Non-contact removal of coadhering and non-coadhering bacterial pairs from pellicle surfaces by sonic brushing and de novo adhesion

Coadhesion between oral microbial pairs is an established factor in the spatiotemporal development and prevalence of mixed-species communities in early dental plaque in vivo. This study compares removal and de novo adhesion of pairs of coadhering and non-coadhering oral actinomyces and streptococci by sonic brushing on salivary pellicles in a non-contact mode as a function of the distance between the brush and the pellicle surface in vitro. First, actinomycetes were adhered to a pellicle surface, after which streptococci suspended in saliva were allowed to adhere. Removal was examined by non-contact, sonic brushing with a wetted brush on a either a wetted or a substratum immersed to a depth of 7 mm. After brushing, de novo adhesion of streptococci to brushed pellicles was studied. For coadhering and non-coadhering pairs, 34% and 9%, respectively, of the adhering bacteria were involved in aggregates comprising more than 10 organisms. Non-contact, sonic brushing removed up to 99% of the adhering bacteria, regardless of the state of immersion of the substratum. Bacterial removal decreased with increasing distance of up to 6 mm between brush and pellicle surface. For the non-coadhering pair, subsequent exposure of pellicles to a streptococcal suspension yielded about 6% of bacteria involved in large aggregates. Alternatively, de novo adhesion of the coadhering streptococcal strain to pellicles brushed on the wetted substratum yielded 31% of bacteria involved in large aggregates, but after brushing the immersed substratum only 12% of the adhering bacteria were found in large aggregates. It is concluded that non-contact sonic brushing, under immersion, removes high percentage of adhering bacterial pairs up to a distance of 6 mm between the brush and the pellicle surface. However, non-contact, sonic brushing with only a thin wet film on the substratum may leave footprints to which streptococci preferentially adhere.     

Differences in mouse strain influence leukocyte and immunoglobulin phenotype response to Porphyromonas gingivalis

This study examined the nature of the infiltrating cells in Porphyromonas gingivalis-induced lesions and immunoglobulins in the serum samples of BALB/c (H-2^d), C57BL6 (H-2^b), DBA/2J (H-2^d) and CBA/CaH (H-2^k) mice. Mice were immunized intraperitoneally with P. gingivalis outer membrane antigens or sham-immunized with phosphate-buffered saline followed by subcutaneous challenge with live organisms 1 week after the final immunization. The resulting skin abscesses were excised 7 days later, cryostat sections cut and an immunoperoxidase method used to detect the presence of CD4⁺ and CD8⁺ T-cell subsets, CD14⁺ macrophages and CD19⁺ B cells. Peroxidase positive neutrophils and IgG1- and IgG2a-producing plasma cells were also identified. Anti P. gingivalis IgG1 and IgG2a subclass antibodies were determined in serum obtained by cardiac puncture. Very few CD8⁺ T cells and CD19⁺ B cells were found in any of the lesions. The percentages of CD4⁺ cells, CD14⁺ cells and neutrophils were similar in lesions of immunized BALB/c and C57BL6 mice, with a trend towards a higher percentage of CD14⁺ cells in sham-immunized mice. The percentage of CD14⁺ cells was higher than that of CD4⁺ cells in immunized compared with sham-immunized DBA/2J mice. The percentages of CD4⁺ and CD14⁺ cells predominated in immunized CBA/CaH mice and CD4⁺ cells in sham-immunized CBA/CaH mice. The percentage of neutrophils in immunized CBA/CaH mice was significantly lower than that of CD14⁺ cells and CD4⁺ cells in sham-immunized mice. IgG1⁺ plasma cells were more dominant than IgG2a⁺ cells in immunized BALB/c, C57BL6 and DBA/2J mice, whereas IgG2a⁺ plasma cells were more obvious in sham-immunized mice. IgG2a⁺ plasma cells were predominant in immunized and sham-immunized CBA/CaH mice. In the serum, specific anti-P. gingivalis IgG2a antibody levels (Th1 response) were higher than IgG1 levels (Th2 response) in sham-immunized CBA/CaH and DBA/2J mice. In immunized BALB/c mice, IgG2a levels were lower than IgG1 levels, while IgG2a levels were higher in immunized C57BL6 mice. In conclusion, this study has shown differences in the proportion of infiltrating leukocytes and in the subclasses of immunoglobulin produced locally and systemically in response to P. gingivalis in different strains of mice, suggesting a degree of genetic control over the response to P. gingivalis.     

Effect of ProRoot MTA mixed with chlorhexidine on apoptosis and cell cycle of fibroblasts and macrophages in vitro

AIM: To compare the percentage of apoptotic cells and the cell cycle profile of fibroblasts and macrophages exposed to either ProRoot mineral trioxide aggregate (MTA) mixed with chlorhexidine (CHX), or exposed to ProRoot MTA mixed with sterile water. METHODOLOGY: Mouse gingival fibroblasts or mouse macrophages were seeded in six-well plates and allowed to attach overnight. Freshly mixed or set (allowed to dry for 24 h) specimens of tooth-coloured (white) ProRoot MTA were prepared with 0.12% CHX gluconate (MTA/CHX) or with sterile water (MTA/H2O). The cells were exposed for 24 h to the MTA specimens, which were placed over permeable membrane inserts to avoid direct contact with the cells. Untreated cells served as controls. Propidium iodide staining followed by flow cytometry was used to evaluate the effects of ProRoot MTA on cell apoptosis and cell cycle. Statistical analyses were performed by one-way anova followed by post-hoc tests with the use of the SigmaStat 2.0 software, and significance was determined at P < or = 0.05. RESULTS: MTA specimens containing CHX induced apoptosis of macrophages and fibroblasts (P < 0.05). In contrast, no change in the proportion of apoptotic cells was observed when sterile water was used to prepare the specimens (P > 0.05). Cell cycle analysis showed that exposure to MTA/CHX decreased the percentage of fibroblasts and macrophages in S phase (DNA synthesis) as compared with exposure to MTA/H2O (P < 0.05). CONCLUSION: This in vitro study demonstrated that the substitution of CHX for sterile water in MTA increases its cytotoxicity. This suggests that the potentially beneficial antimicrobial effect of CHX may be accompanied by an increase in the cytotoxicity of the resulting MTA-based material.     

Tissue destruction in periodontitis: bacteria or cytokines fault?

The pathogenesis of periodontal disease involves the sequential activation of a great variety of components of the host immune response, primarily acting to defend periodontal tissues against bacterial aggression, but also functioning as mediators of tissue destruction. The expression of the disease results from the interaction of host, microbiological agents, and environmental factors. Leukocytes play a critical role in the pathogenesis of the disease, producing different cytokines, chemokines, and other mediators, thus generating a host defense response, as well as inducing tissue inflammation and bone destruction. The aim of this review is to address the role of some inflammatory mediators in response to bacterial aggression in periodontitis.     

Bioreactor cultivation of osteochondral grafts

The clinical utility of tissue engineering depends upon our ability to direct cells to form tissues with characteristic structural and mechanical properties across different hierarchical scales. Ideally, an engineered graft should be tailored to (re)establish the structure and function of the native tissue being replaced. Engineered grafts of such high fidelity would also foster fundamental research by serving as physiologically relevant models for quantitative in vitro studies. The approach discussed here involves the use of human mesenchymal stem cells (hMSC) cultured on custom-designed scaffolds (providing a structural and logistic template for tissue development) in bioreactors (providing environmental control, biochemical and mechanical cues). Cartilage, bone and ligaments have been engineered by using hMSC, highly porous protein scaffolds (collagen; silk) and bioreactors (perfused cartridges with or without mechanical loading). In each case, the scaffold and bioreactor were designed to recapitulate some aspects of the environment present in native tissues. Medium flow facilitated mass transport to the cells and thereby enhanced the formation of all three tissues. In the case of cartilage, dynamic laminar flow patterns were advantageous as compared to either turbulent steady flow or static (no flow) cultures. In the case of bone, medium flow affected the geometry, distribution and orientation of the forming bone-like trabeculae. In the case of ligament, applied mechanical loading (a combination of dynamic stretch and torsion) markedly enhanced cell differentiation, alignment and functional assembly. Taken together, these studies provide a basis for the ongoing work on engineering osreochondral grafts for a variety of potential applications, including those in the craniofacial complex.     

Transverse maxillary distraction with a bone-anchored appliance: dento-periodontal effects and clinical and radiological results

In 29 adult patients presenting with maxillary deficiency, a bone-anchored palatal distractor (Surgi-Tec NV, Brugge, Belgium) was applied after osteotomy of the anterolateral walls of the maxillary sinuses, midpalatal suture, and, eventually, separation of the pterygomaxillary sutures. Expansion proceeded at a rate of 0.33-0.66 mm per day and the device was retained for 4-6 months for consolidation. Active orthodontic therapy was started after 8-10 weeks. The increment of arch width and the perimeter were evaluated using dental casts. Tooth thermal sensitivity and the periodontal side effects of treatment were monitored clinically after distraction, at device removal, and after 1 year. Bone healing was also investigated during the procedure using conventional radiological techniques. This experience confirms that transverse maxillary distraction is an effective technique in adult patients, leading to the formation of new bone. There were no noticeable intraoperative complications, but postsurgical periodontal side effects were documented. The procedure offers advantages over traditional teeth-borne appliances in terms of rapidity of treatment and the absence of mechanical forces acting on the teeth. Further evaluation is required to assess the long-term stability and periodontal consequences of this technique.     

A new "platinum" standard for bone grafting: autogenous stem cells

Autogenous bone has long been considered the gold standard of all bone grafting materials. However, complications have been associated with autogenous bone-harvesting procedures. This article suggests that an alternative approach, grafting with autogenous bone marrow aspirate, may become a new platinum standard. Bone marrow can be extracted from the large flat bones of the body with relative ease and safety, and it provides a rich source of adult stem cells as well as growth factors that facilitate osteogenesis. Mixed with a resorbable matrix or scaffold, bone marrow aspirate has the potential to reconstitute various bony defects in the mouth to reconstruct the severely atrophic maxilla and mandible. The potentiality and plasticity of stem cells have been well documented.