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The authors have addressed all of the reviewers' comments and the manuscript is now ready for publication.
[# PeerJ Staff Note - this decision was reviewed and approved by Vladimir Uversky, a PeerJ Section Editor covering this Section #]
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The authors identified the optimal amount of dye and staining time to identify lymph nodes in rabbit hind limbs. Further biocompatibility of the carbon nanoparticles after 14d was assessed by histology and RT-PCR detection of inflammatory cytokines and markers of apoptosis and fibrosis. The work does not mention that the requirements for the experimental detection of lymph nodes in this study and the selective intraoperative detection of lymph nodes are different. It is therefore necessary to specify the research question that the authors intend to address.
The experimental design overall is good. Bcl-2/Bax are markers of apoptosis, not necrosis.
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- The authors need to highlight the novelty of the findings more clearly.
- The authors should include a graphical representation of the normalized expression data in the main manuscript.
I suggest a revision of the English language to make the reading and comprehension of the article more fluent. The authors used repeted linkers (At present), and the manuscript contains some spelling mistakes.
There is missing data on References: 8, 20, and 30.
In reference 20, it seems that there are two citations instead of one, (it should be clarified).
The study is well laid out and appears to be an alternative for sentinel lymph node mapping.
The authors compared this procedure with other ones already established in clinical practice. In my opinion, it has some benefits but also some limitations being the absortion time one of them. Taking 12 hours for staining significantly the lymph node, it would be interesting if increasing the dose or concentration of CNPs, the time, by contrast, should decrease.
The results derived from this research reveal the feasibility and reproducibility of the assay.
The figures and tables provided show information that clarifies or complements what is described in the text.
The conclusion is a summary of the study results.
Regarding the euthanasia used for the animals, I should recommend another validated method for this species, such as intravenous anesthetic overdose (like pentobarbital), concussion or a blunt blow to the head (only in rabbits weighing less than 5 kg). The 3R principle intends that researchers seek to improve animal welfare by using a painless but effective procedure.
In my opinion, another group with different concentration could be interesting to be done, in order to compare how the concentration affects on the absortion time. It would be interesting to know if higher concentration of the CNPs, the time for a significant staining could be shorter.
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