Msx-1 is suppressed in bisphosphonate-exposed jaw bone analysis of bone turnover-related cell signalling after bisphosphonate treatment.

OBJECTIVES: Bone-destructive disease treatments include bisphosphonates and antibodies against receptor activator for nuclear factor κB ligand (aRANKL). Osteonecrosis of the jaw (ONJ) is a side-effect. Aetiopathology models failed to explain their restriction to the jaw. The osteoproliferative transcription factor Msx-1 is expressed constitutively only in mature jaw bone. Msx-1 expression might be impaired in bisphosphonate-related ONJ. This study compared the expression of Msx-1, Bone Morphogenetic Protein (BMP)-2 and RANKL, in ONJ-affected and healthy jaw bone. MATERIAL AND METHODS: An automated immunohistochemistry-based alkaline phosphatase-anti-alkaline phosphatase method was used on ONJ-affected and healthy jaw bone samples (n = 20 each): cell-number ratio (labelling index, Bonferroni adjustment). Real-time RT-PCR was performed to quantitatively compare Msx-1, BMP-2, RANKL and GAPDH mRNA levels. RESULTS: Labelling indices were significantly lower for Msx-1 (P < 0.03) and RANKL (P < 0.003) and significantly higher (P < 0.02) for BMP-2 in ONJ compared with healthy bone. Expression was sevenfold lower (P < 0.03) for Msx-1, 22-fold lower (P < 0.001) for RANKL and eightfold higher (P < 0.02) for BMP-2 in ONJ bone. CONCLUSIONS: Msx-1, RANKL suppression and BMP-2 induction were consistent with the bisphosphonate-associated osteopetrosis and impaired bone remodelling in BP- and aRANKL-induced ONJ. Msx-1 suppression suggested a possible explanation of the exclusivity of ONJ in jaw bone. Functional analyses of Msx-1- RANKL interaction during bone remodelling should be performed in the future.

Vertical distraction of fibula transplant in a case of mandibular defect caused by shotgun injury.

This case demonstrates the successful aesthetic and functional reconstruction of a complex facial gun-shot injury with extended bone defects and soft tissue destructions using a 3-step procedure. Initially, a reconstruction plate was inserted, later a fibula transplant enabled the basic reconstruction and finally was distructed in a 3rd session. The rationale behind the sequencing of surgical sessions was the extended bony defect and soft-tissue destruction. The main problem in this type of wound is hypoxia or anoxia of the receptor bed for the transplant. A microvascular anastomosized bone transplant is necessary for sufficient oxygen tension in the recipient site. The anatomical dimensional disproportion of the transplanted free fibula graft and the shape of the mandible were corrected prior to the insertion of dental implants by means of vertical distraction.

Establishment and cidofovir sensitivity of a cell line from a heart transplant recipient with multiple cutaneous tumors.

A new cell line, designated as Tuwei00, is described. It originated from an Epstein-Barr virus-positive skin tumor biopsy of a heart transplant recipient, whose numerous cutaneous neoplasms were treated with the antiviral drug cidofovir what caused at least transient remissions. The cell line was established in vitro and maintained for more than 70 passages. Cells of early passages were characterized by a slower growth, the inability to form colonies and a higher sensitivity to cidofovir. After overcoming a crisis, the cells grew faster, to a higher density and were able to form adherent colonies from single cells as well as colonies in soft agar. Chromosome analysis showed diploidy/hyperdiploidy at the earlier and hypodiploidy at the later passages. Sensitivity to cidofovir was distinctly higher in early passages of Tuwei00 cells than in later passages and was characterized by distinct decline of cell survival after long term cidofovir exposure. Established normal human keratinocytes, HaCaT cells, which were checked for comparison, showed a low cidofovir sensitivity similar to late passage Tuwei00 cells.

[Computed tomographic aspects of skull development in a child with cherubism]. / Computertomographische Aspekte der Schädelentwicklung eines Kindes mit Cherubismus.

The clinical picture of cherubism is similar to that of fibrous dysplasia. The initial clinical appearance involves the deformation of the maxillofacial area with orthodontic disorder. Usually it is found in the mandible giving the child a chubby-faced appearance, and it often occurs together with symmetric submandibular lymph node enlargement. This appearance reminds one of the cherubs seen in art. Only histological evidence for cherubism is inconclusive. The presence of multi-nucleated giant cells resembles fibrous dysplasia. A combination of clinical, radiographical and histological findings eventually leads to the correct diagnosis. An example is given of a patient displaying the typical disease process. Over a period of 12 years, we observed the progression of the disease from its initial appearance in a young child, through the full and characteristic display of a cherubic youth, and finally its regression. In conclusion, we advise restraint in planning surgical intervention. The diseases etiology is not entirely dear. The latest research points to genetic defects that lead to failure in the expression of matrix proteins.

Computertomographische Aspekte der Schädelentwicklung eines Kindes mit Cherubismus.

The clinical picture of cherubism is similar to that of fibrous dysplasia. The initial clinical appearance involves the deformation of the maxillo-facial area with orthodontic disorder. Usually it is found in the mandible, giving the child a chubby-faced appearance, and it often occurs together with symmetric submandibular lymph node enlargement. This appearance reminds one of the cherubs seen in art. Only histological evidence for cherubism is inconclusive. The presence of multi-nucleated giant cells resembles fibrous dysplasia. A combination of clinical, radiographical, and histological findings eventually leads to the correct diagnosis.An example is given of a patient displaying the typical disease process. Over a period of 12 years, we observed the progression of the disease from its initial appearance in a young child, through the full and characteristic display of a cherubic youth, and finally its regression.In conclusion, we advise restraint in planning surgical intervention. The disease's etiology is not entirely clear. The latest research points to genetic defects that lead to failure in the expression of matrix proteins.

[Long-term results of the reduction mammaplasty with Revers T-technique]. / Langzeitergebnisse der Mammareduktionsplastik unter Verwendung der T-Schnitttechnik.

Functional and aesthetic aspects make the reduction mammaplasty a favoured surgical procedure. The vertical technique, as well as the technique with central pedicle have clearly gained in importance next to the traditional T-cut technique. We provided questionnaires to a random sample of 110 patients who had been treated according to the method of Strömbeck-Weiner. The results were evaluated and compared with the literature. Our results show that the functional complaints were largely eliminated by the surgery and that the patients experienced a significant improvement in their psychological well being. The applied method produces a satisfactory shape of the breast with programmed ptosis, and to large extent retains the nipple-areola sensitivity. One shortcoming of this procedure is the scar formation which 20 % of the patients found bothering. However, 94 % of the patients would choose the procedure again. We conclude that the reduction mammaplasty with reverse T-technique according to Strömbeck-Weiner continues to be of value for patients with a desired weight reduction of more than 500-600 g and patients over the age of fourty.

A comparative quantitative analysis of laminin-5 in the basement membrane of normal, hyperplastic, and malignant oral mucosa by confocal immunofluorescence imaging.

Laminin-5 (Ln-5) is a heterotrimeric basement membrane (BM) molecule (alpha3beta3gamma2). It is a principal protein constituent of the anchoring filaments, which connect the BM with the hemidesmosomes of the basal keratinocytes and possess a crucial function in keratinocyte adhesion. Confocal immunofluorescence imaging is introduced for a quantitative evaluation of the Ln-5 content in the BM of oral squamous epithelium. The BM of normal oral mucosa was used as a reference (100%) for comparative analysis and showed a nearly uniform Ln-5 immunofluorescence intensity (99-100%). In all hyperplastic lesions of oral mucosa, the Ln-5 immunofluorescence intensity was increased (107-141%). The increased Ln-5 content in the BM of hyperplastic lesions suggests an increased keratinocyte-BM adhesion, possibly resulting in a higher stability of the oral mucosa. In contrast, in the oral squamous cell carcinoma (OSCC) invasive front, the remaining BM segments were characterized by a decrease in Ln-5 immunofluorescence intensity (35-74%). A stronger decrease of Ln-5-linked kerationocyte-BM adhesion correlates with a higher tumor grade. Because in central areas of carcinoma BM segments with a normal Ln-5 content could be demonstrated, the fundamental Ln-5 diminution in BM segments of the invasive front should be considered as an invasion-associated phenomenon.

Fibrillary co-deposition of laminin-5 and large unspliced tenascin-C in the invasive front of oral squamous cell carcinoma in vivo and in vitro.

PURPOSE: Invasion of oral squamous cell carcinoma (OSCC) is associated with laminin-5 (Ln-5) synthesis, focal Ln-5 loss from the basement membrane (BM), and Ln-5 depositions in the stroma beneath invading carcinoma cell complexes. METHODS: The study is focused on the laminin-5 matrix reorganisation within the stroma of the OSCC invasive front outside the basement membrane region as well as in OSCC-fibroblast co-culture in relation to unspliced tenascin-C (Tn-CL) and ED-B+ fibronectin (ED-B+ fn) using confocal laser scanning microscopy. RESULTS: In vivo, Ln-5 was demonstrated as fibrillary deposition in the invasive front. It was co-localised to Tn-CL. In pure OSCC cultures, Ln-5 was synthesised and deposited as a spot-like matrix. Fibrillary structures were not found. In contrast, in the OSCC-fibroblast co-culture, a fibrillary Ln-5 matrix organisation was revealed within the interface of OSCC cell-fibroblast complexes exclusively in co-distribution with Tn-CL and ED-B+ fn. CONCLUSION: At least in vitro, a carcinoma cell-stroma fibroblast interaction is indispensable for fibrillary Ln-5/Tn-CL matrix organisation. Behind the parallels to the initial basement membrane formation in organotypic cultures, the fibrillary multiprotein complexes at the OSCC cell-fibroblast interface are suggested as provisional basement membrane fragments with a possible supportive role for invasive tumour behaviour.

[Tumors of the face in organ transplant patients]. / Tumoren des Gesichts bei organtransplantierten Patienten.

BACKGROUND: The greatly enhanced risk of developing pre-cancerous lesions, basal cell carcinomas, and squamous cell carcinomas in organ transplant recipients is demonstrated on the basis of three of our own cases. DISCUSSION: Immunosuppression is discussed as the primary reason for enhanced tumour incidence and the occurrence of multiple synchronic and metachronic tumours after organ transplantation. UV exposition and oncogenic viruses must be taken into account as further factors for tumour disposition. More aggressive tumour behaviour has to be expected. In consequence, a focused clinical monitoring and a stringent therapy of the skin tumours is absolutely necessary.

[Experimental 5-aminolevulinic acid-induced photodynamic therapy (ALA-PDT) of oral carcinomas. Procedures in treatment of solid tumors and elucidation of cell death]. / Experimentelle 5-Aminolävulinsäure-induzierte photodynamische Therapie (ALA-PDT) oraler Karzinome. Verfahren zur Behandlung solider Tumoren und Aufklärung des Zelltods.

AIMS: Of our own patients suffering from oral squamous cell carcinoma (OSCC), 96% possessed a 5-aminolevulinic acid (ALA)-induced tumor fluorescence. Consequently, the ALA-induced fluorescence of OSCC is suggested as an ideal tool for selective photodynamic therapy (PDT). The aim of the study was to demonstrate selective tumor damage and to analyze the cell death mode (apoptosis vs. necrosis) of OSCC cells in cell culture and in solid, deeply invasive xenotransplants in immunodeficient nude mice using intratumoral laser illumination. MATERIAL AND METHODS: For ALA-PDT, laser light (635 nm, 0.75 W, 10 min, cooled application system) was used intratumorally as well as in cell culture. The therapeutic response was controlled by histology and immunohistochemistry (Ki-67 index). The apoptosis was evaluated by the TUNEL method and in vitro by flow cytometry. Mutations in the apoptosis-controlling p53 gene were investigated by direct genomic sequencing. RESULTS AND CONCLUSIONS: 1. Although all OSCC exhibited an ALA-induced fluorescence in vivo, the evaluation of the cell lines showed differences in intensity of the ALA-induced fluorescence. This points to a different sensitivity of OSCC for ALA-PDT. 2. The use of a cooled laser light application system allowed intratumoral radiation and treatment of deeply invasive OSCC regions. 3. The cytotoxic effect of ALA-PDT in OSCC is evidenced by a diminution of proliferative activity and necrosis but not by apoptosis. 4. Functional mutations within the p53 gene were considered a possible reason for the absence of apoptosis induction by ALA-PDT.

Simultaneous enzyme overlay membrane (EOM)-based in situ zymography and immunofluorescence technique reveals cathepsin B-like activity in a subset of tumour vessels.

The analysis of the invasion front of oral squamous cell carcinoma (OSCC) has revealed a fundamental invasion-associated remodelling of the extracellular matrix as the result of a complex regulated interplay of matrix synthesis and matrix degradation. Cysteine proteinases, in particular cathepsin B, are implicated in tumour invasion in vivo and in vitro and are thought to be important mediators of metastasis. An in situ zymographic assay based on enzyme overlay membranes (EOMs) was established to define the tissue localisation of cathepsin B activity in OSCC. Using confocal laser scanning microscopy we present a double-labelling method for the rapid and reproducible simultaneous detection of cathepsin B-like activity and cellular or extracellular antigens based on an EOM and immunofluorescence technique on frozen sections. Applying this method, cathepsin B-like activity was mainly found in vascular structures within the invasive front of OSCC. Therefore, the results suggest a particular pathogenic role of cathepsin B in tumour angioneogenesis. The method can simply be transferred to other enzymes and can be recommended for more extensive studies of proteolytic activity in human malignancies.

[Laminin-5 in diseases of the oral cavity]. / Laminin-5 bei Erkrankungen der Mundhöhle.

The basement membrane molecule laminin forms a family of proteins. Laminin-5 was identified as key protein in the anchoring filaments of the basement membrane. The anchoring filaments connect the basement membrane to the epithelial cells and together form the epithelial adhesion complex. Disturbances in the epithelial adhesion complex result in subepithelial blistering dermatosis. Autoantibodies against laminin-5 are found in cicatricial pemphigoid. Mutations in genes of laminin-5 with loss of the protein cause epidermolysis bullosa Herlitz. It can be determined during prenatal genetic diagnostic testing. Laminin-5 supports migration of keratinocytes and forms the basis for migrating keratinocytes in oral wound healing. Positive re-epithelialization via laminin-5 has been the subject of experimental studies. Oral squamous cell carcinomas are found to have a focal loss of laminin-5 in conjunction with a loss of the cellular receptor, and increased synthesis of laminin-5 by budding tumor cells has been observed along with deposition of the protein in the stroma. As a result of these observations laminin-5 has been suggested as a route of invasion. Quantitative assessment of this phenomenon and the modulation of laminin-5-tumor cell interaction offer new and hopeful means of describing and affecting the invasive behaviour of oral squamous carcinoma.

Multifocal oncocytic adenomatous hyperplasia of the parotid gland. A case report.

A case report of a multifocal oncocytic adenomatous hyperplasia occurring in a parotid gland is presented. The differential diagnosis of (multicystic) Warthin's tumor is discussed.

Distribution of laminin and fibronectin isoforms in oral mucosa and oral squamous cell carcinoma.

The expression of laminin and fibronectin isoforms varies with cellular maturation and differentiation and these differences may well influence cellular processes such as adhesion and motility. The basement membrane (BM) of fetal oral squamous epithelium contains the laminin chains, alpha2, alpha3, alpha5, beta1, beta2, beta3, gamma1 and gamma2. The BM of adult normal oral squamous epithelium comprises the laminin chains, alpha3, alpha5, beta1, beta3, gamma1 and gamma2. A re-expression of the laminin alpha2 and beta2 chains could be shown in adult hyperproliferative, dysplastic and carcinomatous lesions. In dysplasia and oral squamous cell carcinoma (OSCC), multifocal breaks of the BM are present as indicated by laminin chain antibodies. These breaks correlate to malignancy grade in their extent. Moreover, in the invasion front the alpha3 and gamma2 chain of laminin-5 can immunohistochemically be found outside the BM within the cytoplasm of budding carcinoma cells and in the adjacent stroma. The correlation between the morphological pattern of invasive tumour clusters and a laminin-5 immunostaining in the adjacent stroma may suggest, first, that a laminin-5 deposition outside the BM is an immunohistochemical marker for invasion and second, that OSCC invasion is guided by the laminin-5 matrix. Expression of oncofetal fibronectins (IIICS de novo glycosylated fibronectin and ED-B fibronectin) could be demonstrated throughout the stromal compartment. However, the ED-B fibronectin synthesizing cells (RNA/RNA in situ hybridization) are confined to small stroma areas and to single stroma and inflammatory cells in the invasion front. A correlation of the number of ED-B fibronectin synthesizing cells to malignancy grade could not be seen. ED-B fibronectin mRNA-positive cells seem to be concentrated in areas of fibrous stroma recruitment with a linear alignment of stromal fibro-/myofibroblasts (desmoplasia). Double staining experiments (ED-B fibronectin in situ hybridization and alpha-smooth muscle actin immunohistochemistry) indicated that the stroma myofibroblasts are a preferential source of ED-B fibronectin. In conclusion, in OSCC, a fetal extracellular matrix conversion is demonstrable. Tumour cells (laminin alpha2 and beta2 chain) and recruited stromal myofibroblasts (oncofetal ED-B fibronectin) contribute to the fetal extracellular matrix milieu.

[Partial loss of the auricle: multiphase reconstruction and complete preservation of the helix]. / Partieller Ohrmuschelverlust. Mehrphasige Rekonstruktion unter vollständigem Erhalt der Helix.

The loss of the auricle is a dramatic event in terms of aesthetics of the face. Secondary reconstruction of the ear is difficult and complicated. Therefore, a replantation of the amputated part should be tried. Proper securing and transport of the amputate is essential, in addition to the patient being sent to a specialized clinic. An anatomically correct form of the auricle is necessary for successful reconstruction of the ear. The amputated auricular cartilage is denuded of the skin except for the helical portion and is inserted into a retroauricular pocket. The helix is treated as a composite graft. After 1 to 2 months, the replanted auricular cartilage is elevated, adapted to the remaining ear stump and covered with a skin graft retroauricularly. By means of a representative case, a replantation method is demonstrated which allows complete reconstruction of the helix. In consequence, the replanted ear does not show any deficits compared with the original state.

[Internal gunshot wounds of the maxillofacial area and therapy with reference to minimal invasive technique]. / Steckschuss im Kiefer-Gesichts-Bereich und seine Therapie unter Berücksichtigung minimalinvasiver Technik.

Internal gunshot wounds in the head and neck region are complicated. Two cases of such injuries in the head and neck region are introduced and therapeutic approaches are discussed. If there are no severe additional injuries, the treatment of choice is the endoscopic removal of the projectile. Alternatively, the projectile has to be taken out by conventional surgery, using the approach which induces the least stress for the patient.

Synthesis and protein distribution of the unspliced large tenascin-C isoform in oral squamous cell carcinoma.

The inclusion or omission of the alternatively spliced region in the tenascin-C (Tn-C) mRNA gives rise to the large (Tn-C(L)) or small (Tn-C(S)) variant, respectively. Tn-C(L) is thought to be a typical component of provisional extracellular matrices (ECMs) and is expressed during tumour stroma remodelling. Tn-C(L) synthesis has been studied using RNA/RNA in situ hybridization, and Tn-C(L) protein distribution, using immunohistochemistry (clone BC-2), in 18 oral squamous cell carcinomas (OSCCs) of different grades of malignancy. While the Tn-C(L) protein was demonstrated within the whole stromal compartment regardless of grade of malignancy, the majority of the Tn-C(L) mRNA signal-bearing cells were carcinoma cells. Only a few stromal myofibroblasts were able to synthesize Tn-C(L), as revealed by alpha-smooth muscle actin double staining. In well-differentiated carcinomas (G1), the Tn-C(L) synthesizing carcinoma cells were localized as a single positive cell layer in the tumour stroma interface, particularly in invasive areas. A higher grade of malignancy (G2/G3) is associated with a significantly increased number of Tn-C(L) synthesizing carcinoma cells randomly distributed within the invading tumour areas. Double-staining experiments (Tn-C(L) mRNA ISH/BC-2 immunohistochemistry) indicate that these cells are capable of organizing and depositing a three-dimensional Tn-C(L) matrix. Even though an instructive and/or inductive role of the carcinoma cells in tumour stroma formation cannot be excluded, these results demonstrate that carcinoma cells can directly produce the ECM components of tumour stroma.

[Cell kinetics of oral squamous epithelial carcinomas. Determination of the proliferation index and apoptosis rate with the TUNEL method]. / Zellkinetik oraler Plattenepithelkarzinome. Bestimmung des Proliferationsindex und der Apoptoserate mittels der TUNEL-Methode.

Tumour growth is directly related to the multiplication of tumour cell numbers which can be caused, firstly, by increased proliferation and, secondly, by prolongation of the life span of tumour cells. The aim was the quantification of the increase and decrease (apoptosis) in cell numbers in relation to the cytological grading of oral squamous epithelial cell carcinoma. Proof of apoptosis-specific fragmentation of DNA (double-strand breaks) by the TUNEL method (TdT-mediated dUTP nick end labelling) and of proliferative activity by the monoclonal antibody MIB 1 was found in 41 oral squamous epithelial carcinomas of different grading. With progression of the cytological grading, an increase of proliferative activity and a decrease of apoptotic activity occurs. The apoptosis rates were 13.5% for G1 carcinomas, 8.5% for G2 carcinomas and 6.1% for G3 carcinomas. The results confirm that during progression to higher malignant cellular phenotypes of oral squamous carcinoma, increased proliferative activity as well as reduced apoptosis rates contribute substantially to the multiplication of tumour cell numbers and show that during progression a loss of control of programmed cell death (apoptosis) occurs. However, the high standard deviations of apoptosis rates within the different grades disprove an individual prognostic significance.