Dimensional ridge alterations following socket preservation using a nanocrystalline hydroxyapatite paste: a histomorphometrical study in dogs.

The aim of the study was to evaluate the histological response and dimensional ridge alterations following application of a nanocrystalline hydroxyapatite paste (NHA) into fresh extraction sockets in dogs. Immediately following vertical tooth separation and extraction, NHA was inserted in the extraction socket of the second molar in the lower jaws of 10 dogs. Untreated extraction sites on the opposite side served as controls. Wounds were closed using resorbable sutures after vertical flap elevation. After three and six months, 5 animals were killed. Lingual and buccal bone height, alveolar wall and total bone width 1, 3 and 5mm underneath the top of the crest were evaluated. Histological analysis revealed a high variability of NHA resorption and osteoconductive properties with different rates of material resorption. No statistically significant differences could be observed between the corresponding aspects of test and control sites. Both groups revealed higher alveolar wall resorption on the buccal than on the lingual side at both time periods. NHA does not seem to be useful for socket preservation procedures since it failed to prevent dimensional ridge alterations while revealing osseous integration but unpredictable material resorption. The role of non-resorbed hydroxyapatite remnants for implant placement is unclear and requires further investigation.

A pilot study of Cerasorb and Bio-Oss enhanced new bone formation in animal model.

The aim of this pilot investigation was to develop a new animal model for studying the effects on osteogenesis of agents used in the guided bone regeneration technique. As test material, a mixture of two osseoconductive materials with different physico-chemical characteristics was used. One component of the mixture was Bio-Oss, a bovine hydroxyapatite; the other was Cerasorb, a synthetic tricalcium phosphate. The mixture consisited of 50 volume percent of Bio-Oss and 50 volume percent of Cerasorb. In in vivo pilot experiment, bone wounds were prepared in the proximal third of both femurs of rabbits. A Cerasorb + Bio-Oss mixture was inserted on the test side and the same amount of sterile buffered physiological solution on the control side. After healing for 4 weeks, the bone segments were embedded and cut without decalcification, using the Exact cutting and grinding system. The density of the newly-formed bone was evaluated histomorphometrically. On the Cerasorb + Bio-Oss test side the bone density was almost 1.5 times higher than that on the control side. These results demonstrated that the applied animal model is appropriate for investigation of the effects on osteogenesis of biocompatible graft materials such as Bio-Oss and Cerasorb.

Angiogenesis, vascular endothelial growth factor and platelet-derived growth factor-BB expression, iron deposition, and oxidation-specific epitopes in stented human coronary arteries.

Pathogenesis of in-stent restenosis remains poorly understood because information from human histopathologic studies is scarce. We used an improved saw-grinding and cutting method on methacrylate-embedded samples containing metal stents, which allows in situ hybridization and immunohistochemical analysis of in-stent restenosis. Twenty-one samples were collected 3 hours to 3 years after stenting from 6 patients aged 36 to 81 years. Except in very early samples collected within hours after the stent deployment, neovascularization was present in all segments studied. At advanced stages, extensive neovascularization was located mainly at the luminal side of the stent struts and was only rarely accompanied by inflammatory cells. The neovessels colocalized with vascular endothelial growth factor (VEGF)-A mRNA and protein expression as well as with iron deposits and oxidation-specific epitopes, which imply the presence of chronic oxidative stress. VEGF-A expression was detected in the same areas containing macrophages, endothelial cells, and, to a lesser extent, smooth muscle cells, which also showed platelet-derived growth factor-BB expression. We conclude that in-stent restenosis features neovascularization, VEGF-A and platelet-derived growth factor-BB expression, and iron deposition, which is most probably derived from microhemorrhages. These mechanisms may play an important role in the development of neointimal thickening and could provide useful targets for the prevention and treatment of in-stent restenosis.

Automated segmentation of the optic nerve head for diagnosis of glaucoma.

Glaucoma is the second most common cause of blindness worldwide. Low awareness and high costs connected to glaucoma are reasons to improve methods of screening and therapy. A well-established method for diagnosis of glaucoma is the examination of the optic nerve head using scanning-laser-tomography. This system acquires and analyzes the surface topography of the optic nerve head. The analysis that leads to a diagnosis of the disease depends on prior manual outlining of the optic nerve head by an experienced ophthalmologist. Our contribution presents a method for optic nerve head segmentation and its validation. The method is based on morphological operations, Hough transform, and an anchored active contour model. The results were validated by comparing the performance of different classifiers on data from a case-control study with contours of the optic nerve head manually outlined by an experienced ophthalmologist. We achieved the following results with respect to glaucoma diagnosis: linear discriminant analysis with 27.7% estimated error rate for automated segmentation (aut) and 26.8% estimated error rate for manual segmentation (man), classification trees with 25.2% (aut) and 22.0% (man) and bootstrap aggregation with 22.2% (aut) and 13.4% (man). It could thus be shown that our approach is suitable for automated diagnosis and screening of glaucoma.

Expression and mutation patterns of p53 in benign and malignant salivary gland tumors.

The expression and mutation patterns of p53 were studied in a series of 68 benign pleomorphic adenomas and 237 malignant salivary gland tumors. p53 overexpression (nuclear staining exceeding 10%) was detected in 20% of the malignant salivary gland tumors, with the highest prevalence observed in polymorphous low grade adenocarcinoma, squamous cell carcinoma, and carcinoma ex pleomorphic adenoma and the lowest in adenoid cystic carcinoma and acinic cell carcinoma. In contrast, none of the 68 benign pleomorphic adenomas had nuclear staining exceeding 10%. SSCP and nucleotide sequence analysis of exons 4 to 9 of p53 in 19 malignant tumors revealed 9 mutations in 7 tumors. Our findings indicate that p53 may be a useful marker to help discriminate between benign and malignant salivary gland tumors.

Response of odontoblast-like cells to hydroxyapatite ceramic granules.

In this study the inductive influence of hydroxyapatite ceramic (HAC) granules on preodontoblast differentiation was investigated. Dental papilla cells harvested from upper molar tooth germs were implanted intramuscularly in a pouch created for this purpose. Six months after surgery tooth-root-like bodies had developed with pulp-like cavities in the specimens in which dental papilla cells had been implanted with and without HAC. These bodies consisted of regular tubular dentine in the central part, fibrodentine peripherally and sometimes osteodentine. HAC was haphazardly enclosed in these root-like bodies, whereas the implantation of HAC alone had no effect.

Interface between bone tissue and implants of solid hydroxyapatite or hydroxyapatite-coated titanium implants.

Loaded prestressed implants of dense hydroxyapatite and non-loaded hydroxyapatite-coated titanium implants were placed in edentulous regions of the lower jaw of dogs. After 6 month the jaw specimens were fixed and embedded in methyl-methacrylate. Thin non-decalcified ground sections were made for histology. Although the hydroxyapatite showed histological differences between the coated implants and the prestressed solid ones, both had an extensive apposition of normal lamellar bone on the whole surface of the bone-buried part of the implant. The bone contact was very intimate and without any visible intermediate tissue layer. The tissue response observed forms a good biological base for the clinical application of hydroxyapatite-coated titanium implants.

Development and functionality of isoelastic dental implants of titanium alloys.

Two types of isoelastic endosseous dental implants were produced and their functionality was tested. One type consisted of a porous sintered TiTa30 alloy, the other had a special surface structure consisting of titanium wire loops. Their mechanical properties were optimized by the production parameter (sintering and diffusion bonding, respectively). The functionality was tested after insertion into an artificial jaw which had properties corresponding to the natural mandibular. The elastic properties of both implants were similar to the properties of the bone. In addition the implants have a safe anchorage bone ingrowth. In animal experiments using the implant with surface loops it was observed that the bone entered the loops and even extremely small surface cavities in the wire loops.

De novo bone formation using bovine collagen and platelet-rich plasma.

In order to regenerate critical-size bone defects, a variety of bone substitutes is used in addition to autogenous bone. The regenerative capacity of these bone substitutes is usually compared to the efficacy of autogenous bone known as the "golden standard". Different cytokines influence the regeneration process because of their morphogenic or mitogenic properties. Platelet-rich plasma (PRP), a platelet concentrate, is characterised by having a positive effect on wound healing, reducing bone graft resorption and increasing the density of bone transplants. This experiment was commenced with a view to studying the osseous defect regeneration after placing various combinations of "filler materials" in experimentally created defects in the forehead of adult pigs. Regeneration by means of grafted autogenous bone (Group 1) or a bovine collagen based medical device (Group 4) alone and combined with PRP in two concentrations (Groups 2, 3, 5 and 6) was evaluated by means of microradiography and light microscopy after 2, 4 and 12 weeks. The microradiographic and light microscopic findings showed that autogenous bone in combination with PRP (Groups 2 and 3) had a significant accelerating effect on early bone regeneration (2 weeks). This effect was not evident when PRP was added to the bovine collagen (Groups 5 and 6). When using the collagen alone, significantly higher mineralisation values were achieved after 2 and 4 weeks than when using autogenous bone alone. After a 12-week observation period, the existing differences between the healing processes in the various groups were more or less levelled out. In summary, the results of the study indicate that clinically autogenous bone, as expected, is the ideal defect filler. Combining autogenous bone with PRP did not provide significantly better results. The findings in the groups treated with bovine collagen indicate that its local application mimics the effect of autogenous bone and amplifies bone regeneration when comparing with the control defect.

Mucoepidermoid carcinoma of the salivary glands, Clinical data and follow-up of 52 cases.

Between 1965 and 1993, a total of 52 patients with mucoepidermoid carcinomas underwent surgical treatment. Their TNM stage at the time of initial diagnosis varied (T0:0, T1:24, T2: 19, T3: 2, T4: 7; NO: 46, N1: 4,N2: 2; MO:50, M1: 2). In the majority of patients (n=24) the history of symptoms ranged from more that 0.5 to 2 years without any specificity of features. Radical ablative surgery of the primary tumour is the therapy of choice. In patients suspected of having metastases of the regional lymph nodes, resection of the related lymphatic system has to be included in the therapeutic approach. The prognosis is excellent in patients with a localized manifestation. The patients who died for reasons of tumour metastasis had all been classified as having stage Iii to IV disease at the time of initial diagnosis. Distant metastases are rarely found even decades after surgical therapy. Long-term follow- up is recommended for patients with mucoepidermoid carcinomas.

The congenital basal cell adenoma of salivary glands. Contribution to the differential diagnosis of congenital salivary gland tumours.

Congenital epithelial tumours of the salivary glands are very rare. The Salivary Gland Registry maintained in the Department of Pathology. University of Hamburg, contains only three cases among a total of 6,646 salivary gland tumours from the years 1965-1994. The three cases were classified as congenital basal cell adenoma, two of the parotid gland and one of the submandibular gland. Histologically, the three adenomas were similar in structure to the adult counterpart of basal cell adenoma with solid, trabecular or tubular (duct-like) patterns. In some cystic spaces of the duct-like structures PAS- and Astra blue-positive substances were secreted. On immunocytochemistry, the luminal duct-like cells showed membranous expression of cytokeratins 3, 5, 6, 7, 13 and 19. In the isomorphic basaloid cells of the solid and trabecular cell nests few cells expressed cytokeratin. On the outside of the solid cell nests there were smaller elongated myoepithelial-like cells, which expressed cytokeratin 14 and vimentin. Cytokeratins 1, 2, 4 and 18 were not expressed. The pattern of expression reflects the different stages of maturity of the tumour cells and is related to the development of the salivary glands until the end of the 3rd embryonal month with an arrest of further cell differentiation. No acinic cells, invasive growth, recurrence or metastases were observed. The differential diagnosis includes other congenital salivary gland tumours, such as hybrid basal cell adenoma-adenoid cystic carcinoma, sialoblastoma or embryoma, carcinoma, hamartoma and teratoma.

Unusual choristoma of the parotid gland in a girl. A possible trichoadenoma.

An 8-year-old girl had a painless circumscribed nodule in the right parotid gland for 4 weeks. A tumour (1.3 cm diameter) within the salivary gland parenchyma showed small cystic spaces with horn-like material macroscopically. The tumour tissue contained solid squamous cell formations and cystic spaces limited by multilayered squamous epithelium and covered by layers of ortho- and parakeratotic cells. The cystic spaces contained keratotic lamellae. In some areas pin-like epithelial proliferations were seen. All epithelial cells were characterized by uniform nuclei, and no atypical mitoses were seen. Keratinizated masses with partial calcification were occasionally located in the interstitial tissue and replaced by multinucleated giant cells. The tumour was classified as a choristoma and resembled a trichoadenoma. The ectodermally derived oral and salivary gland epithelium may be the source of skin-like or adnexal tumours.

Light and transmission electron microscopy of the intact interfaces between non-submerged titanium-coated epoxy resin implants and bone or gingiva.

This experiment was aimed at studying the intact tissue/implant interface of non-submerged dental implants with a titanium surface. Epoxy-resin replicas were fabricated from 3.05 x 8 mm cylindrical titanium implants with a plasma-sprayed apical portion and a smooth coronal collar. The replicas were coated with a 90-120-nm-thick layer of pure titanium and autoclaved. The coated replicas were inserted as non-submerged endosseous implants in the edentulous premolar region of dog mandibles and allowed to heal for three months. Jaw sections containing the implants were processed for light and electron microscopic study of the intact tissue/implant interface with and without prior demineralization. Gingival connective tissue fibers were closely adapted to the titanium layer, in an orientation more or less parallel to the implant surface. There was no evidence of any fiber insertions into the surface irregularities of the smooth or rough titanium surface. Undemineralized bone was intimately adapted to the titanium surface without any intervening space. In demineralized sections, the collagen fibers of the bone matrix tended to be somewhat thinner and occasionally less densely packed in the vicinity of the implant surface. However, they extended all the way to the titanium surface, without any intervening fibril-free layer.

The mechanism of pocket formation. A light microscopic study on undecalcified human material.

Two hundred eighteen human teeth with surrounding periodontal tissues in different stages of pathogenesis were studied under light microscopy using the "sawing and grinding" technique. This was done without decalcification and with special emphasis on the dentogingival junction area so as to study the mechanism of pocket formation. Thirty early, 133 established, and 214 advanced lesions were included. No initial lesion was seen. Pocket formation, provoked by microbacterial plaque, seemed to be initiated by the degenerative changes in the second or third cell layers from the innermost cells in the most coronal part of the junctional epithelium facing the microbial plaque. Consequently, an intraepithelial cleavage was formed followed by the degeneration and desquamation of the cells lining the split. This resulted in a deep crevice and gingival pocket formation in both early and established lesions. In advanced lesions, the deep pocket epithelium was exposed for such a long distance adjacent to the plaque and calculus, that the epithelium could be directly affected by toxic bacterial products and mechanical irritation of calculus. Finally, the epithelium became very thin and ulcerated, and a typical periodontal pocket was formed. The disruption of the epithelial barrier along with the concomitant penetration of bacteria and/or their toxic products were considered to be the most significant factors related to the conversion of an established lesion to an aggressive lesion.

Guided bone regeneration utilizing expanded polytetrafluoroethylene membranes in combination with submerged and nonsubmerged dental implants in beagle dogs.

Treatment of partial and total edentulism with submerged and nonsubmerged dental implants which follows the concept of osseointegration has become an accepted treatment modality. With compromised implant sites, practitioners have begun to combine one-stage implants with established techniques including guided bone regeneration. However, the clinical evaluation of this technique is limited. Therefore, the purpose of this study was to evaluate osseointegration and bone regeneration around nonsubmerged or submerged implants placed directly into surgically created osseous defects with or without expanded polytetrafluoroethylene (ePTFE) membranes. A total of 24 implants were placed in the mandibles of 4 beagle dogs and randomly assigned to 1 of 3 treatment groups. In group A, nonsubmerged implants were placed into osseous defects and treated with a poncho style ePTFE membrane. These membranes had a hole punched into the center and were slipped over the nonsubmerged implants. In group B, nonsubmerged implants were placed into osseous defects without an ePTFE membrane. In group C, submerged implants were placed into osseous defects and covered with an ePTFE membrane. Histometric measurements of each treatment group were made to determine percent bone gain or loss along the implant surface. Although a number of membrane removals occurred during the healing period, histological analysis indicated osseous ingrowth and osseointegration around nonsubmerged and submerged implants. An overall comparison of the treatment groups with ANOVA revealed that there were no significant differences between treatment groups, P > or = 0.05. However, when the data were stratified into sites which retained or lost the ePTFE membrane, the percent of bone regeneration was reduced in group A. Therefore, it may be recommended that nonsubmerged implants be placed with a submerged or "semi-submerged" protocol when utilized in conjunction with ePTFE membranes.

Soft tissue reactions to non-submerged unloaded titanium implants in beagle dogs.

The soft tissue reactions to non-submerged unloaded titanium implants were examined. A total of 24 implants were placed in 6 beagle dogs. The implants differed in their crestal area by having either a rough sandblasted, a fine sandblasted, or a polished surface. After 3 months, all implants were firmly anchored in the bone and had no clinical signs of peri-implant inflammation. Undecalcified histologic sections demonstrated that all implants achieved osseointegration with direct bone contact. The epithelial structures showed a peri-implant sulcus with a non-keratinized sulcular epithelium and a junctional epithelium. None of the sections exhibited epithelial downgrowth to the alveolar crest. In the supracrestal area, a direct connective tissue contact to the implant post was observed. An approximately 50 to 100 microns wide zone of dense circular fibers was found close to the implant surface. It was free of blood vessels and resembled closely an inflammation-free scar tissue formation. This zone was surrounded by a looser connective tissue with a 3-dimensional network of collagen fibers running in different directions. No significant differences concerning soft tissue reactions were found between the 3 implant surfaces. In particular, the length of direct connective tissue contact was similar. Concerning bone reactions, a significantly shorter distance from the top of the implant to the most coronal bone-implant contact was observed for rough surfaces. It is concluded that non-submerged unloaded titanium implants achieved a complication-free tissue integration with a dense connective tissue in direct contact to the implant surface in the supracrestal area, and epithelial structures similar to those around natural teeth. The different surface textures did not influence the healing pattern of the soft tissues, but had an influence on the location of the most coronal bone-implant contact.

A comparison of ePTFE membranes alone or in combination with platelet-derived growth factors and insulin-like growth factor-I or demineralized freeze-dried bone in promoting bone formation around immediate extraction socket implants.

The purpose of this study was to compare bone promotion around implants which were augmented with ePTFE membranes alone or in combination with cortical demineralized freeze-dried bone (DFDB) or the combination of platelet derived growth factor-BB (PDGF) and insulin like growth factor I (PDGF/IGF-I). Membranes were placed over titanium implants which had been inserted into fresh extraction sockets with large buccal dehiscences. Twenty-four implants were placed in 4 dogs. At 18 weeks clinical bone height measurements were taken, the animals were sacrificed, and all specimens retrieved for histologic evaluation. Clinically, a significant gain in bone levels was present in both the ePTFE membrane alone group (P < 0.005) and PTFE plus PDGF/IGF-I group (P < 0.01), but not in the PTFE plus DFDB group. Results from histometric measurements revealed an approximately 2-fold increase in the percentage of implant surface in contact with bone, area of bone adjacent to the implant surface, and in the total length of the implant surface in contact with bone in the dehiscence defects treated with ePTFE plus PDGF/IGF-I compared to the defects receiving ePTFE membranes alone (each P < 0.05). The response to the DFDB was highly variable and it did not significantly improve the efficacy of the PTFE membranes for any parameter measured. The distance from the outer surface of the new bone to the implant surface was statistically significant for ePTFE membranes alone and membranes plus PDGF/IGF-I. The results demonstrated that clinically, ePTFE membranes alone or ePTFE membranes with PDGF/IGF-I were equally effective in promoting bone growth around the implants. Histologic measurements demonstrated that sites treated with ePTFE membranes plus PDGF/IGF-I had the highest bone density compared with sites which received ePTFE membranes alone or with ePTFE membranes and DFDB. The results of this study question the use of DFDB and support the use of ePTFE membranes alone or with PDG-F-BB/IGF-I as potential methods of promoting bone formation around dental implants.

Peri-implant bone reactions to immediately loaded implants. An experimental study in monkeys.

BACKGROUND: There are reports which demonstrate that immediately loaded splinted implants can be osseointegrated when they are placed in the anterior part of the lower jaw. The concept of immediate loading has not been well investigated in the posterior mandible. The aim of this study was to evaluate the bone reactions around immediately loaded implants placed in the posterior region of the lower jaw in the monkey model. METHODS: Six adult Macaca fascicularis monkeys were used in this study. A total of 36 implants were placed after extraction of the second premolars, first and second molars, and complete healing of the sockets. Three control (C) group implants were placed in one quadrant of the lower jaw of each monkey. After a delay of 3 months to allow osseointegration to take place, the implants were loaded for 1 month using temporary resin bridges and later for 2 months using metal splinted crowns. In the contralateral region of the lower jaw, 3 test (T) group implants were placed and loaded immediately with the same sequence as carried out for the C implants. Specimens were examined and evaluated histologically after sacrifice. RESULTS: All of the implants were osseointegrated. Compact, cortical bone in contact with the implant surface without any gaps or connective tissue formation was observed. CONCLUSIONS: It was concluded that immediately loaded, splinted implants can be osseointegrated with a similar hard and soft tissue peri-implant response as delayed-loaded implants in the posterior mandible.

Tumour-simulating squamous cell metaplasia (SCM) in necrotic areas of salivary gland tumours.

Squamous cell metaplasia (SCM) adjacent to necrotic areas of salivary gland tumours must be distinguished from other types of SCM (focal SCM in the excretory ducts of salivary glands; necrotizing sialometaplasia; focal SCM within salivary gland tumours) in respect to the tissue structure. Based on the high cellular proliferation, arcade- or cord-like pseudoneoplastic SCM develops with stellate extension in the surrounding tissue and focal inclusion of goblet cell metaplasia. This proliferative SCM resembles the cellular demarcation of radicular dental cysts. In the Salivary Gland Register 8 cases of tumor-simulating SCM could be analysed which clinically and morphologically were suspect of squamous cell or mucoepidermoid carcinoma. Five cases were localized in the parotid gland, 2 cases in the submandibular gland and 1 case in the palatinal glands. Tumour-simulating SCM was developed in pleomorphic adenomas (5 cases) and in multifocal adenomatous oncocytic hyperplasia (3 cases).

Lipomatous pleomorphic adenoma of the parotid gland. Classification of lipomatous tissue in salivary glands.

Lipomatous pleomorphic adenoma is an unusual subtype with a lipomatous stromal component of more than 90% of the tumour tissue. This special type of pleomorphic adenoma must be distinguished from other types of lipomatous tumours or non-tumourous lipomatosis of the salivary glands. Until now only two cases of lipomatous pleomorphic adenoma have been reported in the literature. We report of a 36-year old woman who developed a well circumscribed nodule measuring 3.5 x 2.5 x 2 cm in the right parotid gland. The cut surface was grey-yellowish. Histologically, more than 90% of the tumour tissue was fatty tissue with univacuolar adipocytes. The pleomorphic epithelial elements were duct-like cells forming small lumina and spindle-shaped myoepithelial cell with surrounding mucoid stroma. Components of pleomorphic adenoma were intermingled with mature adipose tissue which was more concentrated in the central portion of the adenoma. Some compressed epithelial cords in the adipose tissue formed a septa-like pattern. The differential diagnosis to other lipomatous tumours (lipoadenoma, lipoma) and to non-tumourous interstitial lipomatosis as well as the possible pathogenesis as metaplastic change or epithelial-mesenchymal transdifferentiation are discussed.