PONE-D-23-00522Bioinspired material: Upper-critical solution temperature (UCST) polymer functionalized nanomedicine for controlled drug release and hypoxia alleviation in tumor therapyPLOS ONE

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ACADEMIC EDITOR: Major revisionPlease ensure that your decision is justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact.

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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: Partly

Reviewer #2: Partly

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

Reviewer #1: Yes

Reviewer #2: 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

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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

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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: This paper presents a type of drug-loaded nanoparticles PA-DOX for the treatment of liver cancer, in which hydrophobic gold nanoparticles are modified with Upper-critical solution temperature (UCST) polymer P(AAm-co-AN), CeO2 and DOX were loaded in the nanosystem. When the system is irradiated by NIR light, the local heating was produced and triggered the phase change of the P(AAm-co-AN) polymer, simultaneously, CeO2 and DOX were released. Released CeO2 can react with internal H2O2 to generate O2 for relieving the hypoxia of tumor and improving the anticancer effect of DOX. This system seems interesting, however, there are many problems in this manuscript, and some results lack of deep discussion. In my opinion, this manuscript needs major revisions before it can be reconsidered for publication in PLOS ONE, and the following concerns should be well addressed in the revision:

1．Please confirm whether hydrogen peroxide and H2O2 in the sentence “CeO2 can decompose hydrogen peroxide to H2O2 and O2” of abstract are the same substance. (Aren't hydrogen peroxide and H2O2 the same substance?)

2．A reference should be cited for the preparation of AuNRs.

3. To check the mean of the sentence“The polymer layer of P(AAm-co-AN) appeared to collapse at low temperatures to confine the release of DOX” in manuscript (page 3, lines 29-30).

4. The written form of PA-DOX should be unified in the manuscript, P(AAm-co-AN)-AuNRs@CeO2-DOX or P(AAm-co-AN)/AuNRs@CeO2-DOX?

5．In formula (2), 2 in H2O should be subscripted; the 4 section, 2 in CeO2 should be subscripted.

6．The drug loading content was calculated to be 7.1%, why is the drug loading content in such a low level?

7．The photothermal performance experiments of PA-DOX should be provided in experiment section.

8．The drug DOX in the nanodrug only releases when the lesion site is irradiated, and the release stops when the temperature in the lesion site drops to the physiological temperature. That means the treatment process is troublesome, otherwise, the drugs loaded cannot be fully utilized. How to explain that?

9. To check the sentence “To investigate the oxygen generation ability of PA-DOX, AuNRs, Au@CeO2 or PA-DOX, respectively, all these samples were dispersed in the H2O2 solution with or without the laser irradiation.” in 3.4 section.

10. All pictures in the manuscript are blurred. Please provide pictures with high resolution.

Reviewer #2: The paper titled “Bioinspired material: Upper-critical solution temperature (UCST) polymer functionalized nanomedicine for controlled drug release and hypoxia alleviation in tumor therapy" reports the Upper-critical solution temperature (UCST) polymer functionalized nanodrugs and the anti-liver cancer activity, in response to near-infrared light release, assisted by gold nanorods and nanoparticles. The nanomase CeO2 catalyzes the decomposition of H2O2 to O2, alleviating tumor anoxia and inhibiting cancer cell migration. The paper is suitable for publication in PLOS ONE, but many questions need to be resolved before final acceptance.

1. Abstract: "CeO2 can decompose hydrogen peroxide to H2O2 and O2..." CeO2 can decompose hydrogen peroxide to H2O and O2... .

2. The first paragraph in Introduction has no close connection with the research content of this paper and should be deleted.

3. “It can improve the antioxidant capacity and reduce the ROS level to prevent these tumor cells from being killed by PTT or chemotherapy ", does it mean that hypoxia can resist PTT? How does hypoxia resit PTT?

4. There should be Spaces between numerals and quantifiers, such as "5 mL". The first letter of the word "Figure" should be capitalized. "4 hours" should be shortened to "4 hours", "30 minutes" should be shortened to "30 minutes", and so on, there are many irregularities in writing.

5. In experiment "2.10", the decomposition of H2O2 should be investigated under the condition of no oxygen.

6. All cell experiments should clearly indicate the drug dosage and the amount of dye used.

7. All figures should be improved their resolution.

8. Draw the synthesis Scheme of nanomaterials in the text.

9. Gel Permeation Chromatography data for P(AAm-co-AN)-CPTD should be provided.

10. Although morphology, surface finish, Zeta potential and size can provide evidence for the synthesis of materials, these characterizations cannot provide evidence for the existence or disappearance of characteristic functional groups. Therefore, infrared spectrograms are still needed for the characterization of AuNR@CeO2 and PA-DOX.

11. The unit of light power should be W/cm2 or mW/cm2

12. Provide experimental data for calculating DOX inclusion rate and drug loading rate.

13. Provide ROS and O2 level experiments in HepG2 cells.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: Reviewers comments.docx

Upper-critical solution temperature (UCST) polymer functionalized nanomedicine for controlled drug release and hypoxia alleviation in hepatocellular carcinoma therapy

PONE-D-23-00522R1

Dear Dr. You,

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.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Shigao Huang

Academic Editor

Additional Editor Comments (optional):

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 #1: All comments have been addressed

Reviewer #2: 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 #1: Yes

Reviewer #2: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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 #1: Yes

Reviewer #2: 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 #1: Yes

Reviewer #2: Yes

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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 #1: This paper presents a type of drug-loaded nanoparticles PA-DOX for the treatment of liver cancer, in which hydrophobic gold nanoparticles are modified with Upper-critical solution temperature (UCST) polymer P(AAm-co-AN), CeO2 and DOX were loaded in the nanosystem. When the system is irradiated by NIR light, the local heating was produced and triggered the phase change of the P(AAm-co-AN) polymer, simultaneously, CeO2 and DOX were released. Released CeO2 can react with internal H2O2 to generate O2 for relieving the hypoxia of tumor and improving the anticancer effect of DOX. This system seems interesting, and the authors have fully answered the questions raised by the reviewer， therefore, recommending acceptance.

Reviewer #2: (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 #1: No

Reviewer #2: No

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