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Background:
The most common failures of dental implants are related to microbial colonization, where usually, these microorganisms form complex structures, called biofilms. While oral biofilms comprise a wide variety of bacterial species, colonization follows a sequence with adhesion of initial colonizers to the acquired pellicle followed by secondary colonization through inter-bacterial adhesion. The anti-biofilm effect of several inorganic or organic agents has been reported in previous studies (Secinti et al., 2011; Zhang et al., 2012; Melo et al., 2013). Reports on the anti-biofilm effect of some synthetic compounds similar to furanones, such as lactams, are widespread in the literature (Defoirdt et al., 2008; Ikonomidis et al., 2008). However, the effect of lactams in dentistry in order to inhibit biofilm formation is unknown.
Aim/Hypothesis:
To assess the anti-biofilm effect of novel lactams on biofilm formation of early colonizer single-species cultures.
Material and Methods:
A group of twenty-four lactams was chosen to be tested individually for each early colonizer bacterial species (Streptococcus oralis, Streptococcus mitis, Streptococcus sanguinis, Streptococcus gordonii and Actinomyces naeslundii). The bacterial strains were inoculated into BHI broth from an isolated colony and incubated overnight at 37 ° C under aerobic conditions. The day after, a solution containing a 1x107 of bacteria was prepared and incubated with the lactams according to CLSI concentrations from ranging from 128 to 0,25 µg/ml. The positive and negative controls were 200 µL of BHI and 200 µL of bacterial suspension. After 24 hours, the fixation of the biofilm was done by ethanol and the fixed biofilms were stained with crystal violet for 15 minutes. The stained biofilms were resuspended using 75 µL of acetic acid 5% for 45 minutes and then the OD was measured by spectrophotometry at 630nm. The data were analysed by two-way Anova analysis and the statistical significance was established at p<0,05.
Results:
Compared to the negative control, the data showed more than 70% of biofilm inhibition for S. oralis in the highest (128 and 64 µg/ml) concentrations for most of the tested lactams (p<0,003). Similar, S. mitis biofilms were inhibited between 60% and 70% (p>0,5). S. sanguinis biofilms could be inhibited around 50% (p<0,05) and A. naeslundi biofilm formation could be inhibited up to 80% (p<0,003). On the other hand, it was noticed that the biofilm formation of S. gordonii was increased at the highest lactam concentrations.
Conclusions and clinical implications:
It can be concluded that the selected lactams can reduce the biofilm formation of some early colonizer strains up to 80%. Although the single species biofilms do not represent the actual complex and dynamic multi-species biofilm in the oral cavity, it can be hypotized that by decreasing the biofilm formation of the initial colonizers, the complex/multi-species biofilm can also be influenced. However further studies with multi-species biofilms are needed to address this hypothesis.
Background:
The most common failures of dental implants are related to microbial colonization, where usually, these microorganisms form complex structures, called biofilms. While oral biofilms comprise a wide variety of bacterial species, colonization follows a sequence with adhesion of initial colonizers to the acquired pellicle followed by secondary colonization through inter-bacterial adhesion. The anti-biofilm effect of several inorganic or organic agents has been reported in previous studies (Secinti et al., 2011; Zhang et al., 2012; Melo et al., 2013). Reports on the anti-biofilm effect of some synthetic compounds similar to furanones, such as lactams, are widespread in the literature (Defoirdt et al., 2008; Ikonomidis et al., 2008). However, the effect of lactams in dentistry in order to inhibit biofilm formation is unknown.
Aim/Hypothesis:
To assess the anti-biofilm effect of novel lactams on biofilm formation of early colonizer single-species cultures.
Material and Methods:
A group of twenty-four lactams was chosen to be tested individually for each early colonizer bacterial species (Streptococcus oralis, Streptococcus mitis, Streptococcus sanguinis, Streptococcus gordonii and Actinomyces naeslundii). The bacterial strains were inoculated into BHI broth from an isolated colony and incubated overnight at 37 ° C under aerobic conditions. The day after, a solution containing a 1x107 of bacteria was prepared and incubated with the lactams according to CLSI concentrations from ranging from 128 to 0,25 µg/ml. The positive and negative controls were 200 µL of BHI and 200 µL of bacterial suspension. After 24 hours, the fixation of the biofilm was done by ethanol and the fixed biofilms were stained with crystal violet for 15 minutes. The stained biofilms were resuspended using 75 µL of acetic acid 5% for 45 minutes and then the OD was measured by spectrophotometry at 630nm. The data were analysed by two-way Anova analysis and the statistical significance was established at p<0,05.
Results:
Compared to the negative control, the data showed more than 70% of biofilm inhibition for S. oralis in the highest (128 and 64 µg/ml) concentrations for most of the tested lactams (p<0,003). Similar, S. mitis biofilms were inhibited between 60% and 70% (p>0,5). S. sanguinis biofilms could be inhibited around 50% (p<0,05) and A. naeslundi biofilm formation could be inhibited up to 80% (p<0,003). On the other hand, it was noticed that the biofilm formation of S. gordonii was increased at the highest lactam concentrations.
Conclusions and clinical implications:
It can be concluded that the selected lactams can reduce the biofilm formation of some early colonizer strains up to 80%. Although the single species biofilms do not represent the actual complex and dynamic multi-species biofilm in the oral cavity, it can be hypotized that by decreasing the biofilm formation of the initial colonizers, the complex/multi-species biofilm can also be influenced. However further studies with multi-species biofilms are needed to address this hypothesis.
Background:
The most common failures of dental implants are related to microbial colonization, where usually, these microorganisms form complex structures, called biofilms. While oral biofilms comprise a wide variety of bacterial species, colonization follows a sequence with adhesion of initial colonizers to the acquired pellicle followed by secondary colonization through inter-bacterial adhesion. The anti-biofilm effect of several inorganic or organic agents has been reported in previous studies (Secinti et al., 2011; Zhang et al., 2012; Melo et al., 2013). Reports on the anti-biofilm effect of some synthetic compounds similar to furanones, such as lactams, are widespread in the literature (Defoirdt et al., 2008; Ikonomidis et al., 2008). However, the effect of lactams in dentistry in order to inhibit biofilm formation is unknown.
Aim/Hypothesis:
To assess the anti-biofilm effect of novel lactams on biofilm formation of early colonizer single-species cultures.
Material and Methods:
A group of twenty-four lactams was chosen to be tested individually for each early colonizer bacterial species (Streptococcus oralis, Streptococcus mitis, Streptococcus sanguinis, Streptococcus gordonii and Actinomyces naeslundii). The bacterial strains were inoculated into BHI broth from an isolated colony and incubated overnight at 37 ° C under aerobic conditions. The day after, a solution containing a 1x107 of bacteria was prepared and incubated with the lactams according to CLSI concentrations from ranging from 128 to 0,25 µg/ml. The positive and negative controls were 200 µL of BHI and 200 µL of bacterial suspension. After 24 hours, the fixation of the biofilm was done by ethanol and the fixed biofilms were stained with crystal violet for 15 minutes. The stained biofilms were resuspended using 75 µL of acetic acid 5% for 45 minutes and then the OD was measured by spectrophotometry at 630nm. The data were analysed by two-way Anova analysis and the statistical significance was established at p<0,05.
Results:
Compared to the negative control, the data showed more than 70% of biofilm inhibition for S. oralis in the highest (128 and 64 µg/ml) concentrations for most of the tested lactams (p<0,003). Similar, S. mitis biofilms were inhibited between 60% and 70% (p>0,5). S. sanguinis biofilms could be inhibited around 50% (p<0,05) and A. naeslundi biofilm formation could be inhibited up to 80% (p<0,003). On the other hand, it was noticed that the biofilm formation of S. gordonii was increased at the highest lactam concentrations.
Conclusions and clinical implications:
It can be concluded that the selected lactams can reduce the biofilm formation of some early colonizer strains up to 80%. Although the single species biofilms do not represent the actual complex and dynamic multi-species biofilm in the oral cavity, it can be hypotized that by decreasing the biofilm formation of the initial colonizers, the complex/multi-species biofilm can also be influenced. However further studies with multi-species biofilms are needed to address this hypothesis.
Background:
The most common failures of dental implants are related to microbial colonization, where usually, these microorganisms form complex structures, called biofilms. While oral biofilms comprise a wide variety of bacterial species, colonization follows a sequence with adhesion of initial colonizers to the acquired pellicle followed by secondary colonization through inter-bacterial adhesion. The anti-biofilm effect of several inorganic or organic agents has been reported in previous studies (Secinti et al., 2011; Zhang et al., 2012; Melo et al., 2013). Reports on the anti-biofilm effect of some synthetic compounds similar to furanones, such as lactams, are widespread in the literature (Defoirdt et al., 2008; Ikonomidis et al., 2008). However, the effect of lactams in dentistry in order to inhibit biofilm formation is unknown.
Aim/Hypothesis:
To assess the anti-biofilm effect of novel lactams on biofilm formation of early colonizer single-species cultures.
Material and Methods:
A group of twenty-four lactams was chosen to be tested individually for each early colonizer bacterial species (Streptococcus oralis, Streptococcus mitis, Streptococcus sanguinis, Streptococcus gordonii and Actinomyces naeslundii). The bacterial strains were inoculated into BHI broth from an isolated colony and incubated overnight at 37 ° C under aerobic conditions. The day after, a solution containing a 1x107 of bacteria was prepared and incubated with the lactams according to CLSI concentrations from ranging from 128 to 0,25 µg/ml. The positive and negative controls were 200 µL of BHI and 200 µL of bacterial suspension. After 24 hours, the fixation of the biofilm was done by ethanol and the fixed biofilms were stained with crystal violet for 15 minutes. The stained biofilms were resuspended using 75 µL of acetic acid 5% for 45 minutes and then the OD was measured by spectrophotometry at 630nm. The data were analysed by two-way Anova analysis and the statistical significance was established at p<0,05.
Results:
Compared to the negative control, the data showed more than 70% of biofilm inhibition for S. oralis in the highest (128 and 64 µg/ml) concentrations for most of the tested lactams (p<0,003). Similar, S. mitis biofilms were inhibited between 60% and 70% (p>0,5). S. sanguinis biofilms could be inhibited around 50% (p<0,05) and A. naeslundi biofilm formation could be inhibited up to 80% (p<0,003). On the other hand, it was noticed that the biofilm formation of S. gordonii was increased at the highest lactam concentrations.
Conclusions and clinical implications:
It can be concluded that the selected lactams can reduce the biofilm formation of some early colonizer strains up to 80%. Although the single species biofilms do not represent the actual complex and dynamic multi-species biofilm in the oral cavity, it can be hypotized that by decreasing the biofilm formation of the initial colonizers, the complex/multi-species biofilm can also be influenced. However further studies with multi-species biofilms are needed to address this hypothesis.