Antibacterial Effects of Curcumin Nanocrystals against Porphyromonas gingivalis Isolated from Patients with Implant Failure

Round 1

Reviewer 1 Report

Dear Authors, 

you made a great work!

However, some improvements are mandatory before acceptance. 

 The paper is an original research study on the Antibacterial effects of curcumin nanocrystals against Porphyromonas gingivalis isolated from patients with implant failure.

The Authors made a great work in terms of methodology and the paper sounds scientific and well written.

However, some improvements are mandatory before acceptance.

The abstract is well written, complete and summary in its various aspects. The keywords are complete and appropriate.

In the introduction:

·         “is attached to the jawbone by osteointegration process” please improve this sentence.

·         “bacteria has reported as one of the leading causes of dental implant failure and, antibiotics are commonly used to control it.” Please explain its effects on periodontal teeth and the relations between these pathologies.

·         in the introduction I suggest to the authors to enrich some aspects by underlining two different issues:

the first is how modern techniques of stem cell stimulation and bone neo-apposition with grafts are progressing, offering great opportunities also in implant re-osseointegration.
“Irastorza I, Luzuriaga J, Martinez-Conde R, Ibarretxe G, Unda F. Adhesion, integration and osteogenesis of human dental pulp stem cells on biomimetic implant surfaces combined with plasma derived products. Eur Cell Mater. 2019 Nov 4;38:201-214. doi: 10.22203/eCM.v038a14.”
“Sayed, M.E.; Mugri, M.H.; Almasri, M.A.; Al-Ahmari, M.M.; Bhandi, S.; Madapusi, T.B.; Varadarajan, S.; Raj, A.T.; Reda, R.; Testarelli, L.; Patil, S. Role of Stem Cells in Augmenting Dental Implant Osseointegration: A Systematic Review. Coatings 2021, 11, 1035. https://doi.org/10.3390/coatings11091035”

Secondly, the implant has been shown to have consistently higher levels of inflammation than the natural tooth, which is why adopting solutions capable of limiting this effect and maintaining the health of peri-implant tissues for a long time is necessary, as indicated by:
“Guarnieri R, Miccoli G, Reda R, Mazzoni A, Di Nardo D, Testarelli L. Sulcus fluid volume, IL-6, and Il-1b concentrations in periodontal and peri-implant tissues comparing machined and laser-microtextured collar/abutment surfaces during 12 weeks of healing: A split-mouth RCT. Clin Oral Implants Res. 2022 Jan;33(1):94-104. doi: 10.1111/clr.13868.”

Materials and methods are clear and well explained. The authors did a great job in the explication of all the variables identified and included in the study.

Results are easy to understand and comprehensive. All the studied characteristics were reported in tables which are clear and concise.

Please check if table 2 and figure 2 both are necessary.

Discussion: this section is complete, and I suggest to write something more with an evaluation of the outcome of different papers present in literature. The overall is comprehensive, concise and complete in its various aspects.

Conclusions are concise and clear.

Bibliography should be formatted respecting the journal’s requirements and no improper citations are evidenced.

Figures and labels are clear and easy to comprehend.

English can be improved.

Author Response

The paper is an original research study on the Antibacterial effects of curcumin nanocrystals against Porphyromonas gingivalis isolated from patients with implant failure.

The Authors made a great work in terms of methodology and the paper sounds scientific and well written. However, some improvements are mandatory before acceptance.

Thanks

The abstract is well written, complete and summary in its various aspects. The keywords are complete and appropriate.

Thanks

In the introduction:

     “is attached to the jawbone by osteointegration process” please improve this sentence.

We improved it.

     “bacteria has reported as one of the leading causes of dental implant failure and, antibiotics are commonly used to control it.” Please explain its effects on periodontal teeth and the relations between these pathologies.

It has been done.

       in the introduction I suggest to the authors to enrich some aspects by underlining two different issues:

the first is how modern techniques of stem cell stimulation and bone neo-apposition with grafts are progressing, offering great opportunities also in implant re-osseointegration.
“Irastorza I, Luzuriaga J, Martinez-Conde R, Ibarretxe G, Unda F. Adhesion, integration and osteogenesis of human dental pulp stem cells on biomimetic implant surfaces combined with plasma derived products. Eur Cell Mater. 2019 Nov 4;38:201-214. doi: 10.22203/eCM.v038a14.”
“Sayed, M.E.; Mugri, M.H.; Almasri, M.A.; Al-Ahmari, M.M.; Bhandi, S.; Madapusi, T.B.; Varadarajan, S.; Raj, A.T.; Reda, R.; Testarelli, L.; Patil, S. Role of Stem Cells in Augmenting Dental Implant Osseointegration: A Systematic Review. Coatings 2021, 11, 1035. https://doi.org/10.3390/coatings11091035”

Secondly, the implant has been shown to have consistently higher levels of inflammation than the natural tooth, which is why adopting solutions capable of limiting this effect and maintaining the health of peri-implant tissues for a long time is necessary, as indicated by:
“Guarnieri R, Miccoli G, Reda R, Mazzoni A, Di Nardo D, Testarelli L. Sulcus fluid volume, IL-6, and Il-1b concentrations in periodontal and peri-implant tissues comparing machined and laser-microtextured collar/abutment surfaces during 12 weeks of healing: A split-mouth RCT. Clin Oral Implants Res. 2022 Jan;33(1):94-104. doi: 10.1111/clr.13868.”

It has been done.

 

Materials and methods are clear and well explained. The authors did a great job in the explication of all the variables identified and included in the study.

Thanks

Results are easy to understand and comprehensive. All the studied characteristics were reported in tables which are clear and concise.

Thanks

Please check if table 2 and figure 2 both are necessary.

We deleted the table 2.

Discussion: this section is complete, and I suggest to write something more with an evaluation of the outcome of different papers present in literature. The overall is comprehensive, concise and complete in its various aspects.

Thanks

Conclusions are concise and clear.

Thanks

Bibliography should be formatted respecting the journal’s requirements and no improper citations are evidenced.

It has been done.

Figures and labels are clear and easy to comprehend.

Thanks


English can be improved.

It has been done.

Reviewer 2 Report

I congratulate the authors for their interesting scientific contribution.

I believe that this manuscript deserves to be considered for publication, after some modifications to improve the understanding of the text.

- Line 29. Please modify the term “attached”.

- Lines 32-33. “… antibiotics are 32 commonly used to control it”. Please provide references for this sentence because it is not clear if and how antibiotic therapy is able to challenge these bacteria.

- The Introduction section is too short. Please implement the section with dentistry-related studies with the use of curcumin (for example: doi: 10.19256/d.cadmos.07.2018.05).

- Line 63. Please define implant failure.

- Line 65. What kind of tooth? Implant supported teeth can be made of different materials. Moreover, the term tooth is not appropriated. 

Author Response

I congratulate the authors for their interesting scientific contribution.

I believe that this manuscript deserves to be considered for publication, after some modifications to improve the understanding of the text.

Thanks

 

- Line 29. Please modify the term “attached”.

It has been done.

 

- Lines 32-33. “… antibiotics are 32 commonly used to control it”. Please provide references for this sentence because it is not clear if and how antibiotic therapy is able to challenge these bacteria.

It has been done.

 

- The Introduction section is too short. Please implement the section with dentistry-related studies with the use of curcumin (for example: doi: 10.19256/d.cadmos.07.2018.05).

- Line 63. Please define implant failure.

- Line 65. What kind of tooth? Implant supported teeth can be made of different materials. Moreover, the term tooth is not appropriated. 

It has been done.

Reviewer 3 Report

Dear Authors,

I am sorry to say that your manuscript does not meet the required standards for publication. You will see that there are major concerns regarding your article prior to publication in such a prestigious journal.

1. First and foremost, English needs to be edited by a professional native speaker, there are numerous spelling errors, grammar and punctuation errors. Please revise.

2. The whole structure needs to be revised, especially the abstract, introduction and material and method sections

3. Please remove the images copied from the internet

Please see the attachment 

Comments for author File: Comments.pdf

Author Response

I am sorry to say that your manuscript does not meet the required standards for publication. You will see that there are major concerns regarding your article prior to publication in such a prestigious journal.

We improved it. We hope the modification can satisfy the respected reviewers.

1. First and foremost, English needs to be edited by a professional native speaker, there are numerous spelling errors, grammar and punctuation errors. Please revise.

It has been done.

2. The whole structure needs to be revised, especially the abstract, introduction and material and method sections

It has been done.

3. Please remove the images copied from the internet

It has been done.

Please see the attachment 

All comments read and improved in the revised version.

Reviewer 4 Report

The authors conducted a study on the potential antibacterial effect that curcumin would have on P.g. isolated from failed implants. For this purpose, they carried out a study on the cultures of isolates from 15 failed implants, where 8 of them were positive for the presence of P.g.

Curcumin, a polyphenol. It is said that curcumin can act as an anti-inflammatory or help fight biliary pathologies or arthritis. Unfortunately, to date, and despite what its proponents claim, studies on the benefits of curcumin are inconclusive. Moreover, curcumin accounts for only 5% of turmeric root.

The authors claim favourable results despite using half the dose that they themselves cite in the literature (Introduction: page 1, line 37 reference 7). Furthermore, on page 7 lines 152-156 references 12,13,14 obtain results with higher concentrations. How do you explain this fact?

Page 2 line: 55. The materials must appear with the name of the manufacturer, city and country. The spray drier does not show the manufacturer.

Page 3, lines 91-97: The authors do not explain how they perform the quantitative determination of MIC, they only explain the procedure. Page 7 lines 180-191: These are not conclusions drawn from the work, they are opinions of the authors that should appear in the discussion.

The authors do not mention any side effects on the use of curcumin, and this should come up in the discussion. For example: cases of hepatitis from consumption of turmeric supplements are not new and are under study.

Conclusions should be written based on the data obtained in this specific study.

References: 59% of the bibliography is more than 5 years old. 18% of the references are more than 10 years old.

Author Response

The authors conducted a study on the potential antibacterial effect that curcumin would have on P.g. isolated from failed implants. For this purpose, they carried out a study on the cultures of isolates from 15 failed implants, where 8 of them were positive for the presence of P.g.

Curcumin, a polyphenol. It is said that curcumin can act as an anti-inflammatory or help fight biliary pathologies or arthritis. Unfortunately, to date, and despite what its proponents claim, studies on the benefits of curcumin are inconclusive. Moreover, curcumin accounts for only 5% of turmeric root.

Thanks. We added some related sentences in discussion section. We did not use turmeric root. We used curcumin nanoparticles. Pure curcumin as the active ingredient isolated from turmeric plant.

The authors claim favourable results despite using half the dose that they themselves cite in the literature (Introduction: page 1, line 37 reference 7). Furthermore, on page 7 lines 152-156 references 12,13,14 obtain results with higher concentrations. How do you explain this fact?

The cited study (Introduction: page 1, line 37 reference 7) used free curcumin against clinically isolated Porphyromonas gingivalis. We used curcumin nanoparticles.

Also, the cited studies (on page 7 lines 152-156 references 12,13,14) used free curcumin against standard bacteria, while we used curcumin nanoparticles against clinically isolated bacteria. We added these reasons to the revised version.

Page 2 line: 55. The materials must appear with the name of the manufacturer, city and country. The spray drier does not show the manufacturer.

It has been done.

Page 3, lines 91-97: The authors do not explain how they perform the quantitative determination of MIC, they only explain the procedure.
The serial dilution method was used. The serial concentration of nanoparticles (50, 25, 12.5 and 6.25 μg/ml concentrations). The visual turbidity detection was used to determine the MIC.

Page 7 lines 180-191: These are not conclusions drawn from the work, they are opinions of the authors that should appear in the discussion.

It has been improved.

The authors do not mention any side effects on the use of curcumin, and this should come up in the discussion. For example: cases of hepatitis from consumption of turmeric supplements are not new and are under study.

We added.

Conclusions should be written based on the data obtained in this specific study.

It has been improved.

References: 59% of the bibliography is more than 5 years old. 18% of the references are more than 10 years old.

We improved them.

Reviewer 5 Report

Dear Authors,

In general Your paper is interesting. However, it require a serious modifications.

First of all the introduction must be rebuild and provide a full view on the dental implant failure aspect. From the present form of Your article readers may understand that bacteria simly are the reason of implant failure. Especially in the cases You presented at pictures, wrong implant positioning in the bone and according to the prostodontic plan could be an initial problem leading to the following issues and further bacteria infiltration. Nevertheless, You need to elaborate and explain the whole cascade of problems that finally can give implant failure. I can suggest these studies as explaning that problems and also potential references to increase the number of references:

1. implant fracture/failure explanation https://doi.org/10.3390/prosthesis3040026

2. You can base on this introduction https://doi.org/10.3390/prosthesis4010004

3. 10.1111/prd.12456

4. 10.1111/jre.12950

Futher in the discussion You also can reach to and mention the problems other than bacteria- I sugest again wrong positionig/wrong prostodontics/too short biologic width- thus without having these issues solved no antibiotics and other antibacterial solution will not help.

Regarding conclusions You can only refer to Your results. "present study confirms that the prepared curcumin nanoparticles can be used as 180 an alternative or a useful adjunct to antibiotics" i would rather say that curcumin in xx% and particle size may have similar activity against P. Gingivalis as xx, yyy, zz...

However You havent mention if curcumin can be use only local or also systemic? That could go to the discussion as well.

"In the future, the loading of curcumin into implant fixture can show some potential 185 benefits. The chemical antimicrobials may be slowly substituted with antibacterial plants 186 such as curcumin. Such a replacement will not only avoid the microbial resistance but will 187 also be a key to decrease the use of chemicals in dentistry. It can not only decrease the 188 implant failure, but also the helpful properties of curcumin for gingival and its optimistic 189 action on bone–implant contact can result in a further stabilization and success of the implant therapy." yhis part is not based on Your results and should be removed.

 

Besides above, all materials and devices should be described with manufacturer/city/state/country.

 

Best regards

 

Reviewer

 

 

Author Response

In general Your paper is interesting. However, it require a serious modifications.

First of all the introduction must be rebuild and provide a full view on the dental implant failure aspect. From the present form of Your article readers may understand that bacteria simly are the reason of implant failure. Especially in the cases You presented at pictures, wrong implant positioning in the bone and according to the prostodontic plan could be an initial problem leading to the following issues and further bacteria infiltration. Nevertheless, You need to elaborate and explain the whole cascade of problems that finally can give implant failure. I can suggest these studies as explaning that problems and also potential references to increase the number of references:

1. Implant fracture/failure explanation https://doi.org/10.3390/prosthesis3040026
2. You can base on this introduction https://doi.org/10.3390/prosthesis4010004
3. 10.1111/prd.12456
4. 10.1111/jre.12950

Further in the discussion you also can reach to and mention the problems other than bacteria- I sugest again wrong positionig/wrong prostodontics/too short biologic width- thus without having these issues solved no antibiotics and other antibacterial solution will not help.

It has been done.

 

Regarding conclusions You can only refer to Your results. "present study confirms that the prepared curcumin nanoparticles can be used as 180 an alternative or a useful adjunct to antibiotics" i would rather say that curcumin in xx% and particle size may have similar activity against P. Gingivalis as xx, yyy, zz...

It has been done.

 

However You have nt mention if curcumin can be use only local or also systemic? That could go to the discussion as well.

It has been done.

"In the future, the loading of curcumin into implant fixture can show some potential 185 benefits. The chemical antimicrobials may be slowly substituted with antibacterial plants 186 such as curcumin. Such a replacement will not only avoid the microbial resistance but will 187 also be a key to decrease the use of chemicals in dentistry. It can not only decrease the 188 implant failure, but also the helpful properties of curcumin for gingival and its optimistic 189 action on bone–implant contact can result in a further stabilization and success of the implant therapy." yhis part is not based on Your results and should be removed.

 We deleted it.

Besides above, all materials and devices should be described with manufacturer/city/state/country.

It has been done.

Round 2

Reviewer 3 Report

Dear authors,

Although the manuscript is improved, there are still multiple issues that need to be addressed prior the publication. There are still multiple grammar errors, reference style is not consistent, multiple error in the reference section (information missing like journal name, volume, pages......)

The weakest point of the manuscript is that you did not performed a sample size calculation. The methodology of your manuscript has no clinical and statistical significance at a such low number of patients.

Please see the attachment. 

Comments for author File: Comments.pdf

Author Response

Although the manuscript is improved, there are still multiple issues that need to be addressed prior the publication. There are still multiple grammar errors, reference style is not consistent, multiple error in the reference section (information missing like journal name, volume, pages......)

We improved the manuscript again according to your valuable comments. Please see the revised version. The modification are highlighted in yellow.

The weakest point of the manuscript is that you did not performed a sample size calculation. The methodology of your manuscript has no clinical and statistical significance at a such low number of patients.

The sample size was determined according to the results of Mombelli et al [1] and considering the first type error (equal to 0.05) and the power of 80%. Then, the number of 15 samples was obtained. And also consider that the study was experimental and not clinical.

 

1. Mombelli, A. and Lang, N.P.J.P., The diagnosis and treatment of peri‐implantitis. 1998, 17, 63-76.

 

Please see the attachment. 

We read all comments of the attached file. We improved the manuscript again according to your valuable comments. Please see the revised version. The modification are highlighted in yellow.

Reviewer 4 Report

The authors have made the suggested improvements, considering acceptable for publication.

Author Response

We improved the manuscript. thanks

Reviewer 5 Report

Dear Authors,

 

paper was improved. However, Still requires some editorial adjustments.

 

Author Response

We improved the manuscript.Thanks