Microorganisms are crucial in the development of pulpal and periapical diseases. The polymicrobial character of the infection in the root canal system is primarily dominated by anaerobes. ¹ One of the frequently isolated among them is E. faecalis. E. faecalis is one of the most resistant micro-organisms and is most difficult to remove from the periapical areas of the tooth through conventional methods. ² About 23–70% E. faecalis can be isolated from infected root canals with symptoms of chronic apical periodontitis. ³

Among Fungi, Candida species are found in about 3–18% of infected root canals. ⁴

According to earlier research, C. albicans and E. faecalis can infiltrate the dentinal tubules to varying depths, hence removal of these bacteria is facilitated by the use of an efficient antimicrobial agent in the root canal. ⁵

Over the years, several materials were tested in an effort to find an ideal obturating material for a primary tooth, such as zinc oxide eugenol, Maisto paste, Triple Antibiotic Paste etc. ⁶ The antimicrobial efficacy of Triple Antibiotic Paste was evaluated for its use as root canal filling material with different modifications by various authors. ^{7 , 8}

In Triple Antibiotic Paste, the antibiotics used are: Ciprofloxacin, metronidazole, and minocycline in 1:1:1 concentration. ⁹

Ciprofloxacin is a second-generation fluoroquinolone antibiotics active against a broad range of bacteria, especially Enterobacteriaceae and Neisseria. ¹⁰ Metronidazole is a nitro imidazole compound having a broad spectrum of anti-anaerobic and anti-protozoal activity. ⁹ Minocycline is a member of the Tetracycline class of broad-spectrum antibiotics, which work by exerting bacteriostatic action on a wide variety of microorganisms. ^{8 , 9}

But, Minocycline causes discoloration of tooth as stated by various authors. ^{11 , 12} For the same reason, Sara Alsubait et al (2020) in their study had replaced minocycline with Cefaclor. Cefaclor is a second generation cephalosporin antibiotic which is mostly effective against most strains of Gram positive aerobes as well as gram negative aerobes. ¹⁰

A broad-spectrum antibiotic called Clindamycin is effective against infections that produce beta-lactamases as well as against aerobic and anaerobic microorganisms. ¹⁰ Prasanna et al (2020) in their study concluded that Clindamycin could inhibit the levels of C albicans by 90% but Metronidazole could not inhibit the C. albicans. ⁸

With respect to above literature, considering the disadvantage associated with Minocycline and reduced antimicrobial efficacy of Metronidazole, the present study has planned to use the 3C paste consisting of Ciprofloxacin, Clindamycin, and Cefaclor as a root canal filling material for primary teeth.

The aim of the present study will be to evaluate and compare the antimicrobial efficacy of 3C paste with conventional Triple Antibiotic Paste against E. faecalis and C. albicans.

The objectives of the study will be to evaluate the antimicrobial efficacy of 3C paste and triple antibiotic paste against E. faecalis and C. albicans; and also to compare the antimicrobial efficacy of 3C paste with Triple Antibiotic Paste against E. Faecalis and C. albicans.

This in vitro study will be conducted at Department of Pediatric and Preventive Dentistry, Sharad Pawar Dental College and Hospital, Datta Meghe Institute of Higher Education and Research, Sawangi Meghe, Wardha in collaboration with the Institution of Pharmaceutical Education and Research, Borgaon (Meghe), Wardha. The present study will be an in vitro experimental microbiological study.

Although no human or animal participants will be involved in this research, as per institutional guidelines, ethical approval for the study was obtained from the institutional ethics committee of Datta Meghe Institute of Higher Education and Research, Wardha (Ref. No. DMIHER (DU)/IEC/2023/564 approved on 02/02/2023).

For the antimicrobial analysis brain heart infusion agar (BHI), Sabouraud’s dextrose agar (SD), Agar plates, Petri dishes , E. faecalis strain , C.albicans strain will be used. For preparation of paste, Ciprofloxacin, Clindamycin, Cefaclor, Metronidazole, Minocycline, Propylene glycol/macrogol will be required. These steps are outlined in full in the sections below.

E. faecalis strain and C.albicans strain and the powder form of antibiotics will be included for the study. However other bacterial strains will be excluded from the study.

Preparation of Triple Antibiotic Paste (control group)

Triple Antibiotic Paste will be prepared by mixing equal proportion of Ciprofloxacin (200 mg), Metronidazole (400 mg), and Minocycline (100 mg) in 1:1:1. Then a part of the solvent (propylene glycol-macrogol) in 1:1 ratio will be dispensed to obtain soft, uniform consistency. ¹³ The resultant paste will be stored in an airtight container.

Preparation of 3C antibiotic paste (experimental group)

3C paste will be prepared in the same manner as Triple Antibiotic Paste. In 3C paste, the antibiotics Ciprofloxacin (200 mg), Cefaclor (100 mg), and Clindamycin (400 mg) (1:1:1) will be mixed with one part of Propylene Glycol-one part of Macrogol in 5:1 (powder: liquid) ratio to get desired consistency as required for the paste.

Procedure

Pure colonies of C. albicans ( ATCC 2091) and E. faecalis ( ATCC 35550) will be grown on Sabouraud’s dextrose (SD) agar and Brain heart infusion (BHI) agar respectively and will be suspended in 5 ml of SD broth and BHI broth for 24 hours at 37°C.

The actively growing microorganisms in broth cultures will have turbidity adjusted to the 0.5 McFarland standard. ^{3 , 5}

The agar well diffusion method will then be followed. Wells of 3-4 mm will be prepared in the respective agar and then all the wells will be loaded with test material. The agar plates will be incubated at 37°C for 24 and 72 hours to achieve good bacterial growth before analysis. The inhibition zones of microbes will be evaluated using digital calliper in millimetre after every 24 and 72 hours. All the procedures will be repeated in triplicates to minimize errors.

Primary outcomes to be measured

The antimicrobial efficacy of newly formulated 3C antibiotic paste will be measured by bacterial growth and will be compared with the Triple Antibiotic Paste.

Data analysis and statistical plan

Statistical analysis will be done using software SPSS Version 23.0. P value will be considered significant for <0.05. Descriptive analysis, i.e., mean distribution of inhibition zones will be recorded. A repeated measures ANOVA for intragroup comparison will be done on application of triplicate test of every group. Independent sample t test will be done for intergroup comparison.

Expected outcome

It is expected that a newly formulated 3C paste will be able to show antimicrobial efficacy and will prove to be the successful as root canal filling material for primary teeth.

Dissemination

This study will be published in an indexed journal.

Not yet started.