Regulation of p53 under hypoxic and inflammatory conditions in periodontium.

OBJECTIVES: Different studies suggest that inflammation as well as hypoxia leads to an increase of p53 protein levels. However, the implication of p53 during oral inflammatory processes is still unknown. The aim of this study was therefore to investigate the effect of hypoxia and inflammation on p53 regulation in human periodontium in vitro and in vivo. MATERIALS AND METHODS: Under hypoxic and normoxic conditions, human primary periodontal ligament (PDL) fibroblasts (n = 9) were stimulated with lipopolysaccharides (LPS) from Porphyromonas gingivalis (P.g.), a periodontal pathogenic bacterium. After different time points, cell viability was tested; p53 gene expression, protein synthesis, and activation were measured using quantitative RT-PCR, immunoblotting, and immunofluorescence. Moreover, healthy and inflamed periodontal tissues were obtained from 12 donors to analyze p53 protein in oral inflammatory diseases by immunohistochemistry. RESULTS: LPS-P.g. and hypoxia initially induced a significant upregulation of p53 mRNA expression and p53 protein levels. Nuclear translocation of p53 after inflammatory stimulation supported these findings. Hypoxia first enhanced p53 levels, but after 24 h of incubation, protein levels decreased, which was accompanied by an improvement of PDL cell viability. Immunohistochemistry revealed an elevation of p53 immunoreactivity in accordance to the progression of periodontal inflammation. CONCLUSIONS: Our data indicate that p53 plays a pivotal role in PDL cell homeostasis and seems to be upregulated in oral inflammatory diseases. CLINICAL RELEVANCE: Upregulation of p53 may promote the destruction of periodontal integrity. A possible relationship with carcinogenesis may be discussed.

Hypoxia and P. gingivalis synergistically induce HIF-1 and NF-κB activation in PDL cells and periodontal diseases.

Periodontitis is characterized by deep periodontal pockets favoring the proliferation of anaerobic bacteria like Porphyromonas gingivalis (P. gingivalis), a periodontal pathogen frequently observed in patients suffering from periodontal inflammation. Therefore, the aim of the present study was to investigate the signaling pathways activated by lipopolysaccharide (LPS) of P. gingivalis (LPS-PG) and hypoxia in periodontal ligament (PDL) cells. The relevant transcription factors nuclear factor-kappa B (NF-κB) and hypoxia inducible factor-1 (HIF-1) were determined. In addition, we analyzed the expression of interleukin- (IL-) 1ß, matrix metalloproteinase-1 (MMP-1), and vascular endothelial growth factor (VEGF) in PDL cells on mRNA and protein level. This was accomplished by immunohistochemistry of healthy and inflamed periodontal tissues. We detected time-dependent additive effects of LPS-PG and hypoxia on NF-κB and HIF-1α activation in PDL cells followed by an upregulation of IL-1ß, MMP-1, and VEGF expression. Immunohistochemistry performed on tissue samples of gingivitis and periodontitis displayed an increase of NF-κB, HIF-1, and VEGF immunoreactivity in accordance with disease progression validating the importance of the in vitro results. To conclude, the present study underlines the significance of NF-κB and HIF-1α and their target genes VEGF, IL-1ß, and MMP-1 in P. gingivalis and hypoxia induced periodontal inflammatory processes.

Impact of radiation history, gender and age on bone quality in sites for orthodontic skeletal anchorage device placement.

AIMS: Stability of orthodontic miniscrew implants is prerequisite to their success and durability in orthodontic treatment. As investigations revealed a positive correlation of miniscrew stability to periimplant bone quality, it has been the aim of this study to analyze the bone structure of resection preparations of human mandibles histologically by investigating the samples according to age, gender and exposure to radiotherapy. METHODS: Inflammation- and tumor-free alveolar bone sections from human mandibles (n = 31) with previously diagnosed carcinoma, chronic osteomyelitis or cysts were analyzed histomorphologically and histomorphometrically as to the dimension of trabeculae in cancellous areas. Group A investigated the impact of a history of radiation therapy, group B of gender and group C contrasted biopsies from individuals aging under 60 or over 60 years. Statistics were performed using the Kruskal-Wallis-test. RESULTS: Radiation, gender and age did not significantly influence bone density. The mean bone density averaged 40.7 ± 15.0% of spongiosa for the total collective with a median age of 58.4 years ± 14.7 years. CONCLUSIONS: Our findings provide new information on bone quality, thus contributing to a more precise evaluation of the parameters affecting and those not affecting miniscrew implant stability. On the basis of these results, the formulation of clinical guidelines for risk assessment of therapeutic approaches in patients prior to insertion of orthodontic skeletal anchorage devices seems to be conceivable.

LPS from P. gingivalis and hypoxia increases oxidative stress in periodontal ligament fibroblasts and contributes to periodontitis.

Oxidative stress is characterized by an accumulation of reactive oxygen species (ROS) and plays a key role in the progression of inflammatory diseases. We hypothesize that hypoxic and inflammatory events induce oxidative stress in the periodontal ligament (PDL) by activating NOX4. Human primary PDL fibroblasts were stimulated with lipopolysaccharide from Porphyromonas gingivalis (LPS-PG), a periodontal pathogen bacterium under normoxic and hypoxic conditions. By quantitative PCR, immunoblot, immunostaining, and a specific ROS assay we determined the amount of NOX4, ROS, and several redox systems. Healthy and inflamed periodontal tissues were collected to evaluate NOX4 and redox systems by immunohistochemistry. We found significantly increased NOX4 levels after hypoxic or inflammatory stimulation in PDL cells (P < 0.001) which was even more pronounced after combination of the stimuli. This was accompanied by a significant upregulation of ROS and catalase (P < 0.001). However, prolonged incubation with both stimuli induced a reduction of catalase indicating a collapse of the protective machinery favoring ROS increase and the progression of inflammatory oral diseases. Analysis of inflamed tissues confirmed our hypothesis. In conclusion, we demonstrated that the interplay of NOX4 and redox systems is crucial for ROS formation which plays a pivotal role during oral diseases.

Comparative analysis of nasal and oral mucosa dendritic cells.

BACKGROUND: Mucosal dendritic cells (DC) play a crucial role in tolerance induction as seen in mucosal immunotherapy of atopic diseases. Nevertheless little is known about the phenotypical differences of oral and nasal mucosal DC (nmDC). Recently, we could show that oral mucosal myeloid CD1a(+) DC (omDC) differ from their skin counterparts especially by the expression of high affinity receptor for immunoglobulin E (IgE; FcepsilonRI). However, expression pattern of FcepsilonRI and phenotypical characteristics of CD1a(+) nmDC have not been elucidated in detailed yet. METHODS: We performed detailed phenotypical comparison of nmDC and omDC of atopic and nonatopic individuals. RESULTS: As reported for omDC, FcepsilonRI on nmDC of atopic donors was elevated and mostly occupied by IgE while FcepsilonRI was present only in low amounts on nmDC of nonatopic donors. Nevertheless, the highest FcepsilonRI expression has been observed on omDC. Furthermore, significant amounts of costimulatory molecules CD40, CD80 and CD86 could be detected on nmDC that expressed more CD80 compared with omDC. Moreover, nmDC displayed less major histocompatability complex (MHC) class I and II molecules than omDC. In addition, nmDC expressed more C-type lectins CD205, CD206 as well as myeloid marker CD11b while omDC displayed increased expression of CD207 and lipopolysaccharide (LPS) receptor CD14. CONCLUSION: Together these data imply that nmDC phenotypical differ from omDC which might result in diverse functional properties and might be of relevance for selecting routes for immunotherapy of atopic diseases. Moreover these data provide a basis for further studies investigating immunological mechanisms underlying mucosal immunotherapy.

Murine synaptosomal lipid raft protein and lipid composition are altered by expression of human apoE 3 and 4 and by increasing age.

Apolipoprotein E (apoE) 4 and aging are risk factors for Alzheimer's disease (AD). Mice expressing human apoE4 and aged wild-type mice show a similarity in the transbilayer distribution of cholesterol in synaptic plasma membranes (SPMs) but differ markedly compared with apoE3 mice and young mice. The largest changes in cholesterol distribution were observed in the SPM exofacial leaflet where there was a doubling of cholesterol. Lipid rafts are thought to be associated with the exofacial leaflet, and we proposed that lipid raft protein and lipid composition would be associated with apoE genotype and age. Lipid rafts were isolated from synaptosomes of different age groups (2, 12, 24 months) of mice expressing human apoE3 and apoE4. Lipid raft markers, alkaline phosphatase (ALP), flotillin-1, cholesterol and sphingomyelin (SM) were examined. Lipid rafts of young apoE4 mice were more similar to older mice as compared with young apoE3 mice in reductions in alkaline phosphatase activity and flotillin-1 abundance. Lipid raft cholesterol and sphingomyelin levels were not significantly different between the young apoE3 and apoE4 mice but cholesterol levels of lipid rafts did increase with age in both genotypes. Results of the present study demonstrate that the two risk factors for Alzheimer's disease, apoE4 genotype and increasing age have similar effects on brain lipid raft protein markers and these findings support the notion that the transbilayer distribution of cholesterol is associated with lipid raft function.

[Preoperative autologous blood donation in orthognathic surgery]. / Eigenblutspende in der Dysgnathiechirurgie.

AIM: To minimize the perioperative risk of transfusion-associated complications, preoperative autologous blood donation is routinely performed in orthognathic surgery. This study critically analyzed the necessity of this procedure with respect to clinical, economic, and juridical issues. MATERIAL AND METHODS: In the Department of Oral and Maxillofacial Surgery, Bonn Medical School, 178 orthognathic surgical procedures were performed from 1994 to 2002. All patients (women, n=101; men, n=77) were informed about the possibility of autologous blood donation. This procedure was carried out in 83%, for single jaw surgery two and for bimaxillary osteotomies three to four units of packed red blood cells were predeposited. Of the patients, 99 underwent monognathic surgery in the lower jaw, 27 patients in the upper jaw, and bimaxillary osteotomy was performed in 52 patients. RESULTS: The average blood loss for the monognathic procedure in the lower jaw was 756 ml, in the upper jaw 858 ml, and in double jaw surgery 1391 ml. In 13 patients the hemoglobin concentration fell below 7.5 g/dl and 12 patients received their predeposited autologous blood units. A total of 57.7% of all donated autologous blood units were discarded. CONCLUSION: Autologous blood donation should be performed from the medical point of view for double jaw surgery and considered in single cases with special risks in monomaxillary osteotomies.

Improvement of the quality of diabetes control and decrease in the concentrations of AGE-products in patients with type 1 and insulin-treated type 2 diabetes mellitus: results from a 10 year-prospective, population-based survey on the quality of diabetes care in Germany (JEVIN).

BACKGROUND: Advanced glycation end (AGE)-products are a complex group of compounds that have been implicated in diabetes related long-term complications. Up to the present only few data exist about serum levels of the AGE-proteins N-epsilon-Carboxymethyllysine (CML) and pentosidine in patients with diabetes mellitus. PATIENTS AND METHODS: In the present 10-year, population-based trial of a selection-free cohort of patients with insulin-treated diabetes mellitus, serum CML and pentosidine levels were examined in correlation to the patients' quality of diabetes control and the prevalence of diabetes related long-term complications. RESULTS: Following the reunification of Germany in 1989 the health care system was decentralised. Up to 1994/95 the relative HbA1c (HbA1c/mean normal) of patients with type 1 diabetes increased (1.65 +/- 0.35 versus 1.52 +/- 0.31, p = 0.002). For patients with type 2 diabetes it remained constant (1.75 +/- 0.4 versus 1.78 +/- 0.31, p = 0.669). During the following period (from 1994/95 to 1999/2000) specialised diabetes care, structured treatment and teaching programmes (TTP), intensified insulin therapy and blood glucose self-monitoring for all patients were broadly implemented. This was accompanied by a substantial improvement in the relative HbA1c of both, patients with type 1 (1.48 +/- 0.3, p<0.0001), and insulin-treated type 2 diabetes mellitus (1.47 +/- 0.25, p<0.0001). During the same period the mean concentration of the AGE-product CML in the sera of patients with type 1 and insulin-treated type 2 diabetes decreased (type 1: 1994/95: 1158.1 +/- 410.0 ng/ml versus 1999/2000: 938.5 +/- 422.4 ng/ml, p<0.0001, type 2: 1994/94: 1244.7 +/- 1231.3 ng/ml versus 1999/2000: 970.9 +/- 458.6 ng/ml, p = 0.007). For pentosidine the same tendency was found for patients with type 1 diabetes (1994/95: 253.6 +/- 280.7 pmol/ml versus 1999/2000: 148.2 +/- 91.4 pmol/ml, p<0.0001). For patients with type 1 diabetes there was a positive correlation between the relative HbA1c-value calculated over the total follow-up period of 10 years and the CML-concentration in 1999/2000 (r = 0.405, p = 0.017). In 1999/2000 a reduced creatinine clearance (

[Molecular biological and immunohistochemical analysis of tp53 in human ameloblastomas]. / Molekularbiologische und immunhistochemische Untersuchung des tp53-Gens in menschlichen Ameloblastomen.

AIM: Ameloblastoma is the most frequent epithelial tumor of the jaws. The spread is locally invasive and it tends to recur. Malignant transformation and occurrence of metastases has been described. Immunohistochemical analysis shows an enhanced expression of P53 protein in ameloblastomas. Mutation of the tp53 tumor suppressor gene was verified in several human tumors. In this study histological sections were analyzed for the expression of P53 protein and the tp53 gene was examined for mutations. MATERIAL AND METHODS: Tumor DNA from 29 patients with an ameloblastoma was examined for mutations in exons 5 to 8 of the tp53 tumor suppressor gene using PCR, SSCP,- and sequential analysis. Histological sections of the tumors were analyzed immunohistochemically for an overexpression of P53 protein. RESULTS: Two tp53 mutations (6.9%) in ameloblastomas were verified for the first time. In all 58.6% of the tumors showed an immunoreactivity for P53 protein. There was a statistically significant positive correlation (Fisher's exact test p<0.0148) between an increased number of P53-positive tumor cells and the appearance of recurrence. DISCUSSION: In the face of the uncertain postoperative behavior of ameloblastomas, the immunohistochemical verification of more than 10% P53-positive tumor cells may give a prognostic indication for a tendency to recurrence.

Diagnosis and management of cervical teratomas.

We present three cases of cervical teratoma, two of which were detected prenatally by ultrasound. The two cases diagnosed prenatally showed rapid early growth, which indicated the aggressive nature of the tumour and assisted the parents' decision to terminate the pregnancy. The third case was undiagnosed prenatally and the child was born after a normal pregnancy and uncomplicated vaginal delivery. She recovered well after excision of the teratoma.

Endodontic and surgical treatment of a geminated maxillary incisor.

AIM: A geminated maxillary incisor required complex multidisciplinary treatment to preserve health and restore aesthetics. This report describes the coordination of coronal division, root extraction, bone grafting, endodontic treatment and orthodontics in achieving treatment goals. It is the intention of this report to show how a difficult case could be managed by properly coordinated multidisciplinary care. SUMMARY: Pulp testing of the geminated central right maxillary incisor gave a normal response. Radiographic investigation indicated a connection of the pulp chambers. Both the mesial and distal root were filled with gutta-percha. The connection between the two root canals was sealed with a flowable dentine-bonded resin. After dividing the crown with a diamond bur, the mesial part of the tooth was removed and the extraction socket was filled with beta-tricalcium phosphate ceramic (Cerasorb). Radiographs taken immediately after surgery and after 6 months showed no periodontal or periapical lesions. No signs of external resorption were identified. The diastema between the central incisors was closed by orthodontic treatment. KEY LEARNING POINTS: Geminated teeth may present aesthetic and functional problems which require multidisciplinary care. Careful clinical and radiographic examination is essential to decide on the fate of the coronal and root halves involved. A proper coordination between endodontic and surgical treatment may result in maintaining one tooth half, even if a midroot connection between the pulp chambers becomes evident. Orthograde endodontic treatment, hemisection and orthodontics may solve the aesthetic problem of a geminated tooth.

Predictive variables for the biological behaviour of basal cell carcinoma of the face: relevance of morphometry of the nuclei.

We did a morphometric analysis of 130 histological sections of basal cell carcinoma (BCC) of the face to find out whether morphometric variables in the structure of the nuclei of BCC cells could serve as predictors of the biological behaviour. We considered the following variables: maximum and minimum diameters, perimeter, nuclear area and five form factors that characterise and quantify the shape of a structure (axis ratio, shape factor, nuclear contour index, nuclear roundness and circumference ratio). We did a statistical analysis of primary and recurring tumours and four histology-based groups (multifocal superficial BCCs, nodular BCCs, sclerosing BCCs and miscellaneous forms) using a two-sided t test for independent samples. Multifocal superficial BCCs showed significantly smaller values for the directly measured variables (maximum and minimum diameters, perimeter and nuclear area). Morphometry could not distinguish between primary and recurring tumours.

[Possible complications in arthroscopic lavage of the temporomandibular joint]. / Mögliche Komplikationen bei der arthroskopischen Lavage des Kiefergelenks.

COMPLICATIONS: Arthroscopy of the temporomandibular joint is an invasive procedure that can entail potential complications. Based on many years of personal experience and taking data from the literature into account, this article discusses functional, surgical, and anesthesiological complications of arthroscopy of the temporomandibular joint. The majority of these complications can be avoided by heeding specific knowledge of topography, careful manipulation, and corresponding surgical experience. INFORMING THE PATIENT: The information given to the patient prior to the intervention should cover not only hemorrhaging, swelling, and wound infection, but also damage to (branches of) the facial nerve, the risk of perforation of the auditory canal as well as the possibility of temporary occlusal disturbances.

Tissue expansion using osmotically active hydrogel systems for direct closure of the donor defect of the radial forearm flap.

Although widely used, the radial forearm flap has been criticized for the poor quality of its donor site. Attempts to avoid donor-site problems have concentrated on the elaboration of the split-thickness and full-thickness skin graft methods of reconstruction. Skin grafts frequently fail over the flexor carpi radialis tendon, leading to chronic skin breakdown or, at best, tendon adhesion. Tissue expansion appears to be a good alternative that allows the use of local tissues to ultimately improve the forearm donor-site appearance. To avoid the disadvantages of traditional silicone balloon expanders (such as pressure peaks, infection, the valve at a distance from the expander, postoperative fillings), an osmotically active system was used. In an 18-month prospective study, 10 osmotically active hydrogel tissue expanders were placed on the forearms of 10 patients. The radial forearm flap was performed for intraoral reconstruction after surgical resection of oral cavity malignancies. The study showed that, in nine out of 10 patients, the expanded skin achieved was sufficient to cover the donor site after raising the forearm flap. Additionally, the expansion-related swelling pressure was well tolerated by the patients, the cosmetic results were very satisfactory, and the incidence of complications was very low. By using osmotically active hydrogel tissue expanders, there is no postoperative filling and no risk of complications arising from defective balloon expanders, filling valves, or missing ports.

[Calcium pyrophosphate arthropathy (pseudogout) of the temporomandibular joint]. / Kalziumpyrophosphatarthropathie (Pseudogicht) des Kiefergelenks.

The temporomandibular joint is rarely affected by crystal deposition disease (gout and pseudogout). Only the temporomandibular joint was affected in a 69-year-old patient with calcium pyrophosphate dihydrate deposition disease (pseudogout). This case is described here. Clinically presenting as painless swelling, computed tomography and magnetic resonance imaging showed signs of a chronic destructive arthritis. The evidence for calcium pyrophosphate dihydrate crystals was provided by histological examination. Diagnostic criteria, differential diagnosis and possibilities for treatment are discussed on the basis of our own experience and the literature.

Type A botulinum toxin for the treatment of hypertrophy of the masseter and temporal muscles: an alternative treatment.

The treatment of hypertrophy of the masseter and temporal muscles has to date been dominated by conservative and surgical measures. Local therapy with type A botulinum toxin permits an alternative method of treatment. After targeted, sometimes electromyographically controlled, intramuscular injection of the affected muscles, marked inactivity atrophy occurred in the muscles of seven patients over the course of 3 to 8 weeks. This atrophy remained constant over a follow-up period of up to 25 months, and no side effects were observed. Because of its minimal invasiveness, this technique seems to have an advantage over conventional surgical therapy. Consequently, treatment with type A botulinum toxin can be regarded as a sensible alternative to surgery in cases of hypertrophy of the masseter and/or temporal muscles.

Fetal cleft lip and palate: sonographic diagnosis, chromosomal abnormalities, associated anomalies and postnatal outcome in 70 fetuses.

OBJECTIVE: The aim of this study was to determine the relationship between facial clefts, associated malformations and chromosomal abnormalities. STUDY DESIGN: Sonograms of 70 fetuses with cleft lip with or without cleft palate were prospectively and retrospectively evaluated in our tertiary referral center for the nature of the cleft lip or palate and for the nature of the associated anomalies. Additionally, karyotyping was performed in 63 of the 70 patients (90%). RESULTS: The frequency of additional anomalies and the mortality rate in this selected population varied with the type of cleft. None of the fetuses presenting an isolated cleft lip had additional anomalies and all survived. All fetuses presenting a median facial cleft had concurrent anomalies (particularly of the central nervous system (90%)) and a fatal outcome. Associated defects were more frequent in fetuses with bilateral clefts (72%) than in those with unilateral clefts (48%). Fetuses with a unilateral cleft lip with or without cleft palate had a better survival rate (52%) than those with a bilateral cleft lip with or without cleft palate (35%). The frequency and type of chromosomal abnormalities varied with the type of cleft. The highest rate of chromosomal abnormalities was found in fetuses with median clefts (82%). CONCLUSIONS: Although no conclusions regarding the prevalence of chromosomal or other anomalies in patients with a cleft lip with or without cleft palate in the general population could be drawn, the study revealed a strong relationship between the type of facial cleft, associated malformations, chromosomal abnormalities and fetal outcome.

[Mandibular hypoplasia. Growth induction by osteochondral transplants or callus distraction?]. / Mandibuläre Hypoplasie. Wachstumsinduktion durch osteochondrale Transplantate oder Kallusdistraktion?

In order to restore function and esthetics in mandibular hypoplasia, transplantation of osteochondral grafts and the distraction osteogenesis technique are used alternatively. In a retrospective study the indication, growth and complication rate of osteochondral graft and distraction osteogenesis cases in mandibular hypoplasia were compared. In total, 12 patients (4-14 years old; 7 female, 5 male) with different grades of mandibular hypoplasia received osteochondral grafts (7 costochondral, 5 iliac crest; observation period 4-9 years). In 5 patients (10-12 years old; 1 female, 4 male), distraction osteogenesis was performed (observation period 10 months to 3 1/2 years). In the results, according to the literature, the range of growth after osteochondral grafting varied considerably, approximate normal growth was noticed in only two cases. On the other hand, all patients treated by distraction osteogenesis showed a stable increase of bone length as expected. After osteochondral grafts, adequate growth is to be expected in the minority of cases and it does not seem predictable in individual cases. Therefore, the indication for osteochondral grafts should be restricted to severe hypoplasia, where the local amount of bone does not allow the use of a distraction device, and to cases where support of the mandibula at the skull base is necessary. In all other cases of mandibular hypoplasia in children, distraction osteogenesis should be discussed as the primary option, even if in the long term a second surgical procedure should be necessary.