Anti-Inflammatory Effects of Vitamin E in Response to Candida albicans

Round 1

Reviewer 1 Report

The novelty and originality of this paper is not questionable, albeit the idea of carry out of in vitro experiment using only affymetrix microarrays, RT pathway arrays and ELISA could be controversial from the potential bias point of view.

The topic of this manuscript falls within the scope of Microorganisms journal, and the addressed content does bring the novel results. The study seems to be scientifically sound, although the methodology is rather confusing and non-standard, which makes the results prone to bias.

Despite the fact that results are considered relevant in providing sufficient and reasonably robust evidence for the anti-inflammatory effect of Vitamin E formulation, in order to avoid over-interpretation, it is my opinion that the authors should revisit the assertion in the methodology/results section. As they know, the are surely other co-factors influencing the obtained results.

Major comments:

1. Why authors did not test purely Vitamin E effect on C.albicans but the complex formulation containing Vitamin E?
2. There is no standardized cytotoxicity method utilized, was it an intentional choice?
3. What is the rationale behind using non-relevant terms: agonists vs antagonists for candinda and formulation respectively?

Minor comments:

1. Did the authors make and any reflections prior to selecting the concentrations of ingredients for investigated formulation?.
3. Considering the results of this study, the author should try to investigate further eg. the synergistic effect of investigated formulation with eg. nanoparticles or other bio-active substances?

Author Response

Reviewer 1

 

1.We thank the reviewer for the comment regarding the results and other potential co-factors that may influence the results. To avoid over-interpretation of the results from the in vitro experiments, we have added in the conclusion section that clinical studies are required to investigate the clinical anti-inflammatory effect of the formulation.

 

Major Comments:

 

1. Why authors did not test purely Vitamin E effect on C. albicans but the complex formulation containing Vitamin E?

The experimental design focused on comparing the vehicle to vehicle + Vitamin E. The complex formulation provides for an extended residence time for the Vitamin E and serves as a delivery vehicle.

 

2. There is no standard cytotoxicity method utilized was it an intentional choice?

Cytotoxicity of the vehicle has been demonstrated and approved by regulatory agencies. FDA guidance has been adhered to and the vehicle was re-formulated in the past to reflect FDA input.

 

3. What is the rationale behind using non-relevant terms: agonists vs. antagonists for Candida and formulation respectively?

Gene expression studies performed to assess effect of additives to formulations commonly use the terminology agonist and antagonist.

 

Minor Comments:

1. Did the authors make any reflections prior to selecting the concentrations of ingredients for investigated formulation?

Yes, different potential additives and varying concentrations were screened prior to performing the experiments with the Vitamin E formulation.

 

2. Considering the results of this study, the author should try to investigate further eg. The synergistic effect of investigated formulation with eg. Nanoparticles or other bioactive substances?

The authors appreciate the reviewer’s suggestion regarding nanoparticles for future studies. Of note, other bioactive substances were screened prior to the investigation of Vitamin E.

Reviewer 1

 

1.We thank the reviewer for the comment regarding the results and other potential co-factors that may influence the results. To avoid over-interpretation of the results from the in vitro experiments, we have added in the conclusion section that clinical studies are required to investigate the clinical anti-inflammatory effect of the formulation.

 

Major Comments:

 

1. Why authors did not test purely Vitamin E effect on C. albicans but the complex formulation containing Vitamin E?

The experimental design focused on comparing the vehicle to vehicle + Vitamin E. The complex formulation provides for an extended residence time for the Vitamin E and serves as a delivery vehicle.

 

2. There is no standard cytotoxicity method utilized was it an intentional choice?

Cytotoxicity of the vehicle has been demonstrated and approved by regulatory agencies. FDA guidance has been adhered to and the vehicle was re-formulated in the past to reflect FDA input.

 

3. What is the rationale behind using non-relevant terms: agonists vs. antagonists for Candida and formulation respectively?

Gene expression studies performed to assess effect of additives to formulations commonly use the terminology agonist and antagonist.

 

Minor Comments:

1. Did the authors make any reflections prior to selecting the concentrations of ingredients for investigated formulation?

Yes, different potential additives and varying concentrations were screened prior to performing the experiments with the Vitamin E formulation.

 

2. Considering the results of this study, the author should try to investigate further eg. The synergistic effect of investigated formulation with eg. Nanoparticles or other bioactive substances?

The authors appreciate the reviewer’s suggestion regarding nanoparticles for future studies. Of note, other bioactive substances were screened prior to the investigation of Vitamin E.

 

Reviewer 2 Report

Dear Authors, 

 

I found your manuscript interesting. However, I have following comments regarding it:

1) Please revise abstract to summarize the studies from scientific perspective rather than methodology

2) Please revise introduction to include practical aspects of your work

3) I could not locate conclusion section in your manuscript.

 

Thank you.

 

Author Response

Reviewer 2

 

1. Please revise the abstract to summarize the studies from the scientific perspective rather than methodology.

The authors thank the reviewer for the suggestion. The abstract has been modified / revised to reflect the helpful comment of the reviewer regarding scientific perspective.

 

2. Please revise introduction to reflect practical aspects of your work.

The authors thank the reviewer for the comment. The manuscript introduction incorporates the overall goal of the study to explore the potential anti-inflammatory properties of a novel dental adhesive formulation that contains Vitamin E as its principal active component for candidiasis. The novel denture adhesive formulation would continue to provide functionality for food occlusion while at the same time reducing inflammation.

 

3. I could not locate conclusion section in your manuscript.

The authors thank the reviewer for this helpful observation. A conclusion section has been added to the paper.

 

Round 2

Reviewer 2 Report

Not applicable

Author Response

Thank you