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[Spontaneous bone marrow micrometastasis of a cerebral glioma. Immunohistochemical diagnosis in a biopsy sample and review of the literature].

A 55 year-old woman was admitted to hospital in January 1981 with transient expressive dysphasia. Past personal history was unremarkable except for a six-month history of renal colic and thrombophlebitis in the veins of the right leg. Computed tomographic scan of the head and carotid angiogram revealed a left calcified temporoparietal tumor. Because of pulmonary embolism it was decided to refute a cerebral biopsy. The patient also declined radiotherapy. In May 1983, a thorough workup revealed an incomplete fracture of the first lumbar vertebra and a diffuse demineralization of the rachis and pelvis. Four weeks later she developed temporal epilepsy and pulmonary embolism. A whole brain irradiation (60 Gy) was performed in August 1983. The patient's condition remained clinically stable until December 1984 when she was readmitted to hospital with a severe weight loss, diffuse osseous pain and pancytopenia. A bone marrow biopsy from the iliac crest showed a diffuse tumor involvement. Peroxidase-antiperoxidase staining using monoclonal antiserum to glial fibrillary acidic protein was strongly positive in numerous tumors cells. The pathological diagnosis was bone marrow metastasis by glioma. She died in March 1985, 4 years and 3 months after the first admission to hospital. Autopsy was not performed. A literature search reveals only 9 cases of extraneural spreading of astrocytomas and glioblastomas in the absence of previous craniotomy with post-mortem examination. The authors also comment on the clinical, pathological and histogenic aspects of extraneural metastasis of gliomas.

Bone Marrow Diseases↗

[Isolated decalcifying algodystrophy of the hip].

Transitory demineralization of the hip, a special form of reflex sympathetic dystrophy, is a rare condition. It should however be distinguished from more severe conditions such as tuberculosis or neoplastic bone disease. The typical symptoms are illustrated from 3 personally observed cases. The patients, usually men between the ages of 30 and 50 years or women in the last months of pregnancy, suffer severe mechanical pain in the hip, with limping and weakness. They usually need crutches for walking. Clinical examination of the hip reveals only slight restriction of passive movements. The erythrocyte sedimentation rate may be slightly elevated. Bone scan discloses hypercaptation, while other laboratory tests are negative. A few weeks after onset of symptoms, pelvic X-rays show extreme demineralization of the femoral head. On the other hand, the joint space and bony contours remain intact. The prognosis is excellent and the symptoms disappear after a few months or after delivery.

Accidents, Traffic↗

Correlation of CT analysis and audiometry in Japanese otosclerosis.

OBJECTIVE: To determine the extent of audiometric correlation with CT findings. METHODS: Forty-four patients (82 ears) with surgically confirmed otosclerosis underwent preoperative CT examination. Based on the computed tomography (CT) findings, the ears were classified into five groups as follows: Group A, the group with no pathological CT findings; Group B1, the group with demineralization localized in the region of the fissula antefenestram; Group B2, the group with demineralization extending towards the cochleariform process from the anterior region of the oval window; Group B3, the group with extensive demineralization surrounding the cochlea; and Group C, the group with thick anterior and posterior calcified plaques. RESULTS: There were 32 ears (39.0%) in Group A, 21 ears (25.6%) in Group B1, 16 ears (19.5%) in Group B2, 7 ears (8.5%) in Group B3, and 6 ears (7.3%) in Group C. The mean bone conduction levels were greater in the order of the extent of demineralization: Groups A, B1-B3 suggesting positive relationship between the cochlear function and the degree of labyrhinthine otosclerosis. CONCLUSION: A good correlation between the preoperative CT findings and audiometry findings suggests that CT with a slice intervals between 0.5 and 1mm could provide useful informations in assuming the extent of otosclerosis in the inner ear.

Adolescent↗

[Effects of maxillary growth of Wistar rats with bilateral artificial cleft palate after premaxillary orthopedic treatment on expression of proliferating cell nuclear antigen(PCNA)].

OBJECTIVE: Being based on the principle of Latham's appliance, this experiment is designed to detect activity of osteoblasts in the maxillary sutures of Wistar rats with bilateral artificial cleft palate by immunohistochemistry technique, and evaluate the effects of this appliance on the growth and development of the maxilla. METHODS: The animal models of male infant Wistar rats with bilateral cleft palate were made by removing some palatal bones, splitting a "V" gap of 1.5 cm wide at the line between premaxilla and segments of maxilla. A sort of appliances, which could be fixed in the mouth of Wistar rats with bilateral cleft palate for correcting protrusion premaxilla was constructed basing on the principle of Latham's appliance. Then the diferent pathological changes of osteoblast proliferation between the experimental group and the two controlled groups were examined. The jugomaxillary sutrues, temporomalar sutures and sphenoipalatine suture were harvested 7, 14 days after premaxillar orthopedic treatment, followed by 4% paraformaldehyde fixing about 1 hour, demineralization with 15% EDTA and 0.5% paraformaldehyde for 48 hours, distilled water washing for a night, dimethylbenzene transparant dealing, and paraffin wax embeding. Proliferating Osteoblasts in all these sutures were investigated using immunohiostochemical technique with monoclonal antibodies of proliferating cell nuclear antigen (PCNA). RESULTS: Seven days after orthopedic treatment, no significant difference was observed between the experimental group and the two controlled groups. While after fourteen days, obvious PCNA-positive expression were observed in cells of all these sutures of the experimental group. CONCLUSION: The distribution of proliferating cells and the degree of cell proliferation change after premaxillary orthopedic treatment. And significant cell proliferation is observed in the experimental group, but there are no significant differences between the two controlled groups.

Animals↗

Automatic quantitative analysis of ore content of human tooth enamel based on image processing.

Introducing the theory of fuzzy set, mathematical morphology and computerized mask fast scanning, we developed the TOOTH.SCA software and method to analyze the effect of fluoride (NaF) on ore content of human tooth enamel automatically and quantitatively. And we obtained some characteristic parameters, such as the depth, the type and the demineralized content of every scathing layer of dental caries. The smallest scale of mask scanning is 0.1 microm x 0.1 microm and the time required to analyze a sample is only 12 s. The applied software and method we built play an important role to the research on the mechanism of pathological changes of teeth and preventing dental caries.

Bone Density↗

Role of cartilage-derived anti-angiogenic factor, chondromodulin-I, during endochondral bone formation.

OBJECTIVE: Cartilage is a typical avasclar tissue that exhibits powerful resistance to angiogenesis or vascular invasion. We previously identified a cartilage-specific 25 kDa glycosylated protein, chondromodulin-I (ChM-I), as anti-angiogenic factor. Taking advantage of ectopic bone formation and xenograft tumour model by human chondrosarcoma cell line OUMS-27, we examined how ChM-I is involved in switching of angiogenesis in cartilage. DESIGN: Gene expression pattern of ChM-I was examined in 4-week-old mice and mouse embryos by northern blot analysis and in situ hybridization. To evaluate the effect of ChM-I on ectopic bone formation, guanidine extracts of demineralized bone matrix were mixed with the ChM-I-bound heparin-Sepharose beads and were implanted onto the fasciae of back muscle of 6-week old nude mice. To analyse the effect of ChM-I on tumour angiogenesis, the level of ChM-I mRNA in cartilaginous tumours was assessed by competitive PCR, and compared with that of articular cartilage. Then, human chondrosarcoma OUMS-27 cells were inoculated into the back of nude mice to form a tumour about 45 mm3 in size. Recombinant ChM-I protein was administrated into OUMS-27 xenograft tumours for the initial 5 days to study its effect against tumour-angiogenesis. RESULTS: ChM-I gene was specifically expressed in cartilage of 4-week-old mice. Eye and thymus were also identified as minor expression sites. However, during endochondral bone development, cartilage changes its character from anti-angiogenic into angiogenic prior to the replacement of calcified cartilage by bone. In embryos, ChM-I mRNA was expressed in proliferative and upper hypertrophic cartilage zones in the developing cartilaginous bone rudiments, but completely abolished in lower hypertrophic and calcified cartilage zones. Purified ChM-I protein apparently inhibited vascular invasion into cartilage induced by the implantation of demineralized bone matrix in nude mice, leading to the inhibition of replacement of cartilage. The level of ChM-I transcripts in the lower-grade chondrosarcomas was substantially reduced to several hundreds or less in the lower-grade chondrosarcomas, compared with that of articular cartilage or other benign cartilage tumours. The local administration of recombinant human ChM-I almost completely blocked tumour angiogenesis and growth in the human chondrosarcoma xenografts in mice. CONCLUSIONS: ChM-I is involved in the anti-angiogenic property of cartilage and its absence creates a permissive microenvironment for vascular invasion into cartilage under physiological and pathological conditions.

Animals↗

[Gingival hypertrophy in I-cell disease (mucolipidosis II). A report of 2 nonfamilial cases. II].

Two nonconsanguineous patients affected by I-cell disease (mucolipidosis II) are reported. I-cell disease, an oligosaccharidosis, is characterized by severe psychomotor retardation, marked shortness of stature, coarse facies, gingival enlargement, generalized bone demineralization, periosteal cloaking of long bones visible in early infancy, a rapid deteriorating course, and death from heart failure or bronchopneumonia, usually by the age of 5 years. This disorder is the result of a deficiency of glycoprotein N-acetylglucosaminylphosphotransferase activity, necessary for proper intracellular processing of lysosomal enzymes. Inheritance is autosomal recessive. It received the name I-cell disease because of several granular inclusions in the cytoplasm of cultured fibroblasts and amniotic fluid cells observed under phase contrast microscopy. These granules represent altered lysosomes. The two patients, reported here, had a very marked gingival hypertrophy and, for this reason, were referred to the Oral Pathology Service of Galliera Hospital. A gingivectomy was performed on patient 2 to improve the mastication, but few months later gingival hypertrophy reappeared.

Child, Preschool↗

Identification of calprotectin, a calcium binding leukocyte protein, in human dental calculus matrix.

Calprotectin is a calcium binding protein produced by leukocytes, macrophages and epithelial cells, and its levels in several tissues increase during infections and in many inflamed areas, suggesting that it may be an indicator of inflammatory activity. Osteopontin is a prominent phosphorylated glycoprotein in bone matrix, having calcium binding capacity. Recently, it has been reported that calprotectin and osteopontin are present in urinary stones (pathological mineralized masses in the body), and that these proteins may be involved in their formation. Dental calculus formed by mineralization of dental plaque is an inflammatory factor which may contribute to periodontal disease. It contains many organic components involved in mineralization. We recently found osteopontin molecules in human dental calculus and suggested that the components of its matrix may be similar to those of urinary stones. In this study, we investigated the presence of calprotectin in human dental calculus by immunohistochemical and immunoblotting analyses using a specific antibody for calprotectin. After fixation and demineralization of dental calculi adhered to tooth roots, sections embedded in paraffin were immunoreacted with the antibody for calprotectin and positive immunostaining for calprotectin was observed. Dental calculus proteins were then extracted with EDTA and separated by electrophoresis on 15% polyacrylamide gels. By immunoblotting analysis, 3 or 4 bands were observed at 11, 14.5, 22-25, 28 or 36.5 kDa and these patterns corresponded to those of calprotectin subunits. When non-immune rabbit serum was used instead of calprotectin-specific antibody as a negative control, no immunoreactivity was observed. These findings indicate that calprotectin is associated not only with antibacterial action but also with calcium binding capacity during dental calculus formation.

Calcium-Binding Proteins↗

Arthritic disorders of the adult radiocarpal joint: anatomic considerations and an evaluation of fifty consecutive abnormal cases.

The anatomy and pathology of the radiocarpal compartment of the adult wrist are described in a study of human cadavers and 50 consecutive patients with radiocarpal joint abnormalities. The most frequently encountered diseases were adult onset rheumatoid arthritis (42) and calcium pyrophosphate deposition disease (22%). Features allowing radiographic diagnosis included the degree of symmetry and the presence of demineralization, sclerosis, joint space narrowing, subchondral cysts and erosions. Evaluation of abnormalities in other compartments of the wrist and the ulnar styloid is mandatory.

Adult↗

Bone mineral density in primary and secondary amenorrhea.

UNLABELLED: Amenorrhea in young women is one of the best clinical indicators for estrogen deficiency, except in the presence of gynecological structural pathology. This study aimed at investigating bone mineral density (BMD) in patients with primary and secondary amenorrhea. Thirty-six patients were enrolled in the study, seven with primary amenorrhea (mean age 24.3 +/- 4.5 yrs.) and twenty-nine with secondary amenorrhea (mean age 31.1 +/- 6.9 yrs.). Eighteen regularly menstruating women (mean age 31.8 +/- 3.7 yrs.) served as controls. BMD was measured at lumbar spine, femoral neck, Ward's triangle and trochanter. RESULTS: BMD was significantly decreased in both primary and secondary hypoestrogen amenorrheic patients. Primary amenorrheic patients were more severely affected with a BMD mean Z score below 80 per cent (osteopenia) at all sites measured. The age of primary amenorrheic women also strongly correlated with degree of demineralization. This should emphasize the importance of early diagnosis and treatment of young amenorrheic patients.

Absorptiometry, Photon↗

Hypophosphatemia and calcium nephrolithiasis.

Our knowledge of phosphate balance under physiological and pathological situations has increased substantially during the last decade thanks to the molecular identification of three dissimilar families of sodium-phosphate cotransport systems, two of them almost exclusively expressed in epithelia whereas the third one has a ubiquitous expression. Intracellular proteins such as NHERF1 (sodium-proton exchanger regulatory factor 1) can interact with phosphate transporters through PDZ domains thus regulating the expression of the transporters at the membrane. Moreover, newly acknowledged paracrine/endocrine peptides, such as fibroblast growth factor 23 (FGF23), also affect the activity of phosphate transporters. Renal phosphate leak, related to invalidation (in the mouse) or to mutations (in humans) of the renal phosphate transporter NPT2a, leads to hypophosphatemia on the one hand, and to nephrolithiasis or bone demineralization on the other hand. Similar features are observed during invalidation of NHERF or in case of overproduction of FGF23. These observations highlight the importance of phosphate homeostasis in common diseases such as renal stones or bone loss.

Animals↗

The TEM characterization of the lamellar structure of osteoporotic human trabecular bone.

The lamellar structure of osteoporotic human trabecular bone was characterized experimentally by means of transmission electron microscopy (TEM). More specifically, the TEM was used to determine if trabecular bone exhibits similar lamellar structural motifs as cortical bone by analyzing unmineralized, mineralized and demineralized bone, and to study the influence of the osteocyte network on the lamellar structure of osteoporotic trabecular bone. Comparison with normal trabecular bone is included. This paper summarizes partial results of a larger study, which addressed the characterization of the hierarchical structure of normal versus osteoporotic human trabecular bone [Rubin, M.A., 2001. Multiscale characterization of the ultrastructure of trabecular bone in osteoporotic and normal humans and in two inbred strains of mice. MS Thesis, Georgia Institute of Technology.] at several structural scales.

Bone and Bones↗

Localization of matrix metalloproteinase 9 (92-kilodalton gelatinase/type IV collagenase = gelatinase B) in osteoclasts: implications for bone resorption.

BACKGROUND: Matrix metalloproteinase 9 (MMP-9, 92-kD gelatinase/type IV collagenase = gelatinase B) is a member of the MMP gene family and implicated in tissue destruction in the various pathophysiologic conditions. Our previous study showed that MMP-9 purified from human fibrosarcoma cells can cleave the cross-link-containing NH(2)-terminal telopeptides of the alpha 2 chain of type I collagen and collagen types III, IV, and V as well as gelatins. EXPERIMENTAL DESIGN: To investigate the role of MMP-9 in bone resorption we have examined its localization in the human bone tissues by immunohistochemistry and in situ hybridization. The enzymic properties were also biochemically studied. RESULTS: Immunohistochemistry using monoclonal antibodies against MMP-1 (interstitial collagenase), MMP-2 (72-kD gelatinase/type IV collagenase = gelatinase A), MMP-3 (stromelysin-1), MMP-9, and tissue inhibitor of metalloproteinases-1 demonstrated that MMP-9 is localized exclusively in osteoclasts of the bone tissues from normal subjects and patients with rheumatoid arthritis or metastatic carcinoma whereas some osteoclasts are also labeled by anti-(MMP-1) antibody. Northern blot and in situ hybridizations of rheumatoid bone tissues using a RNA probe for MMP-9 exhibited strong signals for the mRNA within osteoclasts. MMP-9 depolymerized acid-insoluble polymers of type I collagen and digested collagen fibrils in the demineralized bone. The gelatinolytic activity of the proteinase was optimal at pH 7.5, but 50 to 80% of the full activity was retained at pH 5.5 to 6.0. It was also 90% active in the presence of 100 mM Ca2+. Degradation of acid-soluble and -insoluble type I collagens by MMP-9 was enhanced at higher concentrations of Ca2+. The zymogen of MMP-9 was activated up to approximately 85% of full activity by incubation at pH 2.3. CONCLUSIONS: These results demonstrate that MMP-9 is produced by osteoclasts in the human bone tissues and suggest that it can degrade bone collagens in concert with MMP-1 and cysteine proteinases in the subosteoclastic microenvironment. This proteinase may play a role in the normal bone remodeling and pathologic bone resorption in the human diseases.

Adolescent↗

The use of 2-dimensional CNBr peptide maps for the analysis of crosslinked peptides in bone collagen.

CNBr peptides from insoluble bovine cortical bone collagen were analyzed using a 2-D mapping technique. The major type 1 collagen CNBr peptides were detected by fluorography after general-labelling with 3H-NaBH4 in dimethyl-formamide. These maps were similar to those visualized by coomassie blue staining and demonstrated a proportional decrease of alpha 1CB6. New groups of peptides, different from those normally present in soluble type I collagen were detected. Some of these peptides were slightly larger and more acidic than alpha 1CB6 and were highly labelled when the demineralized bone was specifically labelled for the presence of aldehydes and crosslinks with 3H-NaBH4 in a phosphate buffer, pH 7.4. Based on the size and charge characteristics of these specifically labelled peptides, they were tentatively identified as crosslinking peptides containing different combinations of alpha 1CB6, alpha 1CB0,1 and alpha 1CB5. The specificity of the labelling method using 3H-NaBH4 in phosphate buffer was demonstrated by the detection of other known crosslinked peptides and by the virtual absence of label in alpha 1CB7, CB8, and CB3. We feel that this simple methodological approach developed in these experiments will prove to be very useful in the analysis of collagen crosslinks present in insoluble collagens derived from normal tissues of various ages as well as from pathological states.

Animals↗

[Multiple pathological fractures within the scope of DeToni-Debre-Fanconi syndrome after fumarate therapy in psoriasis].

We report about a rare case of a pathological fracture of the shank following earlier pathological fractures at other locations in a comparatively young female patient with no history of trauma. There were no known diseases other than psoriasis. The shank fracture was treated surgically by osteosynthesis. Osteoporosis, myeloma, or malignancy as causative factors of this fracture could be excluded. Scintigraphy showed an enhancement, especially at the extremities. Other than reactive bone growth, histological examination revealed no further aspects. Laboratory analysis indicated a massive lack of vitamin D3. After transferring the patient to the internal department of our hospital, long-term medication with fumaric acid was determined to be the reason for the osteomalacia of a Fanconi's syndrome. Three months after cessation of these medicaments and treatment with active vitamin D3 metabolites, the patient was free of complaints. The radiographs showed an essential improvement of the demineralization.

Adult↗

Dentin sialoprotein and phosphoprotein induce neutrophil recruitment: a mechanism dependent on IL-1beta, TNF-beta, and CXC chemokines.

Dentin is a reservoir of several potentially active molecules, and dentin sialoprotein (DSP) and dentin phosphoprotein (DPP) are the two major non-collagenous proteins. It has been established that dentin molecules are released as a consequence of osteoclast action during the resorption process. Along with osteoclasts, inflammatory cells seem to play an important role at sites of root resorption. Although the role of dentin molecules in dentinogenesis is well known, their role in pathological processes associated with dentin matrix dissolution is unclear. Recent studies have suggested that dentin components may function as chemotactic and activator signals for inflammatory cells at these sites. Herein we present evidence that demineralized dentin crude extract, DSP, and DPP induced doseand time-dependent neutrophil migration into the peritoneal cavity of mice and that this activity was inhibited by dexamethasone, but not by indomethacin or MK886. The blockade of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) receptors inhibited neutrophil accumulation. The neutrophil migration was also diminished in the absence of the chemokines cytokine-induced neutrophil chemoattractant (KC) and macrophage inflammatory protein-2 (MIP-2), but not in the absence of macrophage inflammatory protein-1alpha (MIP-1alpha). These results demonstrate that dentin induces neutrophil migration via the synthesis of IL-1beta, TNF-alpha, and chemokines and they suggest that dentin matrix proteins may have an active role in inflammatory cell recruitment during pathological processes associated with dentin and bone matrix dissolution.

Animals↗

Cytokine and chemokine response of bone cells after dentin challenge in vitro.

OBJECTIVE: The aim of this study was to characterize the effects of dentin extracts on cytokine, chemokine and nitric oxide (NO) production by primary rat bone cells. STUDY DESIGN: Osteoblastic bone marrow cultures were exposed to particulate (D-part), non-particulate (D-n-part) and demineralized dentin extracts and evaluated for proliferative activity, cell morphology, alkaline phosphatase activity and bone-like nodule formation. Cytokine production was assessed by enzyme-linked immunosorbent assay and NO release by the Griess method. RESULTS: The dentin extracts did not affect osteoblast numbering. Conversely, they up regulated in a dose-dependent manner the production by the osteoblasts of the pro-inflammatory interleukin-1beta (IL-1beta), tumor necrosis factor-alpha, IL-6, cytokine-induced neutrophil chemoattractant-1, and of the anti-inflammatory cytokine, IL-10. The NO production was stimulated only by D-n-part. CONCLUSION: These results demonstrate that dentin induces the production of inflammatory cytokines by osteoblasts and suggest that pro-resorptive pathways might be stimulated when dentin molecules come into contact with bone cells during pathological processes associated with dentin and bone matrix dissolution.

Alkaline Phosphatase↗

[Reflex sympathetic dystrophy: still a poorly defined entity].

The reflex sympathetic dystrophy (algodystrophy) constitutes a large nosological field of which the main characteristics are the appearance of algic and vasomotor symptoms at a segmental level of a limb, in consequence to diverse pathologies (trauma, cardiovascular disease, etc.). The widely accepted theory of a dysregulation of the sympathetic nervous system is nowadays counter-balanced by recent work highlighting the preponderant role of polymodal afferent nerves in the pathophysiology of this disease. The diagnosis, being above-all clinical, is marked by two distinct phases appearing in a variable chronology; a warm phase associating fluctionating pain, stiffness and vasomotor symptoms, and then a cold phase characterized by fibrosis, leading to disabling trophic symptoms. Spontaneous recovery is usual and can be delayed by up to two years, however irreversible sequelae can occur. Paraclinical investigations are necessary to confirm the diagnosis: absence of a biological inflammatory syndrome, early hyperfixation on bone scintography or an abnormality in the MRI signal in the sub-chondral zones. The X-ray shows late local demineralization that is often non-homogenous. The treatment is poorly codified. First-line treatment in France, other than antalgics, often rests on the calcitonins. Intravenous diphosphonates are proposed by some in case of treatment failure. Regional venous blocks are sometimes performed in resistant and disabling forms. Rehabilitation and psychological support have a primordial place throughout the evolution of the illness.

Acute Disease↗