tissue response) considered?

3. Study Design Considerations

To isolate the true effect of silver and heat, the study should ideally include:

A comparison between standard DN, silver-coated DN, and silver-thermal DN to assess individual contributions.

A discussion on whether blinding was possible and how placebo effects were minimized.

If no direct comparison was made, the authors should acknowledge this as a limitation and suggest future studies to address it.

4. Clinical Applicability & Cost-Benefit Considerations

If precise DN is already effective, does the added complexity and potential cost of SNTCT justify its use?

Are there specific clinical scenarios where silver thermal conduction needling would be superior to traditional DN?

Could this method introduce risks or complications that are not present with standard DN?

5. Needling Precision & Local Twitch Responses (LTRs)

The therapeutic effectiveness of DN is highly dependent on precise needling technique and the elicitation of LTRs. The manuscript does not discuss whether silver-thermal needling affects this precision.

The authors should provide detailed descriptions of their needling protocol to ensure consistency with established DN methodologies, such as those described in [https://pmc.ncbi.nlm.nih.gov/articles/PMC3533862/].

If LTRs were monitored, this should be explicitly stated. If not, the authors should acknowledge that this is a limitation of the study.

Minor Issues:

Terminology consistency: Ensure that all references to the intervention (SNTCT, silver-thermal DN, etc.) are consistent throughout the manuscript.

Literature review expansion: Include more references to existing studies comparing different needling techniques.

Clarify study limitations: If the study does not separate silver and heat effects, this should be acknowledged.

Conclusion:

The manuscript must provide a strong, evidence-based justification for incorporating silver and heat into DN. Additionally, the study design should either control for these variables separately or acknowledge this as a limitation. Addressing these concerns will significantly enhance the manuscript’s scientific and clinical relevance.

[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: 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: Dear Author

I am writing to express my sincere gratitude for the modifications you have made to your manuscript. Your diligence in addressing the concerns raised by the reviewers has significantly improved the precision and accuracy of the article.

Reviewer #2: The manuscript has been significantly improved since the initial submission, with clearer explanations and refined methodology. However, a major conceptual issue remains unresolved: the justification for selecting silver needle thermal conduction therapy (SNTCT) over conventional dry needling (DN). This aspect must be explicitly addressed throughout the manuscript to enhance its scientific rigor and clinical relevance.

Major Issues:

1. Justification for Silver Needle Thermal Conduction Therapy (SNTCT)

The manuscript should clearly define why the authors chose this method over conventional DN. The following aspects require detailed discussion:

Why introduce silver and heat to the needle when standard fine needles are already effective for trigger point inactivation?

What specific physiological or biomechanical advantages does silver provide in this context? If its antimicrobial properties or conductivity are relevant, these should be supported with evidence.

How does thermal conduction enhance the therapeutic effect beyond what is achieved with precise DN?

Is there prior research supporting the addition of these variables in needling therapy? If not, a stronger theoretical basis should be provided.

If silver is only used as a thermal conductor, why not use another material?

2. Mechanism of Action & Physiological Effects

The study should clarify the hypothesized mechanisms through which silver and heat alter the effects of DN:

The standard effectiveness of DN is largely attributed to precise intervention and eliciting localized twitch responses (LTRs) in trigger points. How do heat and silver modify this response?

Could thermal conduction alter needle sensitivity, thereby influencing tactile feedback and the ability to localize trigger points?

If heat increases circulation or pain modulation, was this measured or supported by prior research?

Were adverse effects (e.g., risk of burns, altered tissue response) considered?

3. Study Design Considerations

To isolate the true effect of silver and heat, the study should ideally include:

A comparison between standard DN, silver-coated DN, and silver-thermal DN to assess individual contributions.

A discussion on whether blinding was possible and how placebo effects were minimized.

If no direct comparison was made, the authors should acknowledge this as a limitation and suggest future studies to address it.

4. Clinical Applicability & Cost-Benefit Considerations

If precise DN is already effective, does the added complexity and potential cost of SNTCT justify its use?

Are there specific clinical scenarios where silver thermal conduction needling would be superior to traditional DN?

Could this method introduce risks or complications that are not present with standard DN?

5. Needling Precision & Local Twitch Responses (LTRs)

The therapeutic effectiveness of DN is highly dependent on precise needling technique and the elicitation of LTRs. The manuscript does not discuss whether silver-thermal needling affects this precision.

The authors should provide detailed descriptions of their needling protocol to ensure consistency with established DN methodologies, such as those described in [https://pmc.ncbi.nlm.nih.gov/articles/PMC3533862/].

If LTRs were monitored, this should be explicitly stated. If not, the authors should acknowledge that this is a limitation of the study.

Minor Issues:

Terminology consistency: Ensure that all references to the intervention (SNTCT, silver-thermal DN, etc.) are consistent throughout the manuscript.

Literature review expansion: Include more references to existing studies comparing different needling techniques.

Clarify study limitations: If the study does not separate silver and heat effects, this should be acknowledged.

Conclusion:

The manuscript must provide a strong, evidence-based justification for incorporating silver and heat into DN. Additionally, the study design should either control for these variables separately or acknowledge this as a limitation. Addressing these concerns will significantly enhance the manuscript’s scientific and clinical relevance.

**********

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: Yes:  Dr Paul ELHOMSY

Reviewer #2: Yes:  Rostyslav Bubnov

**********

[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 . Please note that Supporting Information files do not need this step.

PONE-D-24-35346R2Quantitative evaluation of therapeutic effect of silver needle thermal conduction therapy on myofascial trigger pointPLOS ONE

Dear Dr. Wang,

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 28 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.

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). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Jiajia Ye

Guest Editor

PLOS ONE

Journal Requirements:

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:

Please address the comments from the reviewers below:

The manuscript has been significantly improved since the initial submission, with clearer explanations and refined methodology. However, a major conceptual issue remains unresolved: the justification for selecting silver needle thermal conduction therapy (SNTCT) over conventional dry needling (DN). This aspect must be explicitly addressed throughout the manuscript to enhance its scientific rigor and clinical relevance.

Major Issues:

1. Justification for Silver Needle Thermal Conduction Therapy (SNTCT)

The manuscript should clearly define why the authors chose this method over conventional DN. The following aspects require detailed discussion:

Why introduce silver and heat to the needle when standard fine needles are already effective for trigger point inactivation?

What specific physiological or biomechanical advantages does silver provide in this context? If its antimicrobial properties or conductivity are relevant, these should be supported with evidence.

How does thermal conduction enhance the therapeutic effect beyond what is achieved with precise DN?

Is there prior research supporting the addition of these variables in needling therapy? If not, a stronger theoretical basis should be provided.

If silver is only used as a thermal conductor, why not use another material?

2. Mechanism of Action & Physiological Effects

The study should clarify the hypothesized mechanisms through which silver and heat alter the effects of DN:

The standard effectiveness of DN is largely attributed to precise intervention and eliciting localized twitch responses (LTRs) in trigger points. How do heat and silver modify this response?

Could thermal conduction alter needle sensitivity, thereby influencing tactile feedback and the ability to localize trigger points?

If heat increases circulation or pain modulation, was this measured or supported by prior research?

Were adverse effects (e.g., risk of burns, altered tissue response) considered?

3. Study Design Considerations

To isolate the true effect of silver and heat, the study should ideally include:

A comparison between standard DN, silver-coated DN, and silver-thermal DN to assess individual contributions.

A discussion on whether blinding was possible and how placebo effects were minimized.

If no direct comparison was made, the authors should acknowledge this as a limitation and suggest future studies to address it.

4. Clinical Applicability & Cost-Benefit Considerations

If precise DN is already effective, does the added complexity and potential cost of SNTCT justify its use?

Are there specific clinical scenarios where silver thermal conduction needling would be superior to traditional DN?

Could this method introduce risks or complications that are not present with standard DN?

5. Needling Precision & Local Twitch Responses (LTRs)

The therapeutic effectiveness of DN is highly dependent on precise needling technique and the elicitation of LTRs. The manuscript does not discuss whether silver-thermal needling affects this precision.

The authors should provide detailed descriptions of their needling protocol to ensure consistency with established DN methodologies, such as those described in [https://pmc.ncbi.nlm.nih.gov/articles/PMC3533862/].

If LTRs were monitored, this should be explicitly stated. If not, the authors should acknowledge that this is a limitation of the study.

Minor Issues:

Terminology consistency: Ensure that all references to the intervention (SNTCT, silver-thermal DN, etc.) are consistent throughout the manuscript.

Literature review expansion: Include more references to existing studies comparing different needling techniques.

Clarify study limitations: If the study does not separate silver and heat effects, this should be acknowledged.

Conclusion:

The manuscript must provide a strong, evidence-based justification for incorporating silver and heat into DN. Additionally, the study design should either control for these variables separately or acknowledge this as a limitation. Addressing these concerns will significantly enhance the manuscript’s scientific and clinical relevance.

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

[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 . Please note that Supporting Information files do not need this step.

PONE-D-24-35346R3

Quantitative evaluation of therapeutic effect of silver needle thermal conduction therapy on myofascial trigger pointPLOS ONE

Dear Dr. Wang,

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 Aug 15 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.

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). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Jiajia Ye

Guest Editor

PLOS ONE

Comments from PLOS Editorial Office:

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

Journal Requirements:

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.

[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 #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 #2: Yes

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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 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 #2: Yes

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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 #2: Yes

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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 #2: The revised manuscript significantly improves on the original submission, offering detailed explanations on the mechanism and application of silver needle thermal conduction therapy (SNTCT) for myofascial pain syndrome (MPS). The use of T2 mapping, sound touch elastography (STE), transmission electron microscopy (TEM), and molecular assays provides a valuable multiparametric approach to objectively assess muscle and mitochondrial recovery. Ethical compliance and experimental methodology are well documented.

However, there remain important issues regarding the characterization of comparative therapies, particularly dry needling (DN), and the limitations of the study design.

Major Strengths

Comprehensive multimodal evaluation including MRI, elastography, EMG, histology, and SIRT3 analysis.

Clear explanation of SNTCT protocol and physiological rationale.

Acknowledgment of study limitations in the discussion section.

Ethically approved and statistically reasonable animal study design.

Key Issues and Recommendations

1. Misrepresentation of Dry Needling

The authors' attempt to differentiate SNTCT from dry needling (DN) is not adequately supported and contains misleading claims. The response (Major Issue 1) suggests that DN is a superficial, acupuncture-point-based technique requiring repeated treatment sessions. This description does not reflect current clinical practice or research evidence.

Modern dry needling—particularly ultrasound-guided dry needling (US-DN)—enables precise targeting of myofascial trigger points (TrPs) under direct anatomical visualization. As shown in publications by Bubnov R. (Bubnov, 2013; https://www.liebertpub.com/doi/abs/10.1089/acu.2013.0973 https://linkinghub.elsevier.com/retrieve/pii/S0003496724543175, etc.), US-DN can be highly effective, reproducible, and minimally invasive, often requiring a single treatment without the need for thermal or material enhancements.

Additionally, the assertion that DN lacks long-term efficacy or acts solely through mechanical disruption without benefit is overly simplistic. DN protocols often elicit local twitch responses (LTRs), a known therapeutic mechanism in TrP inactivation. The role of SNTCT should be discussed as an alternative or adjunct, not a superior replacement, unless comparative data are provided.

Recommendation: The manuscript should be revised to accurately represent DN, particularly ultrasound-guided DN, and provide a balanced comparison. Please acknowledge that:

DN can be anatomically precise.

US guidance enhances safety and efficacy.

DN may not universally require repeated sessions.

Any claimed superiority of SNTCT should be tested in direct comparative studies.

2. Study Design Limitations

The lack of comparator groups (e.g., DN, silver-only, heat-only) prevents conclusions about which component(s) of SNTCT are responsible for observed effects. While acknowledged in the discussion, this limitation is fundamental and should be clearly emphasized in the conclusions.

3. Clinical Translation

Given the strong animal data, the authors should discuss the translational roadmap for SNTCT, including safety, patient selection, cost implications, and potential for integration with ultrasound guidance.

4. Terminology

Please ensure consistent use of terminology (e.g., "silver needle thermal conduction therapy" vs. "SNTCT" vs. "silver-thermal DN").

5. Mitochondrial Mechanism

The link between mitochondrial repair and pain relief is plausible but remains speculative without direct measurement of functional mitochondrial output (e.g., ATP, ROS). This should be acknowledged more explicitly.

Final Recommendation

Minor Revision

The manuscript presents valuable and novel findings. However, it requires clarification and correction of statements about comparative therapies—particularly modern DN techniques—to ensure scientific integrity. A revised conclusion section acknowledging these limitations and a more balanced comparison will significantly strengthen the paper’s clinical relevance and accuracy.

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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 #2: Yes:  Rostyslav Bubnov

**********

[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 . Please note that Supporting Information files do not need this step.

PONE-D-24-35346R4Quantitative evaluation of therapeutic effect of silver needle thermal conduction therapy on myofascial trigger pointPLOS ONE

Dear Dr. Wang,

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 Aug 24 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.

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). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Jiajia Ye

Guest Editor

PLOS ONE

Journal Requirements:

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 :

It is strongly recommended that the authors carefully revise the page and line numbers to ensure that the editor and reviewers can clearly identify the changes made.

Additionally, please note that the suggested references are optional. Authors are encouraged to thoroughly review and assess the relevance of each suggestion, and only cite references that are highly pertinent to the manuscript.

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

[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 . Please note that Supporting Information files do not need this step.

Quantitative evaluation of therapeutic effect of silver needle thermal conduction therapy on myofascial trigger point

PONE-D-24-35346R5

Dear Dr. Wang,

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,

Jiajia Ye

Guest 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 #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 #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 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 #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 #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 #2: This version meets the journal’s publication standards. It offers a well-structured preclinical study with clear scientific rationale, multiparametric objective measurements, and an appropriately framed interpretation of results. The authors have shown responsiveness to peer review and improved the manuscript accordingly.

**********

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 #2: Yes:  Rostyslav Bubnov

**********