PONE-D-20-30549

The effect of maternal HMB supplementation on bone mechanical and geometrical  properties, as well as histomorphometry and immunolocalization of VEGF, TIMP2, MMP13, BMP2 in the bone and cartilage tissue of the humerus of their newborn piglets.

PLOS ONE

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PLOS ONE

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Reviewers' comments:

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Comments to the Author

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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

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

Reviewer #1: Yes

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. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors reported the impacts of HMB ( hydroxy-β-methobutyrate ) supplementation of mothers during pregnancy on the development of the skeletal system of their offspring using sows of the Great White Poland breed. As they mentioned there were some similar studies reported recently using the model while they claimed they focused on humerus bones instead of femur bones. So the novelty is not high for current study. Moreover, it will be interesting to demonstrate the different impacts of HMB on humerus and femur bones if possible. Moreover, the authors tested the expression of some proteins, including VEGF, TIMP2,MMP13, BMP2 using the histomorphometry and immunolocalization methods. As we know, these staining-based methods are not good for quantitively analysis. It is challenging to compare their expression levels based on their current data.

Reviewer #2: General comments

The authors studied the effect of hydroxy-β-methobutyrate maternal supplementation on the immediate postnatal humeral bone, growth plate cartilage and articular cartilage phenotypes of the offspring in a pig model. Supplementation led to greater bone mass and length as well as increases in other architectural variables, collagen structure and protein expression. Some of the positive effects of the supplementation were sex-dependent. This is a careful descriptive study.

Specific comments

1. The ethics statement is incomplete: “If anesthesia, euthanasia, or any kind of animal sacrifice is part of the study, include briefly which substances and/or methods were applied.”

2. Tables showed pooled SEM’s, but it would be much better to include the standard deviation for each individual mean value.

3. Some references to figures in the text did not include the figure number.

4. The Discussion is very long, and, perhaps, could be shortened by not repeating the findings while still providing the comparisons to the literature and interpretations.

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Reviewer #1: No

Reviewer #2: No

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The effect of maternal HMB supplementation on bone mechanical and geometrical  properties, as well as histomorphometry and immunolocalization of VEGF, TIMP2, MMP13, BMP2 in the bone and cartilage tissue of the humerus of their newborn piglets.

PONE-D-20-30549R1

Dear Dr. Blicharski,

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.

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Kind regards,

Xiaofang Wang

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

Reviewer #2: The Discussion is still very long and much of it simply repeats the Results. There may be a way to shorten this, but that is a decision for the Editor.

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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 #1: No

Reviewer #2: No