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PONE-D-24-17211Relationship Between the Extent of Vascular Injury and the Evolution of Surgically Induced Osteochondrosis Lesions in a Piglet Model.PLOS ONE

Dear Dr. Toth,

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Reviewers' comments:

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Comments to the Author

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

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

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: I Don't Know

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

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

Reviewer #1: Yes

Reviewer #2: Yes

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

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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: This is a good researched and well written manuscript describing a novel piglet model of very rare but debilitating disease - osteochondritis dissecans. Authors explore the role of extent of vascular injury in the pathophysiology of OCD. I only have a few minor suggestions for improvement and clarification that should be addressed before publishing this article in PLOS ONE. See my comments below:

1) Intro – ln 43: please flip the order to “both young animals and children, particularly…”. I believe authors refer to children rather ten animals engaged in athletic activities.

2) Methods / MRI – ln 127: It seems that two different flex coils were used to accommodate imaging of larger pigs at 12 weeks. I would recommend stating this fact in the Methods section.

3) Methods / MRI – ln 133: Please provide more info on CE-MRI. What was the contrast dose administered? Were CE images acquired after the acquisition of DESS and T2 images? What was the time delay before contrast injection and start of post-CE MRI?

4) Methods / Data analysis – ln 166: Were pre- and post-contrast MR images registered before calculating the subtraction image? This was in vivo scan. Although the subtracted CE-MR images were analyzed only qualitatively, it would be helpful to describe measures taken to account for potential motion between the pre- and post-CE MRI.

5) Results - ln 212: change “the same” to more fitting “similar”.

6) Results - ln 255: authors probably meant “(piglet #9)” instead of “(piglet #6)”.

7) References #23, #27 and #28 seems incomplete; please check.

8) Figures 2E,F; 3F; 4F; 5F – please change contrast in these images to better depict cartilage and lesions. Figure 3E looks so much better than 2E.

9) Figure 6 - Please add arrows pointing to OCD lesions.

Reviewer #2: This is an interesting and well performed study, which demonstrates that a total vascular interruption to the trochlea ridge of femur will cause larger cartilage lesions than a partial vascular insult. The surgical procedure with a control of the extent of induced ischemia to the growth cartilage of the femoral trochlear is ingenious.

The hypothesis is clear and it is unlikely to expect a different outcome. The physiological laws that an insult to a larger vascular bed should cause a larger infarct seems obvious. But as stated in the discussion (line 308) it is “anecdotal evidence”.

The data shows that the lesions after a partial devascularization is indeed smaller, but also decrease over time which is the most important finding. It would have been even more interesting to follow the piglets longer, with MR images, to see if the lesions eventually would disappear completely in this group and if the larger lesion would have developed into OCD.

Below see a few suggestions/concerns on the manuscript

Specific comments to the authors:

The abstract is clearly written and describes the study. I think a more precise description of the breed of “novel piglet model” could be added already in the abstract.

Suggestion: Line 29-30….Ten 4-week-old Yorkshire piglets underwent…..

The introduction describes the relevant known knowledge of the disease, including accurate references, about osteochondrosis. The author’s hypothesis is clear. They want to demonstrate that partial vascular interruption will cause small OC lesions and that they will decrease in size over time. An interruption of the entire vascularization to the lateral trochlea will cause larger OC lesions that increase in size and develop into OC-manifesta.

Material & Methods.

The description of the material and methods are clear, with a few clarifications suggested.

Line 149: … sectioned into serial 2,0 mm thick slabs that spanned the total width of the trochlea… How many slabs did this result in from each trochlea?

Line 150: Out of these, two 5-µm-thick slabs were processed for histology from each surface of the 2-mm slab. How many histologic sections were evaluated from each trochlea?

The OCM on histology were graded in size (result-section). Small, moderate? Large? Measured?

Data analysis: I am not an expert in this field, hence these analysis and results must be evaluated by another referee. The sample size is small, but the data appear to be consistent and Table 2 clearly shows difference in size between the groups, which is not surprising.

Results:

The description of results are mostly clear. I only have a few questions/concerns.

The weight and exact age of the pigs at the time of surgery is stated in line 189.

How much did the different groups weigh at euthanasia and were all 12,0 weeks?

Is it possible to define the histologic OCM size more precisely?

Small, medium and large sized OCM? Perhaps add a description of the different sizes more clearly in the legends to figure 6, then the reader will understand the different sizes more clearly. The following description could be used in fig. 6: Line 251: large OCM = “most of the lateral trochlear ridge. Line 253: Small= nearly completely surrounded by bone. Line 255: Medium OCM = a substantial portion of the cartilage.

The exact size of the MRI lesions are more precise, since they are measured.

What did the growth cartilage, overlying the OCM look like? Was there resolved vascularization, total disappearance of vessels with chondrification or still a large OCLatens, with necrotic cartilage? Did the control side (not operated) still show vascularization at 12-weeks or a complete lack of vessels or chondrificaiton?

Was the overlying non-vascularized articular cartilage affected at all in the operated side?

Discussion:

The surgical procedure with a control of the extent of induced ischemia to the growth cartilage of the femoral trochlear is ingenious and opens up for more research into OC.

Lines 319-322: The discussion about the different forces (compression and shear) involved in the progression of OCM to OCD is of great importance, and can hopefully be addressed using the presented piglet model.

The authors do discuss the limitations of the study, low number of pigs, the lack of the follow up to healing or development of an OCD and the location of the femoral trochlea instead of the condyles, more predisposed in humans. However in equines, the femoral trochlea is predisposed to OCD with subsequent clinical lameness, hence I think it is of value to also explore this location.

References:

It does include the relevant references

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Reviewer #1: No

Reviewer #2: No

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Relationship Between the Extent of Vascular Injury and the Evolution of Surgically Induced Osteochondrosis Lesions in a Piglet Model.

PONE-D-24-17211R1

Dear Dr. Toth,

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.

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Kind regards,

Andre van Wijnen

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

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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

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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 #1: Yes

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 #1: (No Response)

Reviewer #2: The authors have responded to my questions and added explanations in the text as well as in the figure legends.

I am not an expert in statistics, but to my knowledge it appears sound

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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

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