Anti-Tumor Effect and Neurotoxicity of Ethanol Adjuvant Therapy after Surgery of a Soft Tissue Sarcoma

Round 1

Reviewer 1 Report

In this manuscript, the authors describe the "Anti-tumor effect and neurotoxicity of ethanol adjuvant therapy after surgery of soft tissue sarcoma." While the current version of the manuscript has been significantly improved, it will be helpful if the authors address the minor concerns below:

1) In line 36, the part sentence should be "....challenging issue in orthopedic oncology". The authors should consider revising that part sentence.

2) In line 287, the beginning sentence is ambiguous and should be revised to".... Ethanol cost less and convenient..."

3) While isopropyl alcohol has been used extensively in the field of medicine, it is quite surprising that the authors did not perform a comparative study with ethanol adjuvant to determine their relative efficacies in this study. The authors should consider performing this study (if possible) as it falls within the purview of this study.

In short, this study has a potential to benefit its target audience if the above concerns are addressed.

The English language is good with generally no concerns, except those minor issues of grammar pointed out previously.

Author Response

We would like to thank the reviewer for his/her constructive critique to improve the manuscript. We have made every effort to address the issues raised and to respond to all comments. The revisions are indicated in yellow highlights in the revised manuscript. Please, find next a detailed, point-by-point response to the reviewer's comments. We believe that our revised manuscript presents a clearer account of our study, and we hope that the reviewer will find it suitable for publication in Current Oncology.

Comments to the authors
In this manuscript, the authors describe the "Anti-tumor effect and neurotoxicity of ethanol adjuvant therapy after surgery of soft tissue sarcoma." While the current version of the manuscript has been significantly improved, it will be helpful if the authors address the minor concerns below:

Point 1: In line 36, the part sentence should be "....challenging issue in orthopedic oncology". The authors should consider revising that part sentence.

Response: We would like to thank the reviewer for the insightful comment. In accordance with the reviewer’s suggestion, we have revised this sentence as follows:

“In this regard, ensuring complete cure and preservation of limb function are sometimes a contrary and challenging issue in orthopedic oncology.” (Lines 34–36)

 

 

Point 2: In line 287, the beginning sentence is ambiguous and should be revised to".... Ethanol cost less and convenient..."

Response: We would like to thank the reviewer for the insightful comment. We agree with the reviewer’s comment that the beginning sentence is ambiguous. We have revised this sentence as follows:

“Ethanol is often used in medical fields, costs less, and is convenient. It is medically utilized for the disinfection of the skin and surgical sites [13,14].” (Lines 288–289)

 

Point 3: While isopropyl alcohol has been used extensively in the field of medicine, it is quite surprising that the authors did not perform a comparative study with ethanol adjuvant to determine their relative efficacies in this study. The authors should consider performing this study (if possible) as it falls within the purview of this study.

Response: We would like to thank the reviewer for the insightful comment. As the reviewer suggested, isopropyl alcohol is often used in medical fields. It is also expected to be as effective as ethanol in terms of anti-tumor effect. However, ethanol is often used as adjuvant therapy in the field of malignant bone and soft tissue tumors [References #4, 23, 24], but its efficacy is controversial in basic research. Therefore, it was necessary to first verify the effects of ethanol. Thus, we conducted a basic research using ethanol. Further research using isopropyl alcohol will be necessary in the future. We have discussed this issue as a limitation as follows:

“Eighth, isopropyl alcohol has also been used extensively in the field of medicine. Further research using isopropyl alcohol will be necessary in the future.” (Lines 392–394)

Reviewer 2 Report

In this study, anti-tumor effect of ethanol, as well as its neurotoxicity were assessed. The ethanol adjuvant therapy with 30% ethanol concentration and more demonstrated antitumor effects on HS-SY-Ⅱ, reducing the cell viability, migration function, and invasive ability.  I think the work will be interested in the readers in the scope of anti-tumor therapy and sarcoma.  It seems that it is a revised manuscript.   I recommendate for the acceptence of this paper. 

1, The reason that three different concentrations of ethanol were studied should be elucidated. 

Author Response

We would like to thank the reviewer for his/her constructive critique to improve the manuscript. We have made every effort to address the issues raised and to respond to all comments. The revisions are indicated in yellow highlights in the revised manuscript. Please, find next a detailed, point-by-point response to the reviewer's comments. We believe that our revised manuscript presents a clearer account of our study, and we hope that the reviewer will find it suitable for publication in Current Oncology.

 

 

Comments to the authors
In this study, anti-tumor effect of ethanol, as well as its neurotoxicity were assessed. The ethanol adjuvant therapy with 30% ethanol concentration and more demonstrated antitumor effects on HS-SY-Ⅱ, reducing the cell viability, migration function, and invasive ability.  I think the work will be interested in the readers in the scope of anti-tumor therapy and sarcoma. It seems that it is a revised manuscript.  I recommendate for the acceptence of this paper. 

Point 1: The reason that three different concentrations of ethanol were studied should be elucidated.

Response: We would like to thank the reviewer for the insightful comment. As the reviewer suggested, ethanol concentration was set at 30% followed by 99.5% in this study. Prior to conducting the in vivo experiment, we conducted the in vitro experiment to determine the effective ethanol concentration that would exhibit anti-tumor effects. The MTT assay showed no significant difference in % cell viability between the 30% and 99.5% ethanol concentrations. Wound-healing and invasion assays also showed similar results. Therefore, we did not conduct experiments using ethanol concentrations of 30–99.5% because we thought that the anti-tumor effect was equivalent between these two ethanol concentrations. High-concentration ethanol, specifically 50% or higher, is known to induce neurotoxicity [References #47]. Conversely, ethanol concentrations above 30% demonstrated anti-tumor effects in the vitro experiments. The aim of this study was to determine the effective concentration of ethanol for achieving an anti-tumor effect while avoiding neurotoxicity. We have discussed this issue as a limitation in the revised manuscript as follows:

“Ninth, we did not conduct experiments using ethanol concentrations of 30–99.5%. Wound healing assay and invasion assay also showed similar results. Therefore, we assumed that the anti-tumor effect was equivalent between these two ethanol concentrations. High-concentration ethanol, specifically 50% or higher, is known to induce neurotoxicity [47]. Conversely, ethanol concentrations above 30% demonstrated anti-tumor effects in the vitro experiments. The aim of this study was to determine the effective concentration of ethanol for achieving an anti-tumor effect while avoiding neurotoxicity. Therefore, in this study, experiments were conducted using three different ethanol concentrations” (Lines 394–402)

Author Response File: Author Response.docx