Udo Stratmann

TCP is hardly resorbed and not osteoconductive in a non-loading calvarial model

Tricalciumphosphate (TCP) has been used as a ceramic bone substitute material in the orthopedic field as well as in craniofacial surgery. Some controversies exist concerning the osteoconductive potential of this material in different implantation sites. This study was designed to evaluate the biological response of calvarial bone towards TCP granules under non-loading conditions to assess the potential of TCP as a biodegredable and osteoconductive bone substitue material for the cranial vault. Full-thickness non-critical size defects were made bilaterally in the calvaria of 21 adult Wistar rats. One side was filled by TCP granules, the contralateral side was left empty and used as a control. Animals were sacrified in defined time intervals up to 6 months. Bone regeneration was analyzed with special respect toward the micromorphological and microanalytical features of the material-bone interaction by electron microscopy and electron diffraction analysis. Histologic examination revealed no TCP degradation even after 6 months of implantation. In contrast, a nearly complete bone regeneration of control defects was found after 6 months. At all times TCP was surrounded by a thin fibrous layer without presence of osteoblasts and features of regular mineralization. As far as degradation and substitution are concerned, TCP is a less favourable material tinder conditions of non-loading.

Strain-related bone remodeling in distraction osteogenesis of the mandible.

Distraction osteogenesis has become a mainstay in craniofacial surgery. However, there are several unresolved problems concerning the biology of bone regeneration. We investigated the biomechanical effects of mandibular lengthening in 32 rabbits on a cellular and histologic level. The mandible was subjected to a corticotomy, held in a neutral position for 4 days, and then lengthened at various strain rates and frequencies for 10 days. Radiographic, histologic, and electron microscopic examinations showed a strain-related bone regeneration. Application of physiologic strain rates (2000 microstrains or 0.2 percent) led to a bridging of the artificial fracture exhibiting woven ossification, whereas at 20,000 microstrains trabecular bone formation was demonstrated. In contrast, hyperphysiologic strain magnitudes (200,000 microstrains and 300,000 microstrains) showed a fibrous tissue formation. Multiple strain applications (10 cycles/day versus 1 cycle/day) increased the width of the distraction gap without changing the stage of bone regeneration. The gradual distraction of bone in physiologic magnitudes at higher frequencies seems to be desirable for a bony differentiation and may help to improve clinical applications.

Electrical stimulation influences mineral formation of osteoblast-like cells in vitro

The aim of the present study was to assess the structure of newly formed mineral crystals after electrical stimulation of osteoblast-like cells in vitro. Pulsed electrical stimulation was coupled capacitively or semi-capacitively to primary osteoblast-like cells derived from bovine metacarpals. Computer calculations revealed that the chosen input signal (saw-tooth, 100 V, 63 ms width, 16 Hz repetition rate) generated a short pulsed voltage drop of 100 microV (capacitive coupled mode) and of 350 microV (semi-capacitive coupled mode) across the cell-matrix layer. Stimulated cultures showed an enhanced mineral formation compared to the non stimulated controls. In cultures exposed to capacitively coupled electric fields and in control cultures nodules and mineralized globules were found. Nodules with a diameter of less than 200 nm covered the cell surface, whereas mineral globules with a diameter of up to 700 nm formed characteristic mineral deposits in the vicinity of the cells similar to biomineral formations occurring in mineralizing tissues. In contrast, large rod-shaped crystals were found in cultures stimulated by semi-capacitive coupled electric fields, indicating a non-physiological precipitation process. In conclusion, osteoblasts in culture are sensitive to electrical stimulation resulting in an enhancement of the biomineralization process.

Magnesium in newly formed dentin mineral of rat incisor.

Small amounts of magnesium are always detectable in addition to calcium and phosphorus in mineralized tissues such as dentin or bone. Magnesium has been considered to influence the mineralization process, especially crystal growth. The present study reports on the location and enrichment of magnesium in the newly mineralized dentin by using the high lateral resolution of energy dispersive X‐ray microanalysis combined with scanning transmission electron microscopy. To this end, we have used the continuously growing rat incisor as a model for a collagenous mineralizing system. Dental tissue was dissected free and cryofixed in liquid nitrogen–cooled propane. The distribution of elements was measured in freeze‐dried ultrathin cryosections. The magnesium distribution of the newly formed dentin area near the predentin area was found to be inhomogeneous. In certain small dentin areas, characteristical magnesium enrichments were observed. Further, high magnesium‐to‐phosphate molar ratios were found in these areas, and these were correlated with low calcium‐to‐phosphate molar ratios. Our results support the theory that magnesium is involved in the process of biological apatite crystal formation.

Clinical anatomy and palpability of the inferior lateral pterygoid muscle

STATEMENT OF PROBLEM The intraoral palpation technique of the inferior belly of the inferior lateral pterygoid (ILP) muscle is a standard diagnostic examination method for temporomandibular joint dysfunction syndrome, although different studies have revealed inconsistent results. PURPOSE This study assessed the feasibility of the ILP muscle palpation by a simulated clinical setting. MATERIAL AND METHODS Three dentists performed a bilateral palpation of the ILP muscle in 53 fresh and unfixed human cadavers and decided whether the muscle was palpable or unpalpable. In a second step, it was observed through the dissected infratemporal fossa, whether the examiners finger did or did not touch the ILP muscle by simulating the performed palpation. Palpatory findings were supplemented by 1-dimensional measurements for determination of topographic relations of the ILP muscle within the infratemporal fossa. For statistical analysis, sensitivity, specificity, and negative and positive predictive values of the palpation technique were calculated. Interexaminer agreement was estimated with the kappa value. RESULTS In 86 of 106 dissected specimens, a superficial fascicle of the medial pterygoid muscle was found in direct proximity to the ILP muscle. In these cases, a residual distance of 7.8 +/- 3.2 mm remained between the ILP muscle and buccinator fascia indented by the tip of the examiners finger. In 10 of 20 specimens with an absent superficial fascicle, the finger was able to reach the ILP muscle. CONCLUSION It is recommended that the ILP muscle palpation technique should no longer be considered as a standard clinical procedure because it is nearly impossible to palpate the ILP muscle anatomically and because the risk of false-positive findings (by palpation of the medial pterygoid muscle) is high.

Effect of glycine powder air-polishing on the gingiva

OBJECTIVES Safety and efficacy of glycine powder air-polishing (GPAP) in removing subgingival biofilm have been previously demonstrated. The hypothesis that GPAP results in less gingival erosion than sodium bicarbonate air-polishing (SBAP) or hand-instrumentation was assessed. MATERIAL AND METHODS In each of 10 patients, eight teeth with a residual probing depth of at least 5 mm following initial periodontal therapy were randomly assigned to the following interventions: GPAP (test), SBAP (positive control), hand-instrumentation (positive control), or no treatment (negative control). In each group, gingival biopsies were taken immediately after instrumentation and one 14 days later. Damaged gingival epithelium (GE) was assessed by light microscopy and quantified by a histological score (values 1-4). Differences between groups were evaluated using the marginal homogeneity test. RESULTS GPAP resulted in minor erosions of the GE (scores 1 and 2), whereas positive control specimens displayed moderate to severe erosions (scores 2-4). Differences between GPAP and positive controls were significant (p<0.05). Fourteen days following instrumentation GE under assessment was found to be intact in all groups. CONCLUSION The data indicated that GPAP results in less gingival erosion than SBAP or hand instrumentation, further supporting the safety of this new debridement technique.

The effect of magnitude and frequency of interfragmentary strain on the tissue response to distraction osteogenesis.

PURPOSE Bone regeneration is believed to be partially controlled by the applied local mechanical strain. To test whether the magnitude or frequency of discontinuous traction regulates the tissue response, defined daily strains were applied on mandibular osteotomies using an implanted mechanical distractor. MATERIALS AND METHODS Unilateral mandibular osteotomies were performed in skeletally immature rabbits (n = 36). and distraction was done by applying 2,000, 20,000, 200,000, or 300,000 microstrains once or 10 times (2,000, 20,000 microstrains) per day, respectively. Sham-operated animals (n = 6), serving as controls, underwent frame application and osteotomy without distraction. At the end of the distraction process, the newly formed tissue was evaluated histomorphometrically by the use of a well-defined scoring system of bone-forming indices. RESULTS The highest bone-forming indices were detected in the osteotomized, nondistracted group and in samples exposed to a physiologic strain (2,000 microstrains). Application of hyperphysiologic strains (200,000 and 300,000 microstrains) resulted in the formation of fibrous tissue and decreased bone-forming indices. Using Kruskal-Wallis tests, a statistically significant relationship was found between the bone-forming indices and the applied strain magnitudes. Scanning and transmission electron microscopic examinations showed osteoblastic differentiation and early mineral deposition in samples distracted up to 20,000 microstrains, whereas higher strain magnitudes led to the formation of fibroblast-like cells surrounded by collagen fibrils and only slight mineralization. Multiple strain applications (10 cycles/d vs 1 cycle/d) did not alter the histomorphometric indices or ultrastructural morphology significantly but increased the amount of newly formed tissue. CONCLUSIONS These results suggest that the magnitude and not the frequency of mechanical loading controls the differentiation of bone cells and the subsequent formation of bone tissue.

Reaction of sound and demineralised dentine to Carisolv in vivo and in vitro

OBJECTIVES The aim of the present study was to assess the structural reaction of intact dentine to Carisolv in vivo and in vitro. METHODS For the in vivo study occlusal cavities were prepared in 48 caries-free molars in Wistar rats (age: three months) and Carisolv-solution was placed into the cavities of 24 M for 1, 10 and 20 min. Twenty four contralateral molars served as controls and were treated with an inert liquid containing purified water, carmellose and erythrosin for corresponding periods. For the in vitro study 24 rat molars were resected en bloc and fractured to expose an area of crown dentine area. Molars were demineralised with EDTA for eight weeks to expose the collagenous dentinal matrix. One half of the specimens was then coated with Carisolv-solution for 20 min. The other half served as a control and was coated with an inert solution for 20 min. RESULTS In the in vivo specimens no signs of pulp cell damage were observed in the experimental group. The odontoblastic processes were destroyed in proximity to the floor of the cavity but were intact in the inner portion of dentinal tubules in experimental molars and a mechanical damage was noted in the control molars. In the in vitro specimens no structural discrepancy was detected between the experimental molars and the control molars in collagen fibrils of demineralised dentine. CONCLUSIONS Carisolv causes destruction of cellular components of odontoblastic processes but does not attack healthy collagen fibrils.

Periosteally derived osteoblast-like cells differentiate into chondrocytes in suspension culture in agarose

Pluripotent cells from the periosteal layer adjacent to cortical bone attain an osteoblast‐like phenotype in culture when reaching confluence in monolayer. It is unknown whether such newly differentiated osteoblast‐like cells preserve the chondrogenic potential characteristics for stem cells derived from the periosteum. Primary osteoprogenitor cells derived from bovine metacarpal periosteum were differentiated into alkaline phosphatase‐positive osteoblast‐like cells by an established monolayer culture protocol. After transfer into suspension culture in agarose gels, the cells differentiated into chondrocytes demonstrated by the production of collagen II, but not of collagen I, as well as alkaline phosphatase activity was abated. Contrarily, with continuation of monolayer culture, the cells maintained their osteoblast‐like phenotype and secreted large amounts of collagen I and a minor quantity of collagen III and V. The alkaline phosphatase activity steadily increased during the entire culture period of 2 weeks. Thus, our culture techniques can serve as useful tools to study mechanisms of differentiation by modulating the phenotypic potential of osteogenic cells. The results presented here support the notion that the extracellular environment strongly influences the cell type and its metabolism. Anat Rec 259:124–130, 2000.

The mineralization of mantle dentine and of circumpulpal dentine in the rat: an ultrastructural and element-analytical study

Abstract The purpose of this study was to compare the biomineralization of circumpulpal dentine with that of mantle dentine by ultrastructural and element-analytical techniques. Forty upper second molar germs of 10-day-old albino rats were cryofixed in liquid nitrogen-cooled propane and embedded in resin after freeze drying. Semithin dry sections were cut for analyzing the calcium and phosphorus concentration in initial mantle dentine, at the mineralization front of circumpulpal dentine, in the middle region of circumpulpal dentine and in mantle dentine peripheral to circumpulpal dentine. For the morphological evaluation of mineral deposits we compared ultrathin and unstained sections of cryofixed molars with chemically fixed molars. For both dentine types it was found that they develop via identical steps of mineral formation at collagen fibrils and non-collagenous matrix molecules. In circumpulpal dentine no globular mineral protrusions along the mineralization front (i.e. calcospherites) and no indications of interglobular dentine at the transition from circumpulpal dentine to mantle dentine were present. The von Korff fibres were not only visible in mantle dentine but also in circumpulpal dentine. Matrix vesicles were present only during the formation of an initial coherent layer of mantle dentine and could not be observed during successive formation of mantle dentine and circumpulpal dentine. The element-analytical data did not demonstrate any difference in the mineral content between the two dentine types. Therefore, we conclude that mantle dentine and circumpulpal dentine in the rat molar possess a high degree of structural and chemical similarity and that only the extent of terminal branching of the odontoblast processes gives an approximate estimation of the thickness of mantle dentine.