Hair Regeneration Effects of Lespedeza bicolor Extract In Vivo and In Vitro

Round 1

Reviewer 1 Report

In general, several redrafting and reviewing is required for the whole typescript. 

A. Title:

The title should reflect that this study was performed in vivo and in vitro.

 

B. English Language:

English in the whole manuscript needs to be thoroughly edited.

 

C. Scientific Names:

The scientific name should be written (italicized) in full when first cited , then is abbreviated according to standard format. Need correction in the whole manuscript including figures.

 

D. Abstract:

The word extract shouldn’t be italicized.

qRT-PCR full name

 

E. Introduction:

• See comment B and C.
• Unify the used abbreviations in the whole manuscript.
• Where is reference [8]?

 

F. Materials and Methods

• See comment B and C.
• Don’t repeat the full terms of the previously abbreviated terms.
• Write the full name of RIPA buffer.
• No references were used. Add references where applicable.
• Add description for the extraction steps.
• Why did you use male mice?
• Add a reference for the method of Chase (1954).
• Add references for the used primers.

 

G. Results

• See comment B and C.
• Don’t repeat the full terms of the previously abbreviated terms.
• Why the authors represented results in Figure 1 as %, while the data represent means. It is better to be as folds in comparison to the control (1.0).
• 107%-129.8%???
• Label x and y axis in Figure 2-A, and correct “bcl-2” to “Bcl-2”.
• Label x and y axis in Figure 2-B.
• Why are data in some figures expressed as mean ± SD, while in others as mean ± SEM?
• In Figure 3, was/resin mixture?
• Legend of Figure 3 is too long, and some data is repeated.
• Section 3.4 represented results about water extract, but nothing was mentioned about this extract in the Materials and Methods section.
• qRT-PCR in Figure 6 without previous abbreviation.

 

H. Discussion:

• Don’t use the full terms of the previously abbreviated terms.
• You concluded that “…safe for subcutaneous use, especially for the scalp“. Which part of your results support this conclusion?
• fructus panax ginseng. Present the scientific name correctly.

 

I. References:

• Unify the style of the title. (See Ref. 19 and 22).
• The scientific name should be italicized.

 

Author Response

Reviewer 1

In general, several redrafting and reviewing is required for the whole typescript.

1. Title: The title should reflect that his study was performed in vivo and in vitro.

à I revised the title by reflecting your opinion. “Hair regeneration effects of Lespedeza bicolor extract in vivo and in vitro”

1. English language: English in the whole manuscript needs to be thoroughly edited.

à We conducted English correcting. Attached is the certificate.

1. Scientific names : the scientific name should be written (italicized) in full when first cited, then is abbreviated according to standard format. Need correction in the whole manuscript including figures.

à Thank you. We completed the revision overall by reflecting the opinions of reviewer.

1. Abstract : The word extract shouldn’t be italicized.

qRT-PCR full name

à Thank you for your opinion. I was modified “extract” font type and written qRT-PCR full name.

1. Introduction : See comment B and C.

Unify the used abbreviations in the whole manuscript.

à Thank you for your opinion. I was modified overall manuscript.

1. Materials and Methods:

See comment B and C.

Don’t repeat the full terms of the previously abbreviated terms.

Write the full name of RIPA buffer.

No references were used. Add references where applicable.

Add description for the extraction steps.

Why did you use male mice?

Add a reference for the method of Chase (1954)

Add references for the used primers.

à Thank you for your opinion. I wrote the full name and references (Chase, 1954 and primer accession No. etc).

The C57BL/6 male mouse is an animal model mainly used when conducting hair growth experiments, and experiments were conducted using a male animal model to avoid hair growth caused by estrogen-related hormone secretion.

 

1. Results

See comment B and C.

Don’t repeat the full terms of the previously abbreviated terms.

Why the authors represented results in Figure 1 as %, while the data represent means. It is better to be as folds in comparison to the control (1.0).

107% - 129.8%???

Label x and y axis in figure 2-a, and correct “bcl-2” to “Bcl-2”. Label x and y axis in figure 2-B.

Why are data in some figures expressed as mean ± SD, while in others as mean ± SEM?

In Figure 3, was/resin mixture?

Legend of Figure 3 is too long, and some data is repeated.

section 3.4 represented results about water extract, but nothing was mentioned about this extract in the Materials and Methods section.

qRT-PCR in Figure 6 without previous abbreviation.

à Thank you for your opinion.

Figure 1 is a picture showing the measurement of cell viability in DPCs.

Usually, the control is calculated assuming 100% survival, so it is shown as follows. Therefore, it was confirmed to be 107% when treated with 0.8 ug/ml and 129.8% when treated with 20ug/ml, which mean an increase of 7% and 29.8% when compared to control.

I matched the labels (x axis, y axis)

I modified it to Mean ± SD.

Figure 3, was/resin à wax/resin

Figure 3 legend was revised briefly.

Materials and methods section added “Water extract method”.

 

1. Discussion:

Don’t use the full terms of the previously abbreviated terms.

You concluded that “…safe for subcutaneous use, especially for the scalp”. Which part of your results support this conclusion?

Fructus panax ginseng. Present the scientific name correctly.

 

à Thank you for your opinion.

Overall, we checked and modified whether the previously abbreviated part was marked full term.

In the paper (Part et al., 2011), it was written as “Fructus panax ginseng”, but it has recently been used as “ginseng berry”. I modified it to that word.

 

 

1. References :

          Unify the style of the title (See Ref. 19 and 22).

           The scientific name should be italicized.

è Thank you for your opinion. I modified.

Author Response File: Author Response.docx

Reviewer 2 Report

 

Manuscript «Effects of Lespedeza bicolor extract on hair regeneration» авторов Shin S.-Y., Kwon J.E., Kim S., Lee Y.-G., Park S., Kang S.C. is devoted to the study of L. bicolor extract (LBE) effect on hair regeneration. The authors investigated mechanism of action of LBE on hair regeneration which includes effect on cell proliferation, cytotoxicity and cell cycle regulation. Dose-dependent proliferation of DPC in 6-week-old C57BL/6 male mice was observed in response to LBE treatment (0.8–20 μg/ml). Histological and morphologic as well as cell cycle arrest gene expression studies demonstrated the extension of anagen phase of the hair cell cycle after LBE treatment. The authors showed that LBE containing protocatechuic acid (PCA) may be used as a theraputic agent for enhancing DPCs proliferation and hair regrowth via anagen phase arrest.

The drawbacks of the manuscript:

1. The protocol for HPLC analysis of LBE is not described properly. As we can see from Fig. 4, the HPLC profiles of LBE contain only one major peak and one minor peak. It is rather confusing for this plant extract, which is known to contain a lot of various polyphenolic compounds. I think that HPLC conditions are not relevant for the analysis of this plant extract. I think that it would be much better if the authors had used HPLS-MS technique or more reliable identification of compounds in LBE.
2. I strongly recommend to have the manuscript proofread by a high-skilled English speaker to correct grammar mistakes.

Author Response

Comments and suggestions for authors

Manuscript «Effects of Lespedeza bicolor extract on hair regeneration» авторов Shin S.-Y., Kwon J.E., Kim S., Lee Y.-G., Park S., Kang S.C. is devoted to the study of L. bicolor extract (LBE) effect on hair regeneration. The authors investigated mechanism of action of LBE on hair regeneration which includes effect on cell proliferation, cytotoxicity and cell cycle regulation. Dose-dependent proliferation of DPC in 6-week-old C57BL/6 male mice was observed in response to LBE treatment (0.8–20 μg/ml). Histological and morphologic as well as cell cycle arrest gene expression studies demonstrated the extension of anagen phase of the hair cell cycle after LBE treatment. The authors showed that LBE containing protocatechuic acid (PCA) may be used as a theraputic agent for enhancing DPCs proliferation and hair regrowth via anagen phase arrest.

 

The drawbacks of the manuscript:

1. The protocol for HPLC analysis of LBE is not described properly. As we can see from Fig. 4, the HPLC profiles of LBE contain only one major peak and one minor peak. It is rather confusing for this plant extract, which is known to contain a lot of various polyphenolic compounds. I think that HPLC conditions are not relevant for the analysis of this plant extract. I think that it would be much better if the authors had used HPLS-MS technique or more reliable identification of compounds in LBE.

a : Thank you for valuable indication. As you mentioned, our quantitative condition is not suitable for screening natural product extract. Also there are some literatures which metabolites of this plant were reported. For this reason, it would be better to change the condition or technique (ex. LC/MS) is more reliable identification of compounds in LBE. Despite of these reasons, our quantitative analysis of LBE were focused on active components, protocatechuic acid (PCA). Through our pilot studies, this condition is confirmed optimally detection PCA in the LBE without any interference from other impurities within the shortest time. I hope this response were suitable for your valuable indication.

2. I strongly recommend to have the manuscript proofread by a high-skilled English speaker to correct grammar mistakes.

a: I have received English correction again, and I attach the certificate.

Author Response File: Author Response.docx