Biological Evidence of Improved Wound Healing Using Autologous Micrografts in a Diabetic Animal Model
, Francisco De Assis Leite Souza 1, Álvaro Aguiar Coelho Teixeira 1,2, Leucio Duarte Vieira 5, Fabio Naro 6, Antonio Graziano 4 and Anísio Francisco Soares 1,2
Round 1
Reviewer 1 Report
Palma et al. evaluate the performance of Rigenera technology in diabetic rats here in this research. The authors concluded that the treatment with autologous micrografts (AMG) is useful and effective in treating diabetic wounds by reducing tissue inflammation, which is a key parameter, especially for hard-to-heal wounds. As such, all the wounds treated with AMG healed faster than the non-treated group. This is of high importance nowadays, especially if these chronic wounds get infected, they will never heal, which leads to (co)morbidity.
First of all, it was a pleasure to read the manuscript, though there are minor points to address before publication.
Please, see my detailed list with suggestions:
l. 42: should be 16% of “total body weight”
l. 44: The epidermis is “the superficial layer” not just “the most superficial layer”
l. 45 -51; 53 -58; 98-113: Please add adequate references and as well, all over the introduction, please add more references.
l. 114: Should be “subcutaneous layer” instead of “layer subcutaneous”
l. 148: Should be “leading to chronic wound” instead of “making the wound chronic”
l. 156 – 170: There is something missing between Fig. 2 and the text.
l. 215 – 219: It is not clear how the groups were separated. Maybe the authors should separate the groups with “;” instead of “hyphen” because it is confusing.
l. 217: Control group (C) was not even treated with PBS or saline?
Fig. 3, 4 & 7 would benefit if the authors would re-arrange them exactly as Fig. 9. Or if they could explain what the difference between Fig. 4 and Fig. 7 is, as both are at day 14 and images are quite similar, except the resolution is different.
l. 391-395: I would replace “animal in the C and CM groups had more marking in the fragments” with “wounds/tissues in the C and CM groups had more marking in the fragments”
l. 442, 466: should be just “Figure” not “Figures”
Overall: The authors should re-check for the italics and spaces between units and numbers.
Please, see my detailed list with suggestions:
l. 42: should be 16% of “total body weight”
l. 44: The epidermis is “the superficial layer” not just “the most superficial layer”
l. 45 -51; 53 -58; 98-113: Please add adequate references and as well, all over the introduction, please add more references.
l. 114: Should be “subcutaneous layer” instead of “layer subcutaneous”
l. 148: Should be “leading to chronic wound” instead of “making the wound chronic”
l. 391-395: I would replace “animal in the C and CM groups had more marking in the fragments” with “wounds/tissues in the C and CM groups had more marking in the fragments”
l. 442, 466: should be just “Figure” not “Figures”
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript entitled "Biological Evidence of Improved Wound Healing Using Autologous Micrografts in a Diabetic Animal Model" in which the authors examined model the biological evidence related to the use of the Rigenera® technology, an innovative mechanical procedure to isolate autologous micrografts (AMG) in an animal model of diabetes. They found that animals treated with micrografts showed more favorable results for healing compared to those did not receive treatment, demonstrating a positive participation of the micrografts in the treatment of difficult-to-heal wounds.
The work is the topic is important. However, this paper suffers from many shortcomings that must be modified to be suitable for publication in Diabetology Journal.
Shortcomings:
1- Please add more references in the whole article especially in paragraphs lines 83-93, lines 98-113,…..etc
2- The authors wrote “Research findings on hematoxylin-eosin-…………….at the start of healing (3 days)” in lines 222-223. Did the author collect the samples in day 3 after healing or seven and fourteen days after the artificial wound creation? Please clarify and write the days of collecting the samples.
3- Please add the number of samples and the name of groups not abbreviated in all figure legends.
4- I think if the figures 3-7 were merged in one whole figure like the figure 9 it will be less confusing. Also I recommend to put figure 8 below the HE photos to be one whole figure and put figure 10 below the HE photos in figure 9 to be one whole figure.
5- The authors wrote “Figure 9. Photomicrograph displaying the mouse skin with Mallory’s Trichrome….etc” Was the skin of rat or mouse?
6- Please merge the figures 12-16 in one whole figure.
7- Why did the authors use different number of samples in different study groups? Please justify.
8- The authors wrote “The animals in both the experimental and control groups were sedated with ketamine hydrochloride (80 mg/kg) and xylazine (6.0 mg/kg) intramuscularly”. Did the authors use anesthesia for rats during the lesion creation and injection of Autologous Micrografts?
9- Please clarify if the lesion was performed on which day after DM induction? Was it on day 7 ? Was Autologous Micrografts injected immediately after creation of the wound lesion?
10- Why did the authors collect the skin wound seven and fourteen days after the artificial wounds creation? Is this duration enough for wound healing? Please justify and add reference support your use.
11- Please add this reference in Line 735 to the end of this sentences (TNF-α dose was determined using the ELISA method, according to the manufacturer's instructions (RAB0479-1KT - Sigma-Aldrich).
Mohammad HMF etal. Protective effects of evening primrose oil on behavioral activities, nigral microglia and histopathological changes in a rat model of rotenone-induced parkinsonism. J Chem Neuroanat. 2023 Jan;127:102206. doi: 10.1016/j.jchemneu.2022.102206. Epub 2022 Dec 1. PMID: 36464068.
12- Please add statistical analysis in the end of materials and methods section showing the way of analysis, post hoc test and the used software for analysis…..etc.
13- Please remove these sentences in Acknowledgments section “in this section, you can acknowledge any support given which is not covered……… (e.g., materials used for experiments” and write “Not applicable”.
14- The introduction is very long and the discussion is short. I recommend to use some paragraphs of the introduction to be added in the discussion section.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
The manuscript entitled "Biological Evidence of Improved Wound Healing Using Autologous Micrografts in a Diabetic Animal Model" in which the authors examined model the biological evidence related to the use of the Rigenera® technology, an innovative mechanical procedure to isolate autologous micrografts (AMG) in an animal model of diabetes. They found that animals treated with micrografts showed more favorable results for healing compared to those did not receive treatment, demonstrating a positive participation of the micrografts in the treatment of difficult-to-heal wounds.
The revised manuscript is improved compared to prior revision. All my comments were adequately answered and explained by the authors. Therefore, I consider that the revised manuscript is acceptable and suitable for publication in Diabetology Journal.
Comments for author File: 
Comments.docx
