Effects of Bone Marrow Sparing and TGF-β3 Treatment in Total Body Irradiation of C57BL/6J Mice

Round 1

Reviewer 1 Report

1.     The hypothesis of this manuscript was to investigate TGF-b3 for protection from ARS by decreasing the severity of radiation-induced fibrosis, but in the conclusion part, authors mentioned that in vitro study showed that TGF-b treatment increase a-SMA expression. TGF-b should have positive correction with collagen accumulation, a-SMA expression and fibrosis progression. Authors need to read more and clarify the hypothesis and conclusion.

2.     Authors mentioned that the pilot study showed the TGF-b3 increased the survival after 10.5 Gy of TBI, however, later on, results exhibited that TGF-b didn’t improve the survival after 8.5 GY TBI. Why?

3.     For the cell culture study, authors need to clarify the total dose of irradiation. Line 89.

4.     Survival study, n = 5 was not enough to do any statistical analysis. What’s the body weight of those mice when perform the survival study? Mice need to reach 30 g when perform ARS survival study.

5.     Line 124, please correct the group D treatment detail.

6.     Please correct the Figure 3A solid blue line and dashed blue line.

7.     ARS is acute injury. Sampling time point should be within 3 days, and do the histology evaluation, instead of sampling time point at day 73.

8.     Authors need to include the methods related to Figure 12, collagen content in tissue from mice. What about the kidney injury and kidney fibrosis?

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

-          The treatment protocol is unclear. A figure representing the groups, the treatments, how many survived till the end of the experiments, etc. is necessary to make it understandable.

-          Can you explain the link between TGFb3 and the α-SMA expression in vitro? If α-SMA expression is correlated with fibrosis, does it mean that a treatment with TGFb3 increased the level of fibrosis?

-          What is the effect of TGFb3 on the in vitro radiosensitivity? Any studies that showed that it can increase cell survival, enhance dna damage repair, etc?

-          How did you choose the drug concentrations in vitro and in vivo? Is there any clinical relevance to these concentrations?

-          Is the drug well tolerated by patients? Any reported toxicities in clinical trials?

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

This is an interesting, well written and scientifically sound paper. I have a number of relatively minor corrections to suggest to improve the readability of the paper:

(1)   Abstract: The abstract would benefit from a structured format, for better understanding of challenges, methods and findings.

(2)   The statements in lines 36-38 require references.   

(3)   Methods: cell culture & treatment – add the total dose delivered to cell lines not just the dose rate.

(4)   Methods: treatment – please justify the total dose of 8.5Gy used for irradiation (there is some general info in the Introduction, but the authors should explain their choice in the Methods as well).

(5)   Figure 9 – relabel the two graphs - instead of A and B use (I) and (II)- as it gets confusing with the mice groups also labeled with capital letters A-F.

(6)   Discussion – add a section on the limitations of your study: small lot number; mice-dependent results (including feeding/growing conditions) shown by the different results from your pilot study; one specific dose / dose-rate used for irradiation and a specific radiation quality (photons), no differential analysis of results according to sex, etc.  

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Good revision. Can be accepted with current version.

There are still some of minor grammar issue need to be addressed.

Reviewer 2 Report

The authors have made the requested modifications in the manuscript. in my opinion it is now publishable.

Reviewer 3 Report

All comments have been adequately addressed in the revised version.