It has recently been proposed that three asaccharolytic species forming pigmented colonies on blo... more It has recently been proposed that three asaccharolytic species forming pigmented colonies on blood agar should be transferred from the genus Bacteruides to the new genus Purphyrumunas. In the taxonomy of the genus Bacteruides cellular fatty acid profiles obtained by gas chromatography and mass spectrometry after methylation and derivatization of whole cells have proved to be useful. In this study
Oral surgery, oral medicine, and oral pathology, 1980
Five different methods of transport and cultivation of bacteria from dental root canal specimens ... more Five different methods of transport and cultivation of bacteria from dental root canal specimens were evaluated. Bacteria were recovered from 29 percent of the specimens when they were transported in VMG IV and subcultured in PRAS medium or on the surface of blood agar in an anaerobic box. When a PRAS-peptone-yeast extract broth was used for transport and the specimens were initially subcultured in the broth, bacteria were recovered from 49 percent of the specimens. When fluid thioglycollate medium USP (BBL) and Clausen medium (Oxoid) were used for transport as well as for initial subculture, bacteria were recovered from 58 percent and 47 percent of the specimens, respectively. The high recovery of bacteria by these media could to a significant degree be ascribed to the use of the media for initial subculture of the specimens. Recovery of bacteria by fluid thioglycollate medium USP was as good or even better as with the more elaborate PRAS medium. Fluid thioglycollate medium USP in ...
The existence of commensal or antagonistic relationships between microorganisms in the root canal... more The existence of commensal or antagonistic relationships between microorganisms in the root canals of teeth with apical periodontitis was investigated. Samples were taken from 65 infected human root canals and were analysed according to species, frequency of occurrence and proportion of the total isolated flora. The most frequent species were Fusobacterium nucleatum, Prevotella intermedia, Peptostreptococcus micros, Peptostreptococcus anaerobius, Eubacterium alactolyticum, Eubacterium lentum and Wolinella recta. An odds ratio system was used to calculate positive or negative associations between the isolated bacteria. Strong positive associations were found between F. nucleatum and P. micros, Porphyromonas endodontalis, Selenomonas sputigena and W. recta. There was also a positive association between P. intermedia and P. micros, P. anaerobius and the eubacteria. In general, species of streptococci, Propionibacterium propionica, Capnocytophaga ochracea and Veillonella parvula showed no or negative associations with the other bacteria. The results are consistent with the concept of a special and selective environment occurring in the root canal that is due, in part, to the cooperative as well as antagonistic nature of the relationships between bacteria in the root canal.
Light and electron microscopy were used to analyze nine therapy-resistant and asymptomatic human ... more Light and electron microscopy were used to analyze nine therapy-resistant and asymptomatic human periapical lesions, which were removed as block biopsies during surgical treatment of the affected teeth. The cases that required surgery represented about 10% of all of the cases which received endodontic treatment and root fillings during the period 1977 to 1984. These cases revealed periapical lesions when they were examined 4 to 10 yr after treatment. The biopsies were processed for correlated light and electron microscopy. Six of the nine biopsies revealed the presence of microorganisms in the apical root canal. Four contained one or more species of bacteria and two revealed yeasts. Of the four cases in which bacteria were found, only in one biopsy could they be found by light microscope. In the other three specimens, the bacterial presence could be confirmed only after repeated electron microscopic examination of the apical root canal by serial step-cutting technique. Among the three cases in which no microorganisms could be encountered, one showed histopathological features of a foreign body giant cell granuloma. These findings suggest that in the majority of root-filled human teeth with therapy-resistant periapical lesions, microorganisms may persist and may play a significant role in endodontic treatment failures. In certain instances such lesions may also be sustained by foreign body giant cell type of tissue responses at the periapex of root-filled teeth.
Although the primary etiological factor of periapical lesions is microbial, there are other indep... more Although the primary etiological factor of periapical lesions is microbial, there are other independent factors that can adversely affect the outcome of endodontic treatment. In this communication, we present morphological evidence in support of the role of a foreign body reaction of periapical tissue to root-filling materials. The specimen consisted of a surgical biopsy of an asymptomatic periapical lesion which persisted after a decade of postendodontic follow-up. The biopsy was processed for correlated light and electron microscopy and was analyzed by various microtechniques. The unique feature of the lesion was the presence of vast numbers of large multinucleated cells and their cytoplasmic inclusion bodies. Morphologically, these multinucleated cells resembled foreign body giant cells. They contained characteristic birefringent cytoplasmic inclusions which on electron-probe X-ray microanalysis consistently revealed the presence of magnesium and silicon. The magnesium and silicon are presumably the remnants of a root-filling excess which protruded into the periapex and had been resorbed during the follow-up period. These observations strongly suggest that in the absence of microbial factors, root-filling materials which contain irritating substances can evoke a foreign body reaction at the periapex, leading to the development of asymptomatic periapical lesions that may remain refractory to endodontic therapy for long periods of time.
The antibacterial effect of ultrasonic instrumentation in the treatment of infected root canals w... more The antibacterial effect of ultrasonic instrumentation in the treatment of infected root canals was clinically evaluated. Sodium hypochlorite solution (0.5%) was used as an irrigant, but no antibacterial intracanal dressing was used between the appointments. The ultrasonic technique eliminated the bacteria from the canals more efficiently than hand instrumentation alone. Even though ultrasonication definitely improves the procedure of root canal disinfection, the use of an antibacterial dressing between appointments is necessary to achieve as complete a reduction in bacterial levels as possible.
The fate of DNA from bacteria that infect the root canal but cannot survive is currently unknown,... more The fate of DNA from bacteria that infect the root canal but cannot survive is currently unknown, yet such information is essential in establishing the validity of polymerase chain reaction (PCR)-based identification methods for root canal samples. This in vitro study tested the hypothesis that PCR-detectable DNA from dead bacteria might persist after cell death and investigated the efficiency of sodium hypochlorite (NaOCl) as a field decontamination agent. Using heat-killed Enterococcus faecalis, the persistence of DNA encoding the 16S rRNA gene was monitored by PCR. While most probable number analysis showed an approximate 1000-fold decay in amplifiable template, E. faecalis DNA was still PCR-detectable 1 year after cell death. NaOCl (1%) eliminated amplifiable DNA within 60 seconds of exposure. Our findings also disclosed a previously overlooked problem of concentration-dependent inhibition of the PCR reaction by thiosulfate-inactivated NaOCl. These results highlight the challenges of reliably identifying the authentic living root canal flora with PCR techniques.
Genomic fingerprints from the DNA of 27 strains of Porphyromonas endodontalis from diverse clinic... more Genomic fingerprints from the DNA of 27 strains of Porphyromonas endodontalis from diverse clinical and geographic origins were generated as random amplified polymorphic DNA (RAPD) using the technique of PCR amplification with a single primer of arbitrary sequence. Cluster analysis of the combined RAPD data obtained with three selected 9- or 10-mer-long primers identified 25 distinct RAPD types which clustered as three main groups identifying three genogroups. Genogroups I and II included exclusively P. endodontalis isolates of oral origin, while 7/9 human intestinal strains of genogroup III which linked at a similarity level of 52% constituted the most homogeneous group in our study. Genotypic diversity within P. endodontalis, as shown by RAPD analysis, suggests that the taxon is composed of two oral genogroups and one intestinal genogroup. This hypothesis remains to be confirmed.
To investigate the ability of Actinomyces radicidentis to survive and establish in soft connectiv... more To investigate the ability of Actinomyces radicidentis to survive and establish in soft connective tissue that grew into subcutaneously implanted tissue cages in Sprague-Dawley rats. Known concentrations of A. radicidentis suspension, grown on blood agar and broth cultures, were inoculated into tissue cages in rats. The cage contents were retrieved after 7, 14, and 28 days for culturing and correlative light and transmission electron microscopy. Cell suspensions harvested from both types of cultures showed substantial decline in numbers in tissue cages during the observation period. However, correlative light and transmission electron microscopy revealed numerous aggregates of coccoid bacteria already by 7 days of observation compared with the formation of well established colonies with characteristic actinomycotic features by 14 days after inoculation. These results suggest that the pathogenicity of A. radicidentis is due to its ability to form large aggregates of cells held together by embedding themselves in an extracellular matrix in vital host tissues. Thus, A. radicidentis, like other pathogenic Actinomyces, existing in the protected biofilm-environment can collectively evade destruction and elimination by host defenses, including phagocytosis.
The ability of Bacteroides gingivalis to generate chemotactic activity and the complement fragmen... more The ability of Bacteroides gingivalis to generate chemotactic activity and the complement fragment C5a in human serum was assayed. When fresh serum was incubated with B. gingivalis there was a rapid increase of chemotactic activity of the serum during the first 15 ...
It has recently been proposed that three asaccharolytic species forming pigmented colonies on blo... more It has recently been proposed that three asaccharolytic species forming pigmented colonies on blood agar should be transferred from the genus Bacteruides to the new genus Purphyrumunas. In the taxonomy of the genus Bacteruides cellular fatty acid profiles obtained by gas chromatography and mass spectrometry after methylation and derivatization of whole cells have proved to be useful. In this study
Oral surgery, oral medicine, and oral pathology, 1980
Five different methods of transport and cultivation of bacteria from dental root canal specimens ... more Five different methods of transport and cultivation of bacteria from dental root canal specimens were evaluated. Bacteria were recovered from 29 percent of the specimens when they were transported in VMG IV and subcultured in PRAS medium or on the surface of blood agar in an anaerobic box. When a PRAS-peptone-yeast extract broth was used for transport and the specimens were initially subcultured in the broth, bacteria were recovered from 49 percent of the specimens. When fluid thioglycollate medium USP (BBL) and Clausen medium (Oxoid) were used for transport as well as for initial subculture, bacteria were recovered from 58 percent and 47 percent of the specimens, respectively. The high recovery of bacteria by these media could to a significant degree be ascribed to the use of the media for initial subculture of the specimens. Recovery of bacteria by fluid thioglycollate medium USP was as good or even better as with the more elaborate PRAS medium. Fluid thioglycollate medium USP in ...
The existence of commensal or antagonistic relationships between microorganisms in the root canal... more The existence of commensal or antagonistic relationships between microorganisms in the root canals of teeth with apical periodontitis was investigated. Samples were taken from 65 infected human root canals and were analysed according to species, frequency of occurrence and proportion of the total isolated flora. The most frequent species were Fusobacterium nucleatum, Prevotella intermedia, Peptostreptococcus micros, Peptostreptococcus anaerobius, Eubacterium alactolyticum, Eubacterium lentum and Wolinella recta. An odds ratio system was used to calculate positive or negative associations between the isolated bacteria. Strong positive associations were found between F. nucleatum and P. micros, Porphyromonas endodontalis, Selenomonas sputigena and W. recta. There was also a positive association between P. intermedia and P. micros, P. anaerobius and the eubacteria. In general, species of streptococci, Propionibacterium propionica, Capnocytophaga ochracea and Veillonella parvula showed no or negative associations with the other bacteria. The results are consistent with the concept of a special and selective environment occurring in the root canal that is due, in part, to the cooperative as well as antagonistic nature of the relationships between bacteria in the root canal.
Light and electron microscopy were used to analyze nine therapy-resistant and asymptomatic human ... more Light and electron microscopy were used to analyze nine therapy-resistant and asymptomatic human periapical lesions, which were removed as block biopsies during surgical treatment of the affected teeth. The cases that required surgery represented about 10% of all of the cases which received endodontic treatment and root fillings during the period 1977 to 1984. These cases revealed periapical lesions when they were examined 4 to 10 yr after treatment. The biopsies were processed for correlated light and electron microscopy. Six of the nine biopsies revealed the presence of microorganisms in the apical root canal. Four contained one or more species of bacteria and two revealed yeasts. Of the four cases in which bacteria were found, only in one biopsy could they be found by light microscope. In the other three specimens, the bacterial presence could be confirmed only after repeated electron microscopic examination of the apical root canal by serial step-cutting technique. Among the three cases in which no microorganisms could be encountered, one showed histopathological features of a foreign body giant cell granuloma. These findings suggest that in the majority of root-filled human teeth with therapy-resistant periapical lesions, microorganisms may persist and may play a significant role in endodontic treatment failures. In certain instances such lesions may also be sustained by foreign body giant cell type of tissue responses at the periapex of root-filled teeth.
Although the primary etiological factor of periapical lesions is microbial, there are other indep... more Although the primary etiological factor of periapical lesions is microbial, there are other independent factors that can adversely affect the outcome of endodontic treatment. In this communication, we present morphological evidence in support of the role of a foreign body reaction of periapical tissue to root-filling materials. The specimen consisted of a surgical biopsy of an asymptomatic periapical lesion which persisted after a decade of postendodontic follow-up. The biopsy was processed for correlated light and electron microscopy and was analyzed by various microtechniques. The unique feature of the lesion was the presence of vast numbers of large multinucleated cells and their cytoplasmic inclusion bodies. Morphologically, these multinucleated cells resembled foreign body giant cells. They contained characteristic birefringent cytoplasmic inclusions which on electron-probe X-ray microanalysis consistently revealed the presence of magnesium and silicon. The magnesium and silicon are presumably the remnants of a root-filling excess which protruded into the periapex and had been resorbed during the follow-up period. These observations strongly suggest that in the absence of microbial factors, root-filling materials which contain irritating substances can evoke a foreign body reaction at the periapex, leading to the development of asymptomatic periapical lesions that may remain refractory to endodontic therapy for long periods of time.
The antibacterial effect of ultrasonic instrumentation in the treatment of infected root canals w... more The antibacterial effect of ultrasonic instrumentation in the treatment of infected root canals was clinically evaluated. Sodium hypochlorite solution (0.5%) was used as an irrigant, but no antibacterial intracanal dressing was used between the appointments. The ultrasonic technique eliminated the bacteria from the canals more efficiently than hand instrumentation alone. Even though ultrasonication definitely improves the procedure of root canal disinfection, the use of an antibacterial dressing between appointments is necessary to achieve as complete a reduction in bacterial levels as possible.
The fate of DNA from bacteria that infect the root canal but cannot survive is currently unknown,... more The fate of DNA from bacteria that infect the root canal but cannot survive is currently unknown, yet such information is essential in establishing the validity of polymerase chain reaction (PCR)-based identification methods for root canal samples. This in vitro study tested the hypothesis that PCR-detectable DNA from dead bacteria might persist after cell death and investigated the efficiency of sodium hypochlorite (NaOCl) as a field decontamination agent. Using heat-killed Enterococcus faecalis, the persistence of DNA encoding the 16S rRNA gene was monitored by PCR. While most probable number analysis showed an approximate 1000-fold decay in amplifiable template, E. faecalis DNA was still PCR-detectable 1 year after cell death. NaOCl (1%) eliminated amplifiable DNA within 60 seconds of exposure. Our findings also disclosed a previously overlooked problem of concentration-dependent inhibition of the PCR reaction by thiosulfate-inactivated NaOCl. These results highlight the challenges of reliably identifying the authentic living root canal flora with PCR techniques.
Genomic fingerprints from the DNA of 27 strains of Porphyromonas endodontalis from diverse clinic... more Genomic fingerprints from the DNA of 27 strains of Porphyromonas endodontalis from diverse clinical and geographic origins were generated as random amplified polymorphic DNA (RAPD) using the technique of PCR amplification with a single primer of arbitrary sequence. Cluster analysis of the combined RAPD data obtained with three selected 9- or 10-mer-long primers identified 25 distinct RAPD types which clustered as three main groups identifying three genogroups. Genogroups I and II included exclusively P. endodontalis isolates of oral origin, while 7/9 human intestinal strains of genogroup III which linked at a similarity level of 52% constituted the most homogeneous group in our study. Genotypic diversity within P. endodontalis, as shown by RAPD analysis, suggests that the taxon is composed of two oral genogroups and one intestinal genogroup. This hypothesis remains to be confirmed.
To investigate the ability of Actinomyces radicidentis to survive and establish in soft connectiv... more To investigate the ability of Actinomyces radicidentis to survive and establish in soft connective tissue that grew into subcutaneously implanted tissue cages in Sprague-Dawley rats. Known concentrations of A. radicidentis suspension, grown on blood agar and broth cultures, were inoculated into tissue cages in rats. The cage contents were retrieved after 7, 14, and 28 days for culturing and correlative light and transmission electron microscopy. Cell suspensions harvested from both types of cultures showed substantial decline in numbers in tissue cages during the observation period. However, correlative light and transmission electron microscopy revealed numerous aggregates of coccoid bacteria already by 7 days of observation compared with the formation of well established colonies with characteristic actinomycotic features by 14 days after inoculation. These results suggest that the pathogenicity of A. radicidentis is due to its ability to form large aggregates of cells held together by embedding themselves in an extracellular matrix in vital host tissues. Thus, A. radicidentis, like other pathogenic Actinomyces, existing in the protected biofilm-environment can collectively evade destruction and elimination by host defenses, including phagocytosis.
The ability of Bacteroides gingivalis to generate chemotactic activity and the complement fragmen... more The ability of Bacteroides gingivalis to generate chemotactic activity and the complement fragment C5a in human serum was assayed. When fresh serum was incubated with B. gingivalis there was a rapid increase of chemotactic activity of the serum during the first 15 ...
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