Bacterial colonisation of suture material after routine neurosurgical procedures: relevance for wound infection.

BACKGROUND: Wound healing impairment is a serious problem in surgical disciplines which may be associated with chronic morbidity, increased cost and patient discomfort. Here we aimed to investigate the relevance of bacterial colonisation on suture material using polymerase chain reaction (PCR) to detect and taxonomically classify bacterial DNA in patients with and without wound healing problems after routine neurosurgical procedures. METHODS: Repeat surgery was performed in 25 patients with wound healing impairment and in 38 patients with well-healed wounds. To determine the presence of bacteria, a 16S rDNA-based PCR detection method was applied. Fragments of 500 bp were amplified using universal primers which target hypervariable regions within the bacterial 16S rRNA gene. Amplicons were separated from each other by single-strand conformation polymorphism (SSCP) analysis, and finally classified using Sanger sequencing. RESULTS: PCR/SSCP detected DNA of various bacteria species on suture material in 10/38 patients with well-healed wounds and in 12/25 patients with wound healing impairment including Staphylococcus aureus, Staphylococcus epidermidis, Propionibacterium acnes and Escherichia coli. Microbiological cultures showed bacterial growth in almost all patients with wound healing impairment and positive results in PCR/SSCP (10/12), while this was the case in only one patient with a well-healed wound (1/10). CONCLUSIONS: Colonisation of suture material with bacteria occurs in a relevant portion of patients with and without wound healing impairment after routine neurosurgical procedures. Suture material may provide a nidus for bacteria and subsequent biofilm formation. Most likely, however, such colonisation of sutures is not a general primer for subsequent wound infection.

Development of a flow chamber system for the reproducible in vitro analysis of biofilm formation on implant materials.

Since the introduction of modern dental implants in the 1980s, the number of inserted implants has steadily increased. Implant systems have become more sophisticated and have enormously enhanced patients' quality of life. Although there has been tremendous development in implant materials and clinical methods, bacterial infections are still one of the major causes of implant failure. These infections involve the formation of sessile microbial communities, called biofilms. Biofilms possess unique physical and biochemical properties and are hard to treat conventionally. There is a great demand for innovative methods to functionalize surfaces antibacterially, which could be used as the basis of new implant technologies. Present, there are few test systems to evaluate bacterial growth on these surfaces under physiological flow conditions. We developed a flow chamber model optimized for the assessment of dental implant materials. As a result it could be shown that biofilms of the five important oral bacteria Streptococcus gordonii, Streptococcus oralis, Streptococcus salivarius, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans, can be reproducibly formed on the surface of titanium, a frequent implant material. This system can be run automatically in combination with an appropriate microscopic device and is a promising approach for testing the antibacterial effect of innovative dental materials.

The Peri-Implant and Periodontal Microbiota in Patients with and without Clinical Signs of Inflammation.

Late implant failures, caused by the inflammation of surrounding tissues are a problem in implant dentistry. The path of bacterial transmission from teeth to implants is not completely understood. Therefore, the purpose of this study was to analyze intraindividual bacterial transmission characterizing subgingival microbiomes in teeth and implants, both in healthy subjects and in those with signs of periodontitis or peri-implantitis. Samples of peri-implant and dental sulcus fluid were collected. To identify the predominant microbiota, amplified fragments of bacterial 16S rRNA gene were separated by single strand conformation polymorphism analysis, sequenced and taxonomically classified. A total of 25 different predominant genera were found in the diseased group and 14 genera in the healthy group. Species richness did not differ significantly between implants, neighboring teeth and teeth with largest probing depth in the diseased group. Additionally, no differences between teeth and implants in the healthy group were detected. In contrast, microbial diversity varied between the different sampling points. Species richness is similar in healthy and diseased sites, but the composition of the bacterial community differed within the individual subjects. The underlying analyses strongly suggest that complete transmission from neighboring teeth to implants is unlikely.

Pyrosequencing of supra- and subgingival biofilms from inflamed peri-implant and periodontal sites.

BACKGROUND: To investigate the microbial composition of biofilms at inflamed peri-implant and periodontal tissues in the same subject, using 16S rRNA sequencing. METHODS: Supra- and submucosal, and supra- and subgingival plaque samples were collected from 7 subjects suffering from diseased peri-implant and periodontal tissues. Bacterial DNA was isolated and 16S rRNA genes were amplified, sequenced and aligned for the identification of bacterial genera. RESULTS: 43734 chimera-depleted, denoised sequences were identified, corresponding to 1 phylum, 8 classes, 10 orders, 44 families and 150 genera. The most abundant families or genera found in supramucosal or supragingival plaque were Streptoccocaceae, Rothia and Porphyromonas. In submucosal plaque, the most abundant family or genera found were Rothia, Streptococcaceae and Porphyromonas on implants. The most abundant subgingival bacteria on teeth were Prevotella, Streptococcaceae, and TG5. The number of sequences found for the genera Tannerella and Aggregatibacter on implants differed significantly between supra- and submucosal locations before multiple testing. The analyses demonstrated no significant differences between microbiomes on implants and teeth in supra- or submucosal and supra- or subgingival biofilms. CONCLUSION: Diseased peri-implant and periodontal tissues in the same subject share similiar bacterial genera and based on the analysis of taxa on a genus level biofilm compositions may not account for the potentially distinct pathologies at implants or teeth.

Microbial diversity of peri-implant biofilms on implant fixed bar and telescopic double crown attachments.

One of the principal problems in oral implantation is inflammation of peri-implant hard and soft tissues caused by bacterial biofilms. The purpose of the present study was to evaluate the microbial diversity of peri-implant biofilms on 2 different implant-anchored attachment types in vivo. Samples of peri-implant sulcus fluid were collected from 8 patients with implant-supported bar attachments and 8 patients with implant-anchored telescopic double crown attachments. Samples of sulcus fluid of the adjacent teeth were also collected from the partially edentulous patients with implant fixed telescopic double crowns. The mixed amplicons of 16S rRNA fragments of different bacterial origins were separated by use of single-strand conformation polymorphism analysis to identify the predominant bacterial genera. With 3.5 ± 2.1 different predominant bacterial genera in the sulcus fluid surrounding implant-supported bar attachments and 6.3 ± 3.1 different predominant genera in the sulcular fluid of implant-anchored double crown attachments, the differences were not statistically significant (P = .11). The microbial diversity in the sulcus fluid surrounding the remaining dentition was similar to that of the implant fixed telescopic attachments (6.3 ± 2.1). Aside from host response and other individual factors, the microbial diversity of peri-implant biofilms seems to be impaired by cofactors such as the possibility of cleaning the implant-supported supraconstructions and the different plaque-retaining sites. Nevertheless, these differences do not lead to statistically significant differences in the microbial diversity of peri-implant plaques.

Metagenomic analysis of the peri-implant and periodontal microflora in patients with clinical signs of gingivitis or mucositis.

The long-term success of osseointegrated oral implants is endangered by inflammation of peri-implant hard and soft tissues caused by bacterial biofilms that may have been initiated by bacterial transmission from the adjacent dentition. The present study aimed to compare the bacterial communities at inflamed implant and tooth sites by broad-range PCR techniques to evaluate the etiological processes of peri-implant and periodontal diseases and potential future therapeutic strategies. Eighteen samples of peri-implant and periodontal microflora were collected from nine partially edentulous patients with implant-retained crowns or bridges revealing clinical signs of gingivitis or mucositis. The clinical parameters plaque index (PI), probing depth (PD), and bleeding on probing were recorded. Amplified fragments of bacterial 16S rRNA genes were separated by use of single-strand conformation polymorphism analysis, and sequences were determined to identify the predominant bacterial genera. The clinical parameters PI and PD were significantly different at implants (PI = 0.4 ± 0.7, PD = 3.1 ± 0.6 mm) compared with teeth (PI = 1.8 ± 0.8, PD = 2.5 ± 0.2 mm). A total of 20 different genera were found at the inflamed tooth and implant sites. The microbial diversity of the microflora surrounding the remaining dentition (12.0 ± 3.8) was significantly higher (p = 0.01) than the diversity of the peri-implant microflora at implant-retained crowns or bridges (6.3 ± 2.3). Within the limitations of the present study, the microbial diversity of the investigated implants and teeth with clinical signs of mucositis or gingivitis exhibits substantial differences, demonstrating that transmission of the complete bacterial microflora from teeth to implants could be excluded. Furthermore, broad-range molecular biological detection methods specify bacterial genera and species in the peri-implant and periodontal microflora which were not in the focus of research interests so far.