Honeycomb-like Structured Film, a Novel Therapeutic Device, Suppresses Tumor Growth in an In Vivo Ovarian Cancer Model

Round 1

Reviewer 1 Report

Ohta and colleagues report an exciting and innovative approach to limits tumor proliferation using Honeycomb-like structured films fabricated from biodegradable polymers using mouse model. The study is timely and innovative, and of excellent technical quality. Even though still there is some staff to be clarified/corrected before publication

1) The manufactured 3D scaffolds were used in other types of cancer?

 

2) Pore sizes on the surface of scaffolds have a significant influence on the morphology, proliferation, differentiation dose this also interfere with normal cells as mentioned, no? Maybe add more referencing that validates this beside any toxicity testing for HCFs?

3) Is it easily reproducible to generate the 3D scaffolds? because it has a lot of limitations specially the foaming step (Big concern)

4) Regarding (2.3. Subcutaneous xenograft model and treatment of HCFs in vivo). You need to rewrite it in a way that is clearer for readers I found it a complete mix! One more point you didn't use MG or ECM before injection??

5) Why didn't you inject tumor spheroids instead of liquid state cells?

6) Which kind of Rodent chew you used for this experiment?

7) Justify the selection of mouse model instead of Rats since they are more resistance and can show more realistic results

8) Improve figure 4 quality

 

 

Thank you.

 

Author Response

Thank you for your thoughtful review of our manuscript. Please find our detailed comments regarding your suggestions below.

1) The manufactured 3D scaffolds were used in other types of cancer?

In in vitro, HCFs were used in gall bladder, lung, oral, stomach, colon, pancreatic and ovarian cancers. However, this study is a first report examining the effect of HCFs on tumor growth in an in vivo ovarian cancer model. We did not use HCFs in other types of cancer in an in vivo. We described it in lines 74-77.

2) Pore sizes on the surface of scaffolds have a significant influence on the morphology, proliferation, differentiation dose this also interfere with normal cells as mentioned, no? Maybe add more referencing that validates this beside any toxicity testing for HCFs?

As you suggested, pore sizes on the surface of scaffolds have a significant influence on the morphology, proliferation, differentiation even in normal cell. We described it and provide sufficient references in the Introduction section (lines 63-70).

3) Is it easily reproducible to generate the 3D scaffolds? because it has a lot of limitations specially the foaming step (Big concern).

Yes, it is. We descried it in lines 100-101.

4) Regarding (2.3. Subcutaneous xenograft model and treatment of HCFs in vivo). You need to rewrite it in a way that is clearer for readers I found it a complete mix! One more point you didn't use MG or ECM before injection??

Thank you for your suggestion. We rewrote the methods of xenograft model in the 2.3 section. We did not use MG or ECM because we were able to confirm the tumor formation in SKOV3ip1 and ES-2 cells without MG or ECM.

5) Why didn't you inject tumor spheroids instead of liquid state cells?

Because we were able to confirm the tumor formation by injecting liquid state cells. If the liquid cell injection did not show tumor formation, the plan was to use tumor spheroids.

6) Which kind of Rodent chew you used for this experiment?

Mice used in this study are thymus glandless, externally hairless and immunologically deficient in T-cell function.

7) Justify the selection of mouse model instead of Rats since they are more resistance and can show more realistic results.

We agree with your comments.

8) Improve figure 4 quality.

Thank you for your suggestion. We improved it.

Author Response File: Author Response.docx

Reviewer 2 Report

The authors describe the development and possible utility of polyurethane HCFs in ovarian cancer management. While the study is interesting there are several concerns which should be addressed, these are provided below,

1. The authors should include a description of the HCFs in the abstract and introduction. From the way it is presented it does not get clear what the HCF composition is.

2. The authors should comment on whether this property is specific to polyurethane based HCFs or HCFs formed from other polymers may also perform similarly. The choice of the polymer is not clear, whether it is the ease to be molded into HCFs or specific mechanical properties that motivated the use of urethane for the present manuscript.

3.  In figure 1, it is not clear why the difference in tumor volume (figure 1A) is not significant but the difference in tumor volumes (figure 1B) is so drastic. The authors should provide an appropriate description of the results.

4. For figure 3A, the authors are suggested to provide a quantification analysis of the size for the two cell lines and the pore size for the 3 HCFs prepared.

5. The authors should include discussion about the biocompatibility and biodegradability of the HCFs. Do the HCFs degrade over time or are they intact at the end of the study duration.

Author Response

Thank you for your thoughtful review of our manuscript. Please find our detailed comments regarding your suggestions below.

1. The authors should include a description of the HCFs in the abstract and introduction. From the way it is presented it does not get clear what the HCF composition is.

As you suggested, we included of the HCFs in the Abstract (line 24-26) and Introduction (lines 57-58). The composition of the HCFs is understood from Figure 3.

2. The authors should comment on whether this property is specific to polyurethane based HCFs or HCFs formed from other polymers may also perform similarly. The choice of the polymer is not clear, whether it is the ease to be molded into HCFs or specific mechanical properties that motivated the use of urethane for the present manuscript.

Thank you for your suggestion. In our study, HCFs were fabricated from polyurethane because it has excellent mechanical strength and elasticity. The inhibitory effect of HCFs on tumor growth is due to their porous structure, and similar results are likely to be obtained are fabricated from other chemical compounds of the polymer. We described it in the Discussion section (lines 366-367 and 376-378).

3. In figure 1, it is not clear why the difference in tumor volume (figure 1A) is not significant but the difference in tumor volumes (figure 1B) is so drastic. The authors should provide an appropriate description of the results.

We assume you are referring to Figure 2 since Figure 1 shows the surgical technique and representative photographs. Figure 2A showed the number of days to achieve a tumor size of 10 mm before applying HCFs. Figure 2B showed the tumor weight after applying HCFs.

4. For figure 3A, the authors are suggested to provide a quantification analysis of the size for the two cell lines and the pore size for the 3 HCFs prepared.

We agree with you. We could not assess the SEM images of ES-2 cells morphologies on the flat films and HCFs.

5. The authors should include discussion about the biocompatibility and biodegradability of the HCFs. Do the HCFs degrade over time or are they intact at the end of the study duration.

Thank you for your suggestion. That is an important point. We described it in the discussion section (lines 367-376).

Author Response File: Author Response.docx

Reviewer 3 Report

The manuscript provides further mechanistic details regarding the bioactivity of honeycomb-like films in ovarian cancer cells using a mouse model coupled with analyzing expression patterns and gene ontology. It brings novelty to the field of material science applied as a putative post-surgery treatment. Results were presented, and the researchers provided the advancements and limitations of the results achieved so far. The authors should make minor modifications to improve the introduction's awareness of its use as post-surgery material. The authors should enhance the quality of the figures and tables a lot once it was difficult to identify some items.

Author Response

Thank you for your thoughtful review of our manuscript. Please find our detailed comments regarding your suggestions below.

 The manuscript provides further mechanistic details regarding the bioactivity of honeycomb-like films in ovarian cancer cells using a mouse model coupled with analyzing expression patterns and gene ontology. It brings novelty to the field of material science applied as a putative post-surgery treatment. Results were presented, and the researchers provided the advancements and limitations of the results achieved so far. The authors should make minor modifications to improve the introduction's awareness of its use as post-surgery material. The authors should enhance the quality of the figures and tables a lot once it was difficult to identify some items.

We described awareness of its use as post-surgery material in the Introduction section (lines 80-81). We also enhanced the quality of the figures and tables.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

The authors have appropriately addressed the concerns. The manuscript can now been accepted.