Exploring Water-Soluble South African Tulbaghia violacea Harv Extract as a Therapeutic Approach for Triple-Negative Breast Cancer Metastasis

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

After reviewing the manuscript entitled “Exploring Tulbaghia violacea Harv South African Plant Extract as a Therapeutic Approach for Triple Negative Breast Cancer (TNBC) Metastasis”, some modifications must be considered.

This research describes a study on the efficacy of Tulbaghia violacea in treating TNBC metastasis. The water-soluble extract of T. violacea inhibits the invasion and metastasis of TNBC cells and promotes cell adhesion, suggesting its ability to reduce the spread and progression of TNBC. Furthermore, the extract shows moderate antioxidant activity, protecting tumor cells from oxidative damage.

I find the paper very interesting, and the results obtained are significant and promising. However, a section discussing these data in the context of previous studies on these plant extracts or on the various phytochemical compounds present in them should be improved. This inclusion would better situate the findings within the current scientific landscape and facilitate understanding of their relevance and novelty. Additionally, I recommend improving the quality of the graphs presented in the article, as some appear blurry, which can hinder precise interpretation of the data. In summary, while the study presents very valuable results, these suggested changes could enhance the clarity and impact of the article.

Others:

I would recommend writing the article in third person. Therefore, make changes to eliminate “our”, “we” or others … review the entire manuscript and keep consistency.

Writing in italics in vitro and in vivo.

Named ALL the plants using the International Code of Nomenclature for plants, adding the authority after the binomial name and the family. Example: Tulbaghia violacea Harv. Writing also in italics.

Author Response

Comment 1

A section discussing these data in the context of previous studies on these plant extracts or on the various phytochemical compounds present in them should be improved. This inclusion would better situate the findings within the current scientific landscape and facilitate understanding of their relevance and novelty.

Response

A closing paragraph has been added to the discussion covering these aspects

Comment 2

Additionally, I recommend improving the quality of the graphs presented in the article, as some appear blurry, which can hinder precise interpretation of the data. In summary, while the study presents very valuable results, these suggested changes could enhance the clarity and impact of the article.

Response

The quality of the graphs has been improved where necessary

Comment 3

I would recommend writing the article in third person. Therefore, make changes to eliminate “our”, “we” or others … review the entire manuscript and keep consistency.

Response

The paper has been rewritten in third person

Comment 4

Writing in italics in vitro and in vivo.

Response

This has been corrected throughout the paper

Comment 5

Named ALL the plants using the International Code of Nomenclature for plants, adding the authority after the binomial name and the family. Example: Tulbaghia violacea Harv. Writing also in italics

Response

The plant names have been re-written to in include taxonomic authority

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript entitled "Exploring Tulbaghia violacea Harv South African Plant Extract as a Therapeutic Approach for Triple Negative Breast Cancer (TNBC) Metastasis" was interesting and clear, I consider that the information they have will be of great interest to the readers of this Journal, In general, I only suggest that you reduce the number of references used in this manuscript, especially in paragraphs where two or more references are used, I suggest that you eliminate the oldest ones and only leave the most recent ones (preferably from the last five years). Additionally, I point out the following specific details that you should consider addressing to improve understanding of the manuscript.

L117, L210, L211, L213: "DDPH" correct to 'DPPH'.

L118-123: Is the data you obtained already published? Please provide the reference that supports this sentence.

L131: "the plant material was finely ground" they must explain how they did the grinding, whether with a mortar or a mill (give the characteristics).

L132-133: irrelevant sentence since it is well known what this process is used for. I suggest removing it.

L134: "the dry powder" it is very important that you put the amount that you dissolved in a liter of water.

L135-L140: The wording of how they prepared the extracts is very confusing, I suppose that in the end they obtained powder from two extracts? one aqueous and the other methanolic?

I suggest improving the writing by separating the processes.

L142: "the required amount of dried extract" it is very important that you state the exact amount used.

L143-146: same observation, you must enter the quantities of each one.

L141-153: It is a very confusing paragraph, I suggest you rewrite it to make it easier to understand.

L166: the information in the sentence is incomplete, please review and complete.

L171: "Absorbance was measured at 550 nm." It must describe the characteristics of the equipment used.

L178: It must have a continuous numerical sequence of references, so [181] does not apply in this section, you must correct it, it should be [38].

L201: "Blios" spelling error, correct it to 'Blois'.

L289: "It had very little effect on the ability" The wording should be corrected to 'there was no effect' since the two treatments are statistically equal (not significant), therefore, the numerical values ​​cannot be taken into account to infer some trend.

Figure 3: I suggest that you use the same order of colors and treatments as the two previous figures, so as not to confuse readers (in order blue, red, purple,green).

L312-314: They have an error of judgment, the p-value = 0.0024 of the Levene test indicates that the null hypothesis is rejected, therefore, there is no homogeneity of the variances. With such a large error between samples (error bar of the treated MCF10A) I suggest that they repeat the analysis of this treatment or perform non-parametric statistical methods for this variable or, failing that, do a transformation of the data to be able to process them by ANOVA).

L321-323: There is a discrepancy between the wording against the error bars of figure 3 of the MCF10A treatments, since when observing only figure 3, with those standard error bars of the mean or standard deviation (you must explain which of the two values ​​used to make the bars) it is assumed that there were statistical differences between the two treatments, since both bars are distant from each other (they do not overlap, which would indicate equality). Please verify and correct either the text or the figure.

Table 1: I suggest using the same number of decimal places in the IC50 values. Check table 2 and apply the same suggestion.

L385-406: I suggest that this information be moved to the discussions section.

L427-446: Same case, change it to discussions.

Author Response

Comment 1

In general, I only suggest that you reduce the number of references used in this manuscript, especially in paragraphs where two or more references are used, I suggest that you eliminate the oldest ones and only leave the most recent ones (preferably from the last five years).

Response

The number of references has been reduced

Comment 2.

L117, L210, L211, L213: "DDPH" correct to 'DPPH'.

Response

This has been corrected

Comment 3

L118-123: Is the data you obtained already published? Please provide the reference that supports this sentence.

Response

This data has been submitted in another manuscript which is under review

Comment 4

L131: "the plant material was finely ground" they must explain how they did the grinding, whether with a mortar or a mill (give the characteristics).

Response

This information has been added

Comment 5

L132-133: irrelevant sentence since it is well known what this process is used for. I suggest removing it.

Response

Sentence has been removed.

Comment 6

L134: "the dry powder" it is very important that you put the amount that you dissolved in a liter of water.

Response

The amount has been specified

Comment 7

L135-L140: The wording of how they prepared the extracts is very confusing, I suppose that in the end they obtained powder from two extracts? one aqueous and the other methanolic?

I suggest improving the writing by separating the processes.

Response

The processes have been separated

Comment 8

L142: "the required amount of dried extract" it is very important that you state the exact amount used.

Response

The exact amounts have been entered

Comment 9

L143-146: same observation, you must enter the quantities of each one.

Response

The exact amounts have been entered

Comment 10

L141-153: It is a very confusing paragraph; I suggest you rewrite it to make it easier to understand.

Response

The paragraph has been rewritten

Comment 11

L166: the information in the sentence is incomplete, please review and complete.

Response

The sentence is now complete

Comment 12

L171: "Absorbance was measured at 550 nm." It must describe the characteristics of the equipment used.

Response

This information has been added

Comment 13

L178: It must have a continuous numerical sequence of references, so [181] does not apply in this section, you must correct it, it should be [38].

Response

This has been corrected

Comment 14

L201: "Blios" spelling error, correct it to 'Blois'.

Response

This has been corrected

Comment 15

L289: "It had very little effect on the ability" The wording should be corrected to 'there was no effect' since the two treatments are statistically equal (not significant), therefore, the numerical values ​​cannot be taken into account to infer some trend.

Response

The wording has been changed to reflect this

Comment 16

Figure 3: I suggest that you use the same order of colors and treatments as the two previous figures, so as not to confuse readers (in order blue, red, purple, green).

Response

The colours have been changed

Comment 17

L312-314: They have an error of judgment, the p-value = 0.0024 of the Levene test indicates that the null hypothesis is rejected, therefore, there is no homogeneity of the variances. With such a large error between samples (error bar of the treated MCF10A) I suggest that they repeat the analysis of this treatment or perform non-parametric statistical methods for this variable or, failing that, do a transformation of the data to be able to process them by ANOVA).

Response

The ANOVA has been performed using a logarithmic adjustment of the data

Comment 18

L321-323: There is a discrepancy between the wording against the error bars of figure 3 of the MCF10A treatments, since when observing only figure 3, with those standard error bars of the mean or standard deviation (you must explain which of the two values ​​used to make the bars) it is assumed that there were statistical differences between the two treatments, since both bars are distant from each other (they do not overlap, which would indicate equality). Please verify and correct either the text or the figure.

Response

The text has been corrected

Comment 19

Table 1: I suggest using the same number of decimal places in the IC50 values. Check table 2 and apply the same suggestion.

Response

Data in each table has been reported to one decimal point in table 1 and two decimal points in table 2

Comment 20

L385-406: I suggest that this information be moved to the discussions section

Response

It and a leading statement have been moved to the discussion

Comment 21

L427-446: Same case, change it to discussions

Response

The statement has been moved to the discussion

Reviewer 3 Report

Comments and Suggestions for Authors

Regarding the introduction, there are several articles on T. violacea extracts, many of them with anti-cancer activity, they need to be cited in the introduction. 

There are several details of the methodology that need to be improved, brands and models of equipment used, brands of solvents.

 The results were presented, but the discussion needs to be improved, taking into account that a plant extract is a complex matrix and should not only be compared to pure substances. 

In my opinion, this work only presents results from the aqueous extract of T. violacea and this needs to be clear in the abstract and perhaps in the title.

Comments for author File: Comments.pdf

Author Response

Comment 1

Regarding the introduction, there are several articles on T. violacea extracts, many of them with anti-cancer activity, they need to be cited in the introduction. 

Response

These papers have been included and referenced

Comment 2

There are several details of the methodology that need to be improved, brands and models of equipment used, brands of solvents.

Response

These details have been included

Comment 3

 The results were presented, but the discussion needs to be improved, taking into account that a plant extract is a complex matrix and should not only be compared to pure substances. 

Response

A new paragraph has been added to the end of the section discussing the compounds in the discussion that addresses this issue

Comment 4

In my opinion, this work only presents results from the aqueous extract of T. violacea and this needs to be clear in the abstract and perhaps in the title

Response

The title has been altered and a statement added to the abstract to clarify this

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Most of the considerations have been met, however, there are points that need to be improved. The conclusions in the paper have been excluded.

Comments for author File: Comments.pdf

Author Response

Comment1

The conclusions in the paper have been excluded.

Response

The Conclusion has been added