Integrating Untargeted and Targeted Metabolomics Coupled with Pathway Analysis Reveals Muscle Disorder in Osteoporosis on Orchiectomized Mice

Round 1

Reviewer 1 Report

Untargeted and targeted metabolomics concerning skeletal muscle of male mice with osteoporosis are integrated in this study. It is the first study to focus on the metablolomic variations about male OP mice. However, there are some questions required to be further disscussed.

1. Early muscle disorder process was mentioned in the. How to evaluate early

phase of muscle disorder? In other words, which index were used in this manuscript to reflex muscle status?

2. It is a novel idea to evaluate metabolomic status of muscle and bone in male mice. Sinece multiple studies were foucsed on femal OP mice. Differences of metabolomic status between male and femal mice should be discussed in the manuscript.

3. Too much contented are mentioned in the section of Introduction. Content with little association should be removed.

Author Response

Response to Reviewer 1 Comments

Point 1: Early muscle disorder process was mentioned in the. How to evaluate early phase of muscle disorder? In other words, which index were used in this manuscript to reflex muscle status?

Response 1: Thank you for your question. We added thigh diameter measurements to the manuscript as an additional indicator of muscle status. These measurements help to further illustrate the significance of the thigh diameter decline in ORX mice, which is a commonly used indicator of muscle mass loss. However, we recognize that these measurements provide only a rough estimation of muscle status, and that the metabolomic analysis provides a more comprehensive evaluation of the metabolic changes associated with muscle disorders.

Therefore, we believe that the combination of metabolomic analysis and thigh diameter measurements provides a more comprehensive evaluation of muscle disorders in osteoporosis mice, and supports the findings of the study.

Thank you for your consideration of our manuscript.

Point 2: It is a novel idea to evaluate metabolomic status of muscle and bone in male mice. Sinece multiple studies were foucsed on femal OP mice. Differences of metabolomic status between male and femal mice should be discussed in the manuscript.

Response 2: Thank you for your valuable feedback on our manuscript. We agree with you that the potential differences in metabolomic status between male and female mice in the context of OP are an important consideration and should be discussed in the manuscript.

In response to your comment, we have added a detailed comparison of muscle disorder in OP male mice and in OP female mice, which we believe sheds light on the potential differences in metabolomic status between the two sexes. Specifically, we have discussed the existing literature on the metabolomic profiles of male and female mice with OP and the possible sex-specific differences in the lipid metabolomic pathway disrupted by OP.

Overall, we hope that our revised manuscript addresses your concerns and provides a more comprehensive discussion of the metabolomic status of muscle and bone in male and female OP mice.

Thank you again for your feedback, which has helped us improve our manuscript.

Point 3: Too much contented are mentioned in the section of Introduction. Content with little association should be removed.

Response 3: In response to your comment, we have removed the conclusion of our outcome from the introduction section to streamline the content and improve the flow of the introduction.

We have also taken the opportunity to revisit the entire introduction section to ensure that all of the content is directly relevant to the topic of the paper. We have removed some of the less relevant or tangential information and grouped related information into broader categories to select only the most essential and informative details to include in the introduction.

Furthermore, we have restructured the introduction to follow a more logical and intuitive flow, starting with a broad overview of the topic and gradually moving towards the specific research question and hypothesis.

We believe that these revisions have significantly improved the clarity and focus of the introduction section, and we hope that you find these changes satisfactory.

Reviewer 2 Report

English and grammatical revisions should be considered starting at section 2.1.

Author Response

Response to Reviewer 2 Comments

Point 1: English and grammatical revisions should be considered starting at section 2.1.

Response 1: Thank you for your feedback on our manuscript. We appreciate your comments and suggestions, and we have taken them into consideration as we revise the manuscript.

In response to your comment about the English and grammatical revisions, we have invited a native English speaker to review and modify our language, starting at section 2.1. We agree that it is important to ensure that the language and grammar of the manuscript are clear and concise, and we believe that the modification will help us achieve this goal.

We are committed to improving the quality of our manuscript, and we will carefully consider all of your feedback as we make revisions. Thank you again for your time and attention to our work.

Reviewer 3 Report

This manuscript with the title 'Integrating untargeted and targeted metabolomics coupled 2 with pathway analysis reveals muscle disorder in osteoporosis on orchiectomized mice' is well-written, however, minor revision needs to be addressed before considering publication on Molecules. 

1. The author should give the structures of seven biomarkers in Figure 5.

2. The sentence in the abstract 'However, it has been a limited number of reports about how 21 muscle metabolism is disturbed by OP in male patients. In this work, pathway analysis based on 22 metabolomic research was carried out to fill this gap.' this is not appropriate for this manuscript because this study used only mice, not male patients.

Author Response

Response to Reviewer 3 Comments

Point 1: The author should give the structures of seven biomarkers in Figure 5.

Response 1: Regarding your comment about Figure 5, we have added the structures of the seven biomarkers as you suggested. We agree that this addition helps to clarify the figure and provide a more complete understanding of the metabolites we analyzed. Thank you for your feedback regarding the manuscript.

Point 2: The sentence in the abstract 'However, it has been a limited number of reports about how 21 muscle metabolism is disturbed by OP in male patients. In this work, pathway analysis based on 22 metabolomic research was carried out to fill this gap.' this is not appropriate for this manuscript because this study used only mice, not male patients.

Response 2: Thank you for your valuable feedback on our manuscript. We have carefully considered your comment regarding the sentence in the abstract that stated "However, it has been a limited number of reports about how muscle metabolism is disturbed by OP in male patients. In this work, pathway analysis based on metabolomic research was carried out to fill this gap."

We understand your concern that this statement may have given the impression that our study was conducted on human male patients, while in reality, we used male mice for our experiments. We apologize for any confusion this may have caused.

We have revised the sentence in question to accurately reflect our study design. The revised sentence now reads as follows: "However, there is a limited understanding of how muscle metabolism is disturbed by OP in male. In this work, we conducted pathway analysis based on metabolomic research to address this knowledge gap."

We appreciate your attention to this matter and hope that the revised statement accurately reflects our study's scope and objectives. Thank you again for your feedback and for helping us improve the quality of our manuscript.