[version 3; peer review: 2 approved with reservations, 1 not approved]
No competing interests were disclosed.
Graphene is the thinnest, strongest, and stiffest imaginable material. The biocompatible property of graphene oxide can initiate and facilitate cell adhesion, proliferation, and differentiation of periodontal ligament, osteogenic, and oral epithelial cells. Furthermore, the antibiofilm and anti-adhesion properties of graphene oxide in the prevention of dental biofilm infections, dental caries, and dental erosion as well as for implant surface modification and as an anti-quorum sensing agent. Composites are the most often utilized materials for restoration in the field of dentistry due to adhesive resins' improved mechanical and cosmetic properties. To safeguard the dentin and prevent dental cavities, dentin adhesives are utilized to affix hydrophobic resin composites to hydrophilic dentin tissue.
Dental adhesives have a harder time adhering to dentin because it contains more water and is less mineralized than enamel. This makes the method more sensitive.
As a result, it was chosen to assess and contrast the impact of 5% Hydroxyapatite nanoparticles and 2% Graphene oxide nanoparticles, both separately and together, on the Micro tensile bond strength of 5th generation adhesive.
Graphene oxide is the most versatile form of Graphite in structural and functional configuration. Graphene oxide possesses extraordinary physical, chemical, optical, electrical and mechanical properties. Among the graphene family nanomaterials, the reduced form of Graphite adding the oxygenated functional group to the structure increases the surface area and therefore exhibits enviable excellent interaction ability with metal and ions as well as organic species. Graphene oxide in dentistry has provided outstanding results in antimicrobial action, regenerative dentistry, bone tissue engineering, drug delivery, physicochemical properties, enhancement of dental biomaterials and oral cancer treatment.
Version 3 of this article includes a better organised abstract.
Considering their enhanced aesthetics and increased mechanical qualities, the most used material for restoration in dentistry is dental composite.
Before materials are marketed in dentistry, it is crucial to conduct and analyze mechanical strength tests on them. Through laboratory investigations, these evaluations are carried out in a variety of ways. Using the TBS test method, the bonding strengths of self-adhesive composite resins with various universal bonding methods were examined in this study. The TBS values increased when the adhesive system was used with universal adhesive systems. As a result, our study's null hypothesis was rejected. The fifth-generation resin adhesive's bond integrity, longevity, and contact with dentin were studied in the current study about the addition of nano-HA and different nano-GO particle concentrations (0.5 wt% and 2 wt%). The Hydroxyapatite-graphene oxide nanoparticles 2% group demonstrated greater micro tensile bond strength than the Hydroxyapatite-graphene oxide nanoparticles 0.5% group, which revealed greater than the controls, according to the data. The idea that adding graphene oxide nanoparticles to the bonding agent would strengthen its binding was therefore considered. Adhesives are essential for the clinical outcome of restorations because inadequate restorative adhesive bonds have been linked to secondary caries development, dentin hypersensitivity (DH), and microleakage.
To evaluate the micro tensile bond strength of the 5th generation bonding agent without incorporating any of the nanoparticles using a universal testing machine. To evaluate the micro tensile bond strength of a 5th generation bonding agent incorporating 2% Graphene oxide nanoparticles in isolation using a universal testing machine. To evaluate the micro tensile bond strength of 5th generation bonding agents incorporating 5% Hydroxy apatite nanoparticles in isolation using a universal testing machine. To compare the micro tensile bond strength of 5th generation bonding agent incorporating 2% graphene oxide and 5% Hydroxyapatite nanoparticles in combination using a universal testing machine.
Graphene oxide nanoparticles (Nano Research Lab, Jamshedpur, India) Hydroxyapatite nanoparticles (Nano Research Lab, Jamshedpur, India) 5th generation bonding agent (3M ESPE Adper Single Bond 2) Dentsplyspectrum - composite restoration
Inclusion criteria 84 Maxillary premolars Teeth devoid of restorations Exclusion criteria Teeth with previous root canal treatment. Extensively carious tooth. Abrasion, attrition, fluorosis, or other enamel defects. Teeth with developmental anomalies. Teeth with external and internal resorption.
A total of 84 samples are used in the study divided into 4 groups, 21 in each group corresponding to the 1, 2, 3, and 4 sample sets. Group 1 – 5th generation bonding agent Group 2 – 5th generation bonding agent with graphene oxide nanoparticles. Group 3 – 5th generation bonding agent with hydroxyapatite nanoparticles Group 4 – 5th generation bonding agent with a combination of graphene oxide and hydroxyapatite nanoparticles.
Procedure:
The nano-HA particles will be silanized after being purchased commercially. 5% silane will be added to the 95% adhesive to help the nano-hydroxyapatite stick to it.
Solvent for silanization using acetone. To create a fifth adhesive system that contains 5 wt% nano-Hydroxyapatite particles, the nano-HA particles will be introduced to the experimental adhesive at a 5% concentration (m/m). The experimental adhesive's nano-hydroxyapatite particles will be dispersed using sonication in a centrifuge to encourage homogenization. To allow the solvent to evaporate, the synthesized adhesives will be stored in isolation for 24 hours at 37 degrees C. Due to their short shelf life, the adhesives will thereafter be stored at 4 degrees C and used for 20 days after formulation.
The graphene oxide powder will be combined with 2 mL of ethanol in a microbial and ultrasonically processed for 10 minutes at room temperature. The dentin adhesive will get this mixture. The nano-graphene oxide particles will first be combined with resin before being sonicated in an ultrasonic bath for 10 minutes creating a uniform combination. After sonication, the mixture will be homogenized for 60 seconds at room temperature in an ultrasonic homogenizer using pulse on/off technology. Every time the mixture is used, the mix will be homogenized in an ultrasonic homogenizer to ensure that the graphene oxide will disperse uniformly after storage. Calculations will be done to determine the resin volume in milliliters and milligram weight of nanoparticles. Using the following:
The graphene oxide powder will be combined with 2 mL of ethanol in a microbiological container before being ultrasonically processed for 10 minutes at room temperature. The nano-HA particles were silanized after being purchased commercially. The 95% acetone solvent will receive an addition of 5% silane for silanization to enhance the adherence of nano-hydroxyapatite to the adhesive. Once both the nanoparticles are made, the made solvents will be added to the 5th generation dentin adhesive. To achieve a homogenous mixture, it will be sonicated in an ultrasonicator duration of 10 min. Post-sonication, the mixture will be homogenized in an ultrasonic homogenizer at pulse on/of for 60 s at room temperature. The mixture will be homogenized in the ultrasonic homogenizer before every use to ensure that graphene oxide and hydroxyapatite nanoparticles will disperse uniformly after storage. Calculations will be done to determine the resin volume in milliliters and milligrams weight of the nanoparticles. This will allow the solvent to evaporate, the synthesized adhesives will be stored in isolation for 24 hours at 37 degrees C. Due to their short shelf life, the adhesives will thereafter be stored at 4 degrees C and will be used within 20 days of the formulation.
The investigation will use extracted permanent maxillary premolars. All teeth will be kept in the storage media for a maximum of three months in distilled water. The 1, 2, 3, and 4 sample sets will be represented by the groups of the 84 premolars, which will be randomly divided into those groups. There will be Class 1 cavity preparation. The 3 M Scotchbond Universal Etchant (30 – 40% phosphoric acid) will be applied to all groups for 15 seconds. The surfaces will then be washed and dried by air. The surplus glue will then be removed using dental cotton rolls after adhesive systems have been placed for 15 seconds. The surfaces will then be exposed to light for nearly 20 seconds at an intensity of around 800 mW/cm 2 using a light-curing device. The previously polymerized adhesive layer will be covered with successive 2-mm thick layers using Dentsply Spectrum -restorative material, which will then be light cured for 20 seconds. Following treatment, all of the teeth will be further separated into resin dentin sticks using a diamond disc and water cooling by ISO/TS 11405, with an average cross-sectional area of 1 mm by 1 mm. The samples will then be kept in deionized (DI) water for 24 hours at 37 °C. Next, utilizing the universal testing machine, the created samples will go through a micro-tensile bond strength test. The beams will be secured to a jig with cyanoacrylate adhesive before being put to tensile stresses at a crosshead speed of one millimeter per minute.
Primary Variable - Microtensile bond strength
Sample size formula for difference between two means:
Where;
Zα = level of significance at 5% i.e., 95%
Confidence interval
Zβ is the power of test
δ1
δ2
Pooled std. dev. = (3.81+3.60)/2 = 3.71
n
The total sample size is 60
Study Reference: [
It is expected that among the material used in the study, the 5th generation bonding agent with graphene oxide and nano-hydroxyapatite crystals in combination may give better results as compared to the 5th generation bonding agent with or without incorporation of graphene oxide and nano hydroxyapatite crystals in isolation in routine practice due to its ability of anti-microbial properties, optimal biocompatibility, and increased bond strength.
The focus of our study is to evaluate the increase in the micro tensile bond strength of the 5th generation adhesive system after the incorporation of graphene oxide and hydroxyapatite nanoparticles. Adhesion of the nanoparticles at the dentin interface leading to widespread surface area may further contribute to the increased bond strength, enhancing the mechanical properties.
Not started yet.
Ethical approval received from Datta Meghe Institute of Higher Education and Research, Sawangi, Wardha
IEC reference number- DMIHER (DU)/IEC/2023/584
No data are associated with this article.
I would like to thank my institute, my Guide, HOD, and my colleagues.
Thank you. I hope it will be good research. Authors need more effort to make the points clear to readers. Please improve your writing. I have some suggestions. Hope it helps.
Graphene and graphene oxide are not the same. In the abstract the author misleads graphene and graphene oxide.
“imaginable material” is not scientifically correct.
“Furthermore, the antibiofilm and anti-adhesion -------------- as an anti-quorum sensing agent.” Please make it meaningful.
“Composites are the most often utilized materials for restoration-------cosmetic properties” please mention what composite are you talking about?
Please mention objective and hypothesis.
“Dental adhesives have a harder time ------- more sensitive. What is this? Please write your proposed study design.
“As a result, it was chosen ----------------- 5th generation adhesive.” Is it the result or objective of the study?
Redundancy should be avoided
An overview of clinical problems, existing research, their limitations, possible solutions, and new novel research obligations should be discussed consistently.
Redundancy should be avoided
A consistent flow of proposed methods and their predictable outcomes should be discussed with appropriate reference.
The way the conclusion was written is meaningless. Please remove these and conclude your expected result (hypothesis). You cannot introduce the graphene oxide in the conclusion
The aim of the study should be reflected in the conclusion.
Is the study design appropriate for the research question?
Partly
Is the rationale for, and objectives of, the study clearly described?
No
Are sufficient details of the methods provided to allow replication by others?
Partly
Are the datasets clearly presented in a useable and accessible format?
No
Reviewer Expertise:
Dental biomaterials, restorative dentistry and cariology
I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.
Respected Reviewer,
Thankyou for all the suggestions, I will do all the corrections required the manuscript.
Thanks and Regards.
Dear Reviewer,
Corrections have been done. Kindly go through the author response below -
Graphene and graphene oxide are not the same. In the abstract, the author misidentifies graphene and graphene oxide.
“Imaginable material” is not scientifically correct.
“Furthermore, the antibiofilm and anti-adhesion -------------- as an anti-quorum sensing agent.” Please make it meaningful.
“Composites are the most often utilized materials for restoration-------cosmetic properties” please mention what composite are you talking about?
Please mention objective and hypothesis.
“Dental adhesives have a harder time ------- more sensitive. What is this? Please write your proposed study design.
“As a result, it was chosen ----------------- 5th generation adhesive.” Is it the result or objective of the study?
Redundancy should be avoided
An overview of clinical problems, existing research, their limitations, possible solutions, and new novel research obligations should be discussed consistently.
Redundancy should be avoided
A consistent flow of proposed methods and their predictable outcomes should be discussed with appropriate reference.
The way the conclusion was written is meaningless. Please remove these and conclude your expected result (hypothesis). You cannot introduce graphene oxide in the conclusion
The aim of the study should be reflected in the conclusion.
The conclusion at the end of the text is not coincide with that at the abstract.
- Discussion needs further explanation and the results should be compared with other studies.
- Clear and definitive conclusions should be drawn at the end of the study highlighting the findings of the study.
Is the study design appropriate for the research question?
Partly
Is the rationale for, and objectives of, the study clearly described?
Partly
Are sufficient details of the methods provided to allow replication by others?
No
Are the datasets clearly presented in a useable and accessible format?
Partly
Reviewer Expertise:
My research interest is surface modification of dental implant with a bioactive material to enhance osseointegration. In addition to that, research in the prevention of peri-implantitis and use of anti-bacterial agents that inhibit bacterial adhesion and proliferation.
I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.
-Discussion needs further explanation and the results should be compared with other studies.
Ans- Discussion has been written with context to the references added and their respective studies highlighting the details of the conclusion. As this is the protocol of the study we are conducting so the final results and their co-relation will be discussed in detail in the original study according to the statistical analysis.
So the results drawn from the original study will be resourceful and will aid in putting up the points concerning discussion of the material.
- Clear and definitive conclusions should be drawn at the end of the study highlighting the findings of the study.
Ans- This is the drafted protocol of the original study which is being conducted and therefore the conclusions/ findings of the same will be aligned once the study is completed and will be discussed in detail.
The above mentioned manuscript discussed the effect of adding nano-particle graphene oxide at different concentrations and nano-HA in isolation and in combination on the tensile strength of the 5th generation bonding agent.
Although the manuscript discussed an interesting subject in the field of conservative dentistry, there are certain points that should be addressed before indexing:
- The structure of the study needs further revision - it should follow papers of the scientific research - background, clear aims, objectives, materials and methods, discussion and conclusion.
Abstract: it needs further organization beginning from the background - materials and methods - results and conclusion. Introduction: it needs a comprehensive revision. In my view, the authors mentioned many studies that used graphene oxide with bioactive glasses or silica or nano-hydroxyapatite alone. Authors can use these studies to compare their results with these studies. - Authors need to add a clear hypothesis of the study (null hypothesis). Also, more focusing on the aim of the study and should be moved to the last paragraph of the introduction section. Materials and methods: there is a huge repetition in the preparing the adhesive materials with graphene oxide and nano-HA. The study lacks important investigation such as SEM to demonstrate the incorporation of graphene oxide and HA into the adhesive materials. Discussion: - An explanation for why graphene oxide and nano-hydroxyapatite improved the mechanical bonding of the adhesive materials should be added. - The main theme of the manuscript is the adhesive strength of the adhesive materials. Was there any correlation between improving the adhesive strength and the bacterial plaque? Conclusion: should be brief and more focused about the new findings.
Is the study design appropriate for the research question?
Partly
Is the rationale for, and objectives of, the study clearly described?
Partly
Are sufficient details of the methods provided to allow replication by others?
No
Are the datasets clearly presented in a useable and accessible format?
Partly
Reviewer Expertise:
My research interest is surface modification of dental implant with a bioactive material to enhance osseointegration. In addition to that, research in the prevention of peri-implantitis and use of anti-bacterial agents that inhibit bacterial adhesion and proliferation.
I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.
The structure of the study needs further revision - it should follow papers of the scientific research - background, clear aims, objectives, materials and methods, discussion, and conclusion.
Abstract: it needs further organization beginning from the background - materials and methods - results and conclusion.
Answer- In conclusion, the present protocol provides first-hand knowledge of experimental graphene oxide-based silane primers. Based on the results, we may infer that graphene oxide-based and hydroxyapatite-based - silane primers increase the micro-tensile bond strength.
Further the corrections are made and the abstract has been re-organized.
Introduction: it needs a comprehensive revision. In my view, the authors mentioned many studies that used graphene oxide with bioactive glasses or silica or nano-hydroxyapatite alone. Authors can use these studies to compare their results with these studies.
Answer- Definitely, all those studies will help in tapering down to the conclusion of the study we will be conducting. The various authors cited in the protocol used different combinations depicting improvement in the properties of dental adhesives. So we have mentioned the combination of nano-HA and GO- nanoparticles hypothesizing improved properties.
- Authors need to add a clear hypothesis of the study (null hypothesis). Also, more focus on the aim of the study should be moved to the last paragraph of the introduction section.
Null hypothesis- Both Graphene oxide and hydroxyapatite nanoparticles are hypothesized to be equally effective in increasing the micro-tensile bond strength in the dental adhesives.
Materials and methods: there is a huge repetition in preparing the adhesive materials with graphene oxide and nano-HA.
Answer- The detailed preparation of the material involving various steps that will be used for the in vitro study has been elaborated to standardize and henceforth bridge the gap for future studies to be conducted.
The study lacks important investigation such as SEM to demonstrate the incorporation of graphene oxide and HA into the adhesive materials.
Answer- Our study protocol aims to demonstrate and investigate the microtensile bond strength and so the parameter demands the method to test the strength using a Universal testing Machine.
Discussion: - An explanation for why graphene oxide and nano-hydroxyapatite improved the mechanical bonding of the adhesive materials should be added. - The main theme of the manuscript is the adhesive strength of the adhesive materials. Was there any correlation between improving the adhesive strength and the bacterial plaque?
Answer- According to Carvalho RM et al in 2012, adhesives are essential for the clinical outcome of restorations because inadequate restorative adhesive bonds have been linked to secondary caries development, dentin hypersensitivity (DH), and microleakage. Solhi L et al in 2012, dental composites' diverse mechanical and physical qualities are reinforced by the universal use of nanoparticles. Nobre CMG et al in 2020 and Pepla E et al in 2014, the nano-HA particles have a great ability to connect with the dental tissues, are biocompatible, and serve as a source of remineralizing ions like Ca and P. According to the current study, adding nano-HA particles to the adhesive produces all the advantages of using HA in adhesives. Nano-HA particles have been proposed in the past as a way to improve the mechanical characteristics of adhesive and enhance surface area for adhesion
- the anti-biofilm and anti-adhesion properties of graphene oxide in the prevention of dental biofilm infections
- Adhesion of the nanoparticles at the dentin interface leading to widespread surface area may further contribute to the increased bond strength, enhancing the mechanical properties.
- Since this is the study protocol, the correlation between improving the adhesive strength and the bacterial plaque has not yet been concluded. Although the previous studies cited in the manuscript described the positive results.
Conclusion: should be brief and more focused on the new findings.
Answer- The newer advances focus on the enhancement of the physical, mechanical, and biological properties by addition of the biocompatible materials in various generations of the Bonding agent by use of advancing technologies and subsequent results aiding in improving the understanding about the dental materials and their properties.
Future research using microtensile testing or a study involving dynamic loading instead of static loading may help to assess, in more detail, the graphene oxide sheet-infused experimental silane primers. Assessment of the elastic modulus of the adhesive layer through finite element analysis would also provide more information and evidence. Although this study explored a load of 0.1 wt% of graphene oxide sheets in silane primers, it is equally important to determine the optimal filler loading in future studies to optimize the mechanical strength of the adhesive layer.
1- Proof reading is required, inappropriate use of grammar, punctuation in some sentences
2- Where are the micro tensile bond strength data? there must be some graphs from the universal tensile machine software.
3- I appreciate that this is a method development study. However, since the authors have conducted statistics, I expect the data and the stats available in the main text. I cant find them anywhere! any explanation please ?
Is the study design appropriate for the research question?
Yes
Is the rationale for, and objectives of, the study clearly described?
Yes
Are sufficient details of the methods provided to allow replication by others?
Partly
Are the datasets clearly presented in a useable and accessible format?
Partly
Reviewer Expertise:
Dental Implantology, Nano coatings, Nano Dentistry, Dental Biomaterials, Antibacterial Biomaterials, Oral Surgery
I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.
Proofreading is required, inappropriate use of grammar, and punctuation in some sentences Where are the micro tensile bond strength data? there must be some graphs from the universal tensile machine software. I appreciate that this is a method development study. However, since the authors have conducted statistics, I expect the data and the stats available in the main text. I can't find them anywhere! any explanation, please?
Answer for the first query - The grammar correction and the proofreading have been done. According to the Grammarly app, the version 2 has more than 95% accuracy.
Answer for the second query- This is the protocol and the method of testing microtensile bond strength designed for the study to be conducted further and so the data and graphs related to same will be posted once the study is performed according to this protocol adopted.
Answer for the third query- As this is a research protocol the statistics mentioned here are for sample size calculation which will be used further for original research, the details of which will be published in the original article.
Thank you for your valuable response.
Proofreading is required, inappropriate use of grammar, and punctuation in some sentences Where are the micro tensile bond strength data? there must be some graphs from the universal tensile machine software. I appreciate that this is a method development study. However, since the authors have conducted statistics, I expect the data and the stats available in the main text. I can't find them anywhere! any explanation, please?
Answer for the first query - The grammar correction and the proofreading have been done. According to the Grammarly app, the version 2 has more than 95% accuracy.
Answer for the second query- This is the protocol and the method of testing microtensile bond strength designed for the study to be conducted further and so the data and graphs related to same will be posted once the study is performed according to this protocol adopted.
Answer for the third query- As this is a research protocol the statistics mentioned here are for sample size calculation which will be used further for original research, the details of which will be published in the original article.
Thank you for your valuable response.
Accepted