Round 1

Reviewer 1 Report

This is an interesting paper to investigate the effect of the mixed extract of Kalopanax pictus Nakai and Achyranthes japonica Nakai as a functional raw material for improving osteoarthritis through inflammation inhibition in macrophages and mouse models.

This manuscript was good scientific writing. 

The Introduction part explant well about this topic including Osteoarthritis, inflammation, inflammatory synovium, Kalopanax pictus Nakai, Achyranthes japonica Nakai. However, the authors did not explant the effect of herbal (except Kalopanax pictus Nakai, Achyranthes japonica Nakai) on inflammation and osteoarthritis.

The Method describes all methods performed in this paper from in vitro for RAW264.7 cells study to in vivo for mouse model.

The Results show many findings from HPLC detection liriodendrin and ecdysterone in herbal, inflammation and NF-kB, MPAK pathways in RAW 264.7 cells, to micro-CT and histological analysis.

However, the Western Blot (NF-kB 337 and MPAK pathways) was not confirmed in mouse model. 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

The authors of the paper, ‘The effect of the mixed extract of SHP-47B on the improvement of degenerative osteoarthritis through inflammation inhibition in the MIA-induced mouse model’ have tried to investigate the effect of mixture extract of Kalopanax pictus and Achyranthes japonica (SHP-47B) on inflammatory pathology of osteoarthritis. The paper is a bit lengthy and has explored some aspects which have already been carried out by some researchers. I have some pointers/queries, which, if addressed will make it better.

1.         Title of the paper is somehow imperfect, though lengthy; I suggest that the word ‘SHP-47B’, should be replaced with ‘Kalopanax pictus and Achyranthes japonica’ and may end with the word osteoarthritis. So the title is, ‘The effect of the mixed extract of Kalopanax pictus and Achyranthes japonica’ on the improvement of degenerative osteoarthritis’. This way its meaning is relevant and clear.

2.         Authors have used abbreviations without giving their full form (as it first time appeared in the text) at many places in the text. Correct it, thoroughly for the whole text of the manuscript including the abstract.

3.         Line 63, the words ‘there is’ should be omitted.

4.         Over use and misuse of word, ‘and’ should be avoided. See lines 66-67, 79-80.

5.         Introduction is too lengthy, so shorten it by omitting repetitive and redundant information such as para starting from lines 65-77 exhibits repeated text (has already been mentioned in the first and second paragraphs)

6.         Authors must explain in the introduction section that when extensive research has already been carried out to examine the influence of Kalopanax pictus and Achyranthes japonica extract in experimental mouse models of osteoarthritis, then why this study has been designed. Its novelty is relevant which will attract the attention of the readers. See below the relevant papers

i)          Effect of fermented Achyranthes japonica (Miq.) Nakai extract on osteoarthritis in Korean Journal of Food Science Technology, 2017, 49(1); 104-109.

ii)            Anti-inflammatory and anti-osteoarthritis effects of fermented Achyranthes japonica Nakai (PMID: 22668504)

iii)          Inhibitory Effects of IL-6-Mediated Matrix Metalloproteinase-3 and -13 by Achyranthes japonica Nakai Root in Osteoarthritis and Rheumatoid Arthritis Mice Models (PMID: 34451873)

iv)           Anti-nociceptive and anti-rheumatoidal effects of Kalopanax pictus extract and its saponin components in experimental animals (11801382)

v)            Chondroprotective Effects of a Standardized Extract (KBH-JP-040) from Kalopanax pictus, Hericium erinaceus, and Astragalus membranaceus in Experimentally Induced In Vitro and In Vivo Osteoarthritis Models (PMID: 29543781)

…….and there is a long list

7.            Rewrite the preliminary two sentences of section 2-4 starting from lines 167 to 173.

8.            A sentence mentioning that ‘ethical committee approval of respective institute (belonging to authors or at least the lead author)  has been taken’ with its due number and date must be mentioned in the Methodology section.

9.         Authors have concluded that NO is reduced due to the effect of mixed extract (SHP-47B) in the 3000μM treatment group. I suggest that authors must discuss the contradictory viewpoints of several research reports that either mention that NO is protective against the inflammatory effects through increased blood flow and reduced nerve irritation or exhibit that higher levels of NO mediates the inhibition of cytokines (IL-1 and TNF-α).

Moderate editing of the text for the English language is required.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

The present article evaluates the effect of the mixed extract of SHP-47B on the improvement of degenerative osteoarthritis through inflammation inhibition in the MIA-induced mouse model. The topic is relevant and up-to-date, but certain deficiencies identified in both content and form need to be addressed in relation to the comments below:

You don't have to put a period at the end of a title

Abstract

If a term is used only once, use only the initial long form. For multiple uses, abbreviate at the first mention, but then use only the abbreviated form until the end of the text. Main text is treated separately from the abstract. Please revise.

The abstract should begin with a series of information that forms the background of the article (do not begin directly with the objective of the study, but the information that led to the setting of the objective is important).

Main text

L35- 6 bibliographic resources are not required for information that is not complex in the way it has been presented. Furthermore, the bibliographic style adopted by the journal should be used (if there are consecutive numbers, only the first and the last one will appear, i.e. from one to six [1-6]). Please review this issue throughout the manuscript as in multiple areas multiple resources have been identified that are not needed in such large numbers.

L64- no bullet point within bibliographic indexes [18, 43,.44]. Please revise.

After the general presentation of the pathophysiological mechanism, it is necessary to briefly present the management of the pathology (non-pharmacological, pharmacological, rehabilitative therapy, etc.), for a continuity of information from which to link the potential role that the proposed model may have. I suggest checking and referring to: PMID: 35454333 and PMID: 35386619.

The aim of the paper should be improved from the perspective of describing the contribution to the field under analysis and the elements of scientific novelty presented.

L99-L105 please revise and renumber the appropriate subsections.

Detection at 210 nm is not very specific as most organic compounds absorb at that wavelength. Please comment about this.

The results section should not include bibliographic resources, representing only the work of the research team. Reference to other results, studies, methods should be made in the discussion chapter.

The discussion section should include in the last paragraph references to the strengths and limitations of this study.

A conclusion section is also necessary to briefly present the implications of what has been obtained, as well as future research directions to further validate the results and address the limitations of the present study.

The Patents section is not necessary in this case.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

The authors responded fully to some suggestions, partially to others and off topic in some cases.

The most important things that need to be re-checked are:

- abbreviations used directly in the abstract LPS, MAPK, IL, MMP, PGE2 etc.

-After the overview of the pathophysiological mechanism, it is necessary to briefly present the management of the pathology (non-pharmacological, pharmacological, rehabilitation therapy, etc.), for a continuity of information from which to link the potential role that can play the proposed model may have. I suggest you check and refer to: PMID: 35454333 and PMID: 35386619 - to this suggestion an area of the manuscript has been put in the letter where the purpose of the paper is stated.

-the limitations part of the study should be more specific and not repeat what was done in the study

Author Response

Dear reviewer,

Thank you for your kind words, and you're welcome! I'm glad to hear that the reviewer's suggestions have helped improve the quality of your manuscript. You have actively incorporated the modifications suggested by the reviewer.

Our study aims to develop pharmaceuticals using natural substances. We have utilized two types of natural substances with known efficacy and conducted cell experiments and animal experiments to verify their efficacy against arthritis through various research outcomes. We sincerely hope that our research will be helpful to researchers who are working on developing pharmaceuticals using natural substances. We also plan to address the limitations of our study by conducting further research, and we hope to present more advanced research results in the future.

I wish you the best of luck with your research, and I'm here to assist you anytime you need further support or advice. Keep up the great work!

The revised content is as follows:

Q1: abbreviations used directly in the abstract LPS, MAPK, IL, MMP, PGE2 etc.

► We have reviewed the content at the reviewer's request and added the correct terms for the abbreviations in the abstract section.

Q2: After the overview of the pathophysiological mechanism, it is necessary to briefly present the management of the pathology (non-pharmacological, pharmacological, rehabilitation therapy, etc.), for a continuity of information from which to link the potential role that can play the proposed model may have. I suggest you check and refer to: PMID: 35454333 and PMID: 35386619 - to this suggestion an area of the manuscript has been put in the letter where the purpose of the paper is stated.

► Based on the pathophysiological observations of the disease model, treatment approaches can be divided into direct tumor removal, alleviation of disease symptoms, and disease progression inhibition through medication, as well as rehabilitation therapy. The purpose of our research findings seems to lie in identifying methods that can alleviate or suppress the progression of the disease. In the conclusion section, we have incorporated this information based on the references provided by the reviewer.

Q3: the limitations part of the study should be more specific and not repeat what was done in the study

► In response to the reviewer's request, we have removed repetitive content from the conclusion section and added information regarding the limitations of our study.

Author Response File: Author Response.pdf

Round 3

Reviewer 3 Report

The authors have improved the manuscript based on the suggestions received.