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PubMed1997

Rapid nodule evaluation computer-aided image analysis procedure for bone nodule quantification.

Using bone cell cultures, the effects of drugs on cell activities such as proliferation, differentiation, matrix formation, and mineralization can be explored. To quantify these parameters accurately and quickly, a kinetic reproducible computed image analysis procedure of culture dishes is proposed which could be conjointly used with biochemical analysis of the medium. In the present article, different mathematical procedures coupled either with or without histochemical staining are investigated and analyzed. Using serial cross sections and microradiographies of bone nodules, we demonstrated that the gray-level parameter is well correlated with bone mass and/or the mineralization status of the nodules. The procedure selected is a multistep procedure called rapid nodule evaluation (RNE), which uses a binary reconstruction program with different thresholds. To challenge this RNE procedure with the classical Von Kossa staining and quantification procedure, we cultured the cells in the presence of 10 nmol/L dexamethasone and compared the results using the two procedures. The RNE procedure appeared to be accurate and reproducible, and also has the advantage of speed and dynamic analysis over the classical Von Kossa quantification procedure.

Animals↗

PubMed1996

Calculation of intercrystalline solution composition during in vitro subsurface lesion formation in dental minerals.

Applications of a novel technique to calculate intercrystalline solution composition during enamel demineralization are presented. Bovine tooth enamel blocks and carbonated apatite (CAP) compressed disks were demineralized in an in vitro subsurface lesion system. The demineralization medium was a 0.1 M acetate buffer at pH 4.5, containing calcium, phosphate, and fluoride (0.5 ppm). Mineral samples were demineralized for various times, and fluoride profiles and mineral density profiles of these samples were determined by electron microprobe and X-ray microradiography, respectively. A model independent data analysis (MIDA) technique uses these data along with the differential equations for mass transfer and permits calculation of the local intercrystalline solution composition profiles inside the porous mineral matrix as functions of time and position. The invariance in diffusivity with time as calculated in the analysis was taken as an indicator of the physical reasonableness of the method. Current outcomes suggest that it is the sharp gradient of fluoride concentration in the intercrystalline solution which causes the formation of subsurface lesions. Since the driving force for mineral dissolution is a function of solution composition, a gradient of this driving force is consequently formed. Using a compressed disk of carbonated apatite powder as a model for block enamel excluded the possibility of the existence of a gradient of mineral composition which could also cause a gradient of the driving force for mineral dissolution. An FAP surface complex hypothesis is consistent with the current view that fluoride in the intercrystalline solution has a stronger inhibition effect on the dissolution of mineral than does fluoride in the mineral phase. With the help of the MIDA technique, calculated results indicate that the mechanism of the formation of subsurface lesions is dynamically controlled by the intercrystalline solution composition.

Animals↗

PubMed2002

Effects of sodium hypochlorite solution on root dentine composition.

Sodium hypochlorite (NaOCl) solution,<or=5% w/v available chlorine (abbreviated subsequently to %), is widely used as an irrigant in root canal treatment of teeth, so its effects on dentine are of clinical importance. The effects of approximately 0.5%, 3% and 5% NaOCl solution on the composition of root dentine were studied at ambient temperature. For dentine powder treated for 30 min, depletion of the organic phase was confirmed by infrared spectroscopy. Apatite lattice parameters showed no significant change, but NaCl was also detected by X-ray powder diffraction. The low solubility of apatite mineral in the NaOCl solutions was demonstrated by the constant weight of bulk enamel specimens immersed for seven days. The stability of the mineral phase was confirmed by scanning microradiography (SMR), an X-ray attenuation method employing photon counting. Repeated SMR measurements of the local mineral content of bulk samples of root dentine and a synthetic hydroxyapatite aggregate during exposure to pumped NaOCl solutions for 100 h showed no mineral loss. As predicted from apatite chemistry, reaction of NaOCl with the mineral phase can be excluded as a primary factor in changes in mechanical properties of treated dentine. Effects of retention of NaCl on endodontic sealants requires further investigation.

Journal Article↗

PubMed1999

Real-time measurement of in vitro enamel demineralization in the vicinity of the restoration-tooth interface.

An X-ray attenuation method using photon counting (scanning microradiography) is presented for the real-time study of in vitro demineralization of dental tissues in the vicinity of the restoration-tooth interface. By repeated measurement of mineral content profiles during the course of demineralization, the pattern of lesion development and the rate of mineral loss can be studied. The method is illustrated by comparison of enamel demineralization near a polyacid-modified composite resin restoration, near a bis-GMA/TEGMA composite resin restoration, and in an unrestored control. The method has potential for study of the influence of restorative materials on susceptibility of tooth tissue to demineralization.

Journal Article↗

PubMed2001

Wound healing response in the presence of stromal irregularities after excimer laser treatment.

PURPOSE: To trace the fate of stromal irregularities after excimer laser treatment and to increase our knowledge of the reasons why surface irregularities in the ablation bed cause inferior postoperative results. METHODS: Twelve New Zealand White rabbits received a transepithelial photoablation to a preset depth of 60 microm. An electron microscopy specimen grid was then placed on the denuded stroma and another 20 microm ablation was applied in order to produce surface irregularities. Another six rabbits received a plano transepithelial photoablation to a preset depth of 80 microm. The treated corneas were harvested at various timepoints and differentially further processed for microradiography, hematoxylin-eosin -, hyaluronan (HA)- and leukocyte protein L1 staining. RESULTS: In the grid treated corneas the subepithelial mesh pattern is clearly discernible after 1 week, and after 4 weeks it is replaced by a subepithelial layer containing HA and water. The thinning of this layer between 1 and 12 weeks is statistically significant (p<0.05). After 4 and 8 week the plano treated corneas only exhibit some subepithelial HA- and water accumulation. After 1 day the grid treated corneas show an extensive stromal infiltration of leukocytes. In the plano treated corneas the leukocytes mainly remain on the surface. CONCLUSIONS: During the healing process stromal irregularities are flattened, leaving a homogeneous zone with increased water content. This subepithelial layer is rarefying as new subepithelial tissue is forming. Postablational irregularities induce a more pronounced healing reaction when compared to a smooth ablation surface. Leukocyte infiltration seems to play a role in this process.

Animals↗

PubMed1999

Sinus floor augmentation with simultaneous placement of dental implants using a combination of deproteinized bone xenografts and recombinant human osteogenic protein-1. A histometric study in miniature pigs.

Maxillary sinus floor augmentation with autogenous bone has become a widely accepted procedure in implant dentistry. The use of osteoconductive bone substitutes in this indication is controversial, since their use can lead to a prolonged healing time, inhomogenous ossification, foreign body reaction, migration of particles and low bone-implant contact (BIC). The purpose of this study was to examine whether the combination of an osteoinductive protein (recombinant human osteogenic protein-1 (rhOP-1 = bone morphogenetic protein-7) with natural bovine bone mineral (BioOss) would improve ossification and the bone-implant contact (BIC) in a sinus floor augmentation with simultaneous placement of implants. In this study, the maxillary sinus floors in 5 miniature pigs were augmented with 3 ml BioOss containing 420 micrograms rhOP-1 on the test side and 3 ml BioOss alone on the control side. At the time of augmentation a titanium implant (ITI) was inserted from a laterocaudal direction. After 6 months of healing the sites of augmentation were removed and examined in non-decalcified sections by microradiography, fluorescence microscopy of sequentially labelled specimens and by histometry. On both sides, significant amounts of newly-formed bone were observed. However, on the test sites, the percentage of BIC in the augmented area was 80.0% versus 38.6% on control sites. It can be concluded that the application of bone morphogenetic proteins caused a more rapid and enhanced osseointegration of simultaneously placed implants when compared to the bone substitute alone. Therefore recombinant human osteogenic protein-1 delivered by natural bone mineral has the potential to become a clinical alternative for autogenous bone grafts in sinus floor augmentation.

Animals↗

PubMed2000

Enamel demineralization under driving forces found in dental plaque fluid.

The present study was carried out to examine the kinetics of enamel demineralization in vitro under driving forces for demineralization (i.e., the degree of saturation with respect to enamel, DS(En)) similar to those found in dental plaque fluid. Thin sections of human enamel were exposed at 25 degrees C to lactic acid solutions with DS(En) values (DS(En) = [(Ca2+)5(OH-)(PO4(3-))3/K(En)]1/9; K(En) = 5.5 x 10(-55)) ranging from 0.28 to 0.79. Lesion development was monitored by quantitative microradiography. Enamel mineral loss in solutions with DS(En) values of 0.28, 0.32 and 0.36 was first detected after 3, 3, and 7 wk of continuous exposure, respectively. Consistent with previous findings, subsurface demineralization was observed and rates of mineral loss increased significantly with decreasing DS(En) values. However, no mineral loss was observed in sections of enamel exposed to solutions with DS(En) values of 0.41 and 0.79, even after 11 months. These results suggest that (outer) enamel mineral behaves as a mineral phase that is less soluble than that dictated by the solubility product constant (K(En)) used in this study. Furthermore, these results indicate that the kinetics and general features of the demineralization process are maintained over a wide range of DS(En) values, including conditions that better reflect those found in the oral cavity. These findings are particularly relevant to the assessment of the cariogenic potential of dental plaque fluids.

Cariogenic Agents↗

PubMed1977

Histological analysis of the postnatal development of the nasal septum.

The importance of the growth in the nasal septum for the development of the facial skeleton has often been stressed. An increase in dimension is well-known from radiographs. It was the purpose of the present study to describe the normal pattern of the growth sites of the nasal septum according to age and sex by histological and microradiographical examination of human autopsy material. The nasal septum was removed from 66 boys and 57 girls who had died in accidents. Decalcified sections for conventional histology as well as undecalcified sections for microradiography were prepared. The superior surface and posterior margin of the vomer as well as the vomeroethmoidal and the vomeromaxillary sutures were studied. It was obvious that increase in size of the vomer could mainly be ascribed to apposition on the superior surface and the posterior margin. This was seen until adult age. After the ethmoid bone was ossified, the connection between the vomer and the perpendicular plate of the ethmoid bone was a slightly sinuous suture in which growth occurred until puberty. The growth pattern implied that a forward, downward sliding of the vomer must take place in relation to the ethmoid bone and the cartilaginous septum. Frontal sections revealed that, after the establishment of the suture between the vomer and the perpendicular plate of the ethmoid, a cartilage island was embedded in the vomer surrounded by thin, fenestrated cortical bone blades. This cartilage still persisted in specimens from adults. There is, on the basis of the present findings, no reason to believe that the septal cartilage plays a major role in the forward, downward growth of the maxillary complex in homo.

Adolescent↗

PubMed2001

A continuous culture biofilm model of cariogenic responses.

AIMS: To validate an in vitro model for the analysis of physiological and ecological responses to sugar challenge in bacterial populations, and subsequent changes in enamel mineralization. METHODS AND RESULTS: A seven-organism bacterial consortium was grown in a biofilm mode on enamel and hydroxyapatite (HA) surfaces in a continuous culture system and exposed to repeated sucrose challenges. This produced 'pH-cycling' conditions within the system. Populations on HA surfaces were enumerated. Changes in relative proportions of the different populations, and in the total viable count, were observed, between different treatments. Microradiography of the enamel sections showed increasing demineralization with increasing sucrose concentration. The lesions formed were similar to 'white-spot' lesions found in vivo. Differences in the quality of biofilms formed were also observed using Confocal Laser Scanning Microscopy. CONCLUSION: An in vitro model has been validated for the analysis of both physiological and ecological responses to sucrose challenges in bacterial populations, and subsequent changes in enamel mineralization. SIGNIFICANCE AND IMPACT OF THE STUDY: This model should facilitate the study of changes in bacterial populations in response to application of putative anticaries agents and concomitant changes in enamel mineralization.

Bacteria↗

PubMed1999

The use of gamma irradiation for the sterilization of enamel for intra-oral cariogenicity tests.

The dose of gamma irradiation sufficient to sterilize enamel samples to be used for intra-oral cariogenicity tests (ICT) without alteration to the structure or solubility of the enamel was determined. Each bovine incisor used was cut vertically into eight portions and each portion assigned to one of eight groups. Group 1 was used as control while other groups were subjected to respective doses of gamma irradiation from a cobalt-60 source. Following sterilization, both control and experimental groups were incubated in nutrient broth for 7 days at 37 degrees C under aerobic and anaerobic conditions. Caries-like lesions were subsequently produced in each specimen using acidic buffer solution, and mineral loss quantified using transverse microradiography. Data were analysed statistically. There was no significant difference in mineral loss between the control and experimental groups, but the numerical value of mineral loss was lower in groups irradiated with> 4080 Gy, in addition to full sterilization and enamel discoloration to cream colour observed in these groups. Microbial growth was observed in the control group and in groups irradiated with <4080 Gy but no discoloration. With 4080 Gy full sterilization without discoloration was observed. It was concluded that 4080 Gy of gamma irradiation is optimal for sterilization of enamel samples for use in ICT.

Analysis of Variance↗

PubMed2003

Influence of abrasion in clinical manifestation of human dental erosion.

The influence of abrasion from oral soft tissues on softened enamel lesion remineralization and erosion development was investigated. Using orange juice, softened enamel lesions were produced on 20 human premolars assigned randomly to 10 volunteers. Sections used as control and two test slabs were cut from each tooth. One of the two slabs from each tooth had an appliance built on it, which protected the lesion from abrasion. The two slabs (with/without appliance) were bonded to the palatal surfaces of upper right and left lateral incisor teeth of the participants who chewed sugar-free gum four times daily. After 28-day intra-oral exposure, mineral loss (Delta Z) and lesion depth (ld) in both control and test samples were quantified using transverse microradiography, and the data was analysed by paired t-test. Delta Z was significantly lower in lesions with appliance (protected), but higher in lesions without appliance (unprotected) when compared with control (unexposed). Similar pattern was observed with lesion depth. In unprotected slabs the lesions were abraded resulting in eroded enamel lesions. It was concluded that erosion observed clinically is the combined effect of demineralization of the tooth surface by an erosive agent and abrasion of the demineralized surface by surrounding oral soft tissues and through food mastication. Abrasion from oral soft tissues can contribute to site-specificity of dental erosion.

Adult↗

PubMed2001

High-resolution MRI detects cartilage swelling at the early stages of experimental osteoarthritis.

OBJECTIVE: The progressive early changes in cartilage and subchondral bone in an experimental model of osteoarthritis (OA) were investigated with high-resolution magnetic resonance imaging (MRI) and microradiography. METHODS: Partial medial meniscectomy was performed in the left knee of 16 rabbits. Four normal and four sham-operated additional rabbits were used as controls. Changes in cartilage and subchondral bone were sequentially assessed after surgery with MRI at 0, 2, 4, 6, 8 and 10 weeks, subchondral bone variations quantified postoperatively on microradiographs of sagittal sections at 6 and 10 weeks and the macroscopic alterations graded according to the severity of joint changes. RESULTS: MRI demonstrated a progressive increase in the articular cartilage thickness in the weight-bearing area of the femur at weeks 4, 6 and 8 vs basal. Tibial cartilage thickness only showed a significant increment at week 6. No significant abnormalities were detected on X-rays in subchondral bone when compared to controls. Macroscopically, 4 weeks after the operation OA rabbits had only slight cartilage discoloration. Cartilage eburnation, pitting, superficial erosions and osteophytes were detected at week 6. These abnormalities were more evident at 8 and 10 weeks after meniscectomy. CONCLUSION: The focal increase in cartilage thickness is one of the earliest measurable changes in OA and preceeds subchondral bone remodeling. The measurement of cartilage thickness variations with MRI can be used to follow the course of OA and to evaluate the potential beneficial effect of novel therapies.

Animals↗

PubMedDate not supplied

Cylindrical press-fit osteochondral allografts for resurfacing the equine metatarsophalangeal joint.

OBJECTIVE: To investigate the feasibility of resurfacing the equine fetlock joint using cylindrical, orthotopic, press-fit, osteochondral allografts. STUDY DESIGN: Experimental study. ANIMALS: Ten mature, mixed-breed horses. METHODS: Cylindrical, osteochondral grafts (6.5-mm diameter) were harvested aseptically from cadaveric equine metatarsophalangeal joints. Allografts were transplanted into 6 horses; 4 horses were sham operated. The surgical approach involved creation of a bone block at the origin of the medial collateral ligament and luxation of the metatarsophalangeal joint. Grafts were placed into the medial and lateral metatarsal condyles. Radiographs were taken at 8 and 25 weeks, and lameness was evaluated at 25 weeks. Horses were killed at 25 weeks. Analyses included gross evaluation, microradiography, paravital staining, light microscopy, and cartilage biochemistry. RESULTS: No complications occurred that could be attributed to the surgical procedure. Graft congruency with the surrounding articular cartilage was fair to excellent. Two horses were sound at 25 weeks. Most grafts had more than 90% articular cartilage coverage, and histologic and microradiographic analysis revealed good graft incorporation and articular cartilage survival. Sulphated glycosaminoglycan concentration was decreased in grafted tissue. CONCLUSIONS: We attribute the viability of osteochondral allografts in the equine fetlock to adequate congruency, stable graft fixation, and the use of orthotopic tissue. Host response to the allograft bone tissue did not affect cartilage viability. CLINICAL RELEVANCE: Before clinical use, improvements to instrumentation are required that would decrease damage to grafts and minimize technique-associated incongruencies of the articular surface at the time of grafting. Larger grafts would also likely be required to resurface a greater surface area.

Animals↗

PubMed2001

[Sinus floor augmentation with simultaneous implant insertion using recombinant human osteogenic protein-1].

INTRODUCTION: The purpose of this study was to examine whether the combination of an osteoinductive protein (recombinant human osteogenic protein-(rhOP-1 = bone morphogenetic protein-7) with natural bovine bone mineral (BioOss) would improve ossification and the bone-implant contact (BIC) in a sinus floor augmentation with simultaneous placement of implants. MATERIAL AND METHODS: In this study, the maxillary sinus floors in five miniature pigs were augmented with 3 ml BioOss containing 420 micrograms rhOP-1 on the test side and 3 ml BioOss alone on the control side. At the time of augmentation a titanium implant (ITI) was inserted from a laterocaudal direction. RESULTS: After 6 months of healing the sites of augmentation were removed and examined in non-decalcified sections by microradiography, fluorescence microscopy of sequentially labelled specimens and by histometry. On both sides, significant amounts of newly formed bone were observed. However, on the test sites, the BIC in the augmented area was 80.0% versus 38.6% on control sites. It can be concluded that the application of bone morphogenetic proteins caused a more rapid and enhanced osseointegration of simultaneously placed implants when compared to the bone substitute alone. DISCUSSION: Therefore recombinant human osteogenic protein-1 delivered by natural bone mineral has the potential to become a clinical alternative for autogenous bone grafts in sinus floor augmentation.

Animals↗

PubMed2001

Primary tissue failure of bioprostheses: new evidence from in vitro tests.

BACKGROUND: Primary tissue failure, which is mainly caused by calcification, is still the limiting factor in the long-term outcome of heart valve bioprostheses. Even though the precise nature of this process is not fully understood, in vitro tests have been developed to reproduce and predict calcification for individual bioprostheses. METHODS: In vitro calcification testing was performed by using an accelerated pulsatile valve tester which was adapted for testing stented as well as stentless bioprostheses with physiological fluid dynamics. A total of 84 bioprostheses (porcine, pericardial and stentless porcine of different manufacturers) were cyclically loaded at a test rate of 300/min at 37 degrees C within a rapid calcification fluid with CaxP = 130(mg/dl)2 at pH 7.4. Calcification was assessed by microradiography after 12 x 10(6) cycles. In a previous step, holographic interferometry was performed to identify irregularities of valve leaflets in order to predict later calcification. Selected specimens of calcified bioprostheses underwent histology, transmission (TEM) and scanning (SEM) electron microscopy. Tissue mineralization was investigated by coupling SEM, electron microprobe analysis (EMPA) and X-ray powder diffraction (XRPD) methods. RESULTS: For all tested bioprostheses, a significant calcification was achieved within 4 to 6 weeks of ongoing testing, and the degree of calcification increased with time. A significant correlation between calcification and leaflet irregularities (detected by holographic interferometry) was found (r = 0.80, p = 0.001). Calcification varied between individual bioprostheses, and significant differences were detected for different groups (calculated as percentage of total leaflet area, mean +/- SD): porcine stented (37.3 +/- 12.0%), bovine stented (23.0 +/- 8.9%), porcine stentless (16.2 +/- 7.6%). Histological and ultrastructural investigation showed intrinsic calcification involving both the spongiosa and fibrosa with collagen fibrils, interfibrillar spaces and cells as early sites of calcification. There was clear evidence of apatite crystallization, and observations made with in vitro calcification were quite similar to those occurring with in vivo implanted bioprostheses. CONCLUSION: In vitro tests can reproduce intrinsic calcification of bioprostheses even in the absence of viable biologic host factors. Moreover, degree and sites of calcification have become predictable. This enables the development and evaluation of bioprostheses with reduction of animal experiments. From our results obtained with a broad range of available bioprostheses, stented bovine and stentless porcine valves seem to be superior to conventional stented porcine bioprostheses with regard to leaflet calcification.

Animals↗

PubMed1984

Revascularized segmental diaphyseal bone transfers in the canine. An analysis of viability.

The biologic behavior of free revascularized bone segments was investigated using a unique canine model. Conventional methods to assess bone kinetics included the histologic osteocyte count, tetracycline uptake, appositional bone formation rate, and quantitative microradiography. The percentage of viable osteocytes in the revascularized segment was 52.4%, compared with 66.8% in the normal controls and 28.5% in the nonrevascularized segment. These differences were statistically significant. Both qualitative and quantitative differences were noted regarding deposition of tetracycline. On the basis of this study, we conclude that an apparently successful vascularized bone transfer retains a degree of viability greater than a conventional bone graft but less than that of normal undisturbed bone.

Animals↗

PubMedDate not supplied

[Osseointegration of a laser-structured titanium hemispheric acetabulum in the animal experiment].

Eight hemispherical titanium-cups with a new laser structured surface had been implanted in sheeps. Duration of implantation was eight weeks and one year. Analysis and evaluation of the results were done by post-operative x-rays, x-rays of the specimen, macroradiography, microradiography, fluorescencemicroscopy and histology. A quantitative analysis of the bony ingrowth was performed with a computer-aided system. The 8 weeks-cups showed an osseointegration rate of 28.8%, the one year group of 52.1% of the available surface. Bony ingrowth of the laserstructured titanium surface is possible. The laserstructured titanium surface can be used for the cementless fixation of implants.

Acetabulum↗

PubMed2001

Amelogenin-deficient mice display an amelogenesis imperfecta phenotype.

Dental enamel is the hardest tissue in the body and cannot be replaced or repaired, because the enamel secreting cells are lost at tooth eruption. X-linked amelogenesis imperfecta (MIM 301200), a phenotypically diverse hereditary disorder affecting enamel development, is caused by deletions or point mutations in the human X-chromosomal amelogenin gene. Although the precise functions of the amelogenin proteins in enamel formation are not well defined, these proteins constitute 90% of the enamel organic matrix. We have disrupted the amelogenin locus to generate amelogenin null mice, which display distinctly abnormal teeth as early as 2 weeks of age with chalky-white discoloration. Microradiography revealed broken tips of incisors and molars and scanning electron microscopy analysis indicated disorganized hypoplastic enamel. The amelogenin null phenotype reveals that the amelogenins are apparently not required for initiation of mineral crystal formation but rather for the organization of crystal pattern and regulation of enamel thickness. These null mice will be useful for understanding the functions of amelogenin proteins during enamel formation and for developing therapeutic approaches for treating this developmental defect that affects the enamel.

Amelogenesis Imperfecta↗