PONE-D-23-34554Enhancing chronic wound healing with Thai indigenous rice variety, Kaab Dum: exploring ER stress and senescence inhibition in HaCaT keratinocyte cell linePLOS ONE

Dear Dr. Palipoch,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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We look forward to receiving your revised manuscript.

Kind regards,

Gopinath Ponnusamy Manogaran, PhD

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Partly

Reviewer #3: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Dear Prof Dr/Editor,

Greetings!

Thank you for your invitation to review the manuscript entitled ‘’ Enhancing chronic wound healing with Thai indigenous rice variety, Kaab Dum: exploring ER stress and senescence inhibition in HaCaT keratinocyte cell line’’.

Please, see below my comments:

Introduction:

1. Please, explain the relationship between senescence, inflammation, and delayed wound healing.

Materials and methods

2. Please, mention the ethanol: water (V/V) used for your extract.

3. Please, mention the final concentration of your stock in DMSO.

4. Please, mention the solute for MTT solution. Is it medium or PBS?

5. The concentration of MTT is 0.5 mg per L or ml???

6. What is the concentration of DMSO after addition of your extract to the medium???

7. What is the type of fixative you used in SA-β-gal staining assay?

8. Correct the title of Table 1 ………polished and unpolished……..

Results

9. Please, revise the abbreviation of TGF-β in all result section.

10. In the wound healing assay, the image 3 and 4 of 0 h is not similar in size as others.

Discussion

11. The discussion section containing too long paragraphs.

Reviewer #2: This article presents a study on the therapeutic potential of Kaab Dum rice extract in the context of chronic wound healing. The investigation focused on the effects of the indigenous rice variety on TGF-β-induced HaCaT cells, specifically examining oxidative stress, endoplasmic reticulum (ER) stress, cell cycle arrest, and cellular senescence. The findings suggest a reduction of oxidative stress, inhibition of ER stress, and promotion of wound healing. However, there are areas where the study could be strengthened:

1. A more detailed investigation into the mechanisms by which Kaab Dum rice extract affects wound healing is necessary to fully understand its therapeutic potential.

2. The absence of in vivo studies or animal models is a significant limitation. Such studies are crucial for assessing the physiological relevance of the rice extract in a more complex biological system.

3. Comparing the effectiveness of Kaab Dum rice extract with other treatments for chronic wounds would provide context and further insight.

4. Detailed information about the reagents used in the study should be provided, including the source, stock number, tradename, company, city, and country. This information is critical for the reproducibility of the research.

5. An inconsistency in Figure 5 regarding the cell scratching assay should be addressed to ensure the validity of the results. The scratch area at 0 hr for the groups treated with TGF-β+0.02 mg/ml PKD and TGF-β+0.1 mg/ml PKD does not match the initial scratch area of the other groups.

6. What is the authors' rationale for choosing TGF-β induced HaCaT cells as a model of chronic wound? What type of chronic wound is this? Further justification and discussion are needed.

Reviewer #3: Dear Authors,

Your manuscript is well-structured and informative. Addressing the following comments will enhance your study's clarity and scientific rigor.

1. Antibiotic-Antimycotic Solution: For clarity, kindly mention the composition of the antibiotic-antimycotic solution used in your experiments.

2. MTT Solution Concentration: Clarify whether the MTT solution concentration of 0.5 mg/L should instead be reported as 0.5 mg/mL for accuracy.

3. Transforming Growth Factor-β (TGF-β): Provide the source and product number of the TGF-β used in your study for better reproducibility.

4. Rice Extract Concentrations: Explain the rationale behind using different concentrations for polished and unpolished Kaab Dum rice extracts (0.02 and 0.1 mg/mL for polished; 0.2 and 1 mg/mL for unpolished). What factors influenced this selection?

5. Scratch Wound as Chronic Wound: Justify why the scratch wound model was chosen to represent a chronic wound in your study.

6. % of Control and % of Inhibition: Clarify the meaning of "% of control." Does it refer to "% of inhibition"? Please provide clarity on these percentages for better understanding.

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6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Xuqiang Nie

Reviewer #3: No

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Attachments

Attachment

Submitted filename: Comments.docx

PONE-D-23-34554R1Enhancing chronic wound healing with Thai indigenous rice variety, Kaab Dum: exploring ER stress and senescence inhibition in HaCaT keratinocyte cell linePLOS ONE

Dear Dr. Palipoch,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Apr 19 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Gopinath Ponnusamy Manogaran, PhD

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Partly

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: I appreciate the authors' responses and I have only a couple of residual questions.

1. The LC-ESI-QTOF-MS/MS chromatogram should be marked with the main peak information.

2. A few typographical errors also remain (i.e. the word " TGF-β" on pg 20-21, Table 2, line 4, [M+Na}? ).

Reviewer #3: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Xuqiang Nie

Reviewer #3: No

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[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Enhancing chronic wound healing with Thai indigenous rice variety, Kaab Dum: exploring ER stress and senescence inhibition in HaCaT keratinocyte cell line

PONE-D-23-34554R2

Dear Dr. Palipoch,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Gopinath Ponnusamy Manogaran, PhD

Academic Editor

PLOS ONE

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