[Review of surgical treatment of suprastomal collapse in tracheostomised children]. / Übersichtsarbeit zur chirurgischen Behandlung des suprastomalen Kollapses bei tracheotomierten Kindern.

BACKGROUND: Suprastomal collapse (SSC) is considered a major late complication of paediatric tracheostomy and can be responsible for decannulation failure in up to 20% of tracheostomised children. Depending on the severity of SSC, surgery may be required. Various strategies and techniques are available, of which the treating with airway team should be aware. OBJECTIVE: This article intends to summarise the aetiology of SSC, its classification, clinical presentation, and the gold standard diagnostic and therapeutic algorithms according to the current literature. MATERIALS AND METHODS: A panel of experts reviewed the available literature on SSC. Published evidence on the different surgical techniques and their advantages and disadvantages was reviewed in detail, and a treatment algorithm created. RESULTS: The gold standard diagnostic procedure for SSC is flexible transnasal laryngotracheoscopy in spontaneous breathing followed by microlaryngoscopy (MLS) under general anaesthesia. Two main types of SSC can be differentiated, which differ in terms of surgical treatment. Purely anterior SSC is usually treated by tracheoplasty using an anterior costal cartilage graft (ACCG). Simple closure of the tracheostomy or excision of SSC with a potassium-titanyl-phosphate (KTP) laser are also described as less invasive approaches. For anterolateral SSC, segmental tracheal resection with end-to-end anastomosis or tracheoplasty with ACCG represent promising treatment options. Tracheal reinforcement with absorbable microplates is also discussed in the literature. With both types of SSC and depending on severity and the age of the child, a watch-and-wait strategy should always be considered. CONCLUSION: Dynamic airway endoscopy in spontaneous breathing followed by MLS in general anaesthesia should always be performed before decannulation. It is particularly important to visualise all segments of the airway during spontaneous breathing. The decision regarding the best surgical option for each child is based on the type and localisation of SSC, as well as on the patient's medical and surgical history and age.

Organ Preservation Strategies in Laryngeal Chondrosarcoma.

OBJECTIVES/HYPOTHESIS: Laryngeal chondrosarcoma (LC) is a rare, slowly growing malignancy. The preferred treatment is laryngeal preservation surgery (LPS). Some patients may require multiple interventions or total laryngectomy (TL). We investigated risk factors for retreatment and TL, and assessed the impact of LPS on oncological and functional outcomes. STUDY DESIGN: Case series METHODS: We searched our institution database for LC. Tumor grading, localization, and margin status were tested as predictors of recurrence and organ preservation. RESULTS: We included 21 patients (seven females, mean age 58 ± 12 years). LPS was applied in 20 (95.2%) of them as a primary procedure. Six patients were treated by transoral approach and 14 received "open-neck" LPS. Fifteen (71.4%) were operated only once, while six patients underwent a total of 15 adjunctive procedures. Additional operations were always performed for recurrence of tumors localized within the cricoid plate. The histological grading was G1 in 81% and G2 in 19%. However, two patients with a primary G1 LC showed a G2 recurrence. Reoperations for recurrence were more frequent among patients with G2 in respect to G1 histology (83% vs. 7%, P < .001). Fifty percent of G2 LC and 8% of G1 underwent TL (P < .05). Margin status had no influence on recurrence rate. CONCLUSIONS: Patients with G2 LC have more recurrences requiring surgery and a higher incidence of TL. Cricoid plate localization is relevant for organ preservation. Margin status signals possible disease persistence, without influencing the need for future surgeries. Need for reoperation entails a risk of not being able to maintain organ functionality. LEVEL OF EVIDENCE: 4 Laryngoscope, 132:838-843, 2022.

[Impact of benign parotid gland tumors on whole salivary flow rate]. / Einfluss gutartiger Tumoren der Glandula parotis auf die Gesamtspeichelflussrate.

OBJECTIVE: The impact of benign tumors of the parotid gland on whole salivary flow or sialochemistry is unclear. MATERIAL AND METHODS: A total of 22 patients with benign parotid tumors and 18 healthy controls underwent measurements of unstimulated and stimulated whole saliva flow and sialochemistry (Na+, K+, Ca++, Amylase, and pH). Assessment of xerostomia was performed by means of a visual analogue scale (VAS) and a questionnaire (QoL). RESULTS: Stimulated whole salivary flow was significantly lower in patients with benign parotid tumors in comparison to the control group (2.76 ±â€Š0.96 ml/min vs. 3.85 ±â€Š0.72 ml/min; p = 0.009). However, assessment of unstimulated whole salivary flow, sialochemistry, and subjective parameters (VAS, QoL) showed no significant differences between the patient and control groups (0.73 ±â€Š0.41 ml/min vs. 0.68 ±â€Š0.39 ml/min; p = 1). CONCLUSIONS: Benign salivary gland tumors appear to reduce whole stimulated salivary flow and leave unstimulated whole salivary flow and sialochemistry unchanged. The patients' subjective feelings of dry mouth do not seem to be influenced by the reduction in salivary flow.

Surgery of the major salivary glands and its impact on salivary flow-A review.

OBJECTIVE: The aim of this study was to bring attention to a rather unnoted side effect of salivary gland surgery-reduced salivary flow. METHODS: A systematic PubMed, Cochrane Library, LIVIVO, and Embase databases search was performed to identify relevant articles. RESULTS: Eight studies matched the inclusion criteria. All studies described an association between salivary gland surgery and reduced salivary flow. In five of the eight studies, patients reported on xerostomia after salivary gland surgery. CONCLUSIONS: Head and neck surgeons should inform their patients more accurately about reduced salivary flow and possible xerostomia after salivary gland surgery, and focus more on conservative strategies and minimally invasive techniques. Laryngoscope, 129:2053-2058, 2019.

Investigation of Cellular Function and DNA Integrity during 2D in vitro Culture of Human Salivary Gland Epithelial Cells.

In vitro culture of human salivary gland epithelial cells (SGEC) is still a challenge. A high quantity and quality of cells are needed for the cultivation of 3D matrices. Furthermore, it is known that DNA damage is supposed to be an important factor involved in carcinogenesis. This study investigates cellular function and DNA integrity of human SGEC during 3 passage steps in 2 groups (group 1: n = 10; group 2: n = 9). Cellular function was analyzed by immunofluorescence, transmission electron microscopy (TEM), and quantitative real-time polymerase chain reaction (qPCR). DNA integrity was tested via the comet assay. Immunohistochemistry and qPCR results showed stable α-amylase and pan-cytokeratin levels; TEM revealed functional cells; and no significant DNA damage could be detected in the comet assay during 3 culture steps. The study shows that not only at cellular but also at DNA level human SGEC can be safely quantified over 3 passages for preclinical tissue engineering without loss of differentiation and function.

Development of Human Salivary Gland-Like Tissue In Vitro.

The loss of salivary gland function caused by radiation therapy of the head and neck is a serious condition and it affects a patient's quality of life. The current lack of effective therapies demands new options to be explored. This study tested whether human salivary gland epithelial cells (SGECs) could be successfully cultured on a decellularized porcine gut matrix (SIS-muc) in both mono- and coculture with microvascular endothelial cells (mvECs). By performing immunofluorescence imaging, transmission as well as scanning electron microscopy (SEM), quantitative polymerase chain reaction (qPCR), and an amylase enzyme assay, it was investigated as to what extent the three-dimensional (3D)-cultured cells could maintain their molecular differentiation and the production of working α-amylase (α-AMY) compared with two-dimensional (2D) culture. In both 3D mono- and coculture, SGECs were successfully cultured and formed acinar-like structures. Those findings were confirmed by SEM imaging. Immunofluorescence imaging revealed that 3D-cultured cells expressed α-AMY, Claudin-1 (CL-1), and water channel protein aquaporin-5 (AQP-5). Two-dimensional-cultured cells only were positive for α-AMY. Real time (RT)-qPCR analysis showed that α-AMY relative gene expression was higher in both 3D mono- and coculture than in 2D culture. In α-AMY enzyme assay, cocultured SGECs showed about 25 times increased enzyme activity compared with 2D-cultured cells. In conclusion, the SIS-muc combined with endothelial coculture seems a suitable culture setting for the tissue engineering of functional human salivary gland tissue.

Time-Dependent Toxic and Genotoxic Effects of Zinc Oxide Nanoparticles after Long-Term and Repetitive Exposure to Human Mesenchymal Stem Cells.

Zinc oxide nanoparticles (ZnO-NP) are widely spread in consumer products. Data about the toxicological characteristics of ZnO-NP is still under controversial discussion. The human skin is the most important organ concerning ZnO-NP exposure. Intact skin was demonstrated to be a sufficient barrier against NPs; however, defect skin may allow NP contact to proliferating cells. Within these cells, stem cells are the most important toxicological target for NPs. The aim of this study was to evaluate the genotoxic and cytotoxic effects of ZnO-NP at low-dose concentrations after long-term and repetitive exposure to human mesenchymal stem cells (hMSC). Cytotoxic effects of ZnO-NP were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Furthermore, genotoxicity was evaluated by the comet assay. For long-term observation over 6 weeks, transmission electron microscopy (TEM) was applied. The results of the study indicated cytotoxic effects of ZnO-NP beginning at high concentrations of 50 µg/mL and genotoxic effects in hMSC exposed to 1 and 10 µg/mL ZnO-NP. Repetitive exposure enhanced cyto- but not genotoxicity. Intracellular NP accumulation was observed up to 6 weeks. The results suggest cytotoxic and genotoxic potential of ZnO-NP. Even low doses of ZnO-NP may induce toxic effects as a result of repetitive exposure and long-term cellular accumulation. This data should be considered before using ZnO-NP on damaged skin.

Two-stage autotransplantation of the human submandibular gland: First long-term results.

OBJECTIVES/HYPOTHESIS: Xerostomia is still one of the predominant side effects of radiotherapy (RT). This study investigates long-term results of a new surgical method that spares the submandibular gland from radiation. STUDY DESIGN: Eleven patients with head and neck carcinoma were enrolled in the study. In five patients 6-month follow-up testing, and in two of these patients 12-month follow-up testing was performed. METHODS: The submandibular gland was transplanted to the patients forearm for the time of radiation. Two months after completion of RT, the gland was retransplanted to the neck. Patients saliva flow was tested via the Saxon test, and patients had to answer the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Head and Neck 35 and visual analog scale. RESULTS: Following the two-stage autotransplantation, xerostomia was reduced in the long term due to improved saliva production of the reimplanted gland. CONCLUSIONS: Whether this promising novel approach is a reliable treatment option for RT patients in general should be evaluated in further studies. LEVEL OF EVIDENCE: 4. Laryngoscope, 126:1551-1555, 2016.

Two-stage autotransplantation of human submandibular gland: a novel approach to treat postradiogenic xerostomia.

Xerostomia is a persistent side effect of radiotherapy (RT), which severely reduces the quality of life of the patients affected. Besides drug treatment and new irradiation strategies, surgical procedures aim for tissue protection of the submandibular gland. Using a new surgical approach, the submandibular gland was autotransplanted in 6 patients to the patient's forearm for the period of RT and reimplanted into the floor of the mouth 2-3 months after completion of RT. Saxon's test was performed during different time points to evaluate patient's saliva production. Furthermore patients had to answer EORTC QLQ-HN35 questionnaire and visual analog scale. Following this two-stage autotransplantation, xerostomia in the patients was markedly reduced due to improved saliva production of the reimplanted gland. Whether this promising novel approach is a reliable treatment option for RT patients in general should be evaluated in further studies.

Vascularization of engineered cartilage constructs in a mouse model.

Tissue engineering of cartilage tissue offers a promising method for reconstructing ear, nose, larynx and trachea defects. However, a lack of sufficient nutrient supply to cartilage constructs limits this procedure. Only a few animal models exist to vascularize the seeded scaffolds. In this study, polycaprolactone (PCL)-based polyurethane scaffolds are seeded with 1 × 10(6) human cartilage cells and implanted in the right hind leg of a nude mouse using an arteriovenous flow-through vessel loop for angiogenesis for the first 3 weeks. Equally seeded scaffolds but without access to a vessel loop served as controls. After 3 weeks, a transposition of the vascularized scaffolds into the groin of the nude mouse was performed. Constructs (verum and controls) were explanted 1 and 6 weeks after transposition. Constructs with implanted vessels were well vascularized. The amount of cells increased in vascularized constructs compared to the controls but at the same time noticeably less extracellular matrix was produced. This mouse model provides critical answers to important questions concerning the vascularization of engineered tissue, which offers a viable option for repairing defects, especially when the desired amount of autologous cartilage or other tissues is not available and the nutritive situation at the implantation site is poor.

Nicotine derived genotoxic effects in human primary parotid gland cells as assessed in vitro by comet assay, cytokinesis-block micronucleus test and chromosome aberrations test.

Genotoxic effects of nicotine were described in different human cells including salivary gland cells. Based on the high nicotine concentration in saliva of smokers or patients using therapeutic nicotine patches, the current study was performed to evaluate the genotoxic potential of nicotine in human salivary gland cells. Therefore, primary salivary gland cells from 10 patients undergoing parotid gland surgery were exposed to nicotine concentrations between 1 µM and 1000 µM for 1 h in the absence of exogenous metabolic activation. The acinar phenotype was proven by immunofluorescent staining of alpha-amylase. Genotoxic effects were evaluated using the Comet assay, the micronucleus test and the chromosome aberration test. Cytotoxicity and apoptosis were determined by trypan blue exclusion test and Caspase-3 assay. Nicotine was able to induce genotoxic effects in all three assays. The chromosome aberration test was the most sensitive and increases in numerical and structural (chromatid-type and chromosome-type) aberrations were seen at ≥1 µM, whereas increases in micronuclei frequency were detected at 10 µM and DNA damage as measured in the Comet assay was noted at >100 µM. No cytotoxic damage or influence of apoptosis could be demonstrated. Nicotine as a possible risk factor for tumor initiation in salivary glands is still discussed controversially. Our results demonstrated the potential of nicotine to induce genotoxic effects in salivary gland cells. These results were observed at saliva nicotine levels similar to those found after oral or transdermal exposure to nicotine and suggest the necessity of careful monitoring of the use of nicotine in humans.

Nanoparticle-induced photocatalytic head and neck squamous cell carcinoma cell death is associated with autophagy.

AIM: To characterize molecular mechanisms underlying photocatalytic cell death of head and neck squamous cell carcinoma (HNSCC) by zinc oxide nanoparticles (ZnO-NPs). MATERIALS & METHODS: Human HNSCC-derived FaDu cells were incubated with ZnO-NPs followed by UVA-1 irradiation. Cytotoxicity was assessed by MTT assay and annexin-V propidium iodide test. Autophagy was detected by autophagosome accumulation, conversion of light chain 3 I to II, and lysosomal activity. The generation of reactive oxygen species was measured using the 2',7'-dichlorofluorescein-diacetate test. RESULTS: Apoptosis-independent cytotoxic effects were induced by 0.2- and 2-µg/ml ZnO-NPs and UVA-1. FaDu cells promoted autophagosome formation. Significantly elevated light chain 3 II and reactive oxygen species were seen after the combined application of both ZnO-NPs and UVA-1 as photocatalytic treatment. Autophagy probably mediates cell survival under UVA-1 or ZnO-NP exposure alone but induces self-digestive cell death after combined treatment. CONCLUSION: The effect of autophagy on HNSCC viability after nanoparticle-induced photocatalytic treatment seems to depend on the impact of the physicochemical trigger.

Nitrogen dioxide is genotoxic in urban concentrations.

In the discussion on toxic and genotoxic thresholds of air pollutants such as nitrogen dioxide (NO2), realistically low urban concentration ranges are of major interest. For NO2, the WHO defines the annual limit value as corresponding to 0.02 ppm. In the present study, the toxicity and genotoxicity of NO2 is set at a concentration under this limit value and examined in human nasal epithelium at different exposure durations in vitro. Nasal epithelial mucosa samples of 10 donors were harvested during nasal air passage surgery and cultured as an air-liquid interface. Exposure to 0.01 ppm NO2 or synthetic air as a control was performed for 0.5, 1, 2 and 3 h. Analysis included the caspase-3 ELISA, the single cell microgel electrophoresis (comet) assay and the micronucleus assay. The caspase-3 activity was not influenced by NO2 exposure, DNA strand fragmentation correlated with exposure durations to NO2 at 0.01 ppm NO2, and no cytotoxic effects such as apoptosis, necrosis or disturbances of cell proliferation were present. However, micronucleus induction as a sign of genotoxicity at an exposure duration of 3 h could be shown. Shorter exposures did not induce micronucleus formation. In summary, genotoxicity of NO2 could be demonstrated at a common urban concentration in vitro, but a threshold of NO2 genotoxicity could not be defined based on the present experiments.

Repetitive exposure to zinc oxide nanoparticles induces dna damage in human nasal mucosa mini organ cultures.

Data on the toxicological properties of zinc oxide nanoparticles (ZnO-NPs) is incomplete. ZnO-NPs may enter humans via inhalation or ingestion. The aim of the current study was to evaluate ZnO-NP-induced genotoxicity in three-dimensional (3D) mini organ cultures (MOCs) of human nasal mucosa following repeated exposure to ZnO-NP and regeneration. Nasal MOCs of 10 patients and ZnO-NPs were cultivated for one week and then characterized by electron microscopy. Nasal MOCs were partially covered by ciliated epithelium after one week of cultivation. ZnO-NPs were distributed to the cytoplasm and the nucleus. MOCs were exposed once, twice, or three times to 0.1 or 5 µg/ml of ZnO-NPs for 1 hr per exposure and were then evaluated for cytotoxicity and genotoxicity. MOCs were cultivated for 24 hr after the triple ZnO-NP exposure to allow for regeneration. ZnO-NP exposure did not result in significant cytotoxicity or apoptosis, as determined by trypan blue exclusion and caspase-3 activity, respectively. A significant increase in DNA damage was detected following repetitive exposure compared to single exposure to ZnO-NPs at 5 µg/ml, but not 0.1 µg/ml ZnO-NPs. At both concentrations of ZnO-NP, DNA fragmentation increased after 24 hr of regeneration. In contrast, DNA damage which was induced by the positive control, methyl methanesulfonate, was significantly reduced after 24-hr regeneration. Thus, our results suggest that repetitive exposure to low concentrations of ZnO-NPs results in persistent or ongoing DNA damage.

Cytotoxic, genotoxic and pro-inflammatory effects of zinc oxide nanoparticles in human nasal mucosa cells in vitro.

Despite increasing application of zinc oxide nanoparticles (ZnO-NPs) for industrial purposes, data about potential toxic properties is contradictory. The current study focused on the cyto- and genotoxicity of ZnO-NPs in comparison to ZnO powder in primary human nasal mucosa cells cultured in the air-liquid interface. Additionally, IL-8 secretion as a marker for pro-inflammatory effects was measured. Particle morphology and intracellular distribution were evaluated by transmission electron microscopy (TEM). ZnO-NPs were transferred into the cytoplasm in 10% of the cells, whereas an intranuclear distribution could only be observed in 1.5%. While no cyto- or genotoxicity could be seen for ZnO powder in the dimethylthiazolyl-diphenyl-tetrazolium-bromide (MTT) test, the trypan blue exclusion test, and the single-cell microgel electrophoresis (comet) assay, cytotoxic effects were shown at a ZnO-NP concentration of 50 µg/ml (P<0.01). A significant enhancement in DNA damage was observed starting from ZnO-NP concentrations of 10 µg/ml (P<0.05) in comparison to the control. IL-8 secretion into the basolateral culture medium was increased at ZnO-NP concentrations of 5 µg/ml (P<0.05), as shown by ELISA. Our data indicates cyto- and genotoxic properties as well as a pro-inflammatory potential of ZnO-NPs in nasal mucosa cells. Thus, caution should be taken concerning their industrial and dermatological application. Additionally, further investigation on repetitive NP exposure is needed to estimate the impact of repair mechanisms.

Nanosized titanium dioxide particles do not induce DNA damage in human peripheral blood lymphocytes.

Industrial application of titanium dioxide nanoparticles (TiO(2) -NPs) as an additive in pharmaceutical and cosmetic products is increasing. However, the knowledge about the toxicity of this material is still incomplete and data concerning health and environmental safety and results of recent studies on TiO(2) nanotoxicology are inconsistent. The in vitro geno- and cytotoxicity of TiO(2) -NPs in the anatase crystal phase was evaluated in human peripheral blood lymphocytes from 10 male donors. Initially, transmission electron microscopy (TEM) was performed to describe particle morphology and size, the degree of particle aggregation, and the intracellular distribution. Cells were exposed to nanoparticles in increasing concentrations of 20, 50, 100, and 200 µg/ml for 24 hr. Cytotoxic effects were analyzed by trypan blue exclusion test and the single-cell microgel electrophoresis (comet) assay was applied to detect DNA double-strand breakage. TiO(2) -NPs were sphere shaped with a diameter of 15-30 nm. Despite dispersive pretreatment, a strong tendency to form aggregates was observed. Particles were detected in the cytoplasm of lymphocytes, but also a transfer into the nucleus was seen. The trypan blue exclusion test did not show any decrease in lymphocyte viability, and there was no evidence of genotoxicity in the comet assay for any of the tested concentrations. In conclusion, TiO(2) -NPs reached the cytoplasm as well as the nucleus and did not induce cyto- or genotoxic effects in human peripheral blood lymphocytes. Complement investigations on different human cell systems will be performed to estimate the biocompatibility of TiO(2) -NPs. Environ. Mol. Mutagen.

Zinc oxide nanoparticles induce photocatalytic cell death in human head and neck squamous cell carcinoma cell lines in vitro.

The aim of this study was to determine the photocatalytic effects of zinc oxide (ZnO) NPs in combination with UVA-1 in human head and neck squamous cell carcinoma (HNSCC) cell lines in vitro. NP characteristics and intracellular distribution were described by transmission electron microscopy (TEM). After pre-incubation with ZnO NPs in concentrations of 0.002-20 µg/ml, the HNSCC cell lines HLaC 78 and UD-SCC 7A as well as primary oral mucosa cells (pOMCs) were treated with UVA-1. Cell survival and vitality was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide-(MTT)-assay and fluorescein diacetate test. Apoptosis was assessed by annexin-V propidium iodide flow cytometry. Intranuclear distribution of the rod-shaped particles was observed in 3.5% of HNSCC and in 0.5% of pOMCs. UVA-1 irradiation of 15 min in combination with 0.2 and 2 µg/ml of ZnO NP dispersion was shown to reduce the vitality of cancer cell lines significantly in comparison to cells without NP exposure or UVA-1 treatment only. For HLaC 78, a significant reduction in viable cells was already seen at 10 min of UVA-1 treatment and a ZnO NP concentration of 2 µg/ml. Flow cytometry indicated that cell death occurred primarily through necrosis. In pOMCs, vitality was not influenced either by UVA-1 treatment or ZnO NP exposure up to 2 µg/ml or a combination of both. ZnO NPs showed cytotoxicity at 20 µg/ml without UVA-1. Due to their photocatalytic properties, ZnO NPs may induce cell death in human HNSCC cell lines in vitro. Further studies will evaluate a possible benefit in adjuvant cancer therapy.

Customized tissue engineering for ear reconstruction.

Tissue engineering (TE) of cartilage for reconstructive surgery has proven to be a promising option for obtaining tissue for 3D structures that results in minimal donor site morbidity. Technological advances in this area are important since many defects can only be treated with customized implants. Most TE strategies rely on the use of resorbable 3D scaffolds to guide the growing tissue, with each tissue requiring a specific scaffold that has precisely defined properties depending on the physiological environment. Rapid prototyping (RP) technologies allow the fabrication of scaffolds of various geometric complexities from a variety of materials and as composites, while even allowing the inner architecture of the object to be varied in a defined manner at any given location. Scaffolds can be manufactured using RP techniques directly from computer aided design (CAD) data sources, e.g. via an STL file. The combination of TE and RP serves as the basis for the production of customized implants, for example the cartilage ear framework, and provides new perspectives for autologous ear reconstruction.

Intracellular distribution, geno- and cytotoxic effects of nanosized titanium dioxide particles in the anatase crystal phase on human nasal mucosa cells.

Nanomaterials are defined as substances with at least one dimension smaller than 100nm in size and are used for a multitude of purposes. Titanium dioxide nanoparticles (TiO(2)-NPs) are an important material used as an additive in pharmaceutical and cosmetic products. Due to their high surface-to-mass index, TiO(2) nanoparticles show different physical and chemical characteristics compared to the bulk substance. The knowledge about geno- or cytotoxic effects of TiO(2)-NPs is incomplete since existing studies show contrary results. Human nasal mucosa cells were obtained from 10 donors and exposed to TiO(2)-NPs in increasing concentrations of 10, 25, 50 und 100mug/ml. Transmission electron microscopy (TEM) was applied to document particle morphology and size distribution, the degree of particle aggregation and the distribution of particles in inter- and intracellular spaces. Furthermore, DNA fragmentation and cytotoxicity caused by TiO(2)-NPs were evaluated. DNA strand breakage was detected by single-cell microgel electrophoresis (comet) assay. Cytotoxic effects were analyzed by trypan blue exclusion test and fluorescein diacetate (FDA) assay. TiO(2) particles used in this study were mainly nanosized but also showed a strong tendency to aggregate in spite of sonication of the suspension. Particles entered the cytoplasm in 11% and the cell nucleus in 4%. The trypan blue exclusion test and the FDA assay did not show any loss of cell viability. In the comet assay, there was no evidence of increased DNA damage for TiO(2)-NPs. In this pilot project, no cyto- or genotoxic effects could be shown for TiO(2)-NPs on human nasal epithelial cells. Further investigations will focus on a variety of metal oxide nanoparticles to describe the biocompatibility in the human organism.

Spontaneous bilateral necrosis of the tongue: a manifestation of giant cell arteritis?

Giant cell arteritis is a chronic vasculitis, which involves large- and medium-sized branches of the arteries originating from the aortic arch. This disease is a diagnostic challenge with a wide range of clinical symptoms and findings due to different affected vessels. Classic symptoms are temporally located headache, thickened temporal artery and jaw claudication. Furthermore, visual symptoms like diplopia or loss of vision can occur. The tongue has an excellent blood supply and ischemic ulceration due to giant cell vasculitis is usually unilateral and rarely described in literature. We present a patient with a spontaneous bilateral tongue necrosis and are convinced that this extraordinary case must be deemed to be a manifestation of giant cell arteritis, although it does not completly satisfy the usually used diagnostic criteria formulated by the American College of Rheumatology.