Osteoprotective Effect of the Phytonutraceutical Ormona® on Ovariectomy-Induced Osteoporosis in Wistar Rats
, Helison de Oliveira Carvalho 1, Priscila Faimann Sales 1, Alessandra Ohana Nery Barcellos 1, Kaio Coutinho de Maria 3, Francinaldo Sarges Braga 3, Heitor Ribeiro da Silva 1, Roberto Messias Bezerra 3 and José Carlos Tavares Carvalho 1,*Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis study investigated the Ormona within the biochemical and hormonal parameters, also the bone histopathology and size of trabecular and femoral diaphysis in wistar rats. After reviewing this paper, some problems should not be neglected, and the specific problems are listed as follows.
1. It is recommended to supplement the introduction to Ormona.
2. In line 47-49, the interconnection between menopause and osteoporosis should be stated in detail.
3. In line 49-51, the last sentence “In this condition, the bones become more porous and lose density~” is recommended to be located right after line 46-47.
4. In line 57-61, the specific phytoestrogens in Ormona needs to be supplemented.
5. Reference 9 is missing. It seems like the 10 was moved from introduction to discussion, and missing reference 89-93 in references part.
6. Table 1, the statistical symbols should be rechecked.
7. The format of the reference needs to be rechecked.
8. Line 341, reference 10 has not mentioned before.
9. In line 283-285, the sentence seems to be awkward. In line 288-29, the sentence seems to be awkward.
10. In line 293-296, the sentence is unclear. It is recommended to change the sentence to simple form.
11. In line 299-302, the sentence should be stated clearly. It is recommended to make it into two sentences, such as “The HRT is known to have clear bone-related benefits. In contrast~”.
12. It is recommended to specify the term “clear bone-related benefits” and “uncertain popularity”.
13. In line 318-319, results and evidence on “no increase in weight gain in the OVX group” is unclear. Most of the ovariectomy experiment has the weight gain results, however, in this study was successful without the significant weight gain.
14. In line 331-337, the evidence for serum estradiol level was great. However, please state the reason that the meaning of elevation of serum estradiol concentration in this study. The safety assessments for Ormona should be mentioned.
15. In line 406-412, It is recommended to give an explanation for increase in ALT in ORM. Increasing levels of ALT can mean the toxicity of the materials in the liver.
16. The discussion was not logical and the focus is unclear. All the data are not related to each other also the references. In discussion should contain what is the truly innovated in this research. Too many references seem like not related to the story of the paper.
Comments on the Quality of English LanguageComments on the quality of English language: Moderate corrections are needed.
Author Response
The attached letter contains responses to the observations highlighted by the reviewers.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsThe article titled:"Osteoprotective effect of the phytonutraceutical Ormona® on post-menopausal osteoporosis in Wistar rats" reports the results of the study investigating the influence of Ormona on bone and lipid metabolism in ovariectomized Wistar rats.
The most important methodological issue is that there was no Sham-operated control group, and therfore it s is impossible to assess the significance of reported changes. Without sham-operated control group it is impossible to answer the questions what are the metabolic changes induced by ovariectomy and if they are amleiorated with investigated extract. Eg. the authors reports that there was no significant weight gain in ovariectomized animals, but without sham-operated control it is impossible to say wheather ovariectomy increased the weight gain or not (we are not able to predict whoul would be the weigh gain without ovariectomy).
The authors should include in the article the characteristic of investigated extract.It is mentioned that Ormona dosing was 20mg flavonoids/kg , but the content of flavonoids is unknown. As Ormona increased the serum concentration of estradiol it should be clcarified if Ormona contains estradiol.
Author Response
The attached letter contains responses to the observations highlighted by the reviewers.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript presents an interesting study. Generally, the study is well-designed and the manuscript is well-written. However, I have some issues:
1. Title: in my opinion, the Authors used a model of post-menopausal osteoporosis in rats, and it should be highlighted in the title, or “post-menopaused osteoporosis” should be replaced by “ovariectomized rats”.
2. In the “introduction” section, the properties of Ormona and why the Authors used this phytonutraceutical to treat post-menopausal osteoporosis should be explained.
3. In “Material and methods” there is a lack of information about using certificated materials in calcium analysis and CV%.
4. Discussion: On what basis do Authors find hypercholesterolemia and low calcium absorption in ovariectomized rats?
5. The limitation of the study should be added, e.g. lack of a control group without ovariectomy or SHAM group.
Author Response
The attached letter contains responses to the observations highlighted by the reviewers.
Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsMinor editing of English language required.
Comments on the Quality of English LanguageThe reviewed paper presents interesting data but some points remain to be clarified and supplemented. Please see my comments and suggestions below:
Introduction: The introduction is quite short and would benefit from some supporting data, such as results from other studies. In this section, it is necessary to characterize the components of the investigated phytonutraceutical Ormona®.
Experimental design: Please provide a rationale for the predetermined number of animals (n=5) in the five experimental groups. Additionally, elucidate the methodology employed for sample size calculation in this study.
Results: For Table 1, please include an explanation directly below the table.
Results: For Figures 1-4, explanations denoted as a, b and c, should be replaced with the following annotations: *** p < 0.0001, ** p < 0.01, and * p < 0.05.
Discussion: In this section, please highlight to a greater extent the following themes: implications and novelty of this study.
Discussion: At the end of this section, please add the advantages and limitations of the study.
Conclusions: In this section, please describe the practical aspects of the study performed.
Citation in the manuscript text: Proper citation order is lacking for references numbered 9 and 10. For reference number 10, citations appear in lines 308 and 341, while for reference number 9, there is an absence of citations throughout the manuscript. Please ensure correct and chronological referencing for both of these sources.
Author Response
The attached letter contains responses to the observations highlighted by the reviewers.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors have made a great effort to address all my comments. I now suggest to accept after minor revision.
1. Line 56-63, hormone replacement therapy only need to state once in the manuscripts, line 60, and 63 can be stated in abbreviation HRT.
Comments on the Quality of English LanguageMinor editing of English language required.
Author Response
Answer attached.
Author Response File: Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for AuthorsDear Authors
I connot agree that lack of the apropriate control group is part of 3R activity, as the study without control group has little impact and the conclusions are not strongly supported.
Comments on the Quality of English LanguageDear Editors
Lack of of the appropriete control group makes the study of little value. I do not recommend it for publication
Author Response
Answer attached.
Author Response File: Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe diagnosis of hypercholesterolemia and the information about low calcium absorption are still unclear. The OVX group has a higher cholesterol value compared to (..?), there is no control group and there are no standards to which these results were compared. What about calcium?
The lack of a control group is a serious limitation of this work and should be noted.
The composition of OMRONA should be moved to the Material and Methods part.
Author Response
Answer attached.
Author Response File: Author Response.pdf
Reviewer 4 Report
Comments and Suggestions for AuthorsThe authors have addressed the questions raised in the first round of review. They have made adjustments to the text based on the provided comments. I have only two minor comments:
1. In the Introduction section, please switch the positions of the last two paragraphs so that the research objective is presented at the end.
2. In the Results section, for Figure 4, kindly replace the explanations denoted as a, b, and c with the following annotations: *** p < 0.0001, ** p < 0.01, and * p < 0.05.
Author Response
Answer attached.
Author Response File: Author Response.pdf
