Health Benefits and Safety of Red Pigmented Rice (Oryza sativa L.): In Vitro, Cellular, and In Vivo Activities for Hair Growth Promoting Treatment

Round 1

Reviewer 1 Report (Previous Reviewer 1)

Dear authors. 

The paper presented for review has been previously reviewed and revised.

It deals with an important topic such as alopecia and the research results presented in it can be implemented in practice.

Kindly complete the statistical significance in Figure 2. 

Author Response

Dear, Reviewer. 

Thank you for reviewer’s suggestions. Our manuscript was revised following the suggestions (See attached file).

 

For response to reviewer suggestion:

Reviewer’s comments	Improvement
The paper presented for review has been previously reviewed and revised. It deals with an important topic such as alopecia and the research results presented in it can be implemented in practice. Kindly complete the statistical significance in Figure 2.	Thank you for kindly suggestion. Results in figure 2 were statistical tested with ANOVA, Duncan’s multiple range tests, p < 0.05. (See in figure 2 page 7).

 

Thank you for your kindness.

 

Best regards.

Author Response File: Author Response.docx

Reviewer 2 Report (Previous Reviewer 2)

Dear authors,

Only minor revisions remain regarding the writing of mean values and their respective experimental uncertainty given by the standard error and the italic formatting of symbols for quantities. I took the liberty to highlight the places in the manuscript that in my opinion need correction.

If I am not mistaken, we usually round the standard deviation to one significant digit, except when the leading digit is 1 where a second digit should be kept (and this I did not know before this revision), that will tell you in which decimal place the uncertain digit of your final result lies. In the event that the uncertainty is in a digit that is not significant, report it as such (198.3 ± 0.03, for example).

Comments for author File: Comments.pdf

Author Response

Dear, Reviewer. 

Thank you for reviewer’s suggestions. Our manuscript was revised following the suggestions (See attached file).

For response to reviewer suggestion:

Reviewer’s comments

Improvement

Only minor revisions remain regarding the writing of mean values and their respective experimental uncertainty given by the standard error and the italic formatting of symbols for quantities. I took the liberty to highlight the places in the manuscript that in my opinion need correction. If I am not mistaken, we usually round the standard deviation to one significant digit, except when the leading digit is 1 where a second digit should be kept (and this I did not know before this revision), that will tell you in which decimal place the uncertain digit of your final result lies. In the event that the uncertainty is in a digit that is not significant, report it as such (198.3 ± 0.03, for example).

Thank you for kindly suggestion. -All symbols were improved to be italic formatting (See attached file). Following suggestion: -Most of results were presented in one digit of decimal (as Mean±S.D. => X.x ± x.x) with simple rule of rounding decimals.   - Previous value before revision was shown in comment note (see in attached file). - Data of bioactive compound might be presented in three digits of decimal (x.xxx) because it can more clearly describe to reader (as line 231-238 page 5).

Thank you for your kindness.

 

Best regards.

Author Response File: Author Response.docx

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

Drodzy autorzy.

The work presented is well prepared and deals with a subject that affects millions of people around the world. The problem of alopecia has various causes and the search for new therapeutic methods is justified. The use of a raw material that can be easily obtained, cheap and available is an important feature of this work.

Despite the high level of the work, I have a few comments:

1. at least exemplary photographs of cell cultures are missing.

2. the paper should be supplemented with own results or cited effects on cancer cells. Here, the positive effect on THP1 cell lines, a model of inflammatory and tumour cells - especially their growth promotion - is particularly worrying.

3 Myślę, że warto podkreślić, iż przedstawione wyniki są wstępne i nadal pozostaje wiele do zrobienia, aby ocenić mechanizm działania badanych ekstraktów.

 

Pomimo uwag, wysoko oceniam przedstawione do recenzji prace i życzę dalszych sukcesów w pracy badawczej. 

Author Response

Dear, Reviewer.

Thank you for your suggestions. Our manuscript was revised following the suggestions (See attached file).

 

Reviewer’s comments	Improvement
1. at least exemplary photographs of cell cultures are missing.	Actually, our phase-contrast photographs of cell culture were damaged by computer virus attack. It made my computer to loss many data including picture files.   For our missing information, the title of manuscript was changed to be preliminary study.
2. the paper should be supplemented with own results or cited effects on cancer cells. Here, the positive effect on THP1 cell lines, a model of inflammatory and tumour cells - especially their growth promotion - is particularly worrying.	Our study aimed to investigate cytotoxicity of extract only, did not focus on in vitro cell model for immune modulation.   THP1 cell was treated by natural rice extract (without adding any inflammatory mediator [e.g. PMA] for induce reaction) for determine alive cell (as cytotoxicity).   Anyway, we think that our results on THP1 cell may make other readers to confuse when they read manuscript. Thus, the result of THP1 was removed from manuscript.   Moreover, this study had another results cytotoxic test of Human keratinocyte cell (HCaT cell) that is target area when cosmetic apply to human body.
3 Myślę, że warto podkreślić, iż przedstawione wyniki są wstępne i nadal pozostaje wiele do zrobienia, aby ocenić mechanizm działania badanych ekstraktów.	We agree with the comments. Thus, title of our manuscript was changed to be preliminary study.

I apologize for any inconvenience.  Thank you for your kindness.

Best regards.

Natthawut Thitipramote

 

Author Response File: Author Response.docx

Reviewer 2 Report

Dear authors,

 Your paper intitled “Novel health benefits and safety of red pigmented rice (Oryza sativa L.): In vitro, cellular, and In vivo activities for hair growth promoting treatment” studies and compares four ethanolic extracts of four Thai rice pericarp varieties in their potential as a natural treatment for androgenic alopecia.

Paper strengths: scientific validation of Thai folk medicines resorting to different experimental models among which pertinent primary follicular cells were used; valorisation of native Thai rice sub products, which is pertinent under a circular economy logic; search for natural based products in cosmetics, which is a tendency to stay.

Weaknesses: lack of real novelty, meaning the fact that proanthocyanidins have been recognized as hair growth promotors since 1998; the performed experimental plan was similar to previous published works on natural products, including rice.

Threats: comparing to other similar published works, the amount of work, namely, in terms of variety of samples tested, is not remarkable. This would be ok, if it was compensated by the research depth on the molecular and cell biology events underneath, but it is not the case.

Other opportunities to improve:

1.      The general English writing and the way values, significant digits and units are represented.

2.      I regret the fact that I am not able to understand the units of the biochemical inhibition assay or the reference in those units for the IC50 of finasteride; meaning, for instance,  values above 1 mg FEA/g  means that it performs better than finasteride?  Please improve methodology, for instance the reference for the IC50 value obtained with finasteride.

3.      Please explain the relevance of evaluating toxicity in THP1 cell line in this study.

Please justify why you did not use finasteride in the in vivo studies instead of minoxidil.

 

Author Response

Dear, Reviewer.

Thank you for your suggestions. Our manuscript was revised following the suggestions (See attached file).

 

Reviewer’s comments	Improvement
1. The general English writing and the way values, significant digits and units are represented.	We’re sorry for our mistake.   Manuscript was revised e.g. unit changed from “/g sample” to “/g extract”.
2. I regret the fact that I am not able to understand the units of the biochemical inhibition assay or the reference in those units for the IC50 of finasteride; meaning, for instance,  values above 1 mg FEA/g  means that it performs better than finasteride?  Please improve methodology, for instance the reference for the IC50 value obtained with finasteride.	Method for 5⍺ reductase inhibition was improved by adding more information (see line 158-168 page 4).
3. Please explain the relevance of evaluating toxicity in THP1 cell line in this study.	Our study aimed to investigate cytotoxicity of extract only, did not focus on in vitro cell model for immune modulation.   THP1 cell was treated by natural rice extract (without adding any inflammatory mediator [e.g. PMA] for induce reaction) for determine alive cell (as cytotoxicity).   Anyway, we think that our results on THP1 cell may make other readers to confuse when they read manuscript. Thus, the result of THP1 was removed from manuscript.   Moreover, this study had another results cytotoxic test of Human keratinocyte cell (HCaT cell) that is target area when cosmetic apply to human body.
4 Please justify why you did not use finasteride in the in vivo studies instead of minoxidil.	For in vivo animal testing, we focused on topical treatment on skin and hair for hair growth promoting treatment. For FDA approve, Minoxidil is topical drug whereas Finasteride is oral drug. Thus, minoxidil was used as control in animal testing of this study.

I apologize for any inconvenience.  Thank you for your kindness.

Best regards.

Natthawut Thitipramote

 

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Dear authors,

I think the title now reflects in a more appropriate way the paper content; the methodology was improved and the paper became more coherent and easier to read.

I do have some minor revisions regarding formatting. Having the SI rules as reference, you should improve the presentation of quantities and units throughout your paper.

As examples,

1)     in the abstract, lines 24-29, 1.495±0.163 should be presented as 1.5±0.2; 97.16±1.32 should be presented as 97±1; etc

2)     please, leave always a space between the numerical value and the unit symbol (also valid for %): -20 °C instead of -20°C; 70 % instead of 70%; etc

3)     the unit symbol for hours is h (not hrs) and for minutes is min (not mins)

4)     the symbols for quantities, standard or defined by you, should be in italic except for pH: p < 0.05 instead of p<0.05; TPC  instead of TPC; etc

Author Response

Dear Reviewer,

 

Thank you for suggestions. Our manuscript was revised following the suggestions (See attached file).  .

For response to reviewer suggestion:

Reviewer’s comments

Improvement

1. you should improve the presentation of quantities and units throughout your paper. As examples, 1)  in the abstract, lines 24-29, 1.495±0.163 should be presented as 1.5±0.2; 97.16±1.32 should be presented as 97±1; etc 2)     please, leave always a space between the numerical value and the unit symbol (also valid for %): -20 °C instead of -20°C; 70 % instead of 70%; etc 3)     the unit symbol for hours is h (not hrs) and for minutes is min (not mins) 4)     the symbols for quantities, standard or defined by you, should be in italic except for pH: p < 0.05 instead of p<0.05; TPC  instead of TPC; etc

Manuscript was revised in term of  the presentation of quantities and units following suggestion (see attached file).

I apologize for any inconvenience. 

Thank you for your kindness.

Best regards.

 

Natthawut Thitipramote

Author Response File: Author Response.docx