Influence on Bone-to-Implant Contact of Non-Thermal Low-Pressure Argon Plasma: An Experimental Study in Rats

Round 1

Reviewer 1 Report

Osteointegration is still a relevant problem in implant surgery.

With their animal experiment study the authors assess the influence of Argon Plasma Treatment on the integration.

Introduction:

This part of the manuscript is too unfocussed. The total possibilities of the surface treatments are not in the focus of the manuscript (page #2, line #42-43), as well as titanium aging (page #2, line #51-56).

“Machined Surfaces“ are something other than “Mechanized Surfaces” as mentioned in the title. 18 of 33 references within the introduction are too much in general.

The end of the introduction should provide a hypothesis which is either proven by the authors or not.

Materials &Methods:

Who is manufacturer of the RBM used?

Statistical Analysis:

The statistical analysis is very detailed described but the whole manuscript does not show a single p-value? The decision algorithm to sacrifice the rats should be shown in the material and methods section (page #5, line #187 and 188).

Results:

What is the benefit of providing information as table AND as figure (table 1 and graphic 1, as well as table 2 and graphic 2). Here are no p-values to be detected? The legend of a graphic should be self-explanatory from its information value. They also should contain p-values.

According to which criteria were the analysis intervals defined?

Discussion:

Opening this section with the roughness classification and the bacterial adhesion on surfaces is unfortunate since both topics are not in the focus of the manuscript.

In general, it is no deficit of a study if differences do not show a statistical significance, then the authors should discuss their result critically.

Conclusions:

The conclusions cannot be comprehended with the here presented data. It is noted that more than 50% (17 of 30) of the references are older than 10 years.

In principle the authors discuss a clinically relevant topic. But the here presented evaluation has considerable deficits which lead to the conclusion that a publication of this manuscript in Applied Sciences cannot be supported.

Author Response

Dear Reviewer,

Thank you for your time and comments that certainly improve the quality and clarity of our work.

Following your recommendations, the following modifications were completed:

1. Introduction: This part of the manuscript is too unfocussed. The total possibilities of the surface treatments are not in the focus of the manuscript (page #2, line #42-43), as well as titanium aging (page #2, line #51-56).

Following your recommendation, those parts have been eliminated from introduction.

2. “Machined Surfaces“ are something other than “Mechanized Surfaces” as mentioned in the title.

The term “machined” has been change for “mechanized” all along the manuscript.

 

3. 18 of 33 references within the introduction are too much in general.

With the corrections done for point 1, a total of 8 references has been eliminated from introduction.

4. The end of the introduction should provide a hypothesis which is either proven by the authors or not.

The last paragraph of the introduction in which we discussed our objective has been replaced by a new paragraph in which the working hypothesis is described.

5. Who is manufacturer of the RBM used?

The manufacturer is Ticare, Mozo-Grau, Valladolid, Spain. It has been added in line 87 after the first appearance of the term RBM.

6. The statistical analysis is very detailed described but the whole manuscript does not show a single p-value?

We have added the p values.

7. What is the benefit of providing information as table AND as figure (table 1 and graphic 1, as well as table 2 and graphic 2). Here are no p-values to be detected? The legend of a graphic should be self-explanatory from its information value. They also should contain p-values.

The tables provide all the data if they are of interest to the reader, since including such a large amount of data in the text would be tedious, on the other hand, the graph provides a quick overview of the results. p values have been added in both text and legends, which have been written in greater detail.

 

8. According to which criteria were the analysis intervals defined?

The confidence intervals were calculated for a confidence level of 95% and have been reflected in tables 1 and 2, which have been modified.

9. Discussion: Opening this section with the roughness classification and the bacterial adhesion on surfaces is unfortunate since both topics are not in the focus of the manuscript.

We agree with the referee and we have eliminated everything related to the roughness of the implants and bacterial adhesion. We have proceeded to rewrite the discussion.
We have also added the bibliography suggested by referee 2

10. Conclusions:The conclusions cannot be comprehended with the here presented data. It is noted that more than 50% (17 of 30) of the references are older than 10 years. In principle the authors discuss a clinically relevant topic. But the here presented evaluation has considerable deficits which lead to the conclusion that a publication of this manuscript in Applied Sciences cannot be supported.

We have updated our bibliography and improved the level of obsolescence.
We have rewritten the conclusions to comply with the suggestion of referee 1.

Author Response File: Author Response.pdf

Reviewer 2 Report

This paper is well-written and fits within the scope of the Applied Sciences. However, the paper in the present form requires a few modifications. Some of the lacunae and possible modifications are given below:

• The title is very long and confusing, it needs to be rewritten in a clearer way.
• Resorbable blast media (RBM) acronym needs to be added to the abstract.
• Please add the SD to the figures in the form of error bars.

Author Response

Dear Reviewer,

Thank you for your time and comments that certainly improve the quality and clarity of our work.

Following your recommendations, the following modifications were completed:

 

• The title is very long and confusing, it needs to be rewritten in a clearer way.

Tittle has been changed to: “Influence on Bone to Implant Contact of Non-thermal Low-Pressure Argon Plasma: An Experimental Study in Rats”

 

• Resorbable blast media (RBM) acronym needs to be added to the abstract.

Following your recommendations, it has been added to the abstract.

 

• Please add the SD to the figures in the form of error bars.

SD have been changed for C.I. as suggested by another reviewer. C.I. has been added to the graphics in the form of error bars, as indicated.

Author Response File: Author Response.pdf

Reviewer 3 Report

Authors should improve the discussion section about the osseointegration issue involving recent published literature (e.g. doi: 10.2147/IJN.S162836; doi: 10.3390/ijms19041022; doi: 10.1002/jbm.a.36213; doi: 10.1097/ID.0000000000000268).

Authors also add the statistical analysis in the histogram pictures.

Author Response

Dear Reviewer,

Thank you for your time and comments that certainly improve the quality and clarity of our work.

Following your recommendations, the following modifications were completed:

1. Authors should improve the discussion section about the osseointegration issue involving recent published literature (e.g. doi: 10.2147/IJN.S162836; doi: 10.3390/ijms19041022; doi: 10.1002/jbm.a.36213; doi: 10.1097/ID.0000000000000268).

We accept the reviewer’s recommendation and have rewritten the discussion. In the other hand, we have added 3 of the 4 suggested references in the Introduction part:

1. Zizzari, V.L.; Marconi, G.D.; De Colli, M.; Zara, S.; Zavan, B.; Salini, V.; Fontana, A.; Cataldi, A.; Piatelli, A. In vitro behavior of primary human osteoblast onto microrough titanium surface. Impl Dent. 2015, 24, 377-383.
2. Diomede, F.; D’Aurora, M.; Gugliandolo, A.; Merciaro, I.; Ettore, V.; Bramanti, A.; Piatelli, A.; Gatta, V.; Mazzon, E.; Fontana, A.; Trubiani, O. A novel role in skeletal segment regeneration of extracellular vesicles released from periodontal-ligament stem cells. Int J Nanomed. 2018, 13, 3805-3825.
3. Diomede, F.; D’Aurora, M.; Gugliandolo, A.; Merciaro, I.; Orsini, T.; Gatta, V.; Piatelli, A.; Trubiani, O.; Mazzon, E. Biofunctionalized scaffold in bone tissue repair. Int J Mol Sci. 2018,19, 1022.

 

2. Authors also add the statistical analysis in the histogram pictures.

Following the recommendation of the reviewer 1 and 2, we have modified the legend of the histograms and added the confidence intervals and p values to the tables.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Legend to table 1 remains unclear (line #200-202). Authors write: “No statistically difference was observed between groups or between time intervals (p<0.05)”

1) In my understanding a p<0.05 reflects a "significant difference". If p>0.05 this means "no significant difference"?

 2) It should be writing standard in scientific manuscripts to show every single p-value for every matched pair values

The new Graphic 1 is more informative but we don’t find a labeling of the x-axis. Why is the scaling of the x-axis in the first picture from 60-85 and in the fourth picture 60-90?

 The same misunderstanding in p-value is found in line #217: no significance but p<0.05

 The authors have added p-values in table 2. It is not enough to write p<0.05 for an ambitious scientific manuscript. Every matched pair of results needs an own calculated p-value.

Same distinct presentation is missing in Graphic 2

 However, the authors present one (two) hypothesis/es at the end of their introduction, but discussion and conclusion do not show a relation to these hypotheses.

The authors revised their manuscript very fast which leaves the impression with the reviewer that final editing has not been done in a proper fashion. Especially with the figures one would appreciate a consistent presentation with more accuracy in general

Author Response

Dear reviewer,

Thank you for your new appreciation, which undoubtedly helps in the substantial improvement of this manuscript. Answering to your suggestion, we have developed the following enhancements:

 

POINT 1. Legend to table 1 remains unclear (line #200-202). Authors write: “No statistically difference was observed between groups or between time intervals (p<0.05)”

• In my understanding a p<0.05 reflects a "significant difference". If p>0.05 this means "no significant difference"?

RESPONSE 1: Your understanding is right. Significant differences were considered with p < 0.05, as described in point 2.8 material and methods, so values of p over 0.05 means NO significant differences.

In legend to table 1: “No statistically difference was observed between groups or between time intervals (p<0.05)”, the p<0.05 in brackets means that this value was taken into consideration to establish the statistical differences. The sentence has been changed to avoid misunderstandings for: “No statistical difference was observed between groups or between time intervals taking into account the significance value p<0.05.”

 

POINT 2. It should be writing standard in scientific manuscripts to show every single p-value for every matched pair values

The new Graphic 1 is more informative but we don’t find a labeling of the x-axis. Why is the scaling of the x-axis in the first picture from 60-85 and in the fourth picture 60-90?

The authors have added p-values in table 2. It is not enough to write p<0.05 for an ambitious scientific manuscript. Every matched pair of results needs an own calculated p-value.

Same distinct presentation is missing in Graphic 2

RESPONSE 2: P values have been added to all matched pairs, which presented significant differences. The scale in the graphics has been changed to the same values.

 

POINT 3. However, the authors present one (two) hypothesis/es at the end of their introduction, but discussion and conclusion do not show a relation to these hypotheses.

RESPONSE 3: Conclusions have been modified with the confirmation of our alternative hypothesis.

Author Response File: Author Response.pdf