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A cremated pelvis dating from the first century A.D. showed evidence of osteosclerotic metastasis, presumably secondary to prostate carcinoma. The case demonstrates the importance of microradiography in palaeopathology as well as some of the structural changes seen in cremated bone.
We investigated the ability of synchrotron radiation computed tomography (SR-CT) to demonstrate trabecular microstructure, detail of trabecular surfaces, and mineralization of bones. Eight rat vertebrae, six rat tibiae, and eight minipig vertebrae were scanned using SR-CT at the synchrotron radiation facility Super Photon ring-8GeV (SPring-8). Images obtained using conventional micro-CT, scanning electron microscopy (SEM), and contact microradiography (CMR) were compared with the SR-CT images. SR-CT showed high image quality without visible partial volume effect. Three-dimensional SR-CT revealed shallow concavities in the bone surface, which were considered to correspond to osteoclastic resorption areas, as well as the connectivity, anisotropy, and shape (rod- or platelike) of trabeculae. Two-dimensional SR-CT showed different density along the surface of the trabecular bone, indicating the degree of bone mineralization. In conclusion, SR-CT seems to be a useful tool for delineating trabecular surfaces, evaluating bone mineralization, and revealing precise trabecular structure.
In order to prove osteocytic osteolysis in vivo, human parathyroid hormone (hPTH (1-34), 749 ng/h), or only solvent of the same volume, was continuously administered to 8-month-old rats by an infusion pump for 4 weeks, and then structural changes in osteocytes in the cortical bones of the tibiae were analyzed morphometrically, histologically, and histochemically. Based on contact microradiography (CMR) observations, the osteocyte lacunae in the PTH group tended to be enlarged, compared with those of the control, while the average lacuna area was 137.0 microm2 in the PTH group versus 93.9 microm2 in the control, suggesting evidence of osteocytic osteolysis. Acid phosphatase enzyme histochemical localization was observed in some osteocytes in the PTH group; therefore, lysosome systems may participate in the osteolytic mechanisms. On histological samples stained with hematoxylin-eosin or toluidine blue, the lacunae of the controls were surrounded by narrow areas of matrices both positive for hematoxylin and metachromatic for toluidine blue, while belt-like areas positive for hematoxylin were observed around the PTH-group lacunae. These findings suggested that, after osteocytic osteolysis, regenerated bone matrices may be added to the walls of osteocytes that possess enlarged lacunae.
Surface staining is widely used for histological studies involving undecalcified thick bone sections. Individual osteons, particularly newly formed ones stained with toluidine blue (TB), show various color intensities. We studied the correlations between TB color intensity and the differences in stiffness and degree of mineralization of individual osteons in undecalcified histological sections of goat tibial diaphysis, measured by scanning acoustic microscopy (SAM) and contact microradiography (CMR), respectively. Results showed that all three measurements correlated significantly with each other (r = 0.567 - 0.786; all P < or = 0.01). The TB surface staining intensity of individual osteons correlated better with the reflection coefficient (stiffness index) measured by SAM (r = 0.713) than with the aluminum step-wedge equivalent thickness measured on CMR micrographs (r = 0.567). The aluminum step-wedge equivalent thickness of individual osteons on CMR correlated slightly better with the SAM reflection coefficient (r = 0.786) than with the TB surface staining intensity (r = 0.713). The results of this study suggest that TB surface staining may be used as a simple method for indicating differences in stiffness and degree of mineralization in individual osteons in comparative histological studies.
To determine the effects of a bisphosphonate on the quality of bone morphogenetic protein-(BMP-) induced bone, incadronate was administered to rats in which subcutaneous ectopic bones were induced by recombinant human BMP-2. Incadronate (1 microg/kg/day) was administered to rats carrying the BMP-induced bones three times per week, from the 3rd to 7th week after BMP implantation (incadronate group). Aliquots of phosphate-buffered saline were administered in the same protocol without incadronate to the control group. During the 3rd, 4th, 7th, or 10th week, the BMP-induced bones were removed and observed by contact microradiography (CMR), H&E staining, enzyme histochemistry for tartrate-resistant acid phosphatase (TRAP), and immunohistochemistry for cathepsin K. By 3 weeks when administration of incadronate began, woven bones formed in the periphery of the BMP pellets and osteoclasts were attached to these bones. At 4 weeks, in both the incadronate and control groups, bone formation advanced inward. However, in the incadronate group, poorly calcified areas, corresponding to the remaining BMP pellets, were found in the middle areas of bone formation, whereas osteoclasts decreased when compared with those of the control group. During the 10 weeks, bone marrow was formed and the characteristics of lamellar bones, in which the lacunae of small osteocytes were regularly arranged, were noted in the control group. In contrast, the poorly calcified areas were still present up to 10 weeks in incadronate group in which the osteoclasts were also scarcer than in the control group. These findings suggested that osteoclast-mediated bone resorption was inhibited by incadronate administration, and that immature bones including the BMP pellets remained for a long time, indicating that the process of bone maturation was blocked. The possibility of using incadronate for the purpose of inhibiting osteoclastic bone resorption was confirmed, but further study is needed before clinical application.
INTRODUCTION: Growth factors are still not commercially available for routine clinical use. One possibility might be the application of Colloss, which is supposed to have osteoinductive properties. Therefore, it was the aim of the following animal experimental study to compare Colloss with autogenous bone and to investigate its osteoproductive potential. MATERIAL AND METHODS: In 24 pigs clinically relevant defects were created in the forehead area. Defects were filled with autogenous bone and Colloss. One defect was left unfilled and served as control. After 2, 4, 12 and 26 weeks, six animals were sacrificed and microradiography, light microscopy and immunohistochemistry were carried out using BMP-2 and osteocalcin as antibodies. RESULTS: After 2 weeks the defects filled with the bovine protein complex showed a mineralization of 52.6% +/-4.03 compared to autogenous bone (38% +/-9.9; p=0.068) and the control defect (30% +/-2.94; p=0.05). After 4 weeks the results became assimilated. Light microscopy revealed especially after 2 weeks an enhanced de novo bone formation in the Colloss-group. This was also evident in an early expression maximum of BMP-2 after 2 weeks. Autogenous bone and the control defect showed a later expression maximum. CONCLUSION: Colloss showed an acceleration of bone regeneration in the early phase. Nevertheless, after 26 weeks the results were comparable with the autogenous bone group.
Although diamine silver fluoride (AgF: (NH3)2AgF) stains teeth black, it is known as a very effective agent to prevent the dental caries progress. In order to find another fluoride that has a similar anticariogenic effect without changing tooth color, we prepared ammonium hexafluorosilicate (SiF: (NH4)2SiF6), in which the silver of AgF is replaced with silicon. In this study, the anticariogenic effect of SiF was evaluated using bovine teeth. Fluoride solutions, SiF, AgF, acidulated phosphate fluoride (APF), and sodium fluoride (NaF), were applied to bovine enamel and dentine blocks, and the depth of demineralization was measured after exposure to a demineralizing solution for 24 h. Also, fluoride was applied to a simulated dentine caries specimen to evaluate the caries progress-preventing ability. For the dentine specimens, mineral loss (Delta Z) was also measured with microradiography. We found that SiF treated enamel showed better acid resistance than specimens treated with NaF or APF. AgF treated enamel also showed similar acid resistance, but was stained black. SiF and AgF treated caries-affected dentine showed reduced demineralization when exposed to a demineralization solution for 24 h. Mineral loss (Delta Z) was reduced to 85% and 75%, respectively. Although the acid resistance of the SiF treated teeth was inferior to that of the AgF treated teeth, we consider that SiF has good potential as anticariogenic agent, since it increased acid resistance without changing tooth color.
Osteosclerosis in adults with primary hyperparathyoidism is rare; the usual skeletal manifestation, when presented, is diffuse osteropenia. We describe a patient with generalized osteosclerosis in association with primary hyperparathyroidism. The findings are documented by conventional and fine-detail radiography, absorptiometric bone mineral analysis, quantitative microradiography and histologic examination of bone. The unique features are contrasted with the manifestations recorded in a recently studied group of 87 hyperparathyroid patients. The data presented here support a causal relationship in this patient between parathyroid hormone excess and the development of densely sclerotic bones.
We studied the aortic root (wall, semilunar cusp, septal myocardium) in 37 patients (29 male and 8 female; mean age; 41 years) who died 2 to 4,380 days (mean, 398 days) after heart (n = 34) and heart-lung (n = 3) transplantation. The aim of the study was to assess tissue viability, graft-host biological interaction, and cusp mineralization with time. Study methods included gross inspection and photos of each specimen, microradiography, histology and immunohistochemistry, scanning and transmission electron microscopy, and atomic absorption spectroscopy. There were no cases of valve dysfunction; in particular, cusp tears or perforations never occurred. Only 1 valve, in place for 12 years, had a pin-point calcification visible to the naked eye. Optimal preservation of the tissue components (endothelium, fibroblasts, collagen and elastic fibers, proteoglycans, intrinsic nervous ganglia) was observed at both short and long term. Concomitant acute rejection of valve leaflets and myocardium was seen in 7, mild valve thickening in 14, myxoid degeneration in 4, and graft aortic atherosclerosis in 26. Mineralization was negligible and was not progressive with time. No linear correlation was found between mineralization and number of acute rejections. In conclusion, we observed optimal cusp viability and integrity even at long term, concomitant valve and myocardium rejection with no valvular sequelae, and negligible, non-progressive cusp calcification. Donor-recipient blood group matching, heart-beating donor, and chronic immunosuppression are the reasonable explanations of the optimal durability of the aortic valve after heart transplantation.
Tooth plates of extant lungfishes were examined by contact microradiography, light microscopy and TEM, to elucidate the nature and origin of the hypermineralized surface layer. TEM indicated that the matrix of this layer consisted of organic filamentous material, and no collagen fibres were seen in the matrix. The matrix appeared to be secreted by oral epithelial cells of the inner fold surrounding the surface of the tooth plate, and deposited in the area between the cell membrane and basal lamina-like structure of these epithelial cells. Therefore, it is suggested that the hypermineralized surface layer of lungfish plates is enamel of ectodermal origin.
Using macroscopic, microradiographic and scanning electron-microscopic methods, the effects of increased fluoride exposure on enamel and dentine formation were studied in fluorosed mandibular premolars and molars of roe deer from the heavily industrialized Ruhr area, Germany. Macroscopically, fluorosed teeth were characterized by opaque and stained enamel and in more severe cases also by enamel surface lesions, reduction or loss of enamel ridges on their occlusal surfaces and increased wear. Microradiographically, fluorosed enamel exhibited different degrees of subsurface hypomineralization, in part apparently indicating a fluoride effect during enamel maturation. In some specimens, a pronounced but varying enhancement of the pattern of Retzius lines was observed throughout the enamel, denoting strongly intermittent fluoride exposure during enamel matrix secretion. This variation in exposure was also reflected histologically in dentine, by bands of interglobular dentine and marked accentuation of incremental lines. Microradiography of sections through enamel surface hypoplastic lesions showed the enamel forming the bottom and partly also the walls of the lesions to be highly mineralized. Scanning electron microscopy showed that the outer enamel along the more pronounced hypoplastic lesions consisted of stacked, thin layers of 'aprismatic' enamel, indicating that the ameloblasts in these areas had lost the distal (rod-forming) regions of their Tomes' processes. These observations demonstrate that the origin of enamel hypoplasias in deer clearly differs from that in rodents, where fluoride induces the formation of subameloblastic cysts. The differences in the degree of fluorotic alteration between the teeth of a single tooth row could be related to the developmental sequence of the dentition in roe deer. The roe deer is thus considered to be a very sensitive and useful bioindicator of environmental pollution by fluorides.
Various biologic and non-biologic materials may be embolized to the brain after the use of cardiopulmonary bypass (CPB) pumps during open heart surgery but their relative frequency and importance are uncertain. Among the nonbiologic materials, Antifoam A, which contains organosilicates and silicon, continues to be employed as an additive to prevent frothing. Recent improvements in filtration and oxygenation techniques have clearly reduced the incidence of large emboli and complications like stroke but other neurologic sequelae following open heart surgery are common and in many cases poorly explained. A recently developed histochemical technique for the demonstration of the endothelial alkaline phosphatase (AP) was employed in a post-mortem study of brains from 8 patients and 6 dogs dying within a few days after open heart surgery employing cardiopulmonary bypass perfusion. Brains from 38 patients and 6 dogs who were not subjected to heart surgery were studied as controls with the same technique. The AP-stained slides are suitable for both light microscopic examination of the thick celloidin sections as well as a subsequent processing for high-resolution microradiography. Small capillary and arteriolar dilatations (SCADs) were seen in the test subjects/animals but not controls. SCADs were seen in all parts of the brain. Approximately 50% of the SCADs showed birefringence when examined with polarized light. SCADs are putative embolic phenomena and the exact nature and source of the embolic material is under investigation. A glycolipid component is indicated by preliminary studies. SCADs are difficult to find in routine paraffin sections and most if not all of the offending material seems to be dissolved during processing.(ABSTRACT TRUNCATED AT 250 WORDS)
Class V cavities and wedge-shaped cavities simulating erosions were prepared in thirty extracted, permanent human teeth. After being etched for 2 minutes with the pertinent cleanser, the wedge-shaped cavities were restored with Enamelite or Restodent. The Class V cavities were etched with 50% phosphoric acid and restored with HL-72. Artificial lesions around the fillings were produced by immersing the teeth in an acid-gelatin system. Ground sections of the teeth with the fillings in situ were studied by polarized light microscopy and microradiography. All the teeth showed "outer lesions," whereas the occurrence of a "wall lesion" was a rare exception.
The nature of the enamel defects in a patient with epidermolysis bullosa hereditaria dystrophica was observed by light microscopy, microradiography, and scanning electron microscopy (SEM) so that by these means more detailed information could be obtained. The random distribution of the round shape of the hypoplastic defects of enamel with hypomineralization, in which there is a continuity of the incremental lines, suggested the possibility that some ameloblasts maintained hypoactivity of their secretory function during their total life span.
The purpose of the present investigation was to study the dentin and pulp changes in the two halves of bisected carious teeth in order to determine if they are comparable. The dentin was studied by means of contact microradiography. Hypo- and hypermineralized zones, which were fairly similar in extension and degree of mineralization, were seen in the primary dentin of both halves of the teeth. If secondary dentin had formed, comparable amounts were usually present in the two halves. The pulp was studied, using demineralized 5 mum thick paraffin serial sections. In teeth with deep carious lesions severe inflammation with abscess formation was often observed in one half, whereas only scattered inflammatory cells were present in the other half. It seems questionable, therefore, whether the study of dentin and pulp reactions to caries in different halves of bisected teeth is a suitable method in investigations concerning a possible correlation between these reactions.
In order to study the interface of calcium phosphate bioglass ceramics, cylinders of standard size were implanted in the tibiae of rabbits. The materials were evaluated by radiography, light microscopy and microradiography. Bioceramics with hydroxyapatite surface give rise to a closer contact with new bone than calcium phosphate glass ceramics.
In order to study bone tissue interaction on calcium phosphate glass ceramics and on apatite ceramics, cylinders of standard size were implanted in the tibiae of rabbits. The materials were evaluated by radiography, light microscopy and microradiography. Apatite ceramics gave rise to a very close contact with new bone. Glass with an apatite surface evoked bone growth similar to that for apatite ceramics but with a loose contact while glass without an apatite surface gave rise to porous remodelling of new bone without close contact to the implant.
This study examines the biodegradation behaviour of calcium phosphate macroporous bioceramics consisting of hydroxylapatite or beta-whitlockite implants. The implantations were performed in hard tissue of femora and muscular tissue of dogs for periods of 6 and 12 month. With microradiography and light microscopy, respectively, of ground and thin sections it was shown that hydroxylapatite remained unchanged and beta-whitlockite degraded within 6 month in bone tissue. In the muscular tissue, the biodegradation of beta-whitlockite was slower. In addition, beta-whitlockite gave rise to a cellular response of lymphocytes and plasma cells rather than hydroxylapatite in the soft tissue region. For tissue response hydroxylapatite is more suitable as an implant material than beta-whitlockite.