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Biomedical subjects

G Sundqvist

Publications and source records attributed to G Sundqvist.

At least 19 recordsLinked to original sources

Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment.

OBJECTIVE: The purposes of this study were to determine what microbial flora were present in teeth after failed root canal therapy and to establish the outcome of conservative re-treatment. STUDY DESIGN: Fifty-four root-filled teeth with persisting periapical lesions were selected for re-treatment. After removal of the root filling, canals were sampled by means of advanced microbiologic techniques. The teeth were then re-treated and followed for up to 5 years. RESULTS: The microbial flora was mainly single species of predominantly gram-positive organisms. The isolates most commonly recovered were bacteria of the species Enterococcus faecalis. The overall success rate of re-treatment was 74%. CONCLUSIONS: The microbial flora in canals after failed endodontic therapy differed markedly from the flora in untreated teeth. Infection at the time of root filling and size of the periapical lesion were factors that had a negative influence on the prognosis. Three of four endodontic failures were successfully managed by re-treatment.

Dental Pulp Cavity

Cholesterol crystals as an etiological factor in non-resolving chronic inflammation: an experimental study in guinea pigs.

The presence of cholesterol crystals has been suggested to be a factor interfering with periapical healing after conventional endodontic treatment. This investigation addresses the role of cholesterol crystals in impairing healing by studying the tissue response to the crystals, which were implanted in animals. Pure cholesterol crystals, prepared to a mushy form, were placed in Teflon cages that were implanted subcutaneously in guinea pigs. The cage-contents were retrieved after 2, 4 and 32 wk of implantation and processed for light and electron microscopy. The cages revealed delicate connective tissue that grew in through perforations on the cage-wall. The crystals were densely surrounded by numerous macrophages and multinucleated giant cells, forming a well-circumscribed area of tissue reaction. The cells, however, were unable to eliminate the crystals during an observation period of 8 months. The congregation of macrophages and giant cells, known to be major sources of apical inflammatory and bone resorptive mediators, suggest that accumulation of cholesterol crystals can be a factor in the failure of certain apical periodontitis lesions to resolve after conventional root-filling therapy.

Animals

Gutta-percha-stimulated mouse macrophages release factors that activate the bone resorptive system of mouse calvarial bone.

This study investigated the capacity of macrophages exposed to gutta-percha particles to produce factors affecting bone metabolism. Peritoneal mouse macrophages were isolated and incubated with and without gutta-percha particles, and the supernatants were assessed for bone resorbing activity by adding macrophage-conditioned media to cultures of neonatal mouse calvarial bones. Bone resorption was measured by mineral mobilization (45Ca release) and matrix degradation (3H from [3H]proline labelled bones). The results showed that supernatant from gutta-percha-stimulated macrophages enhanced bone resorption. This effect was related to the amount of gutta-percha, and the concentration and time of exposure to the conditioned media. Stimulated macrophages released enhanced amounts of prostaglandins E2 and I2; however, indomethacin, which inhibits the prostanoid response, had no effect on bone resorbing activity. The stimulatory effect on bone resorption was inhibited by calcitonin, interleukin-1 receptor antagonistic protein, and by antiserum neutralizing mouse interleukin-1alpha(IL-1alpha), but not by anti-IL-1beta. Filtration experiments revealed that the molecules involved in the resorption activity had an apparent molecular weight between 3000 and 30,000 Da. These experiments show that mouse peritoneal macrophages, when exposed to gutta-percha particles, release factors which have a bone resorbing activity that is primarily due to enhanced production of IL-1alpha.

Animals

Restriction fragment-length polymorphism analysis of 16S rDNA from oral asaccharolytic Eubacterium species amplified by polymerase chain reaction.

Restriction fragment-length polymorphism (RFLP) analysis of 16S rDNA amplified by polymerase chain reaction was used to generate restriction profiles of the type strains of oral asaccharolytic Eubacterium species, that is, Eubacterium brachy, Eubacterium exiguum, Eubacterium lentum, Eubacterium minutum, Eubacterium nodatum, Eubacterium saphenum, Eubacterium timidum and 33 asaccharolytic Eubacterium strains isolated from oral sites. The 16S rRNA gene sequences from isolated genomic DNA samples were amplified by polymerase chain reaction (PCR). PCR products were purified and characterized by single digestions with 7 restriction endonucleases. Among the 7 endonucleases, HpaII was found to discriminate the respective reference strains. Twenty-three isolates, out of 33, were assigned to one of the reference species, on the basis of their restriction profiles by digestion with HpaII. The remaining 10 isolates could not be assigned to any of the established species and constituted 4 distinct groups, each of which may be a new species.

DNA, Bacterial

Bacteria inhibit biosynthesis of bone matrix proteins in human osteoblasts.

The effect of extracts from Staphylococcus aureus and Staphylococcus epidermidis on bone matrix production were assessed by analyzing the biosynthesis of osteocalcin and Type I collagen in a human osteoblastic osteosarcoma cell line (MG-63). In MG-63 cells, extracts from Staphylococcus aureus and Staphylococcus epidermidis decreased 1,25(OH)2-vitamin D3 stimulated osteocalcin biosynthesis, and insulin-like growth factor I induced production of Type I collagen in a concentration dependent manner. The basal rate of osteocalcin and Type I collagen formation was unaffected by the bacterial extracts. The inhibitory effect of the bacteria on osteocalcin biosynthesis was seen after 24 hours of treatment and was maintained for at least 96 hours. The extracts of Staphylococcus aureus and Staphylococcus epidermidis enhanced prostaglandin E2 formation in the MG-63 cells. Abolition of the prostaglandin E2 response by treatment with indomethacin and flurbiprofen did not affect bacteria induced inhibition of osteocalcin production. Stimulation of osteocalcin biosynthesis by 1,25(OH)2-vitamin D3 was associated with a decreased rate of cell proliferation. The inhibitory action of the bacterial extracts was not linked to any inhibition of [3H]-thymidine incorporation into deoxyribonucleic acid. These data show that extracts of Staphylococcus aureus and Staphylococcus epidermidis have the ability to inhibit the biosynthesis of bone matrix proteins by a nonprostaglandin and noncytotoxic dependent mechanism and suggest that bone loss in inflammatory processes containing Staphylococcus aureus or Staphylococcus epidermidis may not be caused only by enhanced bone resorption but also by decreased bone formation.

Calcitriol

Influence of infection at the time of root filling on the outcome of endodontic treatment of teeth with apical periodontitis.

This study investigated the role of infection on the prognosis of endodontic therapy by following-up teeth that had had their canals cleaned and obturated during a single appointment. The root canals of 55 single-rooted teeth with apical periodontitis were thoroughly instrumented and irrigated with sodium hypochlorite solution. Using advanced anaerobic bacteriological techniques, post-instrumentation samples were taken and the teeth were then root-filled during the same appointment. All teeth were initially infected; after instrumentation low numbers of bacteria were detected in 22 of 55 root canals. Periapical healing was followed-up for 5 years. Complete periapical healing occurred in 94% of cases that yielded a negative culture. Where the samples were positive prior to root filling, the success rate of treatment was just 68%--a statistically significant difference. Further investigation of three failures revealed the presence of Actinomyces species in each case; no other specific bacteria were implicated in failure cases. These findings emphasize the importance of completely eliminating bacteria from the root canal system before obturation. This objective cannot be reliably achieved in a one-visit treatment because it is not possible to eradicate all infection from the root canal without the support of an inter-appointment antimicrobial dressing.

Bacteria

[A study of post-traumatic stress reactions among war refugees based on medical records. A standard model may support the treatment].

A special project designed to aid Bosnian refugees in Stockholm County was initiated and funded by the County Council in February 1994. The present study, a joint endeavour involving the Bosnian Project, the Centre for Trauma and Torture Diagnostics, and the Red Cross Centre for torture victims, was designed to fulfil the following aims: To characterise the patient population in terms of social situation, trauma history, symptoms and treatment; To review accumulated experience at the three centres; To outline and discuss the treatment; and To suggest new avenues for further research concerning refugees and trauma. To obtain comparable material for this retrospective review of case records at the three centres, a special case chart format was adopted, covering pre-war and pre-treatment information on general background, social situation, health status and medication, as well as trauma history, symptoms and treatment. Most patients reported good pre-war health, but at the time of the study exhibited manifest effects of trauma history; 95 per cent reported mental problems, and 39 per cent medical disease or physical disorders. These refugees were thus characterised by manifest post-traumatic stress symptoms and need of treatment. Despite their relatively short time in Sweden, almost all had permanent residence permits and fixed addresses. The absence of a comparable control group precluded comparison with other refugee or normal populations. The findings suggest the need of co-ordination and co-operation between the special facilities available to refugees, and of a standardised format for case records.

Adult

Bradykinin and thrombin synergistically potentiate interleukin 1 and tumour necrosis factor induced prostanoid biosynthesis in human dental pulp fibroblasts.

The effects of and interactions between the six inflammatory mediators interleukin-1 (IL-1), tumour necrosis factor (TNF), gamma-interferon (INF-gamma), transforming growth factor-beta (TGF-beta), bradykinin (BK) and thrombin on prostanoid biosynthesis in primary cultures of human, dental, pulp fibroblasts were examined. IL-1 alpha, IL-1 beta, TNF-alpha and TNF-beta caused a time- and concentration-dependent enhancement of prostaglandin E2 (PGE2) formation in the fibroblasts. The onset of action was delayed 1-2 h and maximal response was seen after 24 h. In contrast, BK and thrombin caused a burst of PGE2 formation with maximal response after 10 min. BK (1 microM) and thrombin (3 U/ml) synergistically potentiated IL-1 alpha and IL-1 beta stimulated PGE2 formation in 24 h cultures. The effect of BK and thrombin on IL-1 beta enhanced PGE2 formation was seen both at suboptimal and optimal concentrations of IL-1 beta without affecting the sensitivity to IL-1 beta. BK and thrombin also synergistically potentiated the stimulatory effect of TNF-alpha and TNF-beta on PGE2 formation. The synergistic interactions between BK and IL-1 alpha, IL-1 beta and TNF-alpha were seen after 2-4 h of treatment. BK analogues with affinity to BK B2-receptors, but not to BK B1-receptors, were able to synergistically potentiate IL-1 beta and TNF-alpha-enhanced PGE2 production. The synergistic stimulation of PGE2 formation by IL-1, TNF and BK was abolished by indomethacin and flurbiprofen. Preincubation with IL-1 beta and TNF-alpha for 24 h resulted in a substantial amplification of the PGE2 response to a subsequent 24 h challenge with BK in the absence of cytokine. Similarly, when the pulp fibroblasts were preincubated with or without IL-1 beta or TNF-alpha for 24 h and then challenged with exogenous arachidonic acid for 60 min, PGE2 formation was significantly enhanced in cytokine pretreated cells. BK potentiated cytokine induced amplification of the PGE2 response to arachidonic acid. gamma-IFN and TGF-beta did not enhance PGE2 formation, nor did these cytokines potentiate IL-beta or TNF-alpha-induced PGE2 production. These data show that proinflammatory mediators such as BK and thrombin act in concert with IL-1 and TNF in stimulating prostanoid formation in human pulpal fibroblasts and that the action of BK is linked to BK B2-receptors. The results are compatible with the view that enhanced metabolism of arachidonic acid, probably due to increased activation or de novo synthesis of cyclooxy-genase(s), is involved in the mechanism by which IL-1, TNF, BK and thrombin interact.

Bradykinin

Effects of bradykinin and thrombin on prostaglandin formation, cell proliferation and collagen biosynthesis in human dental-pulp fibroblasts.

Bradykinin and thrombin caused a time- and dose-dependent stimulation of prostanoid biosynthesis in human dental-pulp fibroblasts, as assessed by the release of prostaglandin E2 (PGE2) and 6-keto-prostaglandin F1 alpha (the stable breakdown product of prostacyclin). The stimulatory effect of bradykinin and thrombin on PGE2 biosynthesis was maximal within 5-10 min. The concentration of bradykinin producing half-maximal stimulation (EC50) of PGE2 and prostacyclin formation was 10 nM. EC50 for thrombin-induced formation of PGE2 and prostacyclin were 0.05 and 0.2 U/ml, respectively. Bradykinin analogues with affinity to the bradykinin B2 receptor, but not those with affinity to the B1 receptor, caused a burst of PGE2 formation. The stimulatory action of bradykinin and thrombin on PGE2 biosynthesis was abolished by two structurally different cyclo-oxygenase inhibitors and significantly reduced by two corticosteroids. Thrombin dose-dependently enhanced the incorporation of [3H]-thymidine into DNA in pulpal fibroblasts by a mechanism that was unrelated to the effect on prostanoid biosynthesis. Bradykinin did not affect thymidine incorporation. Thrombin, but not bradykinin, stimulated the biosynthesis of type 1 collagen in the pulpal fibroblasts. The stimulatory effect of thrombin on collagen biosynthesis was not affected by cyclo-oxygenase inhibitors. These data show that human dental-pulp fibroblasts are equipped with receptors for bradykinin and thrombin linked to enhanced prostanoid biosynthesis. Occupancy of the thrombin receptors also leads to a prostaglandin-independent stimulation of cell proliferation and collagen biosynthesis.

6-Ketoprostaglandin F1 alpha

Effect of sonicated material from Fusobacterium nucleatum on the functional capacity of accessory cells derived from dental pulp.

Fusobacterium nucleatum-derived components were studied for their effect on the ability of accessory pulpal cells to induce T-lymphocyte proliferation. In initial experiments, spleen cells were used to establish concentration ranges at which testing of the effect of the bacterial components would be suitable. At low concentrations, bacterial substances induced a stimulatory effect on the proliferation rate of spleen cells, but not when purified T-lymphocytes were incubated in the presence of concanavalin A and pulpal accessory cells. At increasing concentrations, a dose-dependent reduction was observed. Pretreatment with the bacterial products for 4 h gave a stimulatory effect in the absence of concanavalin A for both spleen cells and T-lymphocytes. Removal of low-molecular weight components (<3.5 kDa) did not influence the inhibitory effect, whereas preheating the bacterial substances to 100 degrees C eliminated the observed inhibition. Findings show that heat-sensitive, high-molecular-weight components of F. nucleatum can evoke a concentration-dependent stimulatory or suppressive effect on the proliferation rate of T-lymphocytes in the presence of accessory cells derived from the dental pulp.

Analysis of Variance

Tissue reaction to gutta-percha particles of various sizes when implanted subcutaneously in guinea pigs.

Tissue reaction to gutta-percha was studied using subcutaneously implanted Teflon cages in guinea pigs. Gutta-percha was tested in three forms: (i) as large particles prepared by dividing gutta-percha cones into pieces, (ii) as fine particles prepared by ball-milling of gutta-percha, and (iii) as particles produced by dissolving gutta-percha in rosin-chloroform. Gutta-percha evoked two distinct types of tissue response. The large pieces were well encapsulated and the surrounding tissue was free of inflammation. The fine particles evoked an intense, localized tissue response, characterized by the presence of macrophages and multinucleated giant cells. The rosin-chloroform treated gutta-percha induced a similar tissue reaction to that observed with the fine particles of gutta-percha. In addition, cell remnants were present in association with the material, which indicates an initial toxicity to rosin-chloroform treated gutta-percha. These results show that the size and surface character of gutta-percha can determine the type of tissue reaction to the material. The accumulation of macrophages around gutta-percha may be an important factor in the impairment of healing of periapical lesions when teeth are root filled with excess material.

Animals

Taxonomy, ecology, and pathogenicity of the root canal flora.

The bacteria present in infected root canals include a restricted group of species compared with the total flora of the oral cavity. Conditions exist in the root canal that permit the growth of anaerobic bacteria capable of fermenting amino acids and peptides, whereas bacteria that mainly obtain energy by fermenting carbohydrates are restricted by lack of available nutrients. During the course of infection interrelationships develop between microbial species and population shifts are produced as a result of these interactions. Strong associations between certain species are present. These associations are most likely based on nutritional demands and nutritional relationships. The pathogenicity of the polymicrobial root canal flora is dependent on bacterial synergy.

Bacteria, Anaerobic

Radicular cyst affecting a root-filled human tooth: a long-term post-treatment follow-up.

Apical periodontitis is caused primarily by microorganisms residing in the root canals of affected teeth. Nevertheless, there is convincing evidence implicating other independent factors that adversely affect the outcome of conventional root canal therapy. In this paper, morphological evidence is presented in support of the potential role of two endogenous factors that may interfere with post-endodontic healing of the periapex. The specimens consisted of a surgical biopsy of an asymptomatic periapical lesion which persisted for a follow-up period of 44 months. The biopsy was processed for correlated light and electron microscopy. The lesion was characterized by the presence of a large central lumen lined by a stratified squamous epithelium. The most striking feature of the lesion was the presence of vast numbers of cholesterol crystals which congregated in the connective tissue surrounding the cyst cavity. Extensive light and electron microscopic investigation of the apical part of the root canal and the lesion failed to reveal the presence of microorganisms. These findings strongly suggest that intrinsic factors like the accumulation of certain tissue break-down products such as cholesterol crystals, and the cystic condition of the lesion itself, can adversely affect the healing process of the periapex following root canal therapy. Consequently, such apical lesions can remain refractory to conventional endodontic therapy for long periods of time.

Adult

Ecology of the root canal flora.

The root canal represents a special environment in which selective pressures result in the establishment of a restricted group of the oral flora. Population shifts occur over time with obligate anaerobes ultimately dominating the bacterial mix. Bacterial interrelationships and the nutritional supply are key factors in determining the outcome of the infection. Endodontic treatment, apart from directly eliminating bacteria, can completely disrupt the delicate ecology and deprive persisting bacteria of their nutritional source.

Bacteria, Anaerobic

Pathogenicity of Actinomyces israelii and Arachnia propionica: experimental infection in guinea pigs and phagocytosis and intracellular killing by human polymorphonuclear leukocytes in vitro.

Strains of Actinomyces israelii and Arachnia propionica, isolated from clinical cases of failed endodontic therapy, were examined for: (i) their ability to survive and establish themselves in the soft connective tissue that grew into subcutaneously implanted tissue cages in guinea pigs; (ii) cell-surface hydrophobicity; and (iii) phagocytosis and killing by human polymorphonuclear leukocytes (PMNs) under aerobic and anaerobic conditions. Bacteria were inoculated into the tissue cages in guinea pigs and the cage contents were retrieved after 1, 7, 14 and 21 d for culturing and light and electron microscopy. Both bacterial species showed substantial decline in the number of bacteria by day 7 after the inoculation. Thereafter, the A. israelii strain recovered and, by day 21, had started to increase in number. Light and electron microscopy revealed the formation of typical actinomycotic colonies. A. propionica, on the other hand, continued to decline in number during the entire period of experimental infection and did not form colonies. Both strains were hydrophobic, readily phagocytosed and were efficiently killed by human PMNs under aerobic and anaerobic conditions in vitro. These results suggest that the pathogenicity of A. israelii is due to its ability to establish characteristic cohesive colonies consisting of branching filamentous organisms that are enmeshed in an extracellular matrix. It seems that the organisms existing in such colonies can collectively evade destruction and elimination by host phagocytic cells, whereas in vitro suspensions of the bacteria are easily phagocytosed and efficiently killed by PMNs. With respect to A. propionica, further investigations are necessary to understand its pathogenicity.

Actinomyces

Associations between microbial species in dental root canal infections.

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.

Actinomyces

The antimicrobial effect of calcium hydroxide as a short-term intracanal dressing.

The antibacterial effect of calcium hydroxide as a short-term intracanal dressing was clinically evaluated by applying the medicament for 10 minutes or 7 days in root canals of teeth with periapical lesions. The results showed that the 7-day dressing efficiently eliminated bacteria which survived biomechanical instrumentation of the canal, while the 10-minute application was ineffective.

Bacteria, Anaerobic