1. Introduction

Root canal therapy is one of the endodontic branches that deals with the diseases related to the pulp of tooth and its surrounding tissues. Treatments of the infected root canals lead to preservation of natural teeth as a result of function and appearance of the patient’s masticating system ^[1], and aims to eliminate bacteria from infected root canal system and to prevent spreading of infections to the head and neck region ^[2].

Maintenance of bacteria within the root canal can be considered the principal etiologic factor for the development of pulp and periapical lesions ^[3]. Different types of microorganisms such as bacteria, fungi, and probably viruses can infect dental pulp and lead to periodontitis. Additionally, spread of infection to the surrounding area of the root significantly reduces the treatment success. Therefore, microorganisms have a major role in the treatment failure of the teeth roots ^[4].Thus, success rate of endodontic therapy depends on the probable presence of bacteria. In vital teeth where the number of bacteria in the canal is low, success rate is high. In teeth with lesions around the roots and necrotic teeth, success rate decreases due to the high presence of microorganisms inside the canal ^[5].

After biomechanical preparation, resistant microorganisms may remain inside the root canals, or even penetrate in the root canal system during or after the endodontic treatment. These microorganisms and their byproducts are able to promote persistent or secondary infections, leading to treatment failure. In these situations, micro-organisms in the root canal is composed of fewer species than primary infections and Gram-positive bacteria predominate ^[6]. Furthermore, Gram-positive facultative anaerobes bacteria, seem to be remarkably resistant to local antimicrobial agents that are often used in root canal therapy ^[2].

E.faecalis has high prevalence in root-filled teeth associated with periapical lesions. As well as it possesses several virulence factors that contribute to its ability of surviving the effects of conventional root canal therapy ^[7]. Prevalence of this microorganism is 22% to 77% of cases ^[8]. Because of its ability to sustain PH=11/5 and resist calcium hydroxide which is disposed in the canals between treatment sessions. E.faecalis can stay alive while starving for a long time and grow alone without support from coexistence with other bacteria in treated canals ^[9]. As well as, this bacteria is able to invade dentinal tubules and bind to collagen ^[10]. Thus, dentinal tubules infection by bacteria can act as a source for endodontic recurrent infection. Therefore, disinfection of infected canals is more complicated ^[11].

Chemomechanical preparation can be conventionally accepted for effective root canal disinfection ^[12]. Thus, routine irrigation may remove the majority of the bacteria within the canal but may not be very effective to bacteria located deeply within the dentinal tubules ^[13]. Intracanal medication used to supplement this procedure ^[14]. However, the effectiveness of these conventional strategies against microorganisms remains under debate ^[15]. Thus there is the need for new substances and equipment like lasers ^[16].

Lasers have been suggested to assist endodontic treatments to achieve deep root canal disinfection ^[17]. It can eliminate microorganisms existing in main canal, lateral canals and dentinal tubules which may cause pulp and periapical infection ^[16]. Erbium, Chromium: Yttrium-Scandium-Gallium garnet (Er,Cr:YSGG) laser at wavelength 2,780nm, acts through photoablation since this wavelength correlates with the absorption maximum of hydroxyapatite. When irradiated, water contained in the dental hard tissue evaporates instantaneously and ablates the surrounding tissue with only minimal thermal side effects ^[18]. Although this laser used primarily for the preparation of dental hard substances, it also can be used in treatment of root canals of teeth ^[19], and disinfection of root canal system without being hazardous to surrounding structures ^[20]. Recently, in order to improve light distribution inside root canals, modified radial firing tips were developed ^[13] allowing irradiation of narrow or curved root canals ^[21].

Finally, success of root canal treatment depends on complete sterilization of root canals. This objective becomes more difficult to achieve as root canal system is polymicrobial in nature. Among the various microorganisms which are responsible for the failure of the root canal treatment, E.faecalis holds an important position and is also used as a biological marker ^[22].

2. Materials and Methods

Forty single-rooted, human teeth were selected and cleaned with curettes to remove calculus and rests of periodontal tissue. The teeth were cut at the cementoenamel junction with a diamond sectioning disc (Germany). Working length was determined by passing a size 15 K-file (Japan) in the canal until the apex was visible. The root canals were enlarged 1 mm short of the root apex (14mm) up to a size 50 K-file under irrigation with physiological saline. The apical foramen was sealed by composite resin (USA) and the root surface covered with bonding agent to prevent bacterial leakage (Figure 1). Roots were then embedded in silicone impression material, covered with aluminum foil, fixed in stainless steel boxes and sterilized by autoclaving (121ºC, 15 min).

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw figureFigures index

•  NEW
  View larger figure in new window
• NEXT
  View next figure

Figure 1. Prepared roots

Overnight broth culture (grown at 37ºC) of E.faecalis in brain-heart infusion (BHI) broth was prepared (4×107cfu/ml) ^[23] and used to infect the prepared root canals.

The sterilized tooth specimens were inoculated with 10μL of an overnight BHI broth culture of E.faecalis by using a micropipette with a sterile tip. Specimens were kept at 37ºC for 42 hour to allow bacterial growth. Three root canals not infected and not treated were control negative. After incubation, the samples were divided into three groups, consisting of 10 specimens each.

The specimens were irradiated with an Er,Cr:YSGG laser, λ=2.78μm (Figure 2), with radially emitting firing laser tip type RFT3 inside the canals (Figure 3) that mounted on a 90°-angled handpiece. The laser was adjusted for an average output powers 0.75W, 1W and 1.5 W, 20Hz, 10% air flow without water. For each sample, before each irradiation, the tip was disinfected with 70% alcohol. Each sample was treated with one lasing cycle, which comprised five irradiations of 5s duration with a 20s break in between ^[24]. The tip was placed at the working length (14mm in length). Then, the laser was activated, and the root canal was continuously radiated from apical to coronal, in slow, circling movements. By means of this procedure the irradiation of the entire root canal could be ensured.

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw figureFigures index

•  NEW
  View larger figure in new window
• PREV
  View previous figure
• NEXT
  View next figure

Figure 2. Er,Cr:YSGG Laser

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw figureFigures index

•  NEW
  View larger figure in new window
• PREV
  View previous figure
• NEXT
  View next figure

Figure 3. Laser tip RFT3 mounted on a 90°- angled handpiece

The microbiological procedures were performed under a septic conditions. Immediately after treatment, the sample was taken using sterilized no.1 Gates-Glidden bur to allow dentinal filings ^[13], then 10mg of dentine chips transferred into vial contain1ml normal saline. Followed by spreading of 200-μL of this solution on Enteroccocus agar plates, incubated overnight at 37°C, then counted the resulting colonies. After that, the root canal openings were covered with wax and the specimens were incubated at 37°C for 7 days. Throughout this period, sterile BHI broth was added to the canals every 2 days. Seven teeth did not receive bacterial suspension, only sterile BHI broth (negative control group-NC). Seven days after inoculation, microbiological sample was performed using sterilized no. 1 Gates-Glidden bur, then transferred the sample to test tubes containing 1 ml of sterile saline solution as mentioned above followed by counting the number of bacterial colony.

3. Results

Bacteriological test showed that absence of growth was seen in the negative controls. All specimens of the three groups showed bacterial growth prior to the experiment disinfection procedures.

Statistical analysis showed that the antibacterial effect of laser at different powers and selected time was highly significant compared with the untreated group using ANOVA at level (p<0.001), as shown in (Table 1) and (Figure 4, Figure 5).

From the cfu counts of bacteria immediately after irradiation for all specimens, the reduction value was observed at 0.75W, 1W, 1.5W of output powers that were significantly different from the number of bacterial colonies before root canals irradiation with laser. Additionally, the result showed that significant antibacterial difference between the output powers that used in this study as the output power increase the antibacterial effect of laser against E.faecalis also increase. The result improved complete eradication of E.faecalis at irradiation power 1.5W when the sample was taken immediately after irradiation.

When comparison was made between the first(immediately) and the second (after 7 days) microbiological sampling, using ANOVA at level (p<0.001), the results revealed that at all selected output powers there was increased in the number of bacterial colonies after 7 days of laser application in the root canals.

Further comparison from the second microbiological sampling among different output powers, the result indicated that there were significant differences between 0.75W, 1W and 1.5W of output powers in the number of colonies. As well as non significant differences was obtained between the number of E.faecalis counts at 0.75W immediately after irradiation of root canal and 1W after 7 days of root canal irradiation. The results are showed in (Table 1) and (Figure 4).

Table 1. One way analysis of variance for the comparison among different output powers of laser and comparison between the recovery of E.faecalis counts immediately and after seven days of root canal irradiation

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw tableTables index

•  NEW
  View table in a new window

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw figureFigures index

•  NEW
  View larger figure in new window
• PREV
  View previous figure
• NEXT
  View next figure

Figure 4. Bar chart showing the antibacterial effect of laser on the E.faecalis immediately and after 7 days of irradiation

Download as

• PPT
  PowerPoint Slide
• PNG
  Larger image(png format)

Veiw figureFigures index

•  NEW
  View larger figure in new window
• PREV
  View previous figure

Figure 5. Number of bacterial colonies grown on Enterococcus agar (A) before and (B) after laser irradiation

4. Discussion

Successful endodontology relies, to a great extent, on complete cleaning of the root canal because infected dentine and pulpal tissue can endanger therapy outcome ^[24]. Bacterial colonization on pulpal disease and endodontic treatment failures is a well-known fact ^[25]. These bacteria constitute an essential source for the potential reinfection of an endodontically treated root canal ^[26]. Consequently the complete removal of the pathogenic bacteria and their toxic byproducts is of crucial importance for the therapy outcome ^[27].

E.faecalis is a Gram-positive microorganism commonly detected in asymptomatic, persistent endodontic infections. Its ability to compete with other microorganisms, invade dentinal tubules and resist nutritional deprivation ^[7]. It was chosen as the microbiological marker because it has the ability to colonize the root canal in biofilms, representing the in vivo growth condition ^[28]. In addition, it often survives chemomechanical preparation ^[29] because of its resistance to antimicrobial agents and its ability to cause a monoinfection in the root canals ^[30]. Moreover, it is resistant to temperature and physical stress ^[31]. The presence of this microorganism has been related to persistent apical periodontitis ^[32] and also with failure of endodontical treatments ^[33], and therefore, is qualified for this investigation.

Conventional root canal treatment aims at the removal of the infected pulp and dentin layers by using mechanical techniques and bactericidal irrigants. However, these cleansing techniques are only successful to a certain extent ^[27].

Many complications during root canal irrigation, including inadvertent injection of sodium hypochlorite or hydrogen peroxide into the periapical tissues, air emphysema, and allergic reactions to the solutions ^[34]. Additionally, the discrepancy of the penetration depth of micro-organisms and bactericidal rinsing solutions often holds responsible for therapy resistant cases and long-term failures which can be observed in conventional endodontics ^[27]. Many studies have demonstrated that prognosis of endodontic treatment in the teeth with periapical lesions are the same as teeth without any lesions if the root canal therapy carried out to an optimal level ^[35].

Endodontic treatment is costly procedure and for that reason many studies were done to discover new materials and/or methods for the most important step during root canal therapy which was responsible for reinfection of endodonticaly treated teeth. In this study, we evaluated the ability of an Er,Cr:YSGG laser with radially emitting laser tips to eliminate E.faecalis from the infected specimens.

Through the introduction of a new radial-firing fiber tip, the mode of light emission in the root canal has been improved. Owing to the conical shape of the fiber tip, the laser light is emitted in the form of a broad cone with an angle of about 60°, allowing a more uniform coverage of the whole dentinal surface ^[24].

The results in this experiment showed reduction in the number of E.faecalis counts at all out put powers when the samples were taken immediately after the Er,Cr:YSGG laser irradiation. This is comparable with others Schoop etal., 2004 ^[27], Gordon etal., 2007 ^[13], Eldeniz etal., ^[36] and René Franzen, 2009 ^[37].

The bactericidal effect of laser radiation with a wavelength of 2.78μm is based on the high absorption constant for OH⁻ groups and H₂O. The laser destroys E.faecalis when energy is absorbed into the volume of the bacterium ^[37]. Another explanation for the Er,Cr:YSGG laser’s positive effect on E.faecalis could be some degree of conduction of the laser light within the dentinal tubules, resulting in a penetration of laser light inside it. Other factors, such as shock waves or cavitations effects that have been reported ^[38].

A different results was achieved Jha and colleagues ^[39] who concluded that the Er,Cr:YSGG laser was not able to eliminate an E.faecalis infection in root canals and that the laser was completely ineffective in disinfecting root canals. The difference in the results may be attributed to differences in the methodology used in the studies.

The result of this study showed significant antibacterial differences between the output powers which is in accordance with Schoop et al., 2004 ^[27] who showed that laser at a wave length 2.78μm at a power of 1W, generated minor changes in the bacterial count. Increasing the power to 1.5W showed slightly improved bacterial reduction. Similar finding obtaining by Ulrich Schoop, 2007 ^[21] showed and concluded that Er,Cr:YSGG laser was effective in eliminating the gram-positive E.faecalis, at output power of 1 W. When the laser at an output power setting of 1.5W, the result showed no significant difference when compared with the 1 W group. Additionally, study by Wang et al., 2007 ^[40] showed an experiment in straight root canals that were inoculated with E faecalis for 3 weeks found that the Er,Cr:YSGG laser irradiation resulted in a reduction in bacteria of 77% after irradiation at 1W, and a reduction of 96% after irradiation at 1.5W, but there was no significant difference. Moreover, microbiological reduction of E.faecalis was also discussed by René Franzen, 2009 ^[37] and showed reduction of E.faecalis with a minimal laser output power of 0.25 W.

The result of this experiment showed that the antibacterial effect of Er,Cr:YSGG laser is dependent on the output power as the output power increase the effect against E.faecalis also increase. Different studies showed similar results ^{[13, 27]}.

The basic research study showed statistically significant bactericidal effect on E.faecalis with 2.780nm radiation when the sample was taken immediately after laser application in the root canals, where as in the second microbiological sampling after seven days of irradiation the results showed increased in the number of E.faecalis counts at 0.75W, 1W and 1.5W of output powers in one lasing cycle at the selected time.

One possible explanation for the increased number of bacterial colony counts after seven days of irradiation is the ability of the remaining viable E.faecalis that was not exposed to the irradiation to grow rapidly. This could be due to availability of sufficient nutrition in the root canal. Although the group that is exposed to 1.5W showed negative result in the first microbiological sampling but after 7 days the result showed the presence of bacteria in the same group this could be due to the presence of viable bacteria deeply inside the dentinal tubules not subjected to irradiation, this may be due to the time used in this experiment was not enough to kill all the bacteria, therefore, need more than one lasing cycle.

The present findings in accordance with the study by Castelo-Baz etal., 2012 ^[41] who showed that after 7 days, a high microbial growing in laser group. An explanation can be provided by the fact that laser light cannot reach all areas. Furthermore, survival of E.faecalis can be attributed to the high resistance of E.faecalis to heat, because of its cell-wall structure ^[27].

According to the results of current study all the selected output powers reduce the number of bacterial colony of E.faecalis and the most effective power was 1.5W that a negative result in the first microbiological sampling but due to positive result in the second microbiological more time of laser application is required in order to kill all the bacteria that is penetrated deeply in the dentinal tubules and prevent reinfection in the root canals.

The penetration of the laser into the root dentin is governed by several factors. At the wavelength of the Er,Cr:YSGG laser, there is absorption by dentin owing to the presence of hydroxide and interstitial water. On the basis of the fact that each laser pulse is composed of approximately 150 micropulses and each micropulse is responsible for the penetration of this energy of about 3μm into water, depending on fluence, it is possible to achieve expansion of intratubular water and the collapse of water vapor as deep as 1,000μm or more. This effect, known as “micropulse-induced sequential absorption,” with expansion and collapse of water vapor, is capable of producing acoustic waves strong enough to disrupt intratubular bacteria. This penetration of dentin may provide the laser with advantages versus conventional methods of dentin disinfection that has limit antimicrobial activity ^[13].

E.faecalis has the ability to maintain viability in obturated root canals ^[42] and because of the fact that, none of the filling materials were able to prevent the regrowth of E.faecalis completely ^[43]. For that reason, more work needs to be done; to achieve the goal of complete elimination of infection from the root canal of teeth and the clinical applications of this strategy are under evaluation in my Department.

5. Conclusions

1. The study concluded that the wavelength tested study is a suitable for eliminating bacteria from the infected root canals.

2. Ability of remaining viable E.faecalis to grow rapidly in the root canal after 7 days of irradiation with laser.

3. Time used in this experiment was not enough to eliminate E.faecalis completely from the infected root canal. Therefore, more time is required during laser irradiation. For the results to be confirmed further, another clinical studies are necessary.

References

[1]	Trope M. The vital tooth: Its importance in the study and practice of endodontics. Endodon Topics. 5(1): 1. 2003.
	In article		View Article
[2]	Card S. J., Sigurdsson A., Orstavik D., and Trope M. “The Effectiveness of Increased Apical Enlargement in Reducing Intracanal Bacteria,” J. Endod. 28, pp. 77r9-783. 2002.
	In article
[3]	Siqueira JF, Rocas IN, Alves FR, Silva MG. Bacteria in the apical root canal of teeth with primary apical periodontitis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 107(5): 721-726. 2009.
	In article		View Article
[4]	Ruddle C. Cleaning and shaping the root canal system. In Cohen S, Burns RC, editors. Pathways of the Pulp,ed 8, St Louis, Mosby. 2002.
	In article
[5]	Friedman S. Treatment outcome and prognosis of endodontic therapy. In Orstavik D, Pitt-Ford TR, editors: Essential Endodontology. Prevention and Treatment of Apical Periodontitis, London, Blackwell Science Ltd, p 367. 1998.
	In article
[6]	Pinheiro ET, Gomes BPFA, Ferraz CCR, Sousa ELR, Teixeira FB, Souza-Filho JF. Microorganisms from canals of root-filled teeth with periapical lesions. Int Endod J. 36: 1-11. 2003.
	In article		View Article
[7]	Stuart CH, Schwartz SA, Beeson TJ, Owatz CB. Enterococcus faecalis: Its role in root canal treatment failure and current concepts in retreatment. J Endod. 32: 93-98. 2006.
	In article		View Article
[8]	Fouad AF, Zerella J, Barry J, Spangberg LS. Molecular detection of Enterococcus species in root canals of therapy-resistant endodontic infections. Oral Surg OralMed Oral Pathol Oral Radiol Endod. 99: 112-8. 2005.
	In article		View Article
[9]	Figdor D, Davies JK, Sundqvist G. Starvation survival, growth and recovery of Enterococcus faecalis in human serum. Oral Microbiol Immunol. 18: 234-9. 2003.
	In article		View Article
[10]	Love LM. Enterococcus faecalis - a mechanism for its role in endodontic failure. Int Endod J. 34:399-405. 2001.
	In article		View Article
[11]	Love RM, McMillan MD, Park Y, Jenkinson HF. Coinvasion of dentinal tubules by Porphyromonas gingivalis and Streptococcus gordonii depends upon binding specificity of streptococcal antigen I/II adhesion. Infect Immun. 68: 1359-65. 2000.
	In article		View Article
[12]	Carson KR, Goodell GG, McClanahan SB. Comparison of the antimicrobial activity of six irrigants on primary endodontic pathogens. J Endod 31(6): 471-473. 2005.
	In article		View Article
[13]	Gordon, W., Atabakhsh, V. A., Meza, F., Doms, A., Nissan, R., Rizoiu, I., and Stevens, R. H. “The Antimicrobial Efficacy of the Erbium, Chromium: Yttrium-Scandium-Gallium-Garnet Laser with Radial Emitting Tips on Root Canal Dentin Walls Infected with Enterococcus Faecalis,” J. Am. Dent. Assoc. 138(7), 992-1002. 2007.
	In article		View Article
[14]	Siqueira JF, Magalhaes KM, Rocas IN. Bacterial reduction in infected root canals treated with 2.5% NaOCl as an irrigant and calcium hydroxide/camphorated paramonochlorophenol paste as an intracanal dressing. J Endod. 33(6): 667-672. 2007.
	In article		View Article
[15]	Weiger R, Rosendahl R, Lost C. Influence of calcium hydroxide intracanal dressings on the prognosis of teeth with endodontically induced periapical lesions. Int Endod J. 33(3): 219-226. 2000.
	In article		View Article
[16]	Mohammad Asnaashari, Nassimeh Safavi. Disinfection of Contaminated Canals by Different Laser Wavelengths, while Performing Root Canal Therapy. J Lasers Med Sci. 4(1): 8-16. 2013.
	In article
[17]	Gutknecht N, van Gogswaardt D, Conrads G, Apel C, Schubert C, Lampert F. Diode laser radiation and its bactericidal effect in root canal wall dentin. J Clin Laser Med Surg. 18(2): 57-60. 2000.
	In article
[18]	Hibst R, Keller U. Experimental studies of the application of the Er:YAG laser on dental hard substances, part I: measurement of the ablation rate. Lasers Surg Med. 9(4): 338-44. 1989.
	In article		View Article
[19]	De Moor RJ, Blanken J, Meire M, Verdaasdonk R. Laser induced explosive vapor and cavitation resulting in effective irrigation of the root canal. Part 2: evaluation of the efficacy. Lasers Surg Med. 41(7): 520-523. 2009.
	In article		View Article
[20]	Arnabat J, Escribano C, Fenosa A, Vinuesa T, Gay-Escoda C, Berini L etal. Bactericidal activity of erbium, chromium: yttrium-scandium-gallium-garnet laser in root canals. Lasers Med Sci. 25(6): 805-810. 2010.
	In article		View Article
[21]	Ulrich Schoop, Kawe Goharkhay, Johannes Klimscha, Manuela Zagler, Johann Wernisch, Apostolos Georgopoulos, Wolfgang Sperr, Andreas Moritz. The use of the erbium, chromium: yttrium scandium-gallium-garnet lasers in endodontic treatment The results of an in vitro study. JADA. 138(7): 949-55. 2007.
	In article		View Article
[22]	C. M. Sedgley, S. L. Lennan, O. K. Appelbe: Survival of Enterococcus faecalis in root canals ex vivo. International Endodontic Journal. 38: 735-42. 2005.
	In article		View Article
[23]	Garcez AS, Núñez SC, Lage-Marques JL, Jorge AOC, and Ribeiro MS: Efficiency of NaOCl and laser-assisted photosensitization on the reduction of Enterococcus faecalis in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 102(4): 93-98. 2006.
	In article		View Article
[24]	U. Schoop, A. Barylyak, K. Goharkhay, F. Beer, J. Wernisch, A. Georgopoulos, W. Sperr, A. Moritz: The impact of an erbium, chromium: yttrium-scandium-gallium-garnet laser with radial-firing tips on endodontic treatment. Lasers Med Sci. 2007.
	In article
[25]	Sundqvist G. Taxonomy, ecology and pathogenicity of the root canal flora. Oral Surg Oral med Oral Pathol. 78: 522-530. 1994.
	In article		View Article
[26]	Nair PN, Sjogren U, Krey G, Kahnberg KE, Sundqvist G. Intraradicular bacteria and fungi in root-filled, asymptomatic human teeth with therapy-resistant periapical lesions: A long-term light and electron microscopic follow-up study. J Endod. 16(12): 580-588. 1990.
	In article		View Article
[27]	Schoop U, Kluger W, Moritz A, Nedjelik N, Georgopoulos A, Sperr W. Bactericidal effect of different laser systems in the deep layers of dentin. Lasers Surg Med. 35: 111-116. 2004.
	In article		View Article
[28]	Love RM. Enterococcus faecalis-a mechanism for its role in endodontic failure. International Endodontic Journal. 34, 399-405. 2001.
	In article		View Article
[29]	Gomes BP, Pinheiro ET, Sousa EL, et al. Enterococcus faecalis in dental root canals detected by culture and bypolymerase chain reactin analysis. Oral Pathology Oral Radiology and Endodontics. 102, 247-53. 2006.
	In article		View Article
[30]	Nakajo K, Komori R, Ishikawa S, et al. Resistance to acidic and alkaline environments int he endodontic pathogen Enterococcus faecalis. Oral Microbiology and Immunology. 21, 283-8. 2006.
	In article		View Article
[31]	Fabricius L, Dahlen G, Sundqvist G, Happonen RP, Moller AJ. Influence of residual bacteria on periapical tissue healing after chemo-mechanical treatment and root filling of experimentally infected monkey teeth. Eur J Oral Sci. 114: 278-285. 2006.
	In article		View Article
[32]	Haapasalo M, Orstavik D. In vitro infection and disinfection of dentinal tubules. J Dent Res. 66: 1375-1379. 1987.
	In article		View Article
[33]	Love RM. Enterococcus faecalis—a mechanism for its role in endodontic failure. Int Endod J. 34: 399-405. 2001.
	In article		View Article
[34]	Brown DC, Moore BK, Brown CE Jr, Newton CW. An in vitro study of apical extrusion of sodium hypochlorite during endodontic canal preparation. J Endod. 21: 587-591. 1995.
	In article		View Article
[35]	Barbakow FH, Cleaton- Jones PE, Friedman D. Endodontic treatment of teeth with periapical radiolucent areas in a general dental practice. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 51: 552-59. 1981.
	In article		View Article
[36]	Eldeniz AU, Ozer F, Hadimli HH, Erganis O. Bactericidal efficacy of Er,Cr: YSGG laser irradiation against Enterococcus faecalis compared with NaOCI irrigation: an ex vivo pilot study. Int Endod J. 40(2): 112-9.
	In article		View Article
[37]	René Franzen, Marcella Esteves-Oliveira, Jörg Meister, Anja Wallerang, Leon Vanweersch, Friedrich Lampert & Norbert Gutknecht. Decontamination of deep dentin by means of erbium, chromium:yttrium-scandium-gallium-garnet laser irradiation. Lasers Med Sci. 24: 75-80. 2009.
	In article		View Article
[38]	Lizarelli Rde F, Bagnato VS. Micro-morphological analisys of enamel and dentin exposed to picosecond-pulsed Nd: YAG laser [in Portuguese]. Pesqui Odontol Bras. 16(3): 227-33. 2002.
	In article
[39]	Jha D, Guerrero A, Ngo T, Helfer A, Hasselgren G. Inability of laser and rotary instrumentation to eliminate root canal infection. JADA. 137(1): 67-70. 2006.
	In article		View Article
[40]	Wang QQ, Zhang CF, Yin, XZ. Evaluation of the bactericidal effect of Er,Cr:YSGG and Nd:YAG lasers in experimentally infected root canals. J Endod. 33: 830-832. 2007.
	In article		View Article
[41]	Castelo-Baz P, Martín-Biedma B, Ruíz-Piñón M, Rivas-Mundiña B, Bahillo J, Perez-Estévez A, Gude F, De Moor R, Varela-Patiño P. Combined Sodium Hypochlorite and 940nm Diode Laser Treatment Against Mature E.faecalis Biofilms in vitro. J Lasers Med Sci. 3(3): 116-21. 2012.
	In article
[42]	Sedgley CM. The influence of root canal sealer on extended intracanal survival of Enterococcus Faecalis with and without gelatinase production ability in obturated root canals. J Endod. 33: 561-6. 2007.
	In article		View Article
[43]	Su-Jung Shin, Sang-Wook Jee, Jin-Seon Song, Il-Young Jung, Jeong-Heon Cha, Euiseong Kim: Comparison of regrowth of Enterococcus faecalis in dentinal tubules after sealing with gutta-percha or Resilon. Journal of Endodontics (Impact Factor: 2.93). 34(4): 445-8. 2008.
	In article