Dear Dr. Khalid Ahmed,

Please submit your revised manuscript by Apr 07 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'.
• 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,

Sharun Khan

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. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager.

3. 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.

4. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 

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

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

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

Reviewer #1: Partly

Reviewer #2: Partly

**********

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

Reviewer #1: No

Reviewer #2: No

**********

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: No

**********

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

Reviewer #1: Yes

Reviewer #2: No

**********

Reviewer #1: Septic arthritis is a disease characterized by rapid joint damage as well as cartilage degeneration, and there is a growing interest in the use of MSCs for the treatment of septic arthritis due to their accessibility and regenerative potential.This study compared the efficacy, safety, and delivery of BM-MSCs and AD-MSCs in the treatment of septic arthritis. However, there are still several issues that need to be addressed.

1. In this study, the authors set up a total of nine subgroups, including control group, treated group(BM-MSC1,BM-MSC2,AD-MSC1,AD-MSC2), and untreated group(Un-BM1,Un-BM2,Un-AD1,Un-AD2). However, there are some places in the manuscript where the descriptions of the groups are not clear enough, for example, the abstract section may not be accurate in describing the groups. In addition, it seems that the relevant results for the model group are missing from the manuscript.

2. The definitions of treated and untreated are not clear in the manuscripts. Please rephrase for better understanding.

3. Please indicate the generation of MSCs used for the experiment. In addition, please provide a morphologic map of the MSCs and a file in FCS format for flow cytometry.

4. Please describe in more detail the experimental details of the section of “Materials and Methods”. In addition, there are some problems with the logic of the language in this section, please rewrite this section.

5. This study compared the efficacy and safety of BM-MSCs with AD-MSCs. Interestingly, BM-MSC1 appears to be the optimal candidate for the treatment of septic osteoarthritis. However, what does “1”, and “2” mean respectively, as well as please elucidate the potential mechanism that produces this phenomenon.

6. Please provide HE staining and histochemical results of the model group and untreated group(Un-BM1,Un-BM2,Un-AD1,Un-AD2) to better visualize the efficacy of MSCs. Please add a scale bar to the HE staining and histochemistry diagrams or write the magnification in the figure legends.

7. Please add symbols to the bar graphs that can represent statistical differences. In addition, please indicate in the figure legends the sample size of animals used for experimental analyses.

8. The study shown that BM-MSC outperforms AD-MSC in terms of efficacy, safety, and residence time. Please explain these phenomena.

Reviewer #2: The authors aimed to address a significant and relevant biological question and have employed robust analytical techniques; however, there are notable areas that require improvement. The manuscript, in its current form, presents significant methodological and reporting deficiencies that must be addressed. Without clear experimental design details, proper stem cell characterization, statistical clarity, and complete data presentation, the study’s conclusions remain unsubstantiated. I recommend major revisions to improve the manuscript’s scientific validity before it can be reconsidered for publication.

The study has potential relevance but is not presented or documented properly, it’s crucial to address the bare minimum requirements to improve its quality and credibility.

• Clearly state the methodology and ensure that all treatment groups are properly defined.

• Revise for clarity and completeness.

• Ensure that all abbreviations are defined upon first use in the abstract and text (e.g., ADMSC and BMSC).

• Line 54: Verify the reference that claims BM-MSCs as the gold standard.

• Line 68: Add a supporting reference.

• Specify the total number of animals used, including their sex and distribution across groups.

• Clearly mention how many animals were used for stem cell isolation.

• The study lacks characterization of stem cell markers—explain why specific markers for ADMSC and BM-MSC were not used.

• Minimum reporting criteria for stem cell characterization have not been met. Include data to support MSC characterization

• The current description lacks clarity. Introduce a separate heading for Experimental Design.

• Clearly define ADMSC1, ADMSC2, BMMSC1, and BMMSC2, explaining their roles in the study.

• Line 127: Add a reference for the dose of stem cells used.

• Line 164: Specify the statistical methods used, including the software, significance level, and confidence interval.

• Mention the primers used or provide a reference for RNA quantification methods.

• The ISCT criteria for MSC characterization have not been fully met—include additional characterization data to confirm MSC identity and differentiate ADMSC from BM-MSC.

• Clearly mention the histological scoring method used.

• Include necessary references for materials and methods.

• Histopathology Figures:

o Ensure scale bars and magnifications are specified.

o Specify the organ being analyzed in the figure legends.

• Sox9 Expression: The non-specific binding observed in BM-MSC groups needs to be clarified. Provide an alternative figure or additional data for validation.

• Fold Change Calculations:

o Specify the method used for fold change calculations.

o Include details on statistical analysis of fold changes.

• Pain & Inflammation Data:

o The manuscript mentions measurements in Materials & Methods, but results are not provided.

o If included in supplementary material, mention this in the main text and provide relevant details.

• Figures 3, 4, and 7: Mark statistical significance on graphs.

• Using only one pro-inflammatory marker is insufficient to draw reliable conclusions. Consider adding more markers to strengthen the findings.

• Give data of all scores used

• Give data of real time RNA quantification results -CT values

**********

what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy

Reviewer #1: No

Reviewer #2: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step.

Dear Dr. Khalid Ahmed,

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 01 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'.
• 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 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,

Sharun Khan

Academic 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.

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

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: The authors have made significant revisions and incorporated the necessary data, improving the overall quality of the manuscript. The study is now close to being suitable for publication in PLOS ONE, pending a few minor revisions outlined below:

1. (Lines 119–120) There is a minor confusion regarding the differentiation between the treated and untreated groups. If my understanding is correct: Treated = septic arthritis + MSC therapy, Untreated = septic arthritis only (no therapy) and Control = PBS

To improve clarity, consider rephrasing the sentence as:

“The treated groups received MSC therapy (BM-MSCs or AD-MSCs) after arthritis induction, while the untreated groups received no therapy following arthritis induction and thus served as positive controls.”

2. Repetition of procedures related to MSC isolation has been observed section on MSC isolation. Please revise this section to eliminate redundancy and streamline the methodology for clarity and conciseness.

3. Ensure standardization of letter casing. Please ensure proper use of upper and lower case letters within sentences. For instance, terms such as "Positive marker" or "Red of Alizarin red" should follow standard sentence case conventions unless referring to specific proper nouns. Revise the manuscript accordingly to maintain consistency in formatting.

Once these minor revisions are addressed, the manuscript will be suitable for publication.

**********

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

**********

[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

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.

Title: The Effect of Therapeutic Potential and Safety of Bone Marrow-Derived vs. Adipose-Derived Mesenchymal Stem Cells in Aged Mice with Septic Arthritis

This manuscript explores the comparative therapeutic efficacy and safety of bone marrow-derived versus adipose-derived mesenchymal stem cells in an aged murine model of septic arthritis. The topic is timely and clinically relevant, with strengths including the use of multiple outcome measures (histology, cytokine profiling, and safety assessment), as well as a focus on an aged population. However, there are significant concerns regarding clarity in experimental grouping (control vs. untreated) and replication. Some methodological details (differentiation assays, staining protocols, donor mice) are incomplete, and one referenced figure (Fig. 1) is missing. Addressing these issues will greatly improve clarity, reproducibility, and impact of the manuscript.

Reviewer Comments to Authors

1. Lines 111–112: The text states that only the treated groups were replicated. Please clarify what this means. Were the untreated and control groups not replicated? If so, why? Replication should ideally be consistent across all groups to avoid bias.

2. Line 120: It is stated that the untreated groups received no therapy. However, if all four untreated subgroups (Un-BM1, Un-BM2, Un-AD1, Un-AD2) only received PBS, the distinction between them is unclear. Why are they separately labeled as Un-BM1, Un-BM2, etc.? Please justify the rationale.

3. Line 125: Based on the description, the only difference is that the control group received PBS without arthritis induction, while the untreated groups received PBS after arthritis induction. If correct, please state this explicitly to avoid confusion.

4. Line 172: Please clarify whether separate donor mice were used for MSC isolation or whether experimental mice themselves were used as donors. If separate donors were used, how many animals were involved? This information is important for ethical and methodological transparency.

5. Line 219: The manuscript mentions differentiation but does not provide details of the differentiation assay. Please describe the protocols followed for osteogenic, chondrogenic, and adipogenic differentiation, including induction media and culture duration.

6. Lines 219–220: Only Alizarin Red staining for osteogenesis is mentioned. What staining protocols were used for adipogenesis (e.g., Oil Red O) and chondrogenesis (e.g., Alcian Blue or Safranin O)? Please include full details.

7. Line 222 (Figure 1): Figure 1 is referred to in the text but does not appear in the manuscript. Kindly ensure that all referenced figures are included.

8. Line 245: The manuscript states that arthritis was induced and MSCs were injected within 24 hours. How was arthritis confirmed to have developed before treatment administration? Were clinical, histological, or imaging assessments used, or was this timing based on prior references? Please clarify.

9. Lines 248–250: The description of untreated groups remains confusing here. Please clarify exactly what was administered and why these subgroups differ in labeling.

10. Lines 260–261: Please provide details of the protocols for Hematoxylin & Eosin, Safranin O, Masson’s Trichrome, COL2A1, and SOX9 staining/ IHC, including duration, concentrations, and source references.

11. Line 454: The phrase “untreated cell” is unclear. Does this mean cells were administered but not pre-treated, or does it refer to groups that received no MSCs at all? Kindly clarify terminology.

12. Line 687: The phrase should be revised to “used to” for grammatical accuracy.

13. Line 874: Please ensure that “BM-MSC” is abbreviated consistently at its first mention and uniformly used throughout the manuscript.

Please submit your revised manuscript by Nov 02 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'.
• 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 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,

Md Shaifur Rahman, Ph.D

Academic Editor

PLOS ONE

Journal Requirements:

If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. 

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:

Dear Authors,

Thank you for submitting your manuscript to PLOS ONE. We have now received feedback from our reviewers, and their comments are included at the bottom of this letter. The reviewers find the topic of your manuscript to be timely and relevant.

However, the reviewers have raised several concerns. We believe that addressing these points through careful revision will significantly strengthen the manuscript. Therefore, we invite you to submit a revised version of your manuscript that incorporates all of the reviewers' comments. Please note that your revisions should also include a full response to each point raised, detailing the changes made.

Given the nature of the requested revisions, we consider this decision to be an invitation for Minor Revision.

Please submit your revised manuscript sooner (e.g., 4-6 weeks). If you require additional time, please contact us.

We look forward to receiving your revised manuscript.

Sincerely,

Md Shaifur Rahman, PhD

Academic Editor

PLOS ONE

Title: The Effect of Therapeutic Potential and Safety of Bone Marrow-Derived vs. Adipose-Derived Mesenchymal Stem Cells in Aged Mice with Septic Arthritis

This manuscript explores the comparative therapeutic efficacy and safety of bone marrow-derived versus adipose-derived mesenchymal stem cells in an aged murine model of septic arthritis. The topic is timely and clinically relevant, with strengths including the use of multiple outcome measures (histology, cytokine profiling, and safety assessment), as well as a focus on an aged population. However, there are significant concerns regarding clarity in experimental grouping (control vs. untreated) and replication. Some methodological details (differentiation assays, staining protocols, donor mice) are incomplete, and one referenced figure (Fig. 1) is missing. Addressing these issues will greatly improve clarity, reproducibility, and impact of the manuscript.

Reviewer Comments to Authors

1. Lines 111–112: The text states that only the treated groups were replicated. Please clarify what this means. Were the untreated and control groups not replicated? If so, why? Replication should ideally be consistent across all groups to avoid bias.

2. Line 120: It is stated that the untreated groups received no therapy. However, if all four untreated subgroups (Un-BM1, Un-BM2, Un-AD1, Un-AD2) only received PBS, the distinction between them is unclear. Why are they separately labeled as Un-BM1, Un-BM2, etc.? Please justify the rationale.

3. Line 125: Based on the description, the only difference is that the control group received PBS without arthritis induction, while the untreated groups received PBS after arthritis induction. If correct, please state this explicitly to avoid confusion.

4. Line 172: Please clarify whether separate donor mice were used for MSC isolation or whether experimental mice themselves were used as donors. If separate donors were used, how many animals were involved? This information is important for ethical and methodological transparency.

5. Line 219: The manuscript mentions differentiation but does not provide details of the differentiation assay. Please describe the protocols followed for osteogenic, chondrogenic, and adipogenic differentiation, including induction media and culture duration.

6. Lines 219–220: Only Alizarin Red staining for osteogenesis is mentioned. What staining protocols were used for adipogenesis (e.g., Oil Red O) and chondrogenesis (e.g., Alcian Blue or Safranin O)? Please include full details.

7. Line 222 (Figure 1): Figure 1 is referred to in the text but does not appear in the manuscript. Kindly ensure that all referenced figures are included.

8. Line 245: The manuscript states that arthritis was induced and MSCs were injected within 24 hours. How was arthritis confirmed to have developed before treatment administration? Were clinical, histological, or imaging assessments used, or was this timing based on prior references? Please clarify.

9. Lines 248–250: The description of untreated groups remains confusing here. Please clarify exactly what was administered and why these subgroups differ in labeling.

10. Lines 260–261: Please provide details of the protocols for Hematoxylin & Eosin, Safranin O, Masson’s Trichrome, COL2A1, and SOX9 staining/ IHC, including duration, concentrations, and source references.

11. Line 454: The phrase “untreated cell” is unclear. Does this mean cells were administered but not pre-treated, or does it refer to groups that received no MSCs at all? Kindly clarify terminology.

12. Line 687: The phrase should be revised to “used to” for grammatical accuracy.

13. Line 874: Please ensure that “BM-MSC” is abbreviated consistently at its first mention and uniformly used throughout the manuscript.

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

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #2: All comments have been addressed

Reviewer #3: (No Response)

**********

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

Reviewer #2: Yes

Reviewer #3: Partly

**********

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

Reviewer #2: Yes

Reviewer #3: I Don't Know

**********

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

The PLOS Data policy

Reviewer #2: Yes

Reviewer #3: Yes

**********

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

Reviewer #2: Yes

Reviewer #3: Yes

**********

Reviewer #2: The authors have adequately responded to all concerns raised in the previous round of review. The methodology is now clearly described, the data are appropriately presented, and the discussion has been strengthened to highlight the relevance and implications of the findings.

In my opinion, the manuscript is now suitable for acceptance in its current form.

Reviewer #3: Title: The Effect of Therapeutic Potential and Safety of Bone Marrow-Derived vs. Adipose-Derived Mesenchymal Stem Cells in Aged Mice with Septic Arthritis

This manuscript explores the comparative therapeutic efficacy and safety of bone marrow-derived versus adipose-derived mesenchymal stem cells in an aged murine model of septic arthritis. The topic is timely and clinically relevant, with strengths including the use of multiple outcome measures (histology, cytokine profiling, and safety assessment), as well as a focus on an aged population. However, there are significant concerns regarding clarity in experimental grouping (control vs. untreated) and replication. Some methodological details (differentiation assays, staining protocols, donor mice) are incomplete, and one referenced figure (Fig. 1) is missing. Addressing these issues will greatly improve clarity, reproducibility, and impact of the manuscript.

Reviewer Comments to Authors

1. Lines 111–112: The text states that only the treated groups were replicated. Please clarify what this means. Were the untreated and control groups not replicated? If so, why? Replication should ideally be consistent across all groups to avoid bias.

2. Line 120: It is stated that the untreated groups received no therapy. However, if all four untreated subgroups (Un-BM1, Un-BM2, Un-AD1, Un-AD2) only received PBS, the distinction between them is unclear. Why are they separately labeled as Un-BM1, Un-BM2, etc.? Please justify the rationale.

3. Line 125: Based on the description, the only difference is that the control group received PBS without arthritis induction, while the untreated groups received PBS after arthritis induction. If correct, please state this explicitly to avoid confusion.

4. Line 172: Please clarify whether separate donor mice were used for MSC isolation or whether experimental mice themselves were used as donors. If separate donors were used, how many animals were involved? This information is important for ethical and methodological transparency.

5. Line 219: The manuscript mentions differentiation but does not provide details of the differentiation assay. Please describe the protocols followed for osteogenic, chondrogenic, and adipogenic differentiation, including induction media and culture duration.

6. Lines 219–220: Only Alizarin Red staining for osteogenesis is mentioned. What staining protocols were used for adipogenesis (e.g., Oil Red O) and chondrogenesis (e.g., Alcian Blue or Safranin O)? Please include full details.

7. Line 222 (Figure 1): Figure 1 is referred to in the text but does not appear in the manuscript. Kindly ensure that all referenced figures are included.

8. Line 245: The manuscript states that arthritis was induced and MSCs were injected within 24 hours. How was arthritis confirmed to have developed before treatment administration? Were clinical, histological, or imaging assessments used, or was this timing based on prior references? Please clarify.

9. Lines 248–250: The description of untreated groups remains confusing here. Please clarify exactly what was administered and why these subgroups differ in labeling.

10. Lines 260–261: Please provide details of the protocols for Hematoxylin & Eosin, Safranin O, Masson’s Trichrome, COL2A1, and SOX9 staining/ IHC, including duration, concentrations, and source references.

11. Line 454: The phrase “untreated cell” is unclear. Does this mean cells were administered but not pre-treated, or does it refer to groups that received no MSCs at all? Kindly clarify terminology.

12. Line 687: The phrase should be revised to “used to” for grammatical accuracy.

13. Line 874: Please ensure that “BM-MSC” is abbreviated consistently at its first mention and uniformly used throughout the manuscript.

**********

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

Reviewer #3: 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

The Effect of Therapeutic Potential and Safety of Bone Marrow-Derived against Adipose-Derived Mesenchymal Stem Cells in Aged Mice Associated with Septic Arthritis

PONE-D-25-03442R3

Dear Dr. Khalid Ahmed,

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,

Md Shaifur Rahman, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments: