Perillyl alcohol has antibacterial effects and reduces ROS production in macrophages.

Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 µM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 µM (p<0.05) and 250 µM (p<0.01). The POH increased ROS production at both 10 µM and 100 µM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 µM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 µM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.

Perillyl alcohol has antibacterial effects and reduces ROS production in macrophages

Abstract Natural products have emerged as a rich source of bioactive compounds for adjunctive treatments of many infectious and inflammatory conditions, including periodontitis. Among the monoterpenes with significant biological properties, there is the perillyl alcohol (POH), which can be found in several essential oils and has shown immunomodulatory properties in recent studies, which may be interesting in the treatment of non-neoplastic inflammatory disorders. Objective To determine the antibacterial and immune modulatory activities of the POH. Methodology The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the POH for two significant Gram-negative periodontal pathogens were determined by macrodilution and subculture, respectively. Cell proliferation and cytotoxicity in RAW 264.7 macrophages were determined by Trypan Blue and mitochondrial enzymatic activity assay. The modulation of reactive oxygen species (ROS) was analyzed by flow cytometry and expression of TNF and arginase-1 by real-time PCR. Results The POH was effective against P. gingivalis (ATCC 33277) and F. nucleatum (ATCC 25586) with MIC= MBC=1600 μM. No cytotoxicity up to 100 µM was observed on macrophages. The cell proliferation was inhibited from 48 hours at 100 μM (p<0.05) and 250 μM (p<0.01). The POH increased ROS production at both 10 μM and 100 μM (p<0.05) in unstimulated cells. The PMA-induced ROS production was not affected by POH, whereas 100 μM significantly reduced lipopolysaccharide-induced (LPS-induced) ROS. The expression of TNF was not affected by POH in unstimulated cells or in cells polarized to M1 phenotype, whereas both concentrations of POH reduced (p<0.05) the expression of arginase-1 in M2-polarized macrophages. Conclusion The POH has antibacterial activity against periodontal pathogens and reduced proliferation of murine macrophages without significant cytotoxicity at concentrations up to 100 μM. In addition, the POH reduced the LPS-induced ROS and the expression of arginase-1 in M2-polarized macrophages.

Nanofibers containing tetracycline/ß-cyclodextrin: Physico-chemical characterization and antimicrobial evaluation.

This study aimed to compare two nanofiber drug delivery systems that were prepared with an electrospun process and have the potential to serve as adjuvants for the treatment of periodontal disease. The first system was composed of polycaprolactone loaded with tetracycline (TCN) and the second was composed of polycaprolactone loaded with tetracycline/ß-cyclodextrin (TCN:BCD). An antimicrobial diffusion test was performed for each of these sets of nanofibers with the microorganisms, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, both of which contribute to periodontal disease. In vitro release profiles were also obtained, and the nanofibers were characterized by thermal analysis, x-ray powder diffraction, infrared absorption spectroscopy, and scanning electron microscopy. Profiles of the TCN and TCN:BCD nanofibers showed that drug release occurred for up to 14days. However, the TCN:BCD nanofibers appeared to better protect and enhance the biological absorption of TCN due to the formation of a TCN:BCD inclusion complex.

Occurrence and antimicrobial susceptibility of Porphyromonas spp. and Fusobacterium spp. in dogs with and without periodontitis.

The occurrence of Porphyromonas gulae, Porphyromonas macacae, Fusobacterium nucleatum and Fusobacterium canifelinum in subgingival plaque from dogs with and without periodontitis as well as their antimicrobial susceptibility were evaluated. From 50 dogs with periodontitis were identified 38 P. gulae, 8 P. macacae, 26 F. nucleatum and 15 F. canifelinum, and from 50 dogs without periodontitis were identified 15 P. gulae, 12 F. nucleatum and 11 F. canifelinum. All strains were susceptible to most of the antibiotics tested, however, different resistance rates to clarithromycin, erythromycin and metronidazole among strains were observed. The role of P. gulae, P. macacae, F. nucleatum and F. canifelinum in periodontal disease of household pets needs to be defined to a better prevention and treatment of the canine periodontitis.

In vivo antimicrobial effects of endodontic treatment procedures as assessed by molecular microbiologic techniques.

INTRODUCTION: This culture-independent molecular microbiology study evaluated the antimicrobial effects of chemomechanical preparation supplemented by intracanal medication during treatment of teeth with apical periodontitis. METHODS: Samples were taken from 24 necrotic root canals at the baseline (S1), after chemomechanical preparation by using 2.5% NaOCl as the irrigant (S2), and after a 7-day interappointment medication with calcium hydroxide paste in either glycerin (CHG) or camphorated paramonochlorophenol/glycerin (CHPG) (S3). Bacterial, archaeal, and fungal presence was evaluated by polymerase chain reaction (PCR), and bacterial identifications were performed by a closed-ended reverse-capture checkerboard approach targeting 28 candidate endodontic pathogens. RESULTS: All S1 samples were positive for bacteria but negative for both archaea and fungi. Treatment procedures were highly effective in reducing the bacterial levels and number of taxa. Overall, 46% of S2 samples and 62.5% of S3 samples were PCR-negative for bacteria. Specifically, S2 and S3 samples yielded negative PCR results in 50% and 58% of the canals in the CHG group and in 42% and 67% of the canals in the CHPG group, respectively. Except for comparisons with S1 samples, no other statistically significant differences were observed for intragroup and intergroup comparisons involving S2 and S3. Several taxa were still found in S2 and S3 samples, and the most prevalent were Propionibacterium acnes and Streptococcus species. CONCLUSIONS: Bacterial levels and number of taxa were substantially reduced after chemomechanical preparation and intracanal medication. However, presence of detectable levels of persisting bacteria in many cases indicates that the search for more effective antimicrobial treatment strategies should be stimulated.

[In vitro antimicrobial susceptibility of anaerobic bacteria isolated from pleuropulmonary infections]. / Susceptibilidad in vitro de bacterias anaeróbicas en infecciones pleuropulmonares.

BACKGROUND: Aspirative pleuropulmonary infections are usually caused by anaerobic flora of the mouth, mainly Prevotella, Fusobacterium and Peptostreptococcus spp. Penicillin in high doses is the traditional treatment for this type of infections but the rising resistance developed in recent years has induced the empiric use of clindamycin, increasing treatment costs. AIM: To study antimicrobial susceptibility of anaerobic bacteria isolated from pleuropulmonary infections. MATERIAL AND METHODS: Thirty two strains obtained from bronchoalveolar lavage and 15 strains isolated from pleural effusions between 2000 and 2002, were studied. The phenotype of strains was identified using the semiautomated API 20 A method and their susceptibility to penicillin (PNC), clindamycin (CM) and chloramphenicol (CAF) was tested using the E test methods. RESULTS: All the strains were susceptible to CAF, 95% to CM and 74.4% to PNC. The predominant genus was Prevotella, which also exhibited the higher resistance. CONCLUSIONS: As CM and CAF are active "in vitro", high rates of clinical response should be expected. In contrast, PNC is less effective, especially against pigmented Prevotella.

Susceptibilidad in vitro de bacterias anaeróbicas en infecciones pleuropulmonares / In vitro antimicrobial susceptibility of anaerobic bacteria isolated from pleuropulmonary infections

Background: Aspirative pleuropulmonary infections are usually caused by anaerobic flora of the mouth, mainly Prevotella, Fusobacterium and Peptostreptococcus spp. Penicillin in high doses is the traditional treatment for this type of infections but the rising resistance developed in recent years has induced the empiric use of clindamycin, increasing treatment costs. Aim: To study antimicrobial susceptibility of anaerobic bacteria isolated from pleuropulmonary infections. Material and methods: Thirty two strains obtained from bronchoalveolar lavage and 15 strains isolated from pleural effusions between 2000 and 2002, were studied. The phenotype of strains was identified using the semiautomated API 20 A method and their susceptibility to penicillin (PNC), clindamycin (CM) and chloramphenicol (CAF) was tested using the E test methods. Results: All the strains were susceptible to CAF, 95% to CM and 74.4% to PNC. The predominant genus was Prevotella, which also exhibited the higher resistance. Conclusions: As CM and CAF are active "in vitro", high rates of clinical response should be expected. In contrast, PNC is less effective, especially against pigmented Prevotella.

Microbial susceptibility to calcium hydroxide pastes and their vehicles.

The aim of this study was to investigate the susceptibility of some microorganisms commonly isolated from root canals to calcium hydroxide in combination with several vehicles by the agar diffusion method. Stainless-steel cylinders were placed on each inoculated agar medium. The test medications and their controls were placed inside the cylinders. The zones of growth inhibition were measured and recorded after the incubation period for each plate, and the results were analyzed statistically. Enterococcus faecalis was most resistant, whereas the anaerobic Porphyromonas endodontalis was more susceptible to all medications, followed by P. gingivalis and Prevotella intermedial intermedia. Ca(OH)2 + CMCP + glycerin showed significantly larger mean zones of inhibition when compared with the other medications. We conclude that anaerobic Gram-negative bacteria are more susceptible to calcium hydroxide pastes than facultative Gram-positive microorganisms.

Prueba de susceptibilidad antimicrobiana de un material de obturación endodóntica / Antimicrobial susceptibility test of an endodontic filling material

Los estudios y ensayos de laboratorio "in vitro" realizados en el presente trabajo con P.L.R.M. (pasta lentamente reabsorbible de Maisto) confirman un efecto bacteriostático con todas las cepas de microorganismos estudiados: Fusobacterium periodonticum ATCC 33693 Prevotella Melaninogénica ATCC 439822 Porphyromonas endodontalis ATCC 35406 Staphylococcus sp., Streptococcus viridans y Streptococcus Grupo A, en períodos de 0 a 56 días y valores de 1g. hasta 0,5mg/ml (AU)

In vitro antimicrobial activity of propolis and Arnica montana against oral pathogens.

Arnica and propolis have been used for thousands of years in folk medicine for several purposes. They possess several biological activities such as anti-inflammatory, antifungal, antiviral and tissue regenerative, among others. Although the antibacterial activity of propolis has already been demonstrated, very few studies have been done on bacteria of clinical relevance in dentistry. Also, the antimicrobial activity of Arnica has not been extensively investigated. Therefore the aim here was to evaluate in vitro the antimicrobial activity, inhibition of adherence of mutans streptococci and inhibition of formation of water-insoluble glucan by Arnica and propolis extracts. Arnica montana (10%, w/v) and propolis (10%, w/v) extracts from Minas Gerais State were compared with controls. Fifteen microorganisms were used as follows: Candida albicans--NTCC 3736, F72; Staphylococcus aureus--ATCC 25923; Enterococcus faecalis--ATCC 29212; Streptococcus sobrinus 6715; Strep. sanguis--ATCC 10556; Strep. cricetus--HS-6; Strep. mutans--Ingbritt 1600; Strep. mutans--OMZ 175; Actinomyces naeslundii--ATCC 12104, W 1053; Act. viscosus OMZ 105; Porphyromonas gingivalis; Porph. endodontalis and Prevotella denticola (the last three were clinical isolates). Antimicrobial activity was determined by the agar diffusion method and the zones of growth inhibition were measured. To assess cell adherence to a glass surface, the organisms were grown for 18 h at 37 degrees C in test-tubes at a 30 degree angle. To assay water-insoluble glucan formation, a mixture of crude glucosyltransferase and 0.125 M sucrose was incubated for 18 h at 37 degrees C in test-tubes at a 30 degree angle. Arnica and propolis extracts (20 microl) were added to these tubes to evaluate the % of inhibition of cell adherence and water-insoluble glucan formation. The propolis extract significantly inhibited all the microorganisms tested (p < 0.05), showing the largest inhibitory zone for Actinomyces spp. The Arnica extract did not demonstrate significant antimicrobial activity. Cell adherence and water-insoluble glucan formation were almost completely inhibited by the propolis extract at a final concentration of 400 microg/ml and 500 microg/ml, respectively. The Arnica extract showed slight inhibition of the adherence of the growing cells (19% for Strep. mutans and 15% for Strep. sobrinus) and of water-insoluble glucan formation (29%) at these same concentrations. Thus, the propolis extract showed in vitro antibacterial activity, inhibition of cell adherence and inhibition of water-insoluble glucan formation, while the Arnica extract was only slightly active in those three conditions.

Antibacterial effects of endodontic irrigants on black-pigmented gram-negative anaerobes and facultative bacteria.

The antibacterial effect of endodontic irrigants was evaluated against four black-pigmented Gram-negative anaerobes and four facultative anaerobic bacteria by means of the agar diffusion test. All solutions used were inhibitory against all bacterial strains tested. A 4% NaOCl solution provided the largest average zone of bacterial inhibition of this study that was significantly superior when compared with the other solutions, except 2.5% NaOCl (p < 0.05). Based on the averages of the diameters of the zones of bacterial growth inhibition, the antibacterial effects of the solutions could be ranked from strongest to weakest as follows: 4% NaOCl; 2.5% NaOCl; 2% chlorhexidine; 0.2% chlorhexidine, EDTA, and citric acid; and 0.5% NaOCl.

Influence of different vehicles on the antibacterial effects of calcium hydroxide.

This study evaluated the influence of three different vehicles on the antibacterial activity of calcium hydroxide against four bacterial species commonly found in endodontic infections. For this purpose, a broth dilution test using 24-well cell culture plates was performed. Results showed that all pastes were effective in killing the bacteria tested, but at different times. The calcium hydroxide/camphorated paramonochlorophenol/glycerin paste was the most effective against the four bacterial strains tested.