Volume 8, Issue 5, May – 2023 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Comparative Evaluation of Antibacterial Efficacy of

Newer Intracanal Medicament Nitrofurantoin and
Ozonated Oil Plus Calcium Hydroxide on E.faecalis –
An in Vitro Study
1.
Dr. Sadashiv Daokar (Professor & HOD, Department of Conservative Dentistry & Endodontics)
2.
Dr. Aishwarya Ranjalkar (Postgraduate Student, Department of Conservative Dentistry & Endodontics)
3.
Dr. Kalpana Pawar (Professor, Department of Conservative Dentistry & Endodontics)
4.
Dr. Shubhankar Nandkhedkar (Postgraduate student, Department of Conservative Dentistry & Endodontics)
5.
Dr. Kshama Sarak (Postgraduate student, Department of Conservative Dentistry & Endodontics)
6.
Dr. Snehal Tawar (Postgraduate student, Department of Conservative Dentistry & Endodontics)
7.
Dr. Dhanashri Padwal (Postgraduate student, Department of Conservative Dentistry & Endodontics)

Abstract:- Background: Multiple antibacterial agents canal, which will end in reinfection and failure of root canal
have been mixed and used as an intracanal medicament treatment [2]. Enterococcus faecalis (EF), which is a
to eliminate E.faecalis, which has been most frequently facultative bacteria, is the most predominant and most
identified in the cases of failed root canal treatment and resistant microorganism leading to persistent periradicular
periapical lesions. This study is aimed at using newer lesions and eventually endodontic failure [3–5]. It is found
intracanal medicament Nitrofurantoin and Ozonated oil in root canal failures in nearly 24–70% by culturing methods
plus calcium hydroxide against E.faecalis bacteria. Aim: [6–8] and in 67–77% by molecular methods [9– 12]. In
To compare the antimicrobial efficacy of newer other studies, it was retrieved as a major component, about
intracanal medicament Nitrofurantoin and Ozonated oil 90% [13, 14].
plus calcium hydroxide on E.faecalis - an in vitro study.
Materials and methodology: Thirty extracted premolars This microorganism has many special properties that
were collected and instrumented with rotary files (size enables it to survive in root canal and cause reinfection such
25-4% taper). Then, root canals were contaminated with as the ability to tolerate periods of starvation, deeply invade
a bacterial solution of E.faecalis for 21 days, and divided dentinal tubules [15], the ability to adapt to changing
into two groups. In the root canals of group A environment and antimicrobial resistance. For these reasons,
nitrofurantoin intracanal medicament was placed and in a wide variety of antibacterial intracanal medicaments have
group B ozonated oil plus calcium hydroxide intracanal been used.
medicament was placed and sealed for 7 days. Then on
the 8th day, the contaminated root canals were flushed Calcium hydroxide is considered the first choice
with normal saline to remove the medicament. To intracanal medicament owing to its bactericidal property,
evaluate the degree of contamination, the specimens of stability and capacity to stop inflammatory exudates
the dentin chips from the full length of the root canal compared to other medicaments especially in multiple-visit
were harvested using a sterile rotary instrument (size 25- endodontics.[16] These favourable actions of calcium
6% taper) and placed in BHI broth and same were hydroxide are attributed to its alkalinity (12.5– 12.8) and
subjected for dilution and colony count after 24 hours. presence of calcium ions. The former denatures and detoxify
Data was recorded and subjected to statistical analysis. the bacterial enzymes, whereas the later is important for
Results and conclusion: within the limits of the study, tissue mineralization by stimulating the fibronectin gene
Nitrofurantoin showed greater antibacterial efficacy on expression.[17,18] Further, the dissociation of Calcium
E.faecalis when compared to the mixture of Ozonated oil hydroxide in to calcium and hydroxyl ions is another rate-
plus calcium hydroxide as an intracanal medicament. limiting step in making Calcium hydroxide as a dentist's
choice of intracanal medicament.[19] In order to achieve
Keywords:- Nitrofurantoin, Ozonated oil, Calcium effective dissociation of ions, appropriate choice of a vehicle
hydroxide, E.faecalis, antibacterial efficacy is warranted. Studies have shown that the velocity of
dissociation of these ions at the periapical tissues and root
I. INTRODUCTION canal is greatly influenced by the vehicle used to prepare the
paste.[20,21]
There are several factors that may cause a persistent
periradicular infection as a consequence of root canal Oily vehicle-containing paste is employed in clinical
treatment like intraradicular infection, extraradicular situations like chronic long standing lesions and
infection, or foreign body reaction and cysts [1]. These inflammatory root resorption that necessitates gradual,
infections are the result of bacterial infection of the root uniform and slow ionic liberation. In order to achieve

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Volume 8, Issue 5, May – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
optimum results through an endodontic treatment, the (EDTA) for 5 minutes in an ultrasonic bath to remove the
antimicrobial capacity of the pastes used is also taken into smear layer and rinsed in water for a further 30 minutes.[30]
consideration. Utilization of medical ozone in various forms Then, the teeth were stored in saline until used. Each root
like ozonated water, ozonated oil or gas is gaining was dried and three layers of clear nail varnish were placed
popularity among the dentists. This is mainly due to its over all external root surfaces with care not to occlude the
efficacy in achieving faster healing and antimicrobial root canal entrance and teeth were allowed to dry. Then, the
properties.[22] Use of ozonated oil has shown its roots were autoclaved at 121ºC for 15 minutes.
antimicrobial efficacy on the bacterial species associated
with periradicular diseases.[23] C. Root canal infection:
Nitrofurantoin (Nit) is a synthetic nitrofuran compound Each root canal was inoculated with a bacterial
[24]. It is effective against most gram-positive and gram- solution up to the canal entrance using a sterile endodontic
negative organisms [25] and is a well-known antibacterial needle and 2.5 ml sterile syringe. Each canal was sealed
agent widely used as an oral antibiotic treatment for urinary with a dental wax and all samples were incubated in a closed
tract infections (UTIs) [26]. It is also the drug of choice for container at 37ºC for a period of 21 days. The canals were
the treatment of infections caused by multidrug resistant reinoculated with fresh bacterial samples every 3 days.
pathogens [27–29].
Thus the objective of this study is to compare the D. Root canal medication:
antimicrobial efficacy of a newer intracanal medicament After 21 days, the canal contents were aspirated and
Nitrofurantoin and a mixture of Ozonated oil plus calcium each canal was rinsed with saline, then dried with sterile
hydroxide against E.faecalis bacteria. paper points and divided in two groups.

II. MATERIALS AND METHOD In Group A- Nitrofurantoin; Group B- mixture of 1 mg

of Calcium Hydroxide with 2 mL of Ozonated oil were
A. Selection of Tooth specimens: placed using sterile syringe.
Single rooted , Type- I root canal configuration, non-
carious permanent mandibular premolar with no resorption The roots were left for 7 days at 37ºC after sealing with
or anomaly were included in this study. paraffin wax. On the 8th day, to investigate the degree of
infection of the radicular dentin, specimens of the dentin
B. Preparation of teeth: chips from the full length of the root canal were harvested
Thirty extracted lower premolar single-rooted teeth using a sterile rotary Nickel titanium instrument (size 25 6%
were collected and stored in saline until required. The teeth taper) and placed in BHI broth. They were then subjected
were decoronated using a rotary diamond saw with water for colony count after 24 hours. Data were recorded and
irrigation. The roots were divided into two groups of 15 subjected to statistical analysis.
roots each. The root lengths were measured and the working
length was determined as 1 mm less than the file length III. RESULT
when the tip was visible at the apical foramen. Coronal root
structure was removed perpendicular to the long axis of the The results of the present study showed that
root to produce roots with a working length of 15 mm. The Nitrofurantoin showed almost none of Colony Forming
canals were instrumented with engine-driven nickel titanium Units (CFU) in their respective group. Whereas mixture of
rotary files (NEOENDO FLEX) used in a sequential crown Calcium hydroxide and Ozonated oil group showed mean
down technique to a standardized apical size of 25.4% taper CFU- 48. Statistical analysis was carried out using Students
instrument. 2.5% sodium hypochlorite was used as an Unpaired ‘t’ test using SPSS windows 12.0 version. The P
irrigant during instrumentation with a sterile needle and 2.5 value 0.000 show that the difference in the colony count
ml sterile syringe. The roots were then rinsed in water for 30 between Group A and B is highly significant.
minutes, then rinsed in 17% ethylenediaminetetraacetic acid

Table 1: Colony forming units recorded in Group A and B

Group A 0 0 1 2 0 0 1 1 0 0 0 0 0 0 1
Group B 58 18 36 51 46 52 59 17 50 56 58 57 58 56 52

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Volume 8, Issue 5, May – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
limiting factors determining the anti-microbial effect of
calcium hydroxide.[35]
 The study conducted by Kesha Vasavada and Sonali
Kapoor, concluded that the release of calcium ions was
higher in the ozonated olive oil group which is
suggestive of higher mineralization activity post
ozonation. [36]
 The study conducted by Mewan Salahalddin A.
Alrahman , 1 Bestoon Muhammed Faraj,1 and Kawa F.
Dizaye concluded that at a concentration of 25 mg/mL,
Fig 1 : Colonies formed in Group A Nitrofurantoin paste is effective in eradicating E.faecalis
completely when it is used as an intracanal
medicament.[33]
 Zhanel et al. [37] showed that nitrofurantoin is active
against all isolates of E.faecalis found in UTI, showing
their susceptibility to Nitrofurantoin. Butt et al. [38]
found that, for a period of three years, nitrofurantoin was
an effective antibacterial in vitro agent and can be used
for the treatment of enterococcus urinary tract infections,
as they showed that one hundred and twenty-seven
(88%) isolates of enterococci were susceptible to
Nitrofurantoin.
 Abdulla and Abdulla [39] demonstrated that
nitrofurantoin was effective against EF (cultured from
Fig 2: Colonies formed in Group B
UTI) in 97.3%, while ciprofloxacin was effective in only
35.7%. Rahbar et al. [40] found that nitrofurantoin had
IV. DISCUSSION
the lowest resistance rate compared to other antibiotics
like ciprofloxacin against EF (cultured from UTI) (97%
 Many antibacterial agents have been mixed and used as vs. 33.38%, respectively). Toner et al. [41] concluded
an intracanal medicament to eliminate E.faecalis, which that EF had a sensitivity test 100% to Nitrofurantoin.
has been most frequently identified in the cases of failed
 Sorlozano-Puerto et al. [42] showed that for four years,
root canal treatment and periapical lesions. This study
EF had a sensitivity to nitrofurantoin ranging from 95%
aimed at comparatively evaluating the use of newer
to 100%.
intracanal medicament Nitrofurantoin (Nit) and
Ozonated oil plus calcium hydroxide against E.faecalis
 Significance of Study:
bacteria.
The uniqueness of this study lies in the fact that it is
 Nitrofurantoin was selected in this study because of its the first of its kind in vitro study performed to evaluate the
broad spectrum of antibacterial activity and is both antimicrobial efficacy between these two newer intracanal
bactericidal and bacteriostatic against microorganisms medicaments with type- I root canal configuration.
[31]. It is the drug of choice against E.faecalis, and has
been used for an extended period in UTI’s and chronic  Limitations of the Study:
and recurrent infections caused by E.faecalis [32]. The limitation in the present study is that we studied
 Nitrofurantoin has a unique mechanism of action. It the antibacterial effects of Nit only against EF, which is the
denatures bacterial ribosomal proteins after being principal constituent of the microorganisms involved in
reduced by bacterial flavoproteins; this phenomenon is persistent endodontic infections. This study includes
repeated with other bacterial macromolecules. This inadequate sample size and limited duration of evaluation to
results in suppression of many essential processes inside prove the long-term success of these medicaments. Further
the bacteria like aerobic energy metabolism, cell wall in vivo and in vitro studies with these medicaments in all
synthesis, DNA synthesis, protein synthesis, and types of root canal configurations with a larger sample size
synthesis of RNA. [33] and long-term evaluation deems necessary to use them as
 Because of this suppression mechanisms, there is a very materials of choice.
poor possibility of developing bacterial resistance to
Nitrofurantoin. Thus, bacterial resistance to V. CONCLUSION
Nitrofurantoin is very rarely seen since its introduction
and FDA approval in 1953 until now. It is very rare to Thus to conclude, within the limits of the study,
encounter cross-resistance with antibiotics or Nitrofurantoin showed greater antibacterial efficacy on
transferable resistance in bacteria [34]. E.faecalis when compared to the mixture of Ozonated oil
 Calcium hydroxide has wide utility in endodontics owing plus calcium hydroxide as an intracanal medicament.
to its anti-microbial and biological activity. Diffusion of
calcium ions and higher pH levels are said to be the rate

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Volume 8, Issue 5, May – 2023 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
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