Topical Clonidine Accelerates Cutaneous Wound Healing in Diabetic Rats by Regulating the Expression of Related Cytokine Signaling

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript “Topical clonidine accelerates cutaneous wound healing in diabetic rats by regulating the expression of related cytokine signaling” reports data regarding the effects of clonidine in promoting the healing of experimental cutaneous wounds in rats, regulating the expression of related cytokines. The research proposal has scientific validity, with a significant contribution to the area, and fits perfectly within the scope of the journal. Methods and animal care were carefully described and data were judiciously reported. The discussion of the findings in relation to literature reports was also carried out with care. In view of the above, I consider that the article, after a complete spelling review, can be accepted for publication.

Comments on the Quality of English Language

None.

Author Response

Dear Reviewer,

Thank you very much for your positive and encouraging feedback on our manuscript. We greatly appreciate your recognition of the scientific validity and significant contribution of our work to the field.

We have taken note of your suggestion for a complete spelling review, and we will ensure that this is carefully addressed to meet the publication standards.

Once again, thank you for your thoughtful review and constructive comments. We are grateful for the opportunity to improve our manuscript and look forward to its publication.

Sincerely,

Authors

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript is well deigned and easy to read; however, some comments should be addressed as follows:

1. Introduction: the authors should discuss the normal wound healing process, including relevant stages and the obstacle of wound healing in diabetic wound. The novelty of this study should be elucidated at the end of intro.

2. Fig. 5: please illustrate the difference in structures between different groups. Scale bars are invisible. The authors should conduct Masson trichrom stain to investigate the collagen deposition.

3. Conclusion: it should be one paragraph.

 

4. The authors are encouraged to add a schematic illustration, describing the flow of experimental design and significant results.    

Author Response

Dear reviewer, 
We deeply appreciate the time and effort you dedicated to evaluating our work and providing meaningful suggestions for improvement. Your expertise has been invaluable in shaping the last version of our manuscript. Thank you for your commitment to the peer-review process and for helping us enhance the overall quality of our research.
We look forward to the opportunity to address any further comments or concerns you may have and hope to receive your continued support in the future.
Thank you once again for your time, expertise, and valuable input.
The attachment is our point-by-point response for your consideration.
Sincerely,
Authors

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript entitled “Topical clonidine accelerates cutaneous wound healing in diabetic rats by regulating the expression of related cytokine signaling” submitted to Current issues in molecular biology is well structured and very well written. However, there are some issues that must be addressed. Furthermore, interpretation of the results must be done with caution.

1.        Page 2, line 86- 88 “(…)under the influence of AGEs, clonidine did not exert its effect on cell proliferation, as illustrated in Figure 1B. This result indicated that clonidine had a proliferative effect on HaCaT cells.” Indeed, this results does not indicate that clonidine had a proliferative effect on HaCaT cells, neither all manuscript results indicate that since at higher concentrations clonidine had inhibitory effect. Thus, clonidine had proliferative effects at 15 ng/mL.

2.        The authors should specify the number of replicates and independent experiments done in each assay.

3.        Page 3 line 95-96 “within the medium supplemented with AGEs, HaCaT cells responded robustly to clonidine, exhibiting a notable augmentation in the synthesis of both NADH and NADPH.” At figure 2, the viability increases from an average of 20% after exposure to AGEs to an average of 25 to 30% viability after exposure to 5 and 15 ng/ml of clonidine is not a notable augmentation considering also the standard deviation. It is difficult to understand how this difference translate into statistical significance.

4.        At figure 3 caption, authors mention that “The wounds of the rats recovered faster under clonidine treatment than in the no-treatment group.”, however this is only true for the concentration of 15 ng/ml.

5.        At Figure 4 some western blot results seems to be different from the graphs, as the Phosphoto-STAT 3 that seams to be less expressed after exposure to 15 ng/ml but the graph presents a significant increase (fig. 4D); also, Total STAT3 seams to be increased after exposure to 25 ng/ml but the graph did not show an increase.

6.        At Figure 4 caption the authors stated that “(…) (C) Clonidine treatment led to an increase in the expression of vascular growth factors, including Ang-1. Ang-2 and VEGF. (D) The expression of JAK2 and STAT3 was found to increase in response to clonidine treatment”. But again this is only true for the 15 ng/ml concentration.

7.        Figures caption should not be results description, but instead should indicate the methodology, cell line and conditions studied.

8.        The exposure time used for the viability study, with the cell counting kit-8, is not indicated in the methodology section.

Author Response

Dear reviewer, 
We deeply appreciate the time and effort you dedicated to evaluating our work and providing meaningful suggestions for improvement. Your expertise has been invaluable in shaping the last version of our manuscript. Thank you for your commitment to the peer-review process and for helping us enhance the overall quality of our research.
We look forward to the opportunity to address any further comments or concerns you may have and hope to receive your continued support in the future.
Thank you once again for your time, expertise, and valuable input.
The attachment is our point-by-point response for your consideration.
Sincerely,
Authors

Author Response File: Author Response.docx