Human mesenchymal stem cells (hMSCs) are multipotent by nature and are originally isolated from bone marrow. In light of a future application of hMSCs in the oral cavity, a body compartment with varying oxygen partial pressures and an omnipresence of different bacterial species i.e. periodontitis pathogens, we performed this study to gain information about the behavior of hMSC in an anaerobic system and the response in interaction with oral bacterial pathogens. We established a model system with oral pathogenic bacterial species and eukaryotic cells cultured in anaerobic conditions. The facultative anaerobe bacteria Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were studied. Their effects on hMSCs and primary as well as permanent gingival epithelial cells (Ca9-22, HGPEC) were comparatively analyzed. We show that hMSCs cope with anoxic conditions, since 40% vital cells remain after 72 h of anaerobic culture. The Ca9-22 and HGPEC cells are significantly more sensitive to lack of oxygen. All bacterial species reveal a comparatively low adherence to and internalization into hMSCs (0.2% and 0.01% of the initial inoculum, respectively). In comparison, the Ca9-22 and HGPEC cells present better targets for bacterial adherence and internalization. The production of the pro-inflammatory chemokine IL-8 is higher in both gingival epithelial cell lines compared to hMSCs and Fusobacterium nucleatum induce a time-dependent cytokine secretion in both cell lines. Porphyromonas gingivalis is less effective in stimulating secretion of IL-8 in the co-cultivation experiments. HMSCs are suitable for use in anoxic regions of the oral cavity. The interaction with local pathogenic bacteria does not result in massive pro-inflammatory cytokine responses. The test system established in this study allowed further investigation of parameters prior to set up of oral hMSC in vivo studies.
In patients with periodontitis, it is highly likely that local (progenitor) cells encounter pathogenic bacteria. The purpose of this in vitro study was to elucidate how human dental follicle stem cells (hDFSC) react towards a direct challenge with anaerobic periodontal pathogens under their natural oxygen-free atmosphere. HDFSC were compared to human bone marrow mesenchymal stem cells (hBMSC) and differentiated primary human gingival fibroblasts (hGiF), as well as permanent gingival carcinoma cells (Ca9-22). The different cell types were investigated in a co-culture system with Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum). The viability of the cells and pathogens under anaerobic conditions, as well as interactions in terms of adherence and internalization, were examined. Additionally, the release of pro-inflammatory interleukin-8 (IL-8) and anti-inflammatory interleukin-10 (IL-10) was quantified via enzyme-linked immunosorbent assay. The bacteria adhered less efficiently to hDFSC compared to Ca9-22 (P. gingivalis: 0.18% adherence to hDFSC; 3.1% adherence to Ca9-22). Similar results were observed for host cell internalization (F. nucleatum: 0.002% internalization into hDFSC; 0.09% internalization into Ca9-22). Statistically significantly less IL-8 was secreted from hDFSC after stimulation with F. nucleatum and P. gingivalis in comparison with hGiF (F. nucleatum: 2080.0 pg/ml – hGiF; 19.7 pg/ml – hDFSC). The IL-10 response of the differentiated cells was found to be low in relation to their pro-inflammatory IL-8 response. The results indicate that dental stem cells are less prone to interactions with pathogenic bacteria than differentiated cells in an anaerobic environment. Moreover, during bacterial challenge, the stem cell immune response seems to be more towards an anti-inflammatory reaction. For a potential future therapeutic use of hDFSC, these findings support the idea of a save application.
Background: The treatment of acute pain is part of everyday dental practice. Often, these symptoms result from years of patients' inadequate or missing dental routines and lead to a reduction in the quality of life or health of the patients and to high costs for the health care system. Despite the enormous advantages of modern dentistry, many patients avoid going to the dentist. Therefore, the study aimed to determine the reasons and behaviours that cause patients to avoid visits to the dentist. Methods: We conducted semi-structured interviews with patients who had an above-average DMFT index and had been going to the dentist only irregularly for years. The sample participants were recruited from the northern German region of Mecklenburg-Western Pomerania. 20 individual interviews were recorded, transcribed verbatim and coded. We used a qualitative framework approach to code the transcripts in order to establish a consensus among the researchers. Ultimately, through discussions and reviews of the attributes and meaning of the topics, a typology could be established. Results: A typology of patients who avoid the dentist was developed. Four independent characteristic patterns of dentist avoidance could be developed: avoiding the dentist due to "distance" (type A; includes subtype A1 "avoiding the dentist due to negligence" and subtype A2 "dental avoidance due to neutralization"), "disappointment" (type B), "shame" (type C), and "fear" (type D). Using the typology as a generalised tool to determine the minimum and maximum contrasts, it was possible to capture the diversity and multidimensionality of the reasons and behaviours for avoidance. All patients had negative dental experiences, which had led to different avoidance patterns and strategies. Conclusions: The identified avoidance characteristics represent a spectrum of patients from Northern Germany who avoid going to the dentist. This is the first comprehensive study in Germany representing avoidance behaviour of dentist patients in the form of a typology. The results suggest that dentistry also needs qualitative research to better understand patient characteristics and provide direct access to patients who avoid regular dental visits. Thus, the results make a potentially fundamental contribution to the improvement of dental care and enrich its understanding.
Efforts to improve the performance of rotary NiTi instruments by enhancing the properties of NiTi alloy, or their manufacturing processes rather than changes in instrument geometries have been reported. The aim of this study was to compare in-vitro the shaping ability of three different rotary nickel-titanium instruments produced by different manufacturing methods.
Periodontitis is a chronic inflammatory condition of the periodontium, which often leads to tooth loss. Recently, statins have emerged as potent anti-inflammatory agents with pleiotropic effects that can potentially outperform conventional periodontal treatments. However, the clinical application of statins is limited by the lack of suitable drug carriers that fit the periodontal region and provide a controlled local drug release. In this study, we address the critical gap in localized periodontal drug delivery and introduce an ultrasound-assisted technique to encapsulate atorvastatin within alginate microparticles (10–400 µm in diameter)—a simple, scalable, and biocompatible solution. While ultrasound is widely used in polymer synthesis, its application in alginate polymerization remains underexplored. To mimic physiological conditions, particles were incubated in artificial saliva at 37 °C, with drug release being analyzed via high-performance liquid chromatography. A methylcellulose-based hydrogel served as a conventional reference product. Results revealed that alginate particles exhibited at least a 10-fold increase in mean dissolution time compared to the methylcellulose gel, indicating superior stability. Increasing atorvastatin concentration extended the time interval needed for 50% of the drug to be released (t50%) from 1 h to 11 h, maintaining the overall drug diffusion level for several days. Further analysis showed that covalent cross-linking of alginate with divinyl sulfone significantly delayed the initial drug release by 3 h (p < 0.05) due to the additional molecular stabilization. These findings underscore the utility of ultrasonic atomization for the processing of alginate-based formulations. Given the ease of production, biocompatibility, and small size, successfully fabricated alginate particles represent a promising carrier for delivery of statins or other related drugs in clinical dentistry.
1 September 2015 in Current Directions in Biomedical Engineering
Inkjet printing technology has the potential to be used for seeding of viable cells for tissue engineering approaches. For this reason, a piezoelectrically actuated, drop-on-demand inkjet printing system was applied to deliver viable human dental follicle stem cells (hDFSC) of sizes of about 15 μm up to 20 μm in diameter. The purpose of these investigations was to verify the stability of the printing process and to evaluate cell viability post printing. Using a Nanoplotter 2.1 (Gesim, Germany) equipped with the piezoelectric printhead NanoTip HV (Gesim, Germany), a concentration of 6.6 ×106 cells ml−1 in DMEM with 10% fetal calf serum (FCS) could be dispensed. The piezoelectric printhead has a nominal droplet volume of ~ 400 pl and was set to a voltage of 75 V and a pulse of 50 μs while dosing 50 000 droplets over a time of 100 seconds. The volume and trajectory of the droplet were checked by a stroboscope test right before and after the printing process. It was found that the droplet volume decreases significantly by 35% during printing process, while the trajectory of the droplets remains stable with only an insignificant number of degrees deviation from the vertical line. It is highly probable that some cell sedimentations or agglomerations affect the printing performance. The cell viability post printing was assessed by using the Trypan Blue dye exclusion test. The printing process was found to have no significant influence on cell survival. In conclusion, drop-on-demand inkjet printing can be a potent tool for the seeding of viable cells.