ES2420231A1 - Composition for endodontic sealing with antibacterial effect (Machine-translation by Google Translate, not legally binding) - Google Patents

Abstract

Composition for endodontic sealing with antibacterial effect. The present invention describes a composition for endodontic sealing with antibacterial activity and with the capacity to prevent the formation of bacterial films or biofilms after the sealing process in endodontic treatments. It comprises an endodontic sealant to which one or more cationic antiseptics are incorporated in such a way that, without affecting the physical-chemical properties of the sealant, they confer, under physiological conditions, properties that inhibit the formation of biofilms. On the other hand, the addition of these antiseptics to the sealant gives it bactericidal activity. These antimicrobial characteristics, particularly against E. faecalis, are maintained over time.

Description

COMPOSITION FOR ENDODONTIC SEALING WITH ANTIBACTERIAL EFFECT

TECHNICAL SECTOR

The present invention is framed in the field of dental products. More specifically, it is a sealant cement composition with antibacterial effect of application in endodontic treatments.

STATE OF THE TECHNIQUE

The most important objective of endodontic treatment is the elimination of microorganisms from the root canal system and the prevention of reinfection. However, in most cases, complete elimination of bacteria is impossible. [Torabinejad M, Handysides R, Khademi AA, Bakland LK. Clinical implications of the smear layer in endodontics: a review. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2002; 94: 658-66].

The use of root canal filling materials with antibacterial activity can reduce the number of remaining microorganisms [Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod 2009; 35: 1051-5], [Baer J, Maki JS. In vitro evaluation of the antimicrobial effect of three endodontic sealers mixed with amoxicillin. J Endod 2010; 36: 1170-3.], Prevent recurrent infection of the ducts and promote healing of apical and periapical tissues [Leonardo MR, da Silva LA, Tanomaru Filho M Bonifacio KC, Ito IY. In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics.J Endod 2000; 26: 391-4.].

The Enterococcus genus encompasses a group of species morphologically similar to streptococci. They cause very diverse infections and have a growing interest in the case of opportunistic processes. They are associated in pairs and short chains and, although their natural habitat is the intestine, it has been isolated as a normal microbiota in the buccal mucosa and back of the tongue and in periodontal pockets.

Enterococcus faecalis, one of its most common species, is frequently found in teeth with root canal treatment, both in asymptomatic cases and in which such treatment has failed, as well as cases of persistent apical periodontitis [Stuart CH, Schwartz SA, Beeson TJ , Owatz CB. Enterococcus faecalis: its role in root canal treatment failure and current concepts in retreatment. J Endod 2006; 32: 93-8]. This bacterium penetrates the dentinal tubules and grows as biofilm in the walls of the root canals even under conditions of nutrient deficiency, which makes their elimination especially difficult [George S, Kishen A, Song KP. The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis. J Endod 2005; 31: 86772].

Endodontic sealant cements have some inherent antibacterial capacity depending on their chemical composition. There is a wide variety of sealers on the market. Most of them are based on zinc oxide eugenol, plastic resins, calcium hydroxide and silicones. AH Plus is an epoxy resin based endodontic sealant that has good physical properties, adequate dentin adhesion, excellent fluidity and stability in aqueous solution. In addition, it is biocompatible [Neff T, Layman D, Jeansonne BG: In vitro cytotoxicity evaluation of endodontic sealers exposed to heat before assay. J Endod 2002, 28:12.], Is not genotoxic [Leyhausen G, Heil J, Reifferscheid G, Waldmann P, Geurtsen W. Genotoxicity and cytotoxicity of the

epoxy resin-based root channel sealer AH Plus. J Endod 1999; 25: 109-13.], Has good

tissue tolerance, sealing capacity and long-term dimensional stability [Leonardo MR, da Silva LA, Almeida WA, Utrilla LS. Tissue response to an epoxy resin

-

based root canal sealer. Endod Dent Traumatol. 1999; 15: 28]. Its antibacterial capacity, related to its composition, is greatly reduced after setting [Leonardo MR, L.A. Bezerra da Silva, M.T. Filho, D.S. Santana Release of formaldehyde by 4 endodontic sealers Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontics1999; 88: 221-5.] And is smaller than that found for other sealants [Leonardo MR, da Silva LA, Tanomaru Filho M Bonifacio KC, Ito IY. In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics. J Endod 2000; 26: 391-4].

Previous studies have shown that AH Plus is not able to kill bacteria or inhibit the formation of E. faecalis biofilms [Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod 2009; 35: 1051-5] [Baer J, Maki JS. In vitro evaluation of the antimicrobial effect of three endodontic sealers mixed with amoxicillin. J Endod 2010; 36: 1170-3].

As an essential part of endodontic treatment, together with mechanical instrumentation, it is essential to use irrigating solutions that possess, among other properties, antimicrobial activity. Sodium hypochlorite is one of them, although it poses toxicity problems for the periapice, in case of accidental extrusion, and has no residual antimicrobial activity. In this sense, various cationic disinfectants have been successfully tested as irrigating solutions. Chlorhexidine (CHX), is a bisguanide with potent antimicrobial action and proven residual action that is used as an irrigating solution in the treatment of root canals. It is reported that it is able to inactivate most resistant microorganisms

to endodontic treatment in just 15 seconds of contact [Vianna ME, Gomes

BP, Berber VB, Zaia AA, Ferraz CC, from Souza-Filho FJ. In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2004; 97: 79-84]. Cetrimide (CTR), is a cationic surfactant that has demonstrated its ability to eradicate E. faecalis in vitro [Arias-Moliz MT, Ferrer-Luque CM, Gonzalez-Rodriguez MP, Valderrama MJ, Baca P. Eradication of Enterococcus faecalis biofilms by cetrimide and chlorhexidine. J Endod 2010; 36: 87-90.] And ex vivo [Baca P, Junco P, Arias-Moliz MT, Gonzalez-Rodriguez MP, Ferrer-Luque CM. Residual and antimicrobial activity of final irrigation protocols on Enterococcus faecalis biofilm in dentin. J Endod 2011; 37: 363-6. Baca P, Mendoza-Llamas ML, Arias-Moliz MT, Gonzalez-Rodriguez MP, Ferrer-Luque CM. Residual effectiveness of final irrigation regimens on Enteroccus faecalis-infected root canals. J Endod 2011; 37: 1121-3]. Both CHX and CTR are safe at concentrations of use in dentistry. Other bisguanides, such as alexidine, or other cationic detergents such as cetylpyridinium chloride, may in the future be antiseptic of choice in the field of endodontics.

In order to evaluate the antibacterial effect of endodontic sealant cements, the diffusion agar test (TAD) has been widely used. However, due to its limitations, the direct contact test (TCD) is currently considered the most suitable for evaluating the antibacterial activity of dental materials, including sealant cements. The techniques for evaluating the inhibition of biofilm formation, particularly of E. faecalis, on materials are more recent and there is no consensus on the most appropriate, although there are several valid and reliable ones.

Since the bacteria inside the root canal grow as biofilms [Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284: 1318-22], especially E. faecalis, it is important to determine the antibacterial activity of materials capable of inhibiting their formation. In this regard, it is of particular interest to assess the antibacterial activity and ability to inhibit the formation of E. faecalis biofilms from AH Plus, alone, mixed with CHX or CTR, and combined with both.

There is no known reference to sealant cements that completely prevent the formation of biofilms or have a bactericidal effect, especially for E. faecalis.

OBJECT OF THE INVENTION

The object of the present invention is a composition for endodontic sealing with antibacterial activity and with capacity to prevent the formation of bacterial films or biofilms after the sealing process in endodontic treatments. It comprises an endodontic sealant which incorporates one or more cationic antiseptics so that, without affecting the physical-chemical properties of the sealant, they confer, under physiological conditions, antibacterial and bio-film formation inhibiting properties that, in addition, are maintained in the time. On the other hand, the addition of these antiseptics to the sealant confers bactericidal activity in the short and long term, eradicating the bacteria and inhibiting the formation of biofilms, particularly E. faecalis.

Normally, the incorporation of extraneous substances into a sealing cement can cause it to lose its properties. However, the invention is based on the surprising finding, that by mixing the epoxy resin sealant cement with antiseptics of the CHX, CTR or both type, at adequate concentrations, its physical properties were within the required standards, and it remained the antibacterial activity and inhibition of bacterial biofilm formation of E. faecalis effectively for weeks and months.

DESCRIPTION OF THE INVENTION

The present invention describes a composition for endodontic sealing with antibacterial activity and with capacity to prevent the formation of bacterial biofilms or biofilms after the sealing process in the treatment of root canals. The composition comprises an endodontic sealant to which one or more cationic antiseptics is incorporated. In particular, the composition comprises an endodontic sealant and at least one cationic antiseptic, preferably a surfactant and / or a bisguanide.

Preferably, the cationic surfactant used is cetrimide (CTR), always bearing in mind that when associated with anionic agents it can inactivate them.

Preferably, the bisguanide used is chlorhexidine (CHX), and even more preferably a solution of chlorhexidine digluconate.

In a preferred embodiment, the composition comprises an endodontic sealant cement with at least 0.1% CTR.

In a preferred embodiment, the composition comprises an endodontic sealant at

which incorporates between 1 and 3% of CHX (p / p) and between 0.1 and 1% of CTR (p / p). In an even more preferred embodiment the composition comprises an endodontic sealant which is incorporated between 2 and 3% CHX and between 0.1 and 0.5% CTR.

In a more preferred embodiment, the composition comprises an endodontic sealant, and more preferably with 2% CHX and 0.3% CTR.

In another preferred embodiment, the composition is characterized in that the only cationic disinfectant that is incorporated into the endodontic sealant is CTR in a proportion greater than or equal to 0.1% (w / w), more preferably in a proportion greater than or equal to 1% and even more preferably in a proportion of 1%.

In another preferred embodiment, the CHX employed is CHX digluconate.

In another preferred embodiment, the CTR employed is hexadecyltrimethylammonium bromide.

Preferably, the endodontic sealant is a material selected from the group consisting of plastic resins (epoxy resin, methacrylate resins, polyvinyl resins). More preferably, the material is epoxy resin. Even more preferably, the sealant is a two component paste based on an epoxy resin. Even more preferably, the sealant is AH Plus @, consisting of two pastes that are mixed at the time of application: paste A: Bisphenol-A epoxy resin, Bisphenol-F epoxy resin, Calcium tungstenate, Oxide of Zirconium, Silice and Iron Oxide; and paste B: Dibenzyl diamine, Aminoadamantane, Tricyclodecane diamine, Calcium tungstenate, Zirconium oxide, Silice and Silicone oil.

The addition of CTR and CHX to the sealant gives it bactericidal activity, particularly against E. faecalis.

The combination of AH Plus with CHX + CTR has greatly improved its antimicrobial effect. Using 2% CHX TCD combined with 0.3% CTR achieved 100% bacterial death; and 1% of CHX + 0.1% CTR was able to completely inhibit the formation of E. faecalis biofilms. In addition, low concentrations of antiseptic agents are sufficient to obtain this effect without impairing the physical-chemical properties of the sealant. Previous studies with ionomer glass cements to which CHX and CTR incorporated showed that the improved antimicrobial properties were accompanied by a deterioration of their physical properties.

EMBODIMENTS OF THE INVENTION

In order to determine the characteristics of the composition object of this invention, a series of tests were carried out on different compositions based on the commercial product AH Plus. This endodontic sealant was incorporated with CHX digluconate in aqueous solution at different concentrations (1%, 2% and 3%) (Guinama, Alboraya, Espafa), CTR powder at different concentrations (0.1%, 0.2% , 0.3%, 0.4%, 0.5%, 1%, 2% and 3%) (Sigma-Aldrich, Steinheim, Germany), and combinations of both cationic antiseptics, giving rise to 35 study groups.

In addition, AH Plus was used without incorporating antiseptics as a control group in the trial.

The different concentrations and combinations of both antiseptics with the sealant are shown in Tables 1 and 2. The term "absolute inhibition" (AI) was used to indicate 100% inhibition of biofilm formation and the term "eradication" ( E) was used to express 100% bacterial death.

From a subculture of E. faecalis ATCC 29212, a standard McFarland suspension 1 was prepared in BHI and then diluted to obtain an initial bacterial suspension of approximately 1 x 107 colony forming units per milliliter (CFU / ml) that was used for TCD and biofilm determination. The experiments were performed under sterile conditions in a laminar flow chamber (Telstar SA, Bio-II-B model, Tarrasa, Espafa).

Antimicrobial Activity Test

To test the antimicrobial activity of the materials, a methodology based on TCD adapted by Zhang et al. [Zhang H, Shen Y, Ruse ND, Haapasalo

M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod 2009; 35: 1051-5.]. A 96-well microtiter plate was used. To standardize the area of the materials tested, a custom silicone mold was made, to fit inside each well in the plate, leaving a small lateral area to introduce the same amount of material into each of them. The materials were introduced using a sterile needle syringe system (BD DISCARDIT II, Becton Dickinson SA Fraga, Huesca, Espafa) and allowed to set according to the manufacturer's instructions.

Subsequently, the molds were removed from each well and the 96-well microtiter plate was positioned vertically and 10 of the bacterial suspension of E. faecalis was carefully placed on the materials. After incubation at 37 ° C for 1 h in a humid atmosphere, which ensures contact between the bacteria and the material, the plate was placed in a horizontal position; then 220 he of BHI was added to each well and mixed gently with the pipette for 1 minute. Finally, 50 he were taken from each well and serial dilutions were made for sowing and subsequent viable cell counts.

Formation of biofilms on the materials tested

For the study of the formation of E. faecalis biofilms on sealant cements, a methodology based on the Calgary biofilm device (commercially available as MBEC-highthroughput [HTP]; Innovotech, Edmonton, Alberta, Canada), originally described by Ceri et al [Ceri H, Olson ME, Stremick C, Read RR, Morck D, Buret A. The Calgary Biofilm Device: new technology for rapid determination of antibiotic susceptibilities of bacterial biofilms. J Clin Microbiol 1999; 37: 1771-6] and adapted and modified for E. faecalis [Arias-Moliz MT, Ferrer-Luque CM, Espigares-Garcia M, Baca P. Enterococcus faecalis biofilms eradication by root canal irrigants. J Endod 2009; 35: 711-4]. The upper half of the MBEC-HTP is a cover with 96 identical pivots, which in this case were used to make silicone molds. Each material was introduced into the molds to obtain identical material cylinders. A sterile orthodontic wire of 1 cm in length was placed in each cylinder, while the material set, always leaving approximately 3 mm of it outside the material to facilitate its subsequent manipulation. The molds were incubated at 37 ° C in moist medium until completely set.

A 96-well microtiter plate was inoculated with 180 l / well of the E. faecalis suspension in BHI, after which the cylinders of each material were introduced into the wells. The microtiter plate / cylinder assembly was placed on an oscillating table (oscillation model Sw 8 10000-00015; OVAN, Badalona, Espafa), with 5 oscillations per minute, and incubated at 37 ° C for 24 hours. Then, the cylinders were introduced twice in washing plates containing 200 l of 0.9% saline solution for 2 minutes (each time). Then, they were transferred to a 96-well microtiter recovery plate with 200 µl of BHI / well, which was sonicated in a water table sonicator (model 5510E-MT; Branson, Danbury, CT) for 10 minutes to break Biofilm structure and recover bacteria. This procedure was performed in quadruplicate. Serial dilutions of the recovered biofilms were made and seeded in BHI agar that were incubated for viable cell counts.

The results of the antimicrobial activity and the inhibition of the formation of biofilms of E. faecalis were expressed, respectively, as a percentage of bacterial death and a percentage of inhibition of the formation of biofilms, calculated for each material as follows: (1 - [average CFU material tested / average CFU AH Plus] x 100). The term "absolute inhibition" (AI) was used to indicate 100% inhibition of biofilm formation and the term "eradication" (E) to express 100% bacterial death. To compare the efficacy of the different materials, provided that the percentage of death and the percentage of inhibition varied from 100%, the Student's t-test was used, subjecting the data prior to the Anscombe transformation.

Table 1. Percentage of inhibition of E. faecalis biofilms from AH Plus combined with CHX

and / or CTR.

Groups Without CHX CHX 1.0% CHX 2.0% CHX 3.0%

Without CTR -40.816% 1 97.510% 2 99.693% 3 CTR 0.1% 99.853% IA IA IA IA CTR 0.2% IA IA IA IA CTR 0.3% IA IA IA IA CTR 0.4% IA IA IA IA IA CTR 0.5% IA IA IA IA CTR 1.0 % IA IA IA IA CTR 2.0% IA IA IA IA CTR 3.0% IA IA IA IA

CTR: cetrimide, CHX: chlorhexidine. Read horizontally, the same numbers do not show statistically significant differences. AI: absolute inhibition (100%).

Table 2. Mean of the percentage of death of E. faecalis of AH Plus combined with CHX and / or CTR using the modified direct contact test.

Groups Without CHX CHX 1.0% CHX 2.0% CHX 3.0%

Without CTR -88.947% a, 1 90.262% a, 1 91.052% a, 1 CTR 0.1% 97.618% a, 1 98.223% b, 2 98.052% b, 2 98.381% b, 2 CTR 0.2% 98.25% b, 1 98.355 % b, 1 98.5% c, 1 98.855% c, 2 CTR 0.3% 98.565% b, c, 1 98.684% c, 1 EE CTR 0.4% 98.789% c, 1 99.013% c, 1 EE CTR 0.5% 99.868% d , 1 99.940% d, 1 EE CTR 1.0% EE EE CTR 2.0% EE EE CTR 3.0% EE EE

CTR: cetrimide, CHX: chlorhexidine.

10 Read vertically, the same letters do not show statistically significant differences. Read horizontally, the same numbers do not show statistically significant differences

E: eradication.

Previous studies showed that AH Plus alone is not able to kill bacteria or inhibit the formation of E. faecalis biofilms [Slutzky-Goldberg I, Slutzky H, Solomonov M, Moshonov J, Weiss EI, Matalon S. Antibacterial properties of four endodontic sealers. J Endod 2008; 34: 735-8] [Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test test against Enterococcus faecalis. J Endod 2009; 35: 1051-5]. The percentage of death and the percentage of inhibition of the formation of AH Plus biofilms combined with CHX show a direct relationship with the concentration used, although none of the concentrations tested achieved E or IA.

AH Plus mixed only with CTR achieved AI of biofilm formation at a concentration of 0.2%. However, eradication needed a 5-fold concentration of 1%.

The results of the combinations of CTR and CHX with AH Plus indicate a synergistic antimicrobial effect of both antiseptics, given the percentage of death and the inhibition of the formation of obtained biofilms. 0.3% CTR was necessary to eradicate E. faecalis when combined with 2% or 3% CHX, and only 0.1% CTR was needed for AI of biofilm formation when mixed with any CHX concentration .

Chemical physical properties

The antimicrobials tested have been 1 and 2% Chlorhexidine, 0.1, 0.2 Cetrimide,

0.3 and 0.5% and all possible combinations.

The physical tests performed were the following: 1. setting time, 2. fluidity, 3. solubility and 4. radiopacity. The methodology used has followed the standard ANSI / ADA standards for root canal sealing materials

(specification No. 57). All materials were handled according to

manufacturer's instructions The evaluation was performed by an operator who is unaware of the concentrations of the antimicrobials that each cement incorporated. Five replications have been made of each test and study group. The results have been expressed as mean (standard deviation). The results were statistically analyzed using the Tukey test.

1. Setting time:

For each group, 5 wells of 10 mm in diameter and 2 mm thick were filled in perforated in polyethylene plates and fixed in glass plates. The samples were stored in an oven with 95% relative humidity and 37 ° C. At 150 ± 10 seconds after the sealant began mixing, a Gillmore needle with a mass of 100 ± 0.5 g with a flat termination of 2.0 ± 0.1 mm in diameter, was carefully supported on the horizontal surface of each tested sample . The needle tip was cleaned and the test was repeated until the fingerprint on the surface of the cement ceased to be visible. The time from the beginning of the mixing until that moment was considered the setting time. If the results differed by more than ± 5%, the test was repeated.

According to ANSI / ADA standards, it is considered valid when the setting time variation is less than 10% of the time reported by the manufacturer.

2. Fluency:

For the fluency test, according to specification No. 57 of the American Dental Association (REF), the sealants in all the tested groups were homogeneously spatulated and a total of 0.5 ml of each mixture was placed on a glass plate ( 40x 40x5 mm). After 180 ± 5 seconds of the mixture, another plate with a mass of 20 ± 2 g was placed on top of which a load of 100 g was applied to the top of the material. Ten minutes after the beginning of the mixing, the load was removed and the major and minor diameters of the compressed discs were measured using a digital caliber with a resolution of 0.01 mm (Mitutoyo MTI Corporation, Tokyo, Japan).

Two conditions are necessary to validate the test, the first that the difference between

5 the maximum and minimum diameters do not exceed 1mm and that the compressed discs have a uniform shape. If the conditions are not met, the test must be repeated.

According to ANSI / ADA standards, it is considered that the fluidity of an endodontic sealant should obtain a minimum diameter of 20mm.

10 3. Solubility

On a glass plate covered with cellophane paper, polyethylene plates with perforated rings 20 mm in diameter and 1.5 mm thick were placed and filled completely with the sealants evaluated. Nylon threads were introduced into the sealants, and another glass plate covered with 15 cellophane paper was placed on top. All this assembly was incubated in an oven at 370 and relative humidity of 95%, for a period corresponding to three times the setting time indicated by the manufacturer. After this time, the specimens were demoulded, dehumidified for 1 hour, and weighed three times each specimen, on a scale with a precision of 0.0001 g. The samples, suspended by the nylon threads, 20 were introduced into plastic containers containing 7.5 ml of distilled deionized water, taking special care that the sample does not touch the walls of the container. The entire device was incubated in an oven with 95% relative humidity and 37 ° C for one week. After this time, the materials were dried with absorbent paper, dried in a chamber for 24 hours and weighed

new. The weight loss of each sample (difference between the initial and final weight) is expressed as a percentage of the original mass.

According to ANSI / ADA standards, it is considered valid when the weight loss does not exceed 3%.

4. Radiopacity

Five cylindrical samples of each material were made by pouring the mixed cements into polyethylene molds 10 mm in diameter by 1 mm thick, in accordance with ADA specification No. 57. The filled rings were kept in an oven at 370 C until they completely set. The samples were then removed, and the thickness was checked with a digital caliper (Mitutoyo Corp, Tokyo, Japan).

The sealant samples were placed on occlusal radiographs (D-speed; Kodak Comp, Rochester, NY) near a graduated aluminum scalpel from 2 to 16 mm of aluminum (in 2mm increments). Radiographs were taken, using a radiographic unit (Trophy Radiology, Vincennes, France) operating at 60 kV and 10 mA with an exposure time of 0.3 seconds and a focus-film distance of 30 cm. Immediately afterwards, the radiographs were digitized using a digital scanner and imported into Adobe Photoshop 11.1 for Windows (Adobe System Inc. San Jose CA, USA) for the determination of the radiographic density (densitometric analysis) in grayscale. The radiopacity value was determined according to the radiographic density transformed into millimeters of aluminum following the method described by Duarte et al ([Duarte MA, Ordinola-Zapata R, Bernardes RA, Bramante CM, Bernardineli N, Garcia RB, de Moraes IG Influence of calcium hydroxide association on the physical properties of AH Plus. J Endod. 2010; 36: 1048-51].

The mean and standard deviation of the setting time, fluidity, solubility and radiopacity of the epoxy resin resin AH Plus and the combinations tested are shown in Table 3.

Setting time: AH Plus cement, according to the manufacturer, requires at least one

5 minimum of 8 hours and 25 minutes of setting time (505 min). The maximum time is not specified. In our study, the results of cement as well as all combinations with antimicrobials are within the standards required by ANSI / ADA. Overall, the incorporation of CHX and its combinations with CTR reduce the setting time while CTR alone increases it.

10 Fluency: AH Plus cement and all combinations tested meet the requirements of ANSI / ADA since diameters greater than 20 mm are achieved. The incorporation of antimicrobials slightly modifies the diameters obtained with AH Plus. Chlorhexidine increases the fluidity, cetrimide reduces it and the combinations obtain intermediate values and therefore superimposable to AH Plus.

15 Solubility: AH Plus and all its mixtures did not exceed the 3% weight loss percentage required by ANSI / ADA standards. In no case the solubility of the samples reached 1%.

Radiopacity: All the materials tested showed a radiopacity greater than 3 mm required by international standards. The incorporation of

20 antimicrobials slightly modify the radiopacity obtained with AH Plus. CHX decreases it, CTR increases it and the combinations obtain intermediate values and therefore, superimposable to AH Plus.

TABLE 3. Physical Properties of AH Plus and its combinations with Chlorhexidine and Cetrimide. Mean (deviation

standard).

AH Plus

AH Plus + 1% CHX AH Plus + 2% CHX

AH Plus + O.1% CTR AH Plus + O.2% CTR AH Plus + O.3% CTR AH Plus + O.5% CTR

AH Plus + 1% CHX + O.1% CTR AH Plus + 1% CHX + O.2% CTR AH Plus + 1% CHX + O.3% CTR AH Plus + 1% CHX + O.5% CTR

AH Plus + 2% CHX + O.1% CTR AH Plus + 2% CHX + O.2% CTR AH Plus + 2% CHX + O.3% CTR AH Plus + 2% CHX + O.5% CTR

  Time of setting (min)  

890.59 (1.15) a

588.43 (1.09) b

577.70 (1.46) c, d

906.10 (0.38) g 911.35 (0.46) h

911.44 (1.47) h

913.07 (0.79) h

579.65 (0.80) d, e, 1

581.64 (1.15) 1

588.73

 (0.65) b

589.87

 (0.58) b

576.79 (0.56) c

578.81 (0.62) c, d, e

580.64

 (1.06) e, 1

581.44

 (1.53) 1

  Fluency (mm)  

38.85 (0.24) a, g

40.14 (0.30) b, c

40.50 (0.42) c

37.77 (0.22) d

37.03 (0.57) d

36.23 (0.81) e

35.19 (0.32) 1

39.30 (0.21) a, g

39.06 (0.11) a, g

39.05 (0.27) a, g

38.59

 (0.39) to

39.58

 (0.19) b, g

39.29 (0.23) a, g

39.41 (0.24) b, g

39.14 (0.24) a, g   Solubility Radiopacity (% reduction) (mm H)

0.197 (1.15) to 6.80 (0.03) a, c, d

0.111 (0.074) a, b 6.43 (0.04) b

0.09 (0.045) a, b 6.45 (0.07) b

0.046 (0.053) b 7.06 (0.04) 1, g

0.040 (0.039) b 7.08 (0.04) 1, g

0.041 (0.022) b 7.10 (0.71) g

0.097 (0.028) a, b 7.14 (0.05) g

0.098 (0.024) a, b 6.78 (0.08) a, c

0.107 (0.075) a, b 7.05 (0.01) e, 1, g

0.109 (0.025) a, b 6.91 (0.16) c, d, e

0.134 (0.026) a, b 6.81 (0.05) a, c, d

0.080 (0.036) a, b 6.57 (0.03) b

0.098 (0.02) a, b 6.89 (0.05) c, d

0.026 (0.053) c 6.72 (0.02) a

0.088 (0.083) a, b 6.94 (0.07) d, e, 1

* Read vertically, the same letters show that there are no statistically significant differences (Tukey test). "mm Al" indicates millimeters of aluminum.

Claims (20)

one.

Composition for endodontic sealing comprising an endodontic sealant to which one or more cationic antiseptics is incorporated.

2.

Composition for endodontic sealing comprising an endodontic sealant and a cationic antiseptic, preferably a cationic surfactant and a bisguanide.

3.

Composition, according to the preceding claim, wherein the cationic surfactant is Cetrimide (CTR).

Four.

Composition, according to the preceding claim, wherein the bisguanide is Clohexidine (CHX).

5.

Composition for endodontic sealing comprising an endodontic sealant which is incorporated between 2 and 3% CHX (w / w).

6.

Composition for endodontic sealing comprising an endodontic sealant which incorporates at least 0.1% CTR (w / w).

7.

Composition for endodontic sealing comprising an endodontic sealant which incorporates at least 1% CTR (w / w), preferably 1% CTR (w / w).

8.

Composition for endodontic sealing comprising an endodontic sealant which incorporates between 1 and 3% CHX (w / w) and between 0.1 and 1% CTR (w / w).

9.

Composition according to previous claim comprising between 2 and 3% of CHX and between 0.1 and 0.5% of CTR.

10.

Composition according to previous claim that 2% CHX and 0.3% CTR.

eleven.

Composition according to claims 6 6 7, characterized in that the only cationic disinfectant that is incorporated is CTR.

12.

Composition according to any one of the preceding claims wherein the chlorhexidine employed is chlorhexidine digluconate in aqueous solution.

13.

Composition according to any of the preceding claims in which the cetrimide employed is hexadecyltrimethylammonium bromide.

14.

Composition according to any of the preceding claims characterized in that the endodontic sealant mainly comprises a material selected from the group consisting of plastic resins.

fifteen.

Composition according to the preceding claim characterized in that the endodontic sealant mainly comprises epoxy resin, preferably epoxy amine resin.

16.

Use of a composition according to any of the preceding claims

to make an endodontic sealant that prevents the formation of biofilms bacterial after application in the sealing of root canals.

17.

Use according to previous claim in which the bacterial biofilm is

18.

Use according to any of the claims to manufacture a sealant

19.

Use according to any of the claims to manufacture a sealant

formed by E. Faecalis. Endodontic with bactericidal activity. Endodontic with bactericidal activity against E. Faecalis. SPANISH OFFICE OF THE PATENTS AND BRAND Application no .: 201330953 SPAIN Date of submission of the application: 06.25.2013 Priority Date: REPORT ON THE STATE OF THE TECHNIQUE 51 Int. Cl.: See Additional Sheet RELEVANT DOCUMENTS

Category

56 Documents cited Claims Affected

X

EP 2229929 A1 (DENTSPLY DETREY GMBH) 09/22/2010, paragraphs [0008] - [0011]; [0042] - [0043]; example 11. 1-5, 12-14, 16, 18

Y

6-11, 15, 17, 19

X

WO 0016712 A2 (ADVANTAGE DENTAL PRODUCTS INC et al.) 03/30/2000, page 3, line 11 1, 2, 4, 5, 12, 14,

- page 8, line 22; examples.

16, 18

Y

3, 6-11, 13, 15, 17,

19

 X

WO 2007038764 A2 (ESSENTIAL DENTAL SYSTEMS INC et al.) 04/05/2007, paragraphs [0009]  1, 2, 4, 5, 12, 14

[0011]; [0043].

16, 18

Y

3, 6-11, 13, 17, 19

Y

ARIAS-MOLIZ MARIA TERESA et al. Eradication of Enterococcus faecalis biofilms by cetrimide 3, 6-11, 13, 15, 17,

and chlorhexidine..Journal of endodontics United States Jan 2010 01/01/2010 VOL: 36 No: 1 Pags: 87 - 90 ISSN 1878-3554 (Electronic) Doi: doi: 10.1016 / j.joen.2009.10.013 pubmed: 20003941

19

Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application

This report has been prepared • for all claims • for claims no:

Date of realization of the report 29.07.2013

Examiner N. Vera Gutiérrez Page 1/4

REPORT OF THE STATE OF THE TECHNIQUE Application number: 201330953 CLASSIFICATION OBJECT OF THE APPLICATION C08L63 / 00 (2006.01) A61K6 / 08 (2006.01) C08K5 / 19 (2006.01) C08K5 / 31 (2006.01) Minimum documentation sought (classification system followed by classification symbols) C08L, A61K, C08K Electronic databases consulted during the search (name of the database and, if possible, search terms used) INVENES, EPODOC, REGISTRY, CAS, WPI, MEDLINE, EMBASE, BIOSIS, NPL, XPESP State of the Art Report Page 2/4  WRITTEN OPINION Application number: 201330953 Date of Written Opinion: 29.07.2013 Statement

Novelty (Art. 6.1 LP 11/1986)

Claims Claims 6-11, 17, 19 1-5, 12-16, 18 IF NOT

Inventive activity (Art. 8.1 LP11 / 1986)

Claims Claims 1-19 IF NOT

The application is considered to comply with the industrial application requirement. This requirement was evaluated during the formal and technical examination phase of the application (Article 31.2 Law 11/1986).  Opinion Base.- This opinion has been made on the basis of the patent application as published. State of the Art Report Page 3/4  WRITTEN OPINION Application number: 201330953 1. Documents considered.- The documents belonging to the state of the art taken into consideration for the realization of this opinion are listed below.

Document

Publication or Identification Number publication date

D01

EP 2229929 A1 (DENTSPLY DETREY GMBH) 09.22.2010

D02

WO 0016712 A2 (ADVANTAGE DENTAL PRODUCTS INC et al.) 30.03.2000

D03

WO 2007038764 A2 (ESSENTIAL DENTAL SYSTEMS INC et al.) 04.04.2007

D04

ARIAS-MOLIZ MARIA TERESA et al. Eradication of Enterococcus faecalis biofilms by cetrimide and chlorhexidine..Journal of endodontics United States Jan 2010 00/01/2010 VOL: 36 No: 1 Pags: 87 - 90 ISSN 1878-3554 (Electronic) Doi: doi: 10.1016 / j.joen .2009.10.013 pubmed: 20003941 01.2010

2. Statement motivated according to articles 29.6 and 29.7 of the Regulation of execution of Law 11/1986, of March 20, on Patents on novelty and inventive activity; quotes and explanations in support of this statement The invention relates to a composition for endodontic sealing comprising an endodontic sealant which incorporates one or more cationic antiseptics, such as chlorhexidine or cetrimide. Document D01 discloses a dispersive endodontic sealing composition comprising an aqueous phase with a polymer component and a phase dispersed with a radiopaque agent. The composition may include an antiseptic to prevent bacterial infection; These include chlorhexidine and cetyltrimethylammonium bromide (paragraphs [0042], [0043]). In example 11 a sealing composition based on polyethyleneimine containing chlorhexidine (2.09%) is prepared. Document D02 discloses a sealing composition for dental use comprising a polymerizable resin and an antimicrobial agent. In the examples compositions are prepared which include acrylic polymers and chlorhexidine in proportions 1%, 3%, 5%, 6% and 10%. It is cited that the compositions may include other biocidal and antimicrobial agents (page 8, lines 1-7; lines 18-20). Document D03 discloses an endodontic sealant composition based on an epoxy polymer, which may include chlorhexidine (0.01-3%) as an antimicrobial agent (paragraphs [0009] - [0011]; [0043]). In view of the cited documents, it is considered that the invention as defined in claims 1-5, 12-16, 18 is not new (Article 6.1 L.P.). Document D04 discloses a study to assess the efficacy of cetrimide and chlorhexidine antiseptics, alone or in association, in the eradication of Enterococcus faecalis biofilms. For this, dental irrigation solutions are prepared comprising different proportions of cetrimide (0.1% -0.0025%) and chlorhexidine (4% -0.0078%). Compositions containing 0.1% and 0.05% of cetrimide and any of the proportions of chlorhexidine are shown effective at all contact times tested. The associated use of cetrimide and chlorhexidine provides better results than its applications as individual agents. It is considered that the person skilled in the art, in view of the state of the art (in particular in document D02 cites the possibility of including several antiseptic agents in the composition), could incorporate the knowledge disclosed in D04 to any of the compositions sealants that include chlorhexidine or other antiseptics already existing in the state of the art, with a reasonable expectation of success. Therefore, and in the absence of a surprising technical effect associated with the selection of a proportion of cetrimide greater than 0.1%, it is considered that the invention as defined in claims 6-11, 17, 19 of the application no It implies inventive activity (Article 8.1 LP). State of the Art Report Page 4/4