Lili Luschke Bammann

Antimicrobial evaluation of calcium hydroxide in infected dentinal tubules.

The objective of this study was to evaluate the antimicrobial activity of calcium hydroxide in infected dentinal tubules. Four microorganisms, strains of ATCC (Streptococcus faecalis (ATCC-29212), Staphylococcus aureus (ATCC-6538), Bacillus subtilis (ATCC-6633), and Pseudomonas aeruginosa (ATCC-27853)) and one mixture of these were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37 degrees C for 24 h. Sixty-three human maxillary central incisors were prepared and sterilized by autoclaving. Five groups of 12 teeth each were contaminated for 28 days using new 24-h cultures every 72 h, prepared and adjusted to tube 2 of the MacFarland scale (6 x 10(8) cells/ml). Root canals were then irrigated with 5 ml of saline, dried, and completely filled with calcium hydroxide paste. At intervals of 0, 48, and 72 h, and 7 days, dressings were removed and teeth were immersed in 5 ml of BHI and incubated at 37 degrees C for 48 h to observe the growth and multiplication of the microorganisms. Three uninoculated teeth were maintained in a humid environment as an aseptic control. These teeth were immersed in BHI and maintained at 37 degrees C for 7 days to determine microbial growth. Bacterial growth was shown by turbidity of the culture medium and confirmed by seeding these broths on BHI agar at 37 degrees C for 24 h. The positive BHI tubes were selected, and inoculum was spread on the surface of BHI agar, followed by the same incubation conditions. Gram stain was conducted from BHI growth and from colonies growing on solid medium. Calcium hydroxide in infected dentinal tubules showed no antimicrobial effect on S. faecalis, S. aureus, B. subtilis, P. aeruginosa, or on the bacterial mixture used throughout the experiment.

In vitro determination of direct antimicrobial effect of calcium hydroxide.

The objective of this study was to determine in vitro the time required for calcium hydroxide in direct contact with microorganisms to express its antimicrobial effect. The microorganisms used were: Micrococcus luteus (ATCC-9341), Staphylococcus aureus (ATCC-6538), Fusobacterium nucleatum (ATCC-25586), Pseudomonas aeruginosa (ATCC-27853), Escherichia coli, and Streptococcus sp. The strains were cultivated in Brain Heart Infusion (BHI), with the exception of F. nucleatum (BHI-PRAS). Pure and mixed suspensions of the microorganisms were prepared. Paper cones immersed in these substances were covered with calcium hydroxide paste, and after 0, 1, 2, 6, 12, 24, 48, and 72 h and 7 days they were transferred to an appropriate medium to observe the growth and multiplication of the microorganisms. Incubation was conducted at 37 degrees C for 48 h, according to the requirements of oxygen of each microorganism. The antimicrobial effect of calcium hydroxide was shown to occur after 12 h on M. luteus and F. nucleatum, 24 h on Streptococcus sp, 48 h on E. coli, and 72 h on S. aureus and P. aeruginosa. Mixture II (M. luteus + Streptococcus sp + S. aureus) was sensitive to calcium hydroxide antimicrobial potential after 48 h, whereas mixture I (M. luteus + E. coli + P. aeruginosa), mixture III (E. coli + P. aeruginosa), and mixture IV (S. aureus + P. aeruginosa) were inactivated after 72 h of exposure.

Control of microorganisms in vitro by endodontic irrigants

The aim of this study was to determine the minimum inhibitory concentration (MIC) and antimicrobial effectiveness by the direct exposure test of 4 endodontic irrigants [1% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX), 1% calcium hydroxide (Ca(OH)2; prepared with 1 g of Ca(OH)2 and 100 mL of sterile distilled water), a solution of Ca(OH)2 + detergent (HCT20)] for S. aureus, E. faecalis, P. aeruginosa, B. subtilis, C. albicans and a mixed culture. Microbial growth was analyzed by two methods: turbidity of the culture medium that was confirmed by Gram stain and subculture in a specific nutrient broth. In the dilution test, NaOCl solution showed MIC equal to 0.1% for S. aureus, E. faecalis, P. aeruginosa and C. albicans and equal to 1% for B. subtilis and the mixed culture. CHX (2%) presented MIC equal to 0.000002% for S. aureus, 0.02% for E. faecalis, B. subtilis, C. albicans and the mixed culture and 0.002% for P. aeruginosa. Ca(OH)2 solution (1%) showed MIC greater than 1% for all the microorganisms except P. aeruginosa for which it was equal to 1%. Calcium hydroxide solution + detergent showed MIC equal to 4.5 mL for S. aureus, P. aeruginosa, B. subtilis, C. albicans and the mixed culture and greater than 4.5 mL for E. faecalis. In the direct exposure test, NaOCl had better antimicrobial effectiveness for all microorganisms at all times. CHX (2%) was effective for S. aureus, E. faecalis and C. albicans at all times, and ineffective for P. aeruginosa, B. subtilis and the mixed culture. The other solutions showed the worst results.

Antimicrobial potential of ozone in an ultrasonic cleaning system against Staphylococcus aureus

The aim of this study was to evaluate the antimicrobial potential of ozone applied to 3 different solutions in an ultrasonic cleaning system against Staphylococcus aureus. A total of 120 mL of S. aureus were mixed in 6 L of the experimental solutions (sterile distilled water, vinegar and sterile distilled water + Endozime AWpluz) used in a ultrasonic cleaning system (UCS). Ozone was produced by an electric discharge through a current of oxygen and bubbling with flow rate at 7 g/h ozone (1.2%) into the microbial suspensions. Ten mL of each experimental suspension were collected and 5 fold dilutions were made in 9 mL of BHI and incubated at 37 degrees C for 48 h. Bacterial growth was evaluated by turbidity of the culture medium. At the same time, 1 mL of bacterial samples was collected and inoculated in BHIA plates. After incubation at 37 degrees C for 48 h, the number of colony forming units (cfu) per mL on BHIA surface was counted. In dilution test in BHI tubes and in BHIA plates (cfu/mL), bacterial growth was not observed in any of the experimental solutions when ozone was added. Under the tested conditions, it may be concluded that the addition of ozone to a ultrasonic cleaning system containing different experimental solutions resulted in antibacterial activity against S. aureus.