Comparative Studies on the Antioxidant, Antifungal, and Wound Healing Activities of Solenostemma arghel Ethyl Acetate and Methanolic Extracts

Round 1

Reviewer 1 Report

Introduction - major:

• The title is not concise. Please consider adopting it to fit the content of the manuscript. 
• The authors may avoid the use of abbreviations or introduce them properly. 
• The introduction is concise but lacks specificity. Statements such as "... involve...complicated cellular and biomolecular events..." maybe avoided since they neither deliver valuable information that introduces the topic nor give information. 
• The reader is left alone with the question, what are the most relevant fungal species affecting wound healing? What are typical complications when those fungi spread? 
• What is the clinical relevance of fungal infections of wounds? Are any incidences known for different countries?
• How often is wound healing complicated by local fungal infection?
• While the introduction implies somewhat of a medical intention, it seems a veterinarian's use is intended? Please clearly indicate the hypothesis and aim of the study.

 

Methods - major:

The method section requires substantial care of the authors, instrumentation and methods are described incompletely. 

• For example: In section 2.3: the instrumentation is not mentioned. What are the weight/volume ratios for the extraction? How long did the material incubate? What do triplicates describe here?

Further: 

• The aim of the clinical trial and design is unclear to the reviewer. For example, what had been inclusion criteria - had there been exclusion criteria as well? 
• In the author's rat experiment: Had the rats been housed in groups after the surgical procedure? How was the gel of the extract prepared? How long was it left on the wound? Had the rats been anaesthetized for each application? How did the authors ensure that the material remained on the wound? Why did the authors not apply a splinting ring on the wound to avoid healing by wound contraction if they are interested in the molecular effects of the abstracts?

 

Results - major:

• A comparison of the EtOAc and MeOH extracts would be helpful
• The authors stated they performed the extraction in triplicates. Are the differences of the extractions provided somewhere in the manuscript - what were the differences observed?
• It is not clear what Figure 2 presents. The authors should state what is depicted in the graphs. Please provide statistics and single values/data points?
• Figure 3: are there any (technical) replicates/errors known? How had the different fungi been identified? How had the extracts been applied to the fungi? What does "control" indicate? Was there any vehicle control for EtOAc and MeOH employed?
• Histology: it is challenging to interpret the findings on the picture. Some wounds show signs of contraction while others don't; thus, the healing speed will be different. There is also no scale bar provided that would allow assessing whether the zoom is comparable. The IHC staining is hardly visible in the photographs and hard to interpret.
• Clinical experiments on large animals: it is not clear what are reference and control groups here. Were fungi found in the wounds before and/or after the treatment? The wounds seem to be split but all treated with an extract. There is no information on whether other treatments had been performed. In Figure 9, the authors stated that previous treatments had been unsuccessful. It seems that besides treating the wound with the extract, the wound was clinically sanitized completely, which no information can be found in the text. 

Author Response

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Reviewer 2 Report

In the manuscript entitled “Wound healing, antifungal and antioxidant activities of Solenostemma arghel extract” the authors describe the production and characterization of S. arghel VOCs extracted by two organis solvents.

In introduction, the novelty and the purpose of this work can be improved. A series of specific tests were performed and the corresponding results are presented and sustained by figures and tables. However, I would like to make a few remarks: 

1). For antioxidant activity, which is the used standard (positive control)?

2). In addition, in expressing the results of the antioxidant action, in each series of EtAOc and MeOH extracts, there should be a negative control, consisting only of Ethyl acetate or methanol, in the concentrations used to prepare the extracts.

3). Macroscopic images of the wound healing activity, should include an indicative macroscopic scale (figure 4).

4). You wrote that the study on animals (point 2.9.1) was done after the approval of the Faculty of Veterinary Medicine! You should refer to the approval of the Research Ethics Commission, within your University ... "according to the opinion of the ethics commission no ...., from the date ...."

5). In addition, the clinical study was performed after obtaining which approvel? Who approved a clinical trial? Please highlight these important ethical issues!!

 

 

Author Response

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Round 2

Reviewer 2 Report

The paper has been improved! Please just add in the paper, the concentration as a expresion of w/v....not just weight!

Author Response

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