Sustainable Bioactive Composite of Glehnia littoralis Extracts for Osteoblast Differentiation and Bone Formation
Round 1
Reviewer 1 Report
The manuscript entitled as ‘Sustainable bioactive composite of Glehnia littoralis extracts for osteoblast differentiation and bone formation’ seems interesting and can contribute to the health field focusing on Phytochemicals. It is a well-written manuscript. I suggest improvement of image/diagram qualities further.
There are some points below to be addressed:
1. The quality of all figures must be improved, they could be redrawn if possible.
2. It is not clear to understand labels as letters on the column diagrams. They should be explained. Otherwise, it is recommended to be omitted those letters, and variance between sample groups might be compared within the text if possible.
3. Please check the numbering of Tables and Figures in the whole manuscript since there is mislabelling in the current version. For instance, in line 334, the table name was cited as Table 1 within the text, but I think, Table 2 was meant.
4. Please write expressions of in-vitro and in-vivo in italic style. Check also mismatching writing style in the manuscript as in Line 486 and some units as Line 475 etc.
5. The abbreviations, such as ‘DPPH, STOP, and ABTS’ should be explained when it is mentioned for the first time. All other abbreviations should also be checked for the whole manuscript.
Author Response
Editor
Comments
(I) Please check that all references are relevant to the contents of the manuscript.
Author’s response: Thank you for comments. We have revised the manuscript carefully and answered/modified/revised/re-write the manuscript as per the suggestions and guidelines of reviewers and editor comments.
All the references has been revised and are relevent.
Comments
(II) Any revisions to the manuscript should be marked up using the “Track Changes” function if you are using MS Word/LaTeX, such that any changes can be easily viewed by the editors and reviewers.
Author’s response: Thank you for the valuable comments. I modified it as requested (Track Changes) by the editor.
Comments
(III) Please provide a cover letter to explain, point by point, the details of the revisions to the manuscript and your responses to the referees’ comments.
Author’s response: Thank you for the valuable comments. Cover letters inlcuded in the submitted files.
Comments
(IV) If you found it impossible to address certain comments in the review reports, please include an explanation in your appeal. The revised version will be sent to the editors and reviewers.
Author’s response: Thank you for the valuable comments.
Reviewer #1
Comments and Suggestions for Authors
The manuscript entitled as ‘Sustainable bioactive composite of Glehnia littoralis extracts for osteoblast differentiation and bone formation’ seems interesting and can contribute to the health field focusing on Phytochemicals. It is a well-written manuscript. I suggest improvement of image/diagram qualities further.
Author’s response: Thank you for the valuable comments. All the comments made by the reviewers have been addressed carefully, revised the text. We are thankful for your positive comments. Images/diagrams whihc were not clear have been replaced by suitable and improved images.
Comments
There are some points below to be addressed:
1. The quality of all figures must be improved, they could be redrawn if possible.
Author’s response: Thank you for the valuable comments. I modified it as requested by the reviewer. Images/diagrams whihc were not clear have been replaced by suitable and improved images.
Comments
2. It is not clear to understand labels as letters on the column diagrams. They should be explained. Otherwise, it is recommended to be omitted those letters, and variance between sample groups might be compared within the text if possible.
Author’s response: Thank you for the valuable comments. I modified it as requested by the reviewer.
3. Please check the numbering of Tables and Figures in the whole manuscript since there is mislabelling in the current version. For instance, in line 334, the table name was cited as Table 1 within the text, but I think, Table 2 was meant.
Author’s response: Thank you for the valuable comments. I modified it as requested by the reviewer.
4. Please write expressions of in-vitro and in-vivo in italic style. Check also mismatching writing style in the manuscript as in Line 486 and some units as Line 475 etc.
Author’s response: Thank you for the valuable comments. I modified it as requested by the reviewer.
5. The abbreviations, such as ‘DPPH, STOP, and ABTS’ should be explained when it is mentioned for the first time. All other abbreviations should also be checked for the whole manuscript.
Author’s response: Thank you for the valuable comments. I modified it as requested by the reviewer.
Reviewer 2 Report
To begin I would like to thank the authors for their work. However, several points need clarifications for further amelioration of your manuscript.
-The scientific name of the plant must be in Italic
-In vitro must be in italic.
-Avoid the use of abbreviation in the abstract
- As mentioned in material and methods the plant was collected and prepared in 2020 and the article in submitted in 2023 which makes 3 years of difference.
Please mention the storage protocol followed and did you verify that the storage had no influence on the chemical composition.
- line 128 KCH3COO use the number 3 as index
- line 152 correct “compounds”
-Line 306 and unify along the test: ranging from … to ….
-Line 309 the authors declared in material and methods sextion the use of aluminium nitrate while in the results part they said that they used aluminium chloride. The Authors must verify the protocol for flavonoids and correct it.
-Figure 1 use colors to differentiate between the extracts.
-Line 332 the quantified molecules are represented by mg/g of what??
-Line 346 concentration-dependent manner.
-Line 348 IC50 >> IC50
- Antioxidant activity: absence of the positive control and statistical comparison.
- Did the others verified the non-toxicity of the various used extracts if not add a paragraph explaining the non-toxicity of the plant.
-Figure 4 where is the positive control
Author Response
Reviewer #2
Comments and Suggestions for Authors
To begin I would like to thank the authors for their work. However, several points need clarifications for further amelioration of your manuscript.
Author’s response: Thank you for the valuable comments. All the comments made by the reviewers have been addressed carefully, revised the text. We are thankful for your positive comments.
- The scientific name of the plant must be in Italic
Author’s response: Thank you for the valuable comments. We maintained italic form throughout the manuscript.
- In vitro must be in italic.
Author’s response: Thank you for the valuable comments. We maintained italic form throughout the manuscript.
- Avoid the use of abbreviation in the abstract
Author’s response: All the unnesessary abbriviation have been removed from the abstract.
- As mentioned in material and methods the plant was collected and prepared in 2020 and the article in submitted in 2023 which makes 3 years of difference.
Author’s response: Experiment was conducted in the year 2020. We wrote the manuscript in the year 2021. Due to funding problem, we were unable to publish (submit) it to the MDPI (as we need funding for article processing charge for MDPI). 3 years is not experimental gap but it was a gap in the funding from the institute. Kindly understand the situation.
- Please mention the storage protocol followed and did you verify that the storage had no influence on the chemical composition.
Author’s response: Fresh plant material was collected from the field. The extracts obtained from the samples were used immidiately (stored only for few hours at 4 °C) for the analysis. So, we thought that storing for few hours would not effect the chemical compositions of the sample.
- line 128 KCH3COO use the number 3 as index
Author’s response: Thank you for the valuable comments. KCH3COO replaced by KCH3COO
- line 152 correct c
Author’s response: Thank you for the valuable comments. Typo errors corrected as “compounds”
- Line 306 and unify along the test: ranging from … to ….
Author’s response: Thank you for the valuable comments. Correction has been made.
ranging from 4.27 ± 0.03 to 23.29 ± 0.43 mgGAE/g dry sample.
- Line 309 the authors declared in material and methods sextion the use of aluminium nitrate while in the results part they said that they used aluminium chloride. The Authors must verify the protocol for flavonoids and correct it.
Author’s response: Thank you for the valuable comments. Typo error “aluminium chloride” removed from the text.
- Figure 1 use colors to differentiate between the extracts.
Author’s response: Thank you for the valuable comments. different colour used to differentiate the values of various samples.
- Line 332 the quantified molecules are represented by mg/g of what??
Author’s response: Thank you for the valuable comments. The contents of scopoletin, umbelifferone, imperatorin, and cellopterin contained in 1g of sample (dry weight).
- Line 346 concentration-dependent manner.
Author’s response: Thank you for the valuable comments. Necessary correction has been made in the text.
- Line 348 IC50 >> IC50
Author’s response: Thank you for the valuable comments. Necessary correction has been made in the text.
- Antioxidant activity: absence of the positive control and statistical comparison.
Author’s response:
Information about the positive control used in the DPPH and ABTS included in the text. Statistical comparisons have been added to the text.
- Did the others verified the non-toxicity of the various used extracts if not add a paragraph explaining the non-toxicity of the plant.
Author’s response: Following information included in the text regardng the non-toxicity of various extracts.
'In addition, the lactate dehydrogenase (LDH) cytotoxicity assay proved that plant extracts beyond 5 μg/mL were toxic to the cell lines (Figure 4). We used 5 μg/mL of G. littoralis extract in the subsequent experiments to avoid cytotoxicity and promote MC3T3-E1 cell growth”.
- Figure 4 where is the positive control
Author’s response: Thank you for the valuable comments. In cell survival and cytotoxicity, the extract untreated group is the control group. Previous figures has been replaced by new images with proper labelling.
Thank you
Round 2
Reviewer 2 Report
I would like to thank the authors for their work and for taking the reviewers suggestions seriously.