Combination of Meropenem and Zinc Oxide Nanoparticles; Antimicrobial Synergism, Exaggerated Antibiofilm Activity, and Efficient Therapeutic Strategy against Bacterial Keratitis
, Alzhraa Ali Mohamed 1, Galal Yahya 2,*, Aliaa Abdelghafar 2, Mahmoud Saad Abdel-Halim 2, Sameh Saber 3, Mohamed A. Alfaleh 4,5, Asmaa H. Mohamed 1, Fatma Abdelrahman 6, Hoda A. Fathey 1, Gehad H. Ali 1 and Mohamed Abdel-Haleem 1
Round 1
Reviewer 1 Report
This is an interesting study that extends what is known regarding antimicrobial actions of ZnO nanoparticles.
Several minor revisions are needed.
Figure 1 parts b and c should be replaced by a sample table. Part a is not necessary (or could be moved to the supplementary data).
Line 128 The words "highest intense biofilm" need to be changed to a better description, such as thickest biofilm or greatest biofilm biomass.
Line 133. Figure 2 has been truncated. The legend does not need to refer to (a) as there is only one part to this figure.
Page 8. Panel a of Figure 5 needs at least one scale bar. The legend needs to explain that the black areas represent corneal opacity.
Line 286. The information given for the ZnO nanoparticles is insufficient. Include the Sigma catalogue number (e.g. 677450) and also the batch number of the nanoparticles used. The information given, i.e. MFCD00011300, is just the MDL number and is not useful. (see https://www.sigmaaldrich.com/AU/en/product/aldrich/677450).
Line 381. State the total number of rats used.
Line 395. State the number of rats per group.
Line 406. How were data sets checked for normality before parametric statistical analyses were done?
Line 416. This has duplication and should read
"Our results provide support for the concept of improving carbapenem by combining it with ZnO-NPs."
The paper has a large number of language errors and needs considerable editing to address those. This includes using italics for names of bacteria. The discussion has suboptimal paragraph structure; all the one sentence paragraphs need to be merged.
Author Response
Dear editor,
We would like to thank the reviewers for their valuable comments and efforts to improve the quality of our manuscript. In the following, we highlighted the comments and concerns of the reviewers. We addressed responses specific for each comment bellow. Thank you very much.
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Comments of Reviewer #1 and our responses for each comment |
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Comment |
This is an interesting study that extends what is known regarding antimicrobial actions of ZnO nanoparticles. |
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Response |
We thank the reviewer so much for finding our work interesting |
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Comment |
Figure 1 parts b and c should be replaced by a sample table. Part a is not necessary (or could be moved to the supplementary data). |
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Response |
We thank the reviewer for his comment, we agree with him, and we followed his recommendation and replaced the figure |
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Comment |
Line 128 The words "highest intense biofilm" need to be changed to a better description, such as thickest biofilm or greatest biofilm biomass. |
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Response |
Thanks for your comment, and we replaced it |
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Comment |
Line 133. Figure 2 has been truncated. The legend does not need to refer to (a) as there is only one part to this figure. |
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Response |
Thank you for your comment, and we fixed it |
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Comment |
Page 8. Panel a of Figure 5 needs at least one scale bar. The legend needs to explain that the black areas represent corneal opacity. |
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Response |
Thanks so much for careful notification, we inserted the required scale bar, and we completed the figure legend |
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Comment |
Line 286. The information given for the ZnO nanoparticles is insufficient. Include the Sigma catalogue number (e.g. 677450) and also the batch number of the nanoparticles used. The information given, i.e. MFCD00011300, is just the MDL number and is not useful. (see https://www.sigmaaldrich.com/AU/en/product/aldrich/677450). |
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Response |
Thanks so much for your comment, and we corrected it |
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Comment |
Line 381. State the total number of rats used. |
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Response |
We included it |
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Comment |
Line 395. State the number of rats per group. |
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Response |
We included it |
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Comment |
Line 406. How were data sets checked for normality before parametric statistical analyses were done? |
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Response |
Data sets were checked for normality using Excel before parametric statistical analyses |
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Comment |
Line 416. This has duplication and should read "Our results provide support for the concept of improving carbapenem by combining it with ZnO-NPs." |
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Response |
Thanks so much for your comment, and we removed the duplicated sentence |
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Comment |
The paper has a large number of language errors and needs considerable editing to address those. This includes using italics for names of bacteria. The discussion has suboptimal paragraph structure; all the one sentence paragraphs need to be merged. |
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Response |
We respect the reviewer comment, and we worked hard to improve the language of the manuscript |
Reviewer 2 Report
Dear author
Thank you for your effort. Some notes, recommendations and suggestions:
Please modify writing in the method section, similarity is high.
Microorganisms nomenclature should be italic in all the text.
Line 75-81 references jumped from 19 to 25. Please revise references.
Line 96 Beginning of the phrase needs revision
Table S1 needs to be placed in text, it's essential not supplemental
Figure 1 notion needs revision. It looks like two results for the same tested material.
Figure 2 do you mean the columns in green are before treatment and the colorless column represents after treatment? Explain please
Line 192 lethal bacterial biofilm is a claim that definitely requires verification/reference
Line 253 what temperature you kept your bacteria at?
Line 255: Did you immerse the antibiotic with disc, or you bought ready discs, please explain for the reader
Line 271 , line 293, I believe the concentration of the bacteria is 108 not 108. Line 312 the same. Revise
I believe liter notation is used in uppercase not lower case. please refer to author guidelines for submission to MDPI
Line 253 , symbol of celsius.
Line 270, check the space before 200
Line 276 remove repeated word
Line 277 , remove (exactly) please.
The resolution of SEM micrographs can be improved.
I have some questions and recommendation
Could you add MTT test for biofilm?
Could you exclude the synergism against Proteus species? Could not other strains ?
In the introduction part, could you please identify possible application of ZnO nanopartices? Is for wound/ complicated wound or burns? Could you run toxicity test on fibroblast,cells or retina since you tried animal studies as well.
Author Response
Dear editor,
We would like to thank the reviewers for their valuable comments and efforts to improve the quality of our manuscript. In the following, we highlighted the comments and concerns of the reviewers. We addressed responses specific for each comment bellow. Thank you very much.
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Comments of Reviewer #2 and our responses for each comment |
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Comment |
Thank you for your effort |
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Response |
We thank the reviewer so much for his nice words |
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Comment |
Please modify writing in the method section, similarity is high. Microorganisms nomenclature should be italic in all the text. |
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Response |
We thank the reviewer for his comment, we agree with him, and we corrected in the Microorganisms nomenclature in the methods section |
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Comment |
Line 75-81 references jumped from 19 to 25. Please revise references. |
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Response |
Thanks for your comment, and we fixed this error |
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Comment |
Line 96 Beginning of the phrase needs revision |
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Response |
Thank you for your comment, and we fixed it |
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Comment |
Table S1 needs to be placed in text, it's essential not supplemental |
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Response |
we added it |
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Comment |
Figure 1 notion needs revision. It looks like two results for the same tested material |
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Response |
Thanks so much for your comment, and we replaced Figure 1 |
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Comment |
Figure 2 do you mean the columns in green are before treatment and the colorless column represents after treatment? Explain please |
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Response |
Thanks for your comment, and we explained it in the figure caption |
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Comment |
Line 192 lethal bacterial biofilm is a claim that definitely requires verification/reference |
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Response |
We refer in this statement to the serious nature of P. aeruginosa infection itself and we included reference 32 |
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Comment |
Line 253 what temperature you kept your bacteria at? |
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Response |
Usually, we keep frozen stocks of our isolates at -80℃ |
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Comment |
Line 255: Did you immerse the antibiotic with disc, or you bought ready discs, please explain for the reader |
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Response |
We purchased ready-made antibiotic discs from Oxoid Ltd., England |
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Comment |
Line 271 , line 293, I believe the concentration of the bacteria is 108 not 108. Line 312 the same. Revise |
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Response |
Thanks so much for careful notification, and we revised them |
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Comment |
I believe liter notation is used in uppercase not lower case. please refer to author guidelines for submission to MDPI |
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Response |
Thanks so much for careful notification, and we revised all the units according to the guidelines of the journal |
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Comment |
Line 253 , symbol of celsius. Line 270, check the space before 200 Line 276 remove repeated word Line 277 , remove (exactly) please.
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Response |
Thanks so much for careful notification, all fixed. |
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Comment |
The resolution of SEM micrographs can be improved. |
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Response |
We increase the resolution |
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Comment |
Could you add MTT test for biofilm? |
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Response |
We performed the possible assays to quantify the biofilm intensity depending on the available kits and facilities in our lab but that is a good point, we wish we could do it |
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Comment |
Could you exclude the synergism against Proteus species? Could not other strains? |
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Response |
Very interesting point to start another project, I guess we have to consider other pathogens, the limitation will be the side effects of ZnO-NPs when administered systemically |
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Comment |
In the introduction part, could you please identify possible application of ZnO nanopartices? Is for wound/ complicated wound or burns? Could you run toxicity test on fibroblast,cells or retina since you tried animal studies as well. |
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Response |
We thank so much the reviewer for his ideas and recommendations, actually we built our approach on the effect of ZnO-NPs as a biofilm inhibitor and we chose a model for keratitis infection caused by P. aeruginosa, the problem due to limitation of facilities, we could perform some of the recommended experiments. We shortlisted our introduction to focus on biofilm formed by P. aeruginosa and its role in developing resistance against Carbapenems |
