Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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Dr Mohan Z Mani

"Thank you very much for having published my article in record time.I would like to compliment you and your entire staff for your promptness, courtesy, and willingness to be customer friendly, which is quite unusual.I was given your reference by a colleague in pathology,and was able to directly phone your editorial office for clarifications.I would particularly like to thank the publication managers and the Assistant Editor who were following up my article. I would also like to thank you for adjusting the money I paid initially into payment for my modified article,and refunding the balance.
I wish all success to your journal and look forward to sending you any suitable similar article in future"



Dr Mohan Z Mani,
Professor & Head,
Department of Dermatolgy,
Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



Dr Kalyani R
Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
‘Knowledge is treasure of a wise man.’ The free access of this journal provides an immense scope of learning for the both the old and the young in field of medicine and dentistry as well. The multidisciplinary nature of the journal makes it a better platform to absorb all that is being researched and developed. The publication process is systematic and professional. Online submission, publication and peer reviewing makes it a user-friendly journal.
As an experienced dentist and an academician, I proudly recommend this journal to the dental fraternity as a good quality open access platform for rapid communication of their cutting-edge research progress and discovery.
I wish JCDR a great success and I hope that journal will soar higher with the passing time."



Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




Dr. Arunava Biswas

"My sincere attachment with JCDR as an author as well as reviewer is a learning experience . Their systematic approach in publication of article in various categories is really praiseworthy.
Their prompt and timely response to review's query and the manner in which they have set the reviewing process helps in extracting the best possible scientific writings for publication.
It's a honour and pride to be a part of the JCDR team. My very best wishes to JCDR and hope it will sparkle up above the sky as a high indexed journal in near future."



Dr. Arunava Biswas
MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




Dr. C.S. Ramesh Babu
" Journal of Clinical and Diagnostic Research (JCDR) is a multi-specialty medical and dental journal publishing high quality research articles in almost all branches of medicine. The quality of printing of figures and tables is excellent and comparable to any International journal. An added advantage is nominal publication charges and monthly issue of the journal and more chances of an article being accepted for publication. Moreover being a multi-specialty journal an article concerning a particular specialty has a wider reach of readers of other related specialties also. As an author and reviewer for several years I find this Journal most suitable and highly recommend this Journal."
Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2023 | Month : June | Volume : 17 | Issue : 6 | Page : ZC19 - ZC24 Full Version

Comparative Evaluation of Antibacterial Efficacy of Two Bioceramic Root Canal Sealers Incorporated with Novel Silica Doped TiO2 Nanoparticles: An In-vitro Study


Published: June 1, 2023 | DOI: https://doi.org/10.7860/JCDR/2023/63458.18065
Alekhya Mediboyina, Krishna Prasad Parvathaneni, TBVG Raju

1. Postgraduate, Department of Conservative Dentistry and Endodontics, Sree Sai Dental College and Research Institute, Chapuram, Srikakulam, Andhra Pradesh, India. 2. Professor and Head, Department of Conservative Dentistry and Endodontics, Sree Sai Dental College and Research Institute, Chapuram, Srikakulam, Andhra Pradesh, India. 3. Professor, Department of Conservative Dentistry and Endodontics, Sree Sai Dental College and Research Institute, Chapuram, Srikakulam, Andhra Pradesh, India.

Correspondence Address :
Dr. Alekhya Mediboyina,
M. Nagaraju, 19-1-174, Vinayakanagar, Anantapur, Srikakulam-515001, Andhra Pradesh, India.
E-mail: alekhyaprakhyath02@gmail.com

Abstract

Introduction: The success of Root Canal Treatment (RCT), depends on the complete elimination of microorganisms in combination with complete root canal system closure. Complete elimination of bacteria cannot be done by cleaning and shaping alone from lateral canals, isthmuses, and apical deltas. Thus, root canal sealers with ideal physical, biological, and improved antimicrobial characteristics are thus, necessary to avoid reinfections.

Aim: To evaluate the antibacterial efficacy of two Bioceramic (Bio-C) modified root canal sealers, using Silica Doped Titanium Dioxide Nanoparticles (SiTiO2), a unique, extremely effective antibacterial agent on root canal dentin infected with Enterococcus faecalis (E. faecalis).

Materials and Methods: This in-vitro study was conducted in the Department of Conservative Dentistry and Endodontics in collaboration with the Department of Microbiology at Sree Sai Dental College and Research Institute, Andhra Pradesh, India. The study was done in August 2022. The antibacterial efficacy of the two Bio-C sealers was evaluated by counting Colony Forming Units (CFU) and the percentage of live bacteria by a confocal laser scanning microscope. A total of 60 middle thirds of single-rooted teeth incubated with E. faecalis were chosen. Gutta-percha (GP) was used to fill the canals along with sealers in all six groups (group I-VI), namely Mineral Trioxide Aggregate based (MTA Fillapex); MTA Fillapex+SiTiO2 NPs (1% wt); MTA Fillapex +2% wt SiTiO2 NPs; Bio-C; Bio-C+ 1% wt SiTiO2 NPs; Bio-C+ 2% wt SiTiO2 Nanoparticles (NPs), respectively and incubated for seven days, each tooth was divided into two halves longitudinally. Microbiological analysis was conducted on one half, and microscopic analysis on the other half. The six groups were compared using a one-way Analysis of Variance (ANOVA) and intergroup comparison with Tukey’s post-hoc tests.

Results: The addition of SiTiO2 NPs to Bio-C and MTA Fillapex significantly reduced the bacteria, compared to an unmodified sealer (p-value <0.001). MTA Fillapex with SiTiO2 NPs showed higher bacterial viability compared with Bio-C with SiTiO2 NPs.

Conclusion: Loading endodontic sealers with SiTiO2 NPs has a material-dependent impact on the antibacterial properties, that could lower the frequency of secondary infections.

Keywords

Enterococcus faecalis, Mineral trioxide aggregate, Titanium dioxide

The goal of endodontic therapy is to completely eradicate microorganisms, with the aid of mechanical cleaning and shaping, antibacterial irrigants, and sufficient filling of the empty space. However, these procedures do not result in total root canal disinfection. E. faecalis is commonly isolated from teeth with long lasting endodontic infections. The ability of E. faecalis to survive for long periods without nutrition and the presence of several virulence factors contribute to their persistence. Therefore, to enhance their antibacterial activity and prevent reinfection, antimicrobial agents are added to root canal sealers (1). The evolution of novel, efficient antibacterial agents appears to be of utmost important. Silica is a potent and biocompatible antimicrobial agent with no known antibacterial resistance documented. Due of the increasing surface area, SiO2 activity would become more relevant at the nanoscale. Si nanoparticles, according to research by Ghanbari H et al., prevented bacteria from adhering to oral biofilms (2).

Utilising nanoparticulate materials to disinfect root canals has been a subject of recent research (3). Titanium dioxide Nanoparticles (TiO2 NPs) shows an antibacterial effect due to their physicochemical and biological activities in addition to their unique photocatalytic action (4). Previous studies have investigated the antibacterial effect of silica and titanium dioxide nanoparticles incorporated into many biomaterials like composites, irrigants, bleaching agents, and implant surfaces and this has shown, sustained release rates with the ability to reduce bacterial colonisation, biocompatibility, as well as, chemical bonding to dentin (5),(6). There are a few investigations about the antibacterial properties of bioceramic sealers like MTA Fillapex and Bio-C sealer. These have been emphasised because of their alkaline pH, sealing ability, biocompatibility, dimensional stability, the potential to increase root strength by osteogenic constituents like calcium phosphate and calcium silicate [7,8]. However, there is no study in the available literature, assessing the comparative antimicrobial effectiveness of these sealers incorporated with SiTiO2 NPs.

Thus, the present study was performed to compare and evaluate the antibacterial efficiency of Bio-C root canal sealers, that had been modified with a highly loaded, antimicrobial agent SiTiO2 NPs on root canal dentin, that had been infected with E. faecalis.

Material and Methods

This in-vitro study was conducted in the Department of Conservative Dentistry and Endodontics in collaboration with the Department of Microbiology at Sree Sai Dental College and Research Institute, Andhra Pradesh, India. The study was done in August 2022. Ethical approval was obtained from Institutional Ethics Committee (SSDCRI/IEC/2022/5/S1).

Inclusion criteria: Single-rooted teeth with a mature apex were included in the study.

Exclusion criteria: Teeth having fractures, root caries, morphological deformities, or severely curved roots were excluded from the study (Table/Fig 1)(a-f).

Study Procedure

The antibacterial efficacy of two Bio-C root canal sealers incorporated with SiTiO2 NPs were evaluated using microbiological and microscopic analysis (Table/Fig 2), SiTiO2 NPs (Nano Research Laboratory; Jamshedpur, India) were gradually added to Bio-C sealer at either 1% or 2% wt (Angelus, Paraná, Brazil), or MTA Fillapex (Angelus, Londrina, Brazil). These SiTiO2 NP concentrations were determined following a pilot research, that evaluated the flow of materials with concentrations from 5% to 1% wt; the addition of SiTiO2 NPs in the 1%-2% range had no impact on the setting of the materials (9).

Sample preparation: A total of 60 single-rooted teeth were collected and the middle third of each root was obtained by sectioning them using a diamond disc. ProTaper rotary files upto size F3 were used to prepare the root canals. Finally, 5% sodium hypochlorite and 17% Etheylenediamine Tetraacetic Acid (EDTA) were used to irrigate canals and autoclaved for 20 minutes at 121°C.

Root canal infection with Enterococcus faecalis: Canals were infected with E. faecalis (ATCC 29212) in the present study. In a 96-well microtiter plate, prepared root samples were placed. 20 μL of E. faecalis culture with 300 μL of tryptone soya broth was injected 20into each well, then incubated for three weeks anaerobically at 37°C. For three weeks, the growing media was replaced every other day. All the procedures were done under a laminar flow hood.

Confirmation of tooth contamination: The samples were rinsed with 5 mL of sterile saline, following the incubation period. Six root samples (one from each group) were viewed under a Scanning Electron Microscope (SEM) to verify the presence of bacteria and their structure (Table/Fig 3).

Group specification: The remaining 60 samples were divided into six groups (10 in each group) and canals were filled with GP and sealers:

Group I- MTA Fillapex
Group II- MTA Fillapex + SiTiO2 NPs (1% wt)
Group III- MTA Fillapex + SiTiO2 NPs (2% wt)
Group IV- Bio-C
Group V- Bio-C + SiTiO2 NPs (1% wt)
Group VI- Bio-C + SiTiO2 NPs (2% wt)

Samples were incubated anaerobically for seven days at 37°C. After the incubation period, each tooth was split into two halves; both, microscopic and microbiological analyses were conducted on separate halves of the sample.

Microbiological analysis: Gates-Glidden drills were used to obtain dentin powder from the canal. The samples were all transferred to 2 mL of Brain Heart Infusion (BHI) media and incubated for another 24 hours at 37°C. In microcentrifuge, test tubes containing 1 mL BHI, the resulting dentin powder was collected, vortexed, serially diluted, and plated. At 37°C, the plates were incubated for 48 hours. The Colony Forming Unit (CFU) were calculated and converted to their log10 values (10).

Microscopic analysis: Near the pulp canal space, a 1 mm thick portion was obtained from each dentin block using IsoMet 4000 Precision Saw. The root sections were stained with 0.01% acridine orange (green fluorescence) and 0.01% propidium iodide (red fluorescence) and then rinsed with phosphate-buffered saline. Specimens were transferred to glass coverslips and Zeiss Laser Scanning Microscopy (LSM) 880 Airyscan (Carl Zeiss, Oberkochen, Germany) data were analysed immediately to ascertain the proportion of live/dead cells for each treatment. Image J software was utilised to calculate the proportion of live (green fluorescence) and dead (red fluorescence) cells (11).

Flow test: According to standardised International Organisation for Standardisation (ISO) 6876:2012 protocols, the flow of stock and modified sealers was evaluated (12). Applying 0.05±0.005 mL of sealer with a graduated syringe at the center of a glass plate. A second glass plate with an additional mass, a total of 120±2 g, was positioned in the center of the sealer at 180±5 seconds, following the start of the mixing process. After 10 minutes of the initial mixing time, the weight was taken off and the compressed sealer discs’ maximum and minimum diameters were measured in millimeter (mm) using digital calipers. Final flow rate was calculated by subtracting the minor diameter from the major diameter (12).

Statistical Analysis

The statistical analysis was conducted using Statistical Package for Social Sciences (SPSS) version 20.0. One-way ANOVA was used for the analysis, and Tukey’s post-hoc test was used to compare groups. The level of significance was set at (p-value <0.05).

Results

Microbiological analysis: Bio-C+2% SiTiO2 NPs significantly reduced the CFU count, showing a mean of 0.48±0.672 log CFU/mL compared with Bio-C+ GP (3.46±0.614) log CFU/mL and Bio-C +1% SiTiO2 NPs (1.68±0.277) log CFU/mL [Table/Fig-4,5]; MTA Fillapex+2% SiTiO2 NPs showed an improved antimicrobial performance, showing CFU (2.32±0.804) log CFU/mL versus MTA Fillapex + 1% SiTiO2 NPs (4.73±0.660) log CFU/mL and MTA Fillapex + GP (7.46±1.174) log CFU/mL. Bio-C sealer showed a better reduction of CFUs compared with the MTA Fillapex groups (Table/Fig 6).

Microscopic analysis: Confocal images showed an increased percentage of dead bacteria in all the groups. The addition of SiTiO2 NPs to MTA Fillapex and Bio-C sealers showed a significant reduction of live bacteria compared with the unmodified groups (Table/Fig 7),(Table/Fig 8). The addition of 2% SiTiO2 NPs to Bio-C sealer decreased the bacterial viability compared with 2% SiTiO2 NPs to MTA FA at seven days incubation (4.8%±1.6% vs 46.2%±26.8%, p<0.05) (Table/Fig 9),(Table/Fig 10).

Flow test: With flow diameters of 17 mm, MTA Fillapex and Bio-C unmodified sealers, both were within the ISO 6876:2012 flow criteria (13). SiTiO2 NPs considerably reduced the flow of all modified sealers (p-value <0.05). For the MTA Fillapex, but not the Bio-C groups, the decrease in flow was associated with the mass of SiTiO2 NPs present in the materials (p-value <0.05). All changed sealers were within the permissible ISO 6876 flow limits, with the exception of MTA Fillapex 2% wt SiTiO2 NPs (15.7+0.35 mm) (Table/Fig 11),(Table/Fig 12).

Discussion

Improved disinfection of the root canal system may be facilitated by enhancing and extending the antibacterial properties of sealers. However, the effects of incorporating SiTiO2 NPs into a root canal sealer against E. faecalis impregnated into root dentin have never been reported. According to the results, both modified and unmodified sealers significantly reduced the number of bacterial cells in a dose-dependent manner. The highest antibacterial effect was reported in Bio-C + 2% SiTiO2 NPs group followed by MTA Fillapex+2% SiTiO2 NPs, Bio-C +1% SiTiO2 NPs, Bio-C+GP, MTA Fillapex+1% SiTiO2 NPs, and MTA FA+GP. Pair-wise comparisons between the groups showed significant differences (p-value <0.001).

Flow is a crucial component for root canal filling which enables the sealer to enter the imperfections of root canal networks. According to ISO 6876 criteria, all of the sealers provided flow rates that supported earlier research (14),(15). The Bio-C sealer demonstrated the maximum flow rate. The results of the culture-based microbiological research revealed that, when compared to the MTA Fillapex groups, Bio-C with or without SiTiO2 NPs considerably decreased the quantity of bacteria. Literature shows that, in addition to its hydrophilic nature, and its active calcium hydroxide diffusion, for upto 21 days (7), Bio-C sealer can be alkaline, reaching a pH of 10. It is well known that a pH greater than nine can render microbial cell membrane enzymes inactive, which results in a reduction in biological activity or loss of the plasma membrane’s integrity as reported by Allaker RP (16).

The antibacterial activity of the sealers was examined in the present study, while they were in contact with the root dentin. Due to its buffering properties and the binding of certain cationic compounds to dentin, dentin has been demonstrated to impede the antibacterial action of root canal medications, reducing their antimicrobial efficiency (17),(18). E. faecalis was selected for the present study, because it is the most resistance and commonly appeared in the root canals of teeth with post-treatment apical periodontitis (19),(20),(21),(22). Numerous virulence factors have been implicated in the survival of E. faecalis following endodontic treatment. It is also capable of penetrating into dentinal tubules, as soon as, 48 hours after inoculation (23). The gold standard for identifying and quantifying viable bacteria is CFU counting, using culture-based techniques. However, the accuracy of these techniques depends on the transport and culture medium, culture-based approaches are unable to identify live bacteria. The present study uses Confocal Laser Scanning Microscopy (CLSM) along with the fluorescent stains has made it possible to evaluate the structure, distribution, and viability of certain bacteria inside the biofilm (24),(25).

Studies done by Shakya VK et al., (2016) and Mangat P et al., (2020) showed counts for SiTiO2 NPs modified MTA Fillapex were less than the unmodified sealer and higher than Bio-C. It may be presumed that, despite its alkalinity and calcium release, the environment was still conducive to the life of the microorganism. Its proton pump is most likely a major contributing cause to its resistance to alkaline substances (26),(27). When compared to the effects of the unmodified sealers, the addition of SiTiO2 NPs to both sealers (Bio-C and MTA Fillapex) considerably improved the antibacterial effect (p-value <0.001). The deeper penetration of nanoparticulate medications into the microbial cells and dentinal tubules is the primary goal of adding them to endodontic sealers. According to Waltimo T et al., adding SiNPs to dental biomaterials and cement has a favourable antibacterial effect through the release of ionic alkaline species over time and is being considered as a dentine disinfectant to provide an alternative to calcium hydroxide (28). In recent years, researchers have explored the antibacterial properties of TiO2 nanoparticles as an efficient antimicrobial agent against a large variety of species, including bacteria both gram-positive and gram-negative (29),(30),(31). Unique photocatalytic activity of TiO2 can lead to higher reactive oxygen species this might cause oxidative stress and breakdown bacterial cell walls, acting as an antibacterial agent (32),(33).

To the best of author’s knowledge, no prior research has compared these two sealers incorporated with novel antibacterial agent SiTiO2 NPs. Bio-C+2% SiTiO2 showed more significant antimicrobial activity, when compared to other groups followed by Bio-C+1% SiTiO2 group, MTA Fillapex+ 2% SiTiO2 NPs group, Bio-C, MTA Fillapex+1% SiTiO2 and least for the MTA group.

Limitation(s)

In the present study, the maximum incubation period was only seven days, and longer periods of antimicrobial effect of these sealers, may be examined in future research. A sealers, that is effective against one microbe in-vitro might not necessarily be effective against the same bacterium in-vivo. Thus, it is necessary to do further research utilising the same medications for longer periods in clinical trials of all infected canals to validate the results.

Conclusion

Within the confines of the present study’s limitations, it can be concluded that, root canal sealers modified with SiTiO2 NPs shows long term antimicrobial properties in a dose-dependent manner, while maintaining flow compliance with the acceptable ISO standard. SiTiO2 NPs had limited effect on the antimicrobial properties of MTA Fillapex, when compared to the Bio-C sealer. SiTiO2 NPs could potentially reduce bacterial proliferation and the incidence of secondary root canal infections.

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DOI and Others

DOI: 10.7860/JCDR/2023/63458.18065

Date of Submission: Feb 20, 2023
Date of Peer Review: Mar 28, 2023
Date of Acceptance: Apr 24, 2023
Date of Publishing: Jun 01, 2023

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? No
• For any images presented appropriate consent has been obtained from the subjects. No

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Mar 01, 2023
• Manual Googling: Mar 31, 2023
• iThenticate Software: Apr 19, 2023 (12%)

ETYMOLOGY: Author Origin

EMENDATIONS: 8

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